miR-181b与炎症关系实验

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具核梭杆菌通过调控miR-181b在结肠癌细胞中形成炎性微环境的机制

具核梭杆菌通过调控miR-181b在结肠癌细胞中形成炎性微环境的机制研究背景:具核梭杆菌(Fusobacteriumnucleatum,Fn)是口腔常见致病菌,但近年来研究发现,该菌与结直肠癌的发生发展密切相关。

Fn是一种具有侵袭性,黏附性的厌氧菌,同时它也具有促炎性因子释放以及调节肿瘤免疫微环境的能力。

Fn如何通过炎性因子致结直肠癌的发生发展,当中机制尚不明确。

研究表明,当肠上皮细胞受到Fn感染时,炎性因子表达增高,但尚无相关文献从miRNAs的角度研究由此引起的炎性因子表达上调的相关机制。

目的:通过构建Fn感染Caco-2细胞的炎症模型,运用miRNA测序技术分析由此所引起miRNAs基因表达差异;通过生物信息学的方法筛选出表达差异显著的miRNA及其下游靶基因,并对其调控靶基因的机制及引起的生物学效应进行研究,从而初步探索得到Fn感染在促炎症信号通路上相关结直肠癌发生发展的作用机制。

方法:1.构建了Fn感染Caco-2细胞的炎症模型,运用miRNA测序技术分析由此所引起miRNAs表达差异;2.通过qPCR、ELISA、WesternBlot等方法对模型中TNF-α在转录水平及蛋白水平上的变化进行检测;3.使用Transwell实验分析Fn 与Caco-2细胞共培养后诱导产生的趋化因子及其引起的生物学效应;4.采用qPCR方法验证模型中表达差异显著的miR-181b的表达情况;5.通过转染的手段,分别上调及下调Caco-2细胞中miR-181b,再加入Fn感染处理,由此研究在Fn介导下miR-181b与TNF-α之间的相关关系;6.采用生物信息学工具及双荧光素酶检测方法证实miR-181b与TNF-α的靶标关系。

结果:1.miRNA测序结果分析显示,Fn感染Caco-2细胞引起的相关差异miRNA有653个,其中在处理组中低表达645个,高表达8个,其中,miR-181b表达显著降低(p=0.001345),主要涉及炎症、增殖、细胞凋亡等多个信号通路的变化。

血清微小RNA-181b的表达与子痫前期患者白介素-6、白介素-10、白介素-17的相关性

血清微小RNA-181b的表达与子痫前期患者白介素-6、白介素-10、白介素-17的相关性王茹;邓乾葆;张忠霞;黄素静【期刊名称】《中国性科学》【年(卷),期】2022(31)7【摘要】目的观察子痫前期(PE)患者血清微小RNA(miR)-181b的表达,并分析其与白介素(IL)-6、IL-10、IL-17的关系。

方法选取2018年10月至2020年12月海南医学院第二附属医院收治的96例PE患者作为研究对象,并根据病情程度分为非重度PE(N-PE)组和重度PE(S-PE)组,各48例。

询问并记录患者的基线资料;检测患者入院时血清miR-181b及血清IL-6、IL-10、IL-17等炎性因子水平;分析血清miR-181b的表达与PE患者IL-6、IL-10、IL-17的关系,并进一步探究血清miR-181b与PE病情的关系及其对评估S-PE发生风险的价值。

结果S-PE组患者入院时血清miR-181b、IL-6、IL-17水平均高于N-PE组,血清IL-10水平低于N-PE 组,差异具有统计学意义(P<0.05);相关性检验发现,血清miR-181b的表达与PE患者IL-6、IL-17水平呈正相关(r>0,P<0.05),与血清IL-10水平呈负相关(r<0,P<0.05);回归分析发现,入院时血清miR-181b的过表达与PE患者病情存在一定关系,其可作为S-PE发生的风险因子(P<0.05);绘制受试者工作特征(ROC)曲线,入院时血清miR-181b的表达评估S-PE发生风险的曲线下面积(AUC)为0.896,有一定的评估价值。

结论血清miR-181b的表达与PE患者IL-6、IL-17水平呈正相关,与血清IL-10水平呈负相关,且其过表达与PE患者病情存在一定关系,可作为S-PE发生的风险因子。

【总页数】5页(P79-83)【作者】王茹;邓乾葆;张忠霞;黄素静【作者单位】海南医学院第二附属医院产科【正文语种】中文【中图分类】R711【相关文献】1.血清干扰素-γ、白介素-4、白介素-6及白介素-10在肾癌患者治疗中的表达及意义2.寻常型银屑病患者血清白介素17、白介素23的表达与临床相关性研究3.白介素6、白介素8和白介素10及肿瘤坏死因子α在重症急性胰腺炎中的表达及与预后的相关性研究4.慢性荨麻疹患者血清白介素-4、白介素-6、白介素-8、白介素-10及IFN-γ的检测及临床意义5.白介素-17、白介素-35在稽留流产患者血清中的表达及临床意义因版权原因，仅展示原文概要，查看原文内容请购买。

miR-181b-5p促进肺癌干细胞样细胞发生上皮间质转化中的作用及机制研究的开题报告

miR-181b-5p促进肺癌干细胞样细胞发生上皮间质转化中的作用及机制研究的开题报告研究背景和意义：肺癌是全球常见的恶性肿瘤之一，其发病率和死亡率居高不下。

肺癌干细胞样细胞（CSCs）是一类具有自我更新和分化能力的肺癌细胞，其具有长期存活能力和抗药性，并参与肺癌转移和复发。

因此，研究CSCs的生物学特性和分子机制对于探究肺癌发生发展的新机制以及寻找治疗肺癌的新靶点具有重要意义。

miRNAs是一类长度为20-22个核苷酸，可以调控基因表达的小分子RNA。

近年来的研究表明，miRNAs能够影响CSCs的生成和维持。

miR-181b-5p是miR-181家族的一员，其在多种肿瘤中都有表达异常。

然而，miR-181b-5p在肺癌CSCs的功能和机制方面的研究还很有限。

因此，在本研究中我们将探究miR-181b-5p在肺癌CSCs中的作用及其机制，以期为肺癌的治疗提供新的理论支持。

研究内容和方法：本研究将使用肿瘤干细胞筛选试验（tumor sphere assay）来鉴定肺癌干细胞样细胞，并通过实时荧光定量PCR来检测miR-181b-5p的表达情况。

接着，将使用miR-181b-5p mimics和inhibitor进行miR-181b-5p的上调和下调。

通过Western blot和Immunofluorescence技术来研究miR-181b-5p对E-cadherin、N-cadherin、Vimentin等上皮间质转化相关蛋白的影响，并采用流式细胞术来分析CSCs的增殖和凋亡情况。

研究预期：本实验将研究miR-181b-5p在肺癌CSCs发生上皮间质转化中的作用及其机制，为探索肺癌发生发展的新机制提供新的理论支持，有望为开发新的肺癌治疗策略提供新思路。

食管鳞状细胞癌患者血浆mirna-181ab的表达及诊断价值

食管鳞状细胞癌患者血浆miRNA-181a/b的表达及诊断价值研究生董翠翠导师贺付成河南郑州 450052中文摘要背景与目的食管癌(esophageal cancer, EC)是世界八大常见肿瘤之一，EC主要包括两大常见病理类型：食管鳞状细胞癌(esophageal squamous cell carcinoma, ESCC)和食管腺细胞癌(esophageal adenocarcinoma, EAC)；EC起病隐匿，死亡率居全世界肿瘤第六位。

据2012年公布的数据显示每年新增EC患者约456000例死亡人数高达400000例，五年生存率仅有20%左右。

EC的发病具有地域性， ESCC主要集中在亚洲地区； EAC在西方欧洲国家高发。

EC起病隐匿，肿瘤分期程度直接关系到患者的预后，早期发现是改善EC患者预后的关键。

microRNAs是由18-20个核苷酸组成的非编码RNA分子，由非编码基因的内含子编码。

microRNAs广泛稳定存在于真核动物体内及植物体内。

microRNAs 主要通过与靶基因mRNA的3'非编码区特异性互补结合从而抑制靶基因转录和降解靶基因，最终影响蛋白质的翻译，调控细胞的增殖、分化、凋亡等过程，起到了类似癌基因和抑癌基因的作用。

miRNA-181a/b是miRNA-181基因家族中两个重要组成成分。

miRNA-181家族与多种肿瘤的发生有关。

miRNA-181a在ESCC转移淋巴结中的表达水平高于癌组织，且miRNA-181a对人食管鳞癌TE11细胞的转移、迁移和侵袭能力有影响。

但有关miRNA-181家族在ESCC患者血浆中表达水平方面的报道并不多见。

本实验采用qRT-PCR技术对ESCC患者和健康对照组血浆中的miRNA-181a/b含量进行相对定量，并分析miRNA-181a/b与ESCC患者的临床病理分期、淋巴结转移是否存在相关性，探讨miRNA-181a/b是否可以作为预测ESCC的生物标志物。

miR-181b在胆管癌进展中的功能及分子机制研究

miR-181b在胆管癌进展中的功能及分子机制研究摘要：miR-181b是一种重要的微RNA分子，在胆管癌的进展中扮演着极其重要的角色。

本文旨在系统性地探究miR-181b在胆管癌进展中的功能及其分子机制。

首先，通过文献研究，发现miR-181b在胆管癌的肿瘤增殖、细胞迁移和侵袭能力以及化疗耐药性等重要生物学过程中发挥了重要作用。

接着，本文探究了miR-181b与多种关键蛋白质、信号通路之间的相互作用，以及miR-181b的来源、调控机制等方面。

研究结果表明，miR-181b通过调节多种靶基因、信号通路，影响胆管癌的发生、进展及治疗效果，并可能成为未来胆管癌治疗的潜在靶点。

本文综述了miR-181b在胆管癌进展中的重要作用及其分子机制，有利于深入研究胆管癌发病机理、指导治疗及提高预后。

关键词：miR-181b；胆管癌；进展；分子机制；靶点MiR-181b in the Progression of Cholangiocarcinoma: Function and Molecular MechanismsAbstract:MiR-181b is an important microRNA molecule that playsa crucial role in the progression of cholangiocarcinoma. This article aims tosystematically explore the function and molecularmechanisms of miR-181b in the progression of cholangiocarcinoma. Initially, through literature research, it was found that miR-181b plays a critical role in important biological processes of cholangiocarcinoma, such as tumor proliferation, cell migration, invasion ability, and chemotherapeutic resistance. Subsequently, this article explored the interactions between miR-181b and various key proteins, signaling pathways, as well as the source andregulatory mechanisms of miR-181b. The results showed that miR-181b affects the occurrence, progression, and treatment of cholangiocarcinoma by regulating multiple target genes and signaling pathways and may become a potential target for future cholangiocarcinoma treatment. This review summarizes the important roleof miR-181b and its molecular mechanism in the progression of cholangiocarcinoma, which is conducive to further study of the pathogenesis of cholangiocarcinoma, guide treatment, and improve prognosis.Keywords: miR-181b; cholangiocarcinoma; progression; molecular mechanism; targetmiRNAs have been shown to play a critical role in the regulation of multiple target genes and signaling pathways in the development and progression ofcholangiocarcinoma. Among them, miR-181b has been identified as an important regulator that controls the progression of cholangiocarcinoma through various mechanisms.One of the most important functions of miR-181b is its ability to regulate the expression of multiple target genes involved in cholangiocarcinoma. For instance, miR-181b has been shown to promote cell proliferation, migration, and invasion by targeting tumor suppressor genes, such as TIMP3, PTEN, and C/EBPα, which regulate cell cycle progression, apoptosis, and angiogenesis. In addition, miR-181b can inhibit the expression of other target genes, such as NDRG2 and TSPYL5, which are involved in the regulation of cell adhesion and transformation.Another critical mechanism by which miR-181b contributes to cholangiocarcinoma progression is through its interaction with various signaling pathways, including the Wnt/β-catenin, PI3K/Akt, and NF-κB pathways. For instance, miR-181b has been shown to promote the activa tion of the Wnt/β-catenin pathway by targeting GSK-3β, a negative regulator of this pathway. Similarly, miR-181b can activate thePI3K/Akt pathway by targeting PTEN and suppress the NF-κB pathway by targeting p65 and TAK1.The dysregulation of miR-181b expression has been observed in various human cancers, including cholangiocarcinoma. Several studies have shown that the downregulation of miR-181b expression is associated with increased tumorigenesis, metastasis, and poor prognosis in cholangiocarcinoma patients. Therefore, miR-181b could serve as a potential target for future cholangiocarcinoma treatment.In summary, miR-181b plays a critical role in the progression of cholangiocarcinoma by regulating multiple target genes and signaling pathways. Further studies are needed to elucidate its pathogenic mechanisms and explore its potential as a therapeutic target for cholangiocarcinomaAdditionally, there are other potential target genes and signaling pathways that miR-181b may regulate in cholangiocarcinoma. For example, previous studies have shown that miR-181b targets and downregulates SMAD7, a negative regulator of TGF-β signaling, leading to activation of TGF-β/SMAD signaling and promotion of cholangiocarcinoma cell proliferation and migration (Xiong et al., 2018). Other potential target genes of miR-181b in cholangiocarcinoma include MMP9, RAC1, NF-κB, and AKT (Zhang et al., 2016; Ren et al., 2019;Yang et al., 2019; Zhu et al., 2020).Furthermore, the potential clinical applications of miR-181b in cholangiocarcinoma diagnosis and prognosis are worth exploring. Previous studies have shown that miR-181b is differentially expressed in cholangiocarcinoma tissues and serum samples, and its expression level is associated withclinicopathological features and survival outcomes of cholangiocarcinoma patients (Xiang et al., 2018; Zhang et al., 2019; Zhu et al., 2020). Therefore, miR-181b could serve as a potential diagnostic and prognostic biomarker for cholangiocarcinoma, providing valuable information for clinical decision-making and personalized treatment.In conclusion, miR-181b is a key regulator of cholangiocarcinoma progression, and its dysregulation contributes to tumorigenesis, metastasis, and poor prognosis of cholangiocarcinoma. Understanding the molecular mechanisms underlying miR-181b-mediated regulation of target genes and signaling pathways in cholangiocarcinoma will provide insights into the pathogenesis of this devastating disease and identify potential therapeutic targets for future development of cholangiocarcinoma treatmentIn conclusion, miR-181b has emerged as an important player in the pathogenesis of cholangiocarcinoma. Dysregulation of miR-181b expression has been reported in various studies, and is associated with the progression, metastasis, and poor prognosis of cholangiocarcinoma. The mechanisms by which miR-181b regulates its target genes and signaling pathways in cholangiocarcinoma are complex, and require further investigation.Several target genes and signaling pathways have been identified to be regulated by miR-181b in cholangiocarcinoma. These include the PI3K/AKT/mTOR pathway, the Wnt/β-catenin pathway, and the NF-κB pathway. Targeting these pathways and pathways may provide a promising approach for the development of new therapeutics for cholangiocarcinoma.Recent studies have also highlighted the potential of miRNAs as diagnostic and prognostic biomarkers for cholangiocarcinoma. MiR-181b may serve as a potential biomarker for the early detection and prognosis of cholangiocarcinoma.In conclusion, miR-181b is a promising therapeutic target and a potential diagnostic and prognostic biomarker for cholangiocarcinoma. Future studies areneeded to further elucidate the roles of miR-181b in the pathogenesis of cholangiocarcinoma and to develop novel therapeutic strategies for the treatment of this devastating diseaseIn conclusion, miR-181b has been found to play important roles in the development and progression of cholangiocarcinoma through its regulation of various signaling pathways and target genes. Targeting miR-181b could provide a potential therapeutic strategyfor cholangiocarcinoma treatment. Moreover, miR-181b shows promise as a diagnostic and prognostic biomarker for cholangiocarcinoma. Further research is needed to fully understand the underlying mechanisms andclinical implications of miR-181b in cholangiocarcinoma。

miRNA_181b在牛腺垂体中的表达规律及其调控作用研究_英文_

Study on the Expression Laws and Regulatory Func-tions of miRNA-181b in the Anterior Pituitary of Cat-tleXU Cheng 1，XUE Wen-zhi 1，ZHAO Mao-xin 1，CHEN Xin 1，LIU Ying 1，YU Wang-yang 1，XU Yan-li 1，MA Teng-he 1，GAO Yan 2，ZHANG Jia-bao 2，YUAN Bao 2*1．College of Animal Science and Veterinary Medicine ，Jilin University ，Changchun 130062；2．Laboratory Animal Center ，Jilin University ，Changchun 130062Abstract ［Objective ］The research aimed to discuss the differential expression quantity of miRNA-181b in mature （18-month-old ）and immature （one-month-old ）cattle's anterior pituitary and its regulation function．［Method ］cDNA library of miRNA in mature （18-month-old ）and immature（one-month-old ）cattle's anterior pituitary were established．After Solexa high-throughput sequencing of miRNA in the cDNA library ，miRNA inanterior pituitary of bulls was identified．miRNA-181b with differential expression were selected from the sequencing results．By real-time quanti-tative RT-PCR ，the expression laws of miRNA-181b in the anterior pituitary of Yanbian Cattle in different growth period was validated．And thetarget genes of miRNA-181b were forecast by using TargetScanS prediction software．［Result ］The expression quantity of miRNA-181b hadgreat difference in cattle's anterior pituitary different growth periods．The expression quantity of miRNA-181b in anterior pituitary of one-month-oldcattle was 4．05times as that in 18-month-old cattle．The binding of miRNA-181b with 838－844bases in 3'untranslated region of FSH βgenewas specific and the binding base sites were UGAAUGUA．［Conclusion ］This research provided the theoretical basis for the transcription regula-tion research of FSH β．Key words miRNA ；Real-time quantitative RT-PCR ；Differential expressionReceived ：May 3，2011Accepted ：July 12，2011Supported by National Natural Science Foundation of China （30972100）and Jilin Science and Technology Development Pro-gram （201101015）．*Corresponding author．E-mail ：yuanbao1982@163．commicroRNA （miRNA ）are a kind of non-coding small ribo-nucleic acid （RNA ）molecules from the eukaryotic own ge-nomes with the length of about 19－25nt ［1］，which were pro-duced from their precursors by enzyme digestion．microRNAs mainly inhibit the mRNA translation of target genes or degrade target genes by acting on in the three prime untranslated re-gions （3'-UTR ）of mRNA of target genes ［2－4］．The studies in-dicated that miRNAs regulated more than one third of protein-coding genes and their recognization and functional analysis have been one of research hotpoints in the modern molecular biology field ［5－6］．Anterior pituitary is the most important endo-crine gland in animals ，which can secrete many kinds of hor-mones．And these hormones play important roles in the me-tabolism ，growth ，development ，procreation and other processes of animals．The reproductive performance of ani-mals is directly or indirectly regulated and influenced by vari-ous kinds of hormones that were secreted by anterior pituita-ry ［7－8］．In view of this ，we studied the expression laws of miRNA in anterior pituitary of Yanbian Cattle in different sta-ges and discussed the influences of various miRNA on the re-production and development of bulls in different stages ，ai-ming at providing scientific basis for the molecular mechanism of the reproduction and breeding of bulls for Yanbian Cattle．Materials and MethodsAnimal tissue materialsYanbian Cattle were purchased from Gudonghe YanbianCattle Breeding Limited Company．The cattles were slaugh-tered in Benfu Beef Cattle Slaughterhouse according to the slaughtering standards and the anterior pituitary tissues of im-mature （one-month-old ）and sexual －mature （18-month-old ）Yanbian Cattle bulls （three immature bulls and three sexual －mature bulls ）were collected．Main reagents and instrumentsMain reagents TRIzol （purchased from Takara Company ）；Taq DNA polymerase （5U/μl ）；dNTPs （2．5mmol /L ）；RT-PCR Kit ；SYBR Green realtime PCR master mix plus （Japan Toyobo Co．，Ltd．）；other reagents are analytical reagents．Instruments Realplex quantitative PCR system （GermanyEppendorf Company ）；UV 2102PC-type ultraviolet spectro-photometer （UNICO （Shanghai ）Instruments Co．，Ltd．）；Gel imaging system （Alpha Innotech Company ）．Extraction of total RNA from anterior pituitary Total RNA was extracted from the anterior pituitary of Yanbian Cattle ac-cording to TRIzol manual．And the extraction results of total RNA were identified by using agarose gel electrophoresis．Identification of total RNA from the anterior pituitarycDNA library was constructed by RT-PCR using RT-PCRKit．The system of RT-PCR （total volume of 10μl ）was asfollows ：1μl of total RNA ，0．5μl of Primer Mix ，2μl of 5ˑRT Buffer ，0．5μl of RT Enzyme Mix ，6μl of DEPC H 2O．The mixture was bathed for 15minitues at 37ħand then heated by water bath for 5minitues at 98ħto obtain cDNA library of Yanbian Cattle．Because prolactin only can be expressed in the anterior pituitary ，the primers （L ：ATCCTGGTATCACTAA-CAAAGC ；R ：GCCCAAACAACCCTAACG ）were designed ac-cording to the sesquences of mRNA so as to further validate total RNA by PCR ［9］．The components of PCR system with the total volume of 25μl were as follows ：2．0μl of 10ˑbuffer ，1．0μl of 10mmol /L dNTP ，0．5μl of 10pmol /μl forward primerAgricultural Science ＆Technology ，2011，12（7）：1031－1034Copyright 2011，Information Institute of HAAS．All rights reserved．Agricultural Biotechnologyand0．5μl of10pmol/μl reverse primer，1．0μl of cDNA，0．5μlof5U/μl Taq polymerase（containing20mmol/L MgCl2），19．5μl of sterilized deionized water．PCR was performed under the following conditions：pre-denaturing at94ħfor3min；35cycles of denaturing at94ħfor30s，annealing at60ħfor30s and extending at72ħfor30s；extending at72ħfor5min．The amplification products were preserved at4ħand identified by using1．5%agarose gel electrophoresis．High-throughput sequencing analysis of total RNA from the an-terior pituitaryThe validated total RNA was analyzed in Shenzhen Branch of Beijing Liuhe Gene Technology Co．，Ltd．by using Solexa high-throughput sequencing analysis．RT-PCR of miRNA-181bTaking the above RNA extracted from the anterior pituita-ry of cattle as template，reverse transcription primer（GTCG-TATCCAGTGCGTGTCGTGGAGTCGGCAATTGCACTGGATA CGACTTCCCT）were designed for RT-PCR according to the specific sequences of miRNA-181b．The componnents of RT-PCR system（total volume of10μl）were as follows：1μl of to-tal RNA，0．5μl of RT Primer，2μl of5ˑRT Buffer，0．5μl ofRT Enzyme Mix，6μl of DEPC H2O．The components were mixed and heated at37ħfor15min and then heated at98ħfor5min．The products of RT-PCR were obtained and pre-served at－20ħ．Detection of miRNA-181b by real-time fluorescent quantitative PCRAccording to the specific sequences of miRNA-181b，the forward primer（GAACAAACAAAGCAGACGG）and reverse primer（TGCGTGTCGTGGAGTC）were designed for real-time fluorescent quantitative PCR［10－11］．The components of real-time fluorescent quantitative PCR system were as fol-lows：0．5μl of miRNA-181b cDNA，0．5μl forward primer and0．5μl reverse primer，12．5μl SYBR dye，11μl of dH2O．PCR conditions were as follows：pre-denaturing at95ħfor one minute；40cycles of denaturing at95ħfor15s，annea-ling at60ħfor15s and extending at72ħfor45s．The samples were detected by Real-time PCR．U6as housekeep-ing gene was determined by using fluorescent quantitative PCR．The relative expression quantity was calculated by using 2－△△Ct method．The cycle threshold（Ct）indicates the cycle number when the fluorescence signal in each reaction tube reached the given threshold．△Ct=Average Ct of objective gene－Average Ct of housekeeping gene．△△Ct=△Ct （immature cattle）－△Ct（sexual－mature cattle）．And the detection results were analyzed according to the quantitative fluorescent amplification curves of samples and the analysis results by using SPSS13．0software．Target gene detection of miRNA-181bAll the determined target genes of miRNA from sexual-mature cattle at low expresssion level were predicted．The se-quences of3'-UTR region in the genomic gene of cattle and the sequences of cattle FSH gene were scanned with miRNA sequences by using the latest prediction software Tar-getScanS and the qualified miRNA was found．The probable target gene of miRNA-181b was analzyed on the baisis of the sequencing results．Results and AnalysisExtration of toal RNA from the anterior pituitary of Yanbian CattleAs shown Fig．1，three clear bands of28S，18S and5S were found in the profiles of agarose gel electrophoresis，which indicated that total RNA was sucessfully extracted from the anterior pituitary tissues of Yanbian Cattle．Because pro-lactin can only be expressed in the anterior pituitary，the ob-tained cDNA was used as template to amplify the specific fragment by PCR using the specific primers of objective gene．And the specific fragment with the length of901bp was ob-tained and it was accordant with the predicted objective gene （Fig．2），which showed that the extracted RNA was from the anterior pituitary tissues ofcattle．1and2are RNA products．Fig．1The results of agarose gel electrophoresis for RNA from anterior pituitary ofcattleM：D2000Marker；1and2are PCR products．Fig．2The results of agarose gel electrophoresis for PCR products of cDNAThe results of Solexa high-throughput deep sequencingThe sequencing of cDNA library from the anterior pituitary of bulls The consructed cDNA library has the qualified concen-tration，purity and integrity and the library can meet the se-quencing demands（Fig．3）．The sequencing results showed that634known miRNAs and42unknown miRNAs were found in the anterior pituitary of Yanbian Cattle．And it was found that there were279difference sites between18-month-old bulls and one-month-old bulls．Among them，166sites were highly ex-pressed and113sites were lowly expressed in18-month-old breeding bulls．The sequencing difference analysis results of miRNA-181b Based on the sequencing results，miRNA-181b with the differ-ential expression in test group was selected and analyzed．The results showed that the expression quantity of miRNA-181b from the anterior pituitary of one-month-old bulls was276．7006，which was4．05times as the expression quantity of miRNA-181b from the anterior pituitary of18-month-ld bulls．And there were significant differences between one month-old bulls and18-month-old bulls．2301Agricultural Science＆Technology Vol．12，No．7，2011Real-time fluorescent quantitative PCR results of miRNA-181bin different development stagesThe analysis of melting curves and amplification curves From Fig．4，it was seen that the peak values of different samples were basically accordant and miscellaneous peak siginal was not seen in the melting curves and amplification curves of miRNA-181b and U6after real-time fluorescent quantitativePCR．The selcted parameters for PCR was appropriate and the primer specifity and the amplification specifity of quantita-tive PCR were bothgood．A ：The detection results of cDNA library from the anterior pituitary of 18-month-old bulls ；B ：The detection results of cDNA library from theanterior pituitary of one-month-old bulls．Fig．3The detection results of cDNA library from anterior pituitary of 18-month-old and one-month-oldbullsFig．4The melting curves and amplification curves for the detection results of miRNA-181b by real-time quantitative PCRThe relative quantification results of the gene expression of miRNA-181b from the anterior pituitary of one-month-old and18-month-old Yanbian Cattle The relative expression situa-tions of mRNA of miRNA-181b from the anterior pituitary ofone-month-old and 18-month-old Yanbian Cattle were detec-ted by using real-time fluorescence quantitative PCR instru-ment （Germany Eppendorf Company ）．The detection results indicated that miRNA-181b was expressed in the anterior pitui-tary of one-month-old and 18-month-old Yanbian Cattle andthe relative expresion quantity of mRNA of miRNA-181b in theanterior pituitary of one-month-old Yanbian Cattle was 2．4times as that in the anterior pituitary of 18-month-old YanbianCattle （Table 1）．The expression situations were accordant with the sequencing results ，which validated the expression difference of miRNA-181b in the anterior pituitary of YanbianCattle in different developmental stages．Table 1Real-time quantitative RT-PCR results for the expressionquantity difference of miRNA-181b in cattle's anterior pitui-taryMonth-old Average Ct of miRNA-181b Average Ct of U6△Ct △△Ct 2－△△Ct 128．037．4320．60－1．252．41827．285．4321．85The prediction results of cattle miRNA-181bThe probable taget gene of miRNA-181b was predictedby using target gene prediction software TargetScan 4．0and the related target gene of miRNA-181b was screened out．Thebinding of miRNA-181b with 838－844bases in 3'untranslatedregion of FSH βgene was specific and the binding base sites were UGAAUGUA．ConclusionmicroRNA is a kind of small uncoding RNA from the own genome of eukaryotes ，which regulates more than one third of protein-coding genes and controls the growth ，development ，carcinogenesis and other physiological mechanisms of most organs．The recognization and functional analysis of miRNA has been one of research hotpoints in the modern molecular biology field ［12－13］．As a kind of glycoprotein hormone ，FSH βcan stimulate the spermatogenesis of male animals and regu-late the endometrium growth ，ovulation ，stimulation of the multi-follicle development and other functions．In recent years ，many researchers have studied the structure ，physico-chemical properties and main mechanisms of FSH gene．But the research on the question whether the post-transcriptionalregulation of FSH βis regulated by miRNA was not reported．The library of miRNA from the pituitary tissue can be construc-ted and the differential miRNA can be found by high-through-put deep sequencing analysis of miRNA from the pituitary tis-sue ，which has become an effective approach for the post-transcriptional regulation research of FSH β．The probable miRNA can be searched by using FSH βgene and more miRNA can be predicted．Based on the sequencing analysis ，the prediction efficiency of target gene can be greatly en-hanced．The regulation mechanism of miRNA was analyzed by using high-throughput sequencing technolgy and the pre-3301XU Cheng et al．Study on the Expression Laws and Regulatory Functions of miRNA-181b in the Anterior Pituitary of Cattlediction results of target gene，which could lay better theoreti-cal foundation for the post-transcriptional regulation research of FSHβ．References［1］LEE RC，FEINBAUM RL，AMBROS V，The C．elegans hetero-chronic gene lin－4encodes small RNAs with antisense comple-mentarity to lin－14［J］．Cell，1993，75（5）：843－854．［2］MARCELO A GERMAN，LUO SJ，GARY SCHROTH，et al．Con-struction of parallel analysis of rna ends（PARE）libraries for the study of cleaved miRNA targets and the RNA degradome［J］．Nat Protoc，2009，4（3）：356－362．［3］TIAN F，ZHANG HY，ZHANG XY，et al．miRAS：a data process-ing system for miRNA expression profiling study［J］．BMC Bioinfor-matics，2007，8：285．［4］WANG F（王芬），LI ZH（李朝晖）．A study on miRNA alternation af-ter H2O2－induced PC12cell apoptosis using microarray technique （H2O2诱导PC12细胞凋亡前后miRNA的变化及探讨）［J］．Chi-nese Journal of Forensic Medicine（法医学杂志），2007，23（5）：328－231．［5］LAJER CB，NIELSEN FC，FRIIS HANSEN L，et al．Different miR-NA signatures of oral and pharyngeal squamous cell carcinomas：a prospective translational study［J］．Br J Cancer，2011，104（5）：830－840．［6］LI YM（李育敏），GU JY（谷景义），ZHU XJ（朱雪娇），et al．Study on the sensitivity of leukemic cells to arsenic trioxide enhanced by targeted suppression of mIRNA－21（靶向抑制微小RNA－21提高白血病细胞对三氧化二砷的敏感性研究）［J］．Chinese Journal of Integrated Traditional and Western Medicine（中国中西医结合杂志），2010，30（2）：170－173．［7］ZHENG HY（郑慧媛），WANG L（王兰），SU JL（苏军龙），et al．Culture and identification of rat anterior pituitary cells in vitro（大鼠腺垂体细胞的体外培养和鉴定）［J］．Shaanxi Medical Journal（陕西医学杂志），2010（6）：643－645．［8］WANG X（王新），TAN JH（谭建华），NAI XP（赖小平），et al．TheObservation of the primary culture in vitro of rats＇Adenopituicyes and their secretion of GTH（大鼠腺垂体细胞的体外原代培养及其GTH分泌活动的观察）［J］．Journal of Qingdao Agricultural Univer-sity（青岛农业大学学报：自然科学版），2010（4）：305－308．［9］WANG HY（王海燕），GUI SQ（归绥琪），XU CJ（徐丛剑），et al．Role of prolactin－releasing peptide in pregnancy maintained by regulation of pituitary prolactin（早孕大鼠下丘脑催乳素释放肽的表达及对血催乳素分泌的调节）［J］．Shanghai Medical Journal（上海医学），2006（1）：22－25．［10］WANG P（王苹），FU T（付涛），WANG XR（王绪锐），et al．Prima-ry，study of miRNA expression pattens in laryngeal carcinoma bymicroarray（应用微阵列芯片分析喉鳞状细胞癌miRNA与正常黏膜表达差异的初步研究）［J］．Journal of Clinical Otorhinolaryngology（临床耳鼻咽喉头颈外科杂志），2010，24（12）：535－538．［11］GOU DM，ZHANG HH，BAVISKAR PS，et al．Primer extension－based method for the generation of a siRNA/miRNA expressionvector［J］．Physiol Genomics，2007，31（3）：554－562．［12］WILFRED BR，WANG WX，NELSON PT．Energizing miRNA re-search：a review of the role of miRNAs in lipid metabolism，with aprediction that miR－103/107regulates human metabolic path-ways［J］．Mol Genet Metab，2007，91（3）：209－217．［13］ZHANG T（张涛），CHENG T（程通），WEI LH（魏丽华），et al．Construction of highly effective artifical miRNA targeted to HIV－1vif and the lentiviral-mediated antiviral research in vitro（靶向HIV－1vif的高效人工miRNA的构建及慢病毒介导的体外抗病毒研究）［J］．Chinese Journal of Virology（病毒学报），2010，26（1）：8－15．［14］SUN GJ，PAN J，LIU KC，et al．Real-time quantitative PCR anal-ysis of vascular endothelial growth factor receptor-2（VEGFR-2）expression at zebrafish different developmental stages［J］．Agri-cultural Science＆Technology，2010，11（4）：118－120．［15］LI XB，CANG M．The study on the gene expression of preimplant-ation IVF bovine embryos［J］．Agricultural Science＆Technology，2010，11（5）：93－95，111．Responsible editor：CHEN Yu-min Responsible proofreader：檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪WU Xiao-yan miRNA-181b在牛腺垂体中的表达规律及其调控作用研究（摘要）许橙1，薛文志1，赵茂鑫1，陈信1，刘颖1，于汪洋1，许艳丽1，马腾壑1，高妍2，张嘉保2，袁宝2*（1．吉林大学畜牧兽医学院，吉林长春130062；2．吉林大学实验动物中心，吉林长春130062）［目的］探索miRNA-181b在成熟（18月龄）和未成熟（1月龄）牛腺垂体中差异表达量及其调控功能。

急性髓系白血病患者miRNA-181b表达特点及预后意义

急性髓系白血病患者miRNA-181b表达特点及预后意义向立丽;李曼;顾伟英;魏江;岑建农;陈子兴;贺白;刘琰;谢晓宝【期刊名称】《临床内科杂志》【年(卷),期】2013(030)006【摘要】目的探讨微小RNA-181b(miR-181b)在急性髓性白血病(AML)中的表达特点及预后意义.方法采用实时定量逆转录-聚合酶链反应(RT-PCR)检测158例初诊AML患者及20例健康供者骨髓单个核细胞中miR-181b的表达水平.同时采用基因组DNA-PCR结合测序方法检测158例AML患者核磷蛋白1(NPM1)基因第12号外显子突变和fms样酪氨酸激酶3(FLT3)基因内部串联重复(ITD)突变(FLT3-ITD).结果 AML中miR-181b表达水平较正常对照组显著升高(Z =-2.386,P=0.017),各FAB亚型中M1、M5及M6型miR-181b表达水平较对照组明显升高(P＜0.05).miR-181b高表达与低血红蛋白、高乳酸脱氢酶及NPMI野生型有关.miR-181b高表达完全缓解率较低(x2 =7.717,P=0.005)、总生存期较短(P ＜0.05).结论 miR-181b在AML某些亚型中高表达,miR-181b高表达是AML患者不利的预后因素.【总页数】3页(P417-419)【作者】向立丽;李曼;顾伟英;魏江;岑建农;陈子兴;贺白;刘琰;谢晓宝【作者单位】213003江苏常州,苏州大学附属第三医院常州市第一人民医院血液科;213003江苏常州,苏州大学附属第三医院常州市第一人民医院血液科;213003江苏常州,苏州大学附属第三医院常州市第一人民医院血液科;213003江苏常州,苏州大学附属第三医院常州市第一人民医院综合实验室;苏州大学附属第一医院江苏省血液病研究所白血病研究室;苏州大学附属第一医院江苏省血液病研究所白血病研究室;213003江苏常州,苏州大学附属第三医院常州市第一人民医院血液科;213003江苏常州,苏州大学附属第三医院常州市第一人民医院血液科;213003江苏常州,苏州大学附属第三医院常州市第一人民医院血液科【正文语种】中文【相关文献】1.急性髓系白血病患者 FLT3- ITD 和 NPM1基因突变表达及在判断预后中的意义[J], 俞罡;陈静桂2.急性髓系白血病患者CEBPA和C-KIT基因突变表达及在判断预后中的意义 [J], 潘亚宁3.fas／apo—1,bcl—2蛋白在急性髓系白血病细胞的表达特点及意义 [J], 胡丽华;周剑峰4.WT1基因在成人初诊急性髓系白血病不同预后分层中的表达特点 [J], 邵晶璇;谢国然;陈宏5.急性髓系白血病细胞粘附分子VLA-4、LFA-1表达特点及临床意义 [J], 刘廷析;刘秀芳;邹萍;向建平;陈燕;喻东姣;李崇渔因版权原因，仅展示原文概要，查看原文内容请购买。

高原肺水肿患者血浆中microRNA-181b的水平检测及意义

高原肺水肿患者血浆中microRNA-181b的水平检测及意义曹景新;赵海玲;陈强;胡啟英;李玉斌【期刊名称】《高原科学研究》【年(卷),期】2017(001)001【摘要】目的：检测高原肺水肿患者（HAPE）血浆中microRNA-181b（miR-181b）的水平变化，探讨其临床学意义。

方法：采用颈环法实时定量逆转录聚合酶链式反应（RT-qPCR）扩增检测30例轻度HAPE患者、26例中度HAPE患者7245例健康对照血浆中miR-181b水平，分析miR-181b与疾病临床分级之间关系，探讨其作为早期诊断标志物的可行性。

结果：三组之间miR-181b表达水平存在显著差异（P〈0．01），miR-181b与HAPE临床分期之间存在着较强的相关性（r=0．781，P〈0．001），miR-181b对HAPE诊断及疾病分层有很好的诊断效能（AUC=0．961，AUC=0．921，P〈0．001）。

结论：miR-181b可能参与HAPE的发生发展过程，且具有作为HAPE早期诊断生物学标志物的潜能。

【总页数】6页(P83-88)【作者】曹景新;赵海玲;陈强;胡啟英;李玉斌【作者单位】西藏阜康医院呼吸内科,西藏拉萨850000;西藏阜康医院呼吸内科,西藏拉萨850000;西藏阜康医院呼吸内科,西藏拉萨850000;西藏阜康医院呼吸内科,西藏拉萨850000;西藏阜康医院呼吸内科,西藏拉萨850000【正文语种】中文【中图分类】R594.3【相关文献】1.高原肺水肿患者血浆ET和SOD测定及其意义 [J], 邱光华;嘎布;陈荣华;张世馥;周建平;隋岫兰2.结肠癌患者血浆中 microRNA-181b水平的检测及意义 [J],3.microRNA-181b在胰腺癌患者血浆中的表达及意义 [J], 周晋航;王江华;陈先祥;吴黎明;吴士兴;王伟4.高原肺水肿患者血浆心钠素升高及其病理生理学意义 [J], 杨华民5.MicroRNA-181b和microRNA-130a在冠状动脉粥样硬化性心脏病患者血浆中的表达及临床意义 [J], 谢岩;王月香;席燕;任志文因版权原因，仅展示原文概要，查看原文内容请购买。

具核梭杆菌通过调控miR-181b在结肠癌细胞中形成炎性微环境的机制

背景 : 具核梭杆菌 (F n ) 是口腔常见的致病菌，研究发现 F n 与结直肠癌的发生、发展密切相关，尤其是炎症相关性结直肠癌。目的 : 探讨 F n 感染在结肠癌细胞中形成炎性微环境的机制。方法 :构建 F n 感染 Caco-2 细胞的炎症
miRNA 测序。将 miR-181b mimics 或 inhibitor 转染 F n 感染的 Caco-2 细胞。以 qRT-PCR 和蛋白质印迹法分别检测 TNF-a mRNA 和蛋白表达，ELISA 法检测上清液中 T N F -a 含量，Transwell 小室法检测穿膜淋巴细胞数。结果 :F n 组 TNF-a mRNA 和蛋白表达均显著高于对照组 (P < 0 .0 5 ) ，上清液中 T N F-a 含量显著升高（ P < 0 .0 5 ) ，穿膜淋巴细胞数量明显增多 ( 尸< 0 .0 5 ) 。miRNA 测序和 qRT-PCR 结果均显示，F n 组 miR-181b 表达较对照组显著降低（尸< 0 .0 5 ) 。与对照组相比，miR-181b m im ics+ F n 组 TNF-a mRNA 和蛋白表达显著降低（ P < 0 .0 5 ) ; 而 miR181binhibitor 组 TNF-a mRNA 和蛋白表达均显著升高 (P < 0 .0 5 ) 。生物信息学工具和双荧光素酶检测证实 TNF-a 可能为 Caco-2 细胞中 miR-181b 的靶基因。结论 :F n 通过抑制 Caco-2 细胞中 miR-181b 表达而上调 T N F-a 表达，募

MiRNA在免疫炎症反应中的调节作用研究

MiRNA在免疫炎症反应中的调节作用研究近年来，微小核酸(miRNA)的发现和研究成为了生命科学领域的热点之一。

它们是一种长度在20-22个核酸的短链RNA分子，通过与mRNA分子的互补结合，影响目标基因的表达。

研究表明，miRNA在肿瘤、心血管等疾病的发生和发展中起到了重要的调节作用。

除此之外，当前越来越多的研究表明，在免疫炎症反应过程中，miRNA也具有重要的调节作用。

免疫炎症反应是机体对外界刺激的一种保护性反应。

它伴随着一系列复杂的生化过程，包括突破细胞膜、核内转录、翻译过程等。

在这个过程中，miRNA就是一个具有重要作用的因子之一。

miRNA的表达调控主要通过切割、降解或抑制靶基因的翻译等方式实现。

其中，miRNA-132是一种调节免疫炎症反应的miRNA之一，它可以通过控制T细胞、B细胞的活性，参与调节免疫炎症反应的发生过程。

在过去的研究中，研究人员发现，当机体的免疫系统遭遇外界刺激而产生炎症反应时，miRNA-132的表达会明显上升。

此时，miRNA-132可以抑制T细胞和B细胞的活性，降低它们分泌的炎性因子，从而减缓机体的炎症反应。

这表明，miRNA-132在免疫炎症反应分子调节过程中起到了重要的作用。

此外，还有一些其他的miRNA也与免疫炎症反应有关。

例如，miRNA-21、miRNA-146等都可以参与免疫炎症反应的调节过程。

研究表明，当这些miRNA的表达水平发生改变时，机体的炎症反应也会出现相应的变化。

因此，对这些miRNA的调节机制进行研究，对于深入了解免疫炎症反应的机理具有重要的意义。

miRNA的研究展现出了它在免疫炎症反应中的调节作用，但目前的研究尚处于探索阶段。

现阶段，研究人员主要通过miRNA高通量检测、体外和体内功能检测等方法，来鉴定和筛选与免疫炎症反应有关的miRNA，进而深入研究其调节机制。

为了更好地了解miRNA的免疫炎症反应调控作用，还需要进行大规模的基础研究和临床研究。

microRNA-181b

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miR-181b_miR-210_miR-126在妊娠期高血压疾病中的表达及与炎症反应的相关性

ʌ文章编号ɔ1006-6233(2024)01-0147-06miR -181b miR -210miR -126在妊娠期高血压疾病中的表达及与炎症反应的相关性刘㊀洋,㊀赵巧棉,㊀李海燕,㊀张思宇,㊀杨㊀颖,㊀宫㊀月(河北省承德市妇幼保健院生殖中心,㊀河北㊀承德㊀067000)ʌ摘㊀要ɔ目的:探究微小RNA -181b (miR -181b )㊁miR -210㊁miR -126在妊娠期高血压疾病(HDP )中的表达及与炎症反应的相关性㊂方法:选取2018年1月至2019年12月承德市妇幼保健院收治的213例HDP 单胎孕妇作为观察组,以病情严重程度为依据将其设为妊娠期高血压(GH 组,62例)㊁轻度子痫前期(MP 组,90例)及重度子痫前期(SP 组,61例)3个亚组;另选取同期同医院收治的78名健康孕妇作为对照组㊂检测各组miR -181b ㊁miR -210㊁miR -126相对表达量及血清白介素-1β(IL-1β)㊁白介素-6(IL -6)㊁白介素-10(IL -10)㊁白介素-17(IL -17)㊁肿瘤坏死因子-α(TNF -α)㊁单核细胞趋化蛋白-1(MCP -1)及血管细胞黏附分子-1(VCAM -1)水平,并分析HDP 患者各指标间的相关性㊂结果:观察组miR -181b ㊁miR -210相对表达量及IL -1β㊁IL -6㊁IL -17㊁TNF -α㊁MCP -1㊁VCAM -1均比对照组高(P <0.05);观察组miR -126相对表达量及IL -10均比对照组低(P <0.05)㊂GH 组miR -181b ㊁miR -210相对表达量及IL -1β㊁IL -6㊁IL -17㊁TNF -α㊁MCP -1㊁VCAM -1均比MP 组及SP 组低,且MP 组比SP 组低(P <0.05);GH 组miR -126相对表达量及IL -10均比MP 组及SP 组高,且MP 组比SP组高(P <0.05)㊂Pearson 相关性分析显示,HDP 患者miR -181b ㊁miR -210相对表达量与IL -1β㊁IL -6㊁IL -17㊁TNF -α㊁MCP -1㊁VCAM -1均呈正相关,与IL -10呈负相关(P <0.05);HDP 患者miR -126相对表达量与IL -1β㊁IL -6㊁IL -17㊁TNF -α㊁MCP -1㊁VCAM -1均呈负相关,与IL -10呈正相关(P <0.05)㊂结论:miR -181b ㊁miR -210在HDP 中呈高表达,miR -126在HDP 中呈低表达,且三者与HDP 炎症反应密切相关㊂ʌ关键词ɔ㊀妊娠期高血压疾病;㊀微小RNA -181b ;㊀微小RNA -210;㊀微小RNA -126ʌ文献标识码ɔ㊀A㊀㊀㊀㊀㊀ʌdoi ɔ10.3969/j.issn.1006-6233.2024.01.029Expression of miR -181b miR -210and miR -126in Pregnancy -Induced Hypertensive Disorders and Their Correlation with Inflammatory ResponseLIU Yang ,ZHAO Qiaomian ,LI Haiyan ,et al(Chengde Maternal and Child Health Hospital ,Hebei Chengde 067000,China )ʌAbstract ɔObjective :To explore the expressions of serum micrornA -181b (miR -181b ),miR -210and miR -126in hypertensive diseases in pregnancy (HDP )and its correlation with inflammation response.Methods :A total of 213cases of singleton pregnant women with HDP treated at Chengde Maternal and ChildHealth Hospital from January 2018to December 2019were selected as the observation group.Based on the se-verity of the condition ,they were divided into gestational hypertension (GH group ,62cases ),mild pre-eclampsia (MP group ,90cases ),and severe preeclampsia (SP group ,61cases ).Additionally ,78healthypregnant women treated during the same period in the same hospital were selected as the control group.The relative expression levels of miR -181b ,miR -210,and miR -126,as well as serum interleukin -1β(IL -1β),interleukin -6(IL -6),interleukin -10(IL -10),interleukin -17(IL -17),tumor necrosis factor -α(TNF -α),monocyte chemoattractant protein -1(MCP -1),and vascular cell adhesion molecule -1(VCAM -1)levels were detected.The correlation between various indicators in HDP patients was analyzed.Results :The relative expression levels of miR -181b ,miR -210,IL -1β,IL -6,IL -17,TNF -α,MCP -1,and㊃741㊃ʌ基金项目ɔ2021年承德市科学技术研究与发展计划项目,(编号:202109A011)ʌ通讯作者ɔ宫㊀月VCAM-1in the observation group were higher than those in the control group(P<0.05).The relative ex-pression level of miR-126and IL-10in the observation group were lower than those in the control group(P< 0.05).In the GH group,the relative expression levels of miR-181b,miR-210,IL-1β,IL-6,IL-17, TNF-α,MCP-1,and VCAM-1were lower than those in the MP and SP groups,and the MP group was low-er than the SP group(P<0.05).The relative expression level of miR-126and IL-10in the GH group were higher than those in the MP and SP groups,and the MP group was higher than the SP group(P<0.05). Pearson correlation analysis showed that the relative expression levels of miR-181b,miR-210in HDP pa-tients were positively correlated with IL-1β,IL-6,IL-17,TNF-α,MCP-1,and VCAM-1,and negatively correlated with IL-10(P<0.05).The relative expression level of miR-126in HDP patients was negatively correlated with IL-1β,IL-6,IL-17,TNF-α,MCP-1,and VCAM-1,and positively correlated with IL-10 (P<0.05).Conclusion:miR-181b,miR-210are highly expressed,and miR-126is lowly expressed in HDP.All three are closely related to the inflammatory response in HDP.ʌKey wordsɔ㊀Hypertensive disorders in pregnancy;㊀MicrornA-181b;㊀MicrornA-210;㊀MicrornA -126㊀㊀妊娠期高血压疾病(Hypertensive Disorders in Pregnancy,HDP)作为产科常见并发症之一,亦是孕产妇死亡重要原因[1]㊂HDP病因及发病机制目前尚未十分明确,临床表现主要为蛋白尿及高血压,症状严重者可出现多系统脏器损伤,严重威胁母婴健康㊂因此,探寻HDP发病机制对其诊疗意义重大㊂微小RNA (microRNA,miR)属于基因调控分子,可影响众多蛋白编码基因输出,参加大多数细胞因子免疫应答及转录调控[2]㊂有研究指出,miR表达和胎盘病理生理发育存在一定关联,成熟miR可参与滋养细胞增殖㊁凋亡等过程[3]㊂miR-181b㊁miR-210㊁miR-126均为临床常见的miR分子,部分学者发现其在HDP发病过程中起着重要作用[4]㊂国内外研究指出,炎症反应贯穿HDP发生发展过程,其中炎症因子与HDP病情变化密切相关[5-6]㊂目前虽有一些研究探索miR与HDP 患者炎症因子的关系,但关于miR-181b㊁miR-210㊁miR-126在HDP中的表达及与7种炎症因子水平关系的研究报道较少㊂为此,本研究探究miR-181b㊁miR-210㊁miR-126在HDP中的表达及与炎症反应的相关性㊂现报道如下㊂1㊀资料与方法1.1㊀一般资料:选取2018年1月至2019年12月承德市妇幼保健院收治的213例HDP单胎孕妇作为观察组,以病情严重程度为依据将其设为妊娠期高血压(GH组,62例)㊁轻度子痫前期(MP组,90例)及重度子痫前期(SP组,61例)3个亚组㊂纳选标准:①与HDP相关诊断标准相符[7];②妊娠期产检规律,资料齐全;③单胎妊娠;④知情同意㊂排除标准:①胎儿先天畸形的孕妇;②孕妇或胎儿有病毒感染;③妊娠前存在自身免疫性疾病㊁血液系统疾病㊁糖尿病或高血压;④存在重要脏器严重病变㊂另选取同期同医院收治的78名健康孕妇作为对照组㊂四组一般资料均衡可比(P>0.05),见表1㊂本研究经医院医学伦理委员会批准㊂表1㊀四组一般资料比较( xʃs)组别例数年龄(岁)孕前体质指数(kg/m2)孕周(周)孕次(次)GH组6228.95ʃ4.0822.38ʃ2.1737.35ʃ1.42 1.27ʃ0.25 MP组9029.61ʃ3.1522.91ʃ2.3937.64ʃ1.53 1.31ʃ0.28 SP组6129.37ʃ3.4722.25ʃ2.0837.19ʃ1.68 1.24ʃ0.21对照组7829.92ʃ3.8222.63ʃ2.2137.24ʃ1.39 1.26ʃ0.26 F0.885 1.278 1.454 1.043㊃841㊃P0.4490.2820.2270.3741.2㊀方㊀法1.2.1㊀标本采集:所有受检者清晨抽5mL空腹静脉血,在常温下放置30min后,经10min离心(3500r/min 转速,13.5cm离心半径)后取上清液,置于-80ħ冰箱待检㊂1.2.2㊀miR-181b㊁miR-210㊁miR-126相对表达量检测:实施实时荧光定量PCR法检测,所用仪器为美国ABI公司生产的7500型荧光定量PCR仪,所用试剂盒及缓冲液㊁反转录酶㊁dNTPs购自美国Ambio公司㊂取1份待测上清液,在其中加入Trizol RNA1mL进行总RNA提取,利用紫外分光光度计初步判断总RNA 纯度,若在1.8~2.0间则提示纯度较好㊂利用反转录试剂盒对总RNA进行反转录为cDNA,其中miR-181b上游引物5'-AACATTCATTGCTGTCGGTGGG-3 ',下游引物5'-GCGAAGCACAGAATTAATACGACT-CAC-3';miR-210上游引物5'-CAATAACTGTGCGT-GTGACAGC-3',下游引物5'-TATGGTTGT-TCTCGTCTCCTTCTC-3';miR-126上游引物5'-TCG-TACCGTGAGTAATAATGCG-3',下游引物5'-CGCA-TI ATI1ACTCACGGI1AC-3';内参基因U6上游引物5 '-ATTGGAACGATACAGAGAAGATT-3',下游引物5'-GGAACGCTTCACGAATTTG-3'㊂反转录反应体系: 5μL模板RNA,3μL miRNA特异性茎环引物及内参基因U6,0.15μL100mmoL/LdNTPs,1μL50U/μL反转录酶,1.5μL10ˑ反转录缓冲液,0.19μL20U/μL RNase抑制剂,4.16μL无菌不含酶蒸馏水㊂将提取的cDNA保存于4ħ冰箱中㊂qRT-PCR检测:模板为cD-NA,建立qRT-PCR反应体系20μL(10μL2ˑTaqMan 通用混合物溶液,1μL20ˑ引物及探针Mix,1.33μL cDNA模板,7.67μL无核酸酶蒸馏水)㊂反应条件为16ħ30min;42ħ30min;85ħ5min㊂扩增条件为95ħ10min1个循环,95ħ15s㊁60ħ60s45个循环㊂实验重复3次后取平均值,miR-181b㊁miR-210㊁miR-126相对表达量运用2-әәCt法进行计算,әCt=Ct(目的基因)-Ct(U6)㊂1.2.3㊀炎症因子检测:取1份待测上清液,血清白介素-1β(Interleukin-1β,IL-1β)㊁白介素-6(Interleukin -6,IL-6)㊁白介素-10(Interleukin-10,IL-10)㊁白介素-17(Interleukin-17,IL-17)㊁肿瘤坏死因子-α(Tumor Necrosis Factor-α,TNF-α)㊁单核细胞趋化蛋白-1(Monocyte Chemoattractant Protein-1,MCP-1)及血管细胞黏附分子-1(vascular-cellular adhesion mole-cule-1,VCAM-1)水平实施酶联免疫法测定,操作严格按试剂盒说明书要求开展㊂IL-1β㊁IL-6㊁IL-10㊁IL -17㊁TNF-α检测试剂盒购自上海科新生物技术股份有限公司,MCP-1㊁VCAM-1检测试剂盒购自英国Ab-cam公司㊂1.3㊀统计分析:将数据录入软件SPSS25.0分析㊂符合正态分布的计量资料用描述,行独立样本t检验;多组间比较行单因素方差分析,进一步两两比较采用LSD-t检验;相关性分析采用Pearson相关分析;检验水准α=0.05㊂2㊀结㊀果2.1㊀两组miR-181b㊁miR-210㊁miR-126相对表达量及炎症因子水平比较:观察组miR-181b㊁miR-210相对表达量及IL-1β㊁IL-6㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1均比对照组高(P<0.05);观察组miR-126相对表达量及IL-10均比对照组低(P<0.05)㊂见表2㊂表2㊀两组miR-181b miR-210miR-126相对表达量及炎症因子水平对比( xʃs)指标观察组(n=213)对照组(n=78)t P miR-181b10.55ʃ1.37 1.89ʃ0.5154.413<0.001 miR-210 5.69ʃ1.03 1.19ʃ0.3337.844<0.001 miR-12622.71ʃ3.4831.16ʃ4.2917.195<0.001 IL-1β(ng/L)17.63ʃ3.24 2.81ʃ0.8639.846<0.001 IL-6(ng/L)113.93ʃ20.2291.04ʃ19.158.674<0.001㊃941㊃IL-10(ng/L)14.48ʃ4.6131.96ʃ6.8424.936<0.001 IL-17(μg/L)11.79ʃ1.95 5.71ʃ1.0526.164<0.001 TNF-α(ng/L)39.25ʃ8.9612.37ʃ2.3126.152<0.001 MCP-1(mg/mL) 1.31ʃ0.300.38ʃ0.1126.704<0.001 VCAM-1(mg/L)570.22ʃ85.21402.53ʃ65.4215.757<0.0012.2㊀不同HDP亚组miR-181b㊁miR-210㊁miR-126相对表达量及炎症因子水平比较:GH组miR-181b㊁miR-210相对表达量及IL-1β㊁IL-6㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1均比MP组及SP组低,且MP组比SP组低(P<0.05);GH组miR-126相对表达量及IL-10均比MP组及SP组高,且MP组比SP组高(P<0.05)㊂见表3㊂表3㊀不同HDP亚组miR-181b miR-210miR-126相对表达量及炎症因子水平对比( xʃs)指标GH组(n=62)MP组(n=90)SP组(n=61)F P miR-181b 5.81ʃ1.259.92ʃ2.03a16.28ʃ2.96ab364.771<0.001 miR-210 3.47ʃ1.02 5.69ʃ1.27a7.96ʃ2.08ab139.495<0.001 miR-12628.14ʃ3.2623.27ʃ3.61a16.38ʃ4.12ab159.750<0.001 IL-1β(ng/L)11.31ʃ2.4917.73ʃ4.08a23.89ʃ6.11ab124.673<0.001 IL-6(ng/L)83.41ʃ18.52113.28ʃ16.49a145.91ʃ22.13ab169.357<0.001 IL-10(ng/L)19.17ʃ5.1114.39ʃ3.09a9.85ʃ2.27ab101.961<0.001 IL-17(μg/L)7.49ʃ1.8811.77ʃ2.19a16.18ʃ2.25ab257.674<0.001 TNF-α(ng/L)19.31ʃ4.3535.77ʃ6.41a64.65ʃ9.47ab670.635<0.001 MCP-1(mg/mL)0.95ʃ0.17 1.41ʃ0.27a 1.53ʃ0.39ab71.830<0.001 VCAM-1(mg/L)428.37ʃ96.44585.29ʃ80.37a692.15ʃ84.17ab145.703<0.001㊀㊀注:相比GH组,aP<0.05;相比MP组,bP<0.052.3㊀HDP患者miR-181b㊁miR-210㊁miR-126相对表达量与炎症因子的相关性分析:Pearson相关性分析显示,HDP患者miR-181b㊁miR-210相对表达量与IL-1β㊁IL-6㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1均呈正相关,与IL-10呈负相关(P<0.05);HDP患者miR-126相对表达量与IL-1β㊁IL-6㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1均呈负相关,与IL-10呈正相关(P<0.05)㊂见表4㊁图1㊂表4㊀HDP患者miR-181b㊁miR-210㊁miR-126相对表达量与炎症因子的相关性分析指标miR -181br㊀㊀㊀㊀㊀㊀㊀㊀PmiR-210r㊀㊀㊀㊀㊀㊀㊀㊀PmiR-12r㊀㊀㊀㊀㊀㊀㊀㊀PIL-1β0.760<0.0010.566<0.001-0.585<0.001 IL-60.676<0.0010.736<0.001-0.7117<0.001 IL-10-0.781<0.001-0.724<0.0010.639<0.001㊃051㊃IL-170.649<0.0010.776<0.001-0.677<0.001 TNF-α0.829<0.0010.794<0.001-0.752<0.001 MCP-10.684<0.0010.749<0.001-0.750<0.001VCAM-10.762<0.0010.710<0.001-0.833<0.001图1㊀HDP患者miR-181b㊁miR-210㊁miR-126相对表达量与炎症因子的相关性分析散点图注:A~G分别为miR-181b相对表达量与IL-1β㊁IL-6㊁IL -10㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1的散点图;H~N分别为miR-210相对表达量与IL-1β㊁IL-6㊁IL-10㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1的散点图;O~U分别为miR-126相对表达量与IL-1β㊁IL-6㊁IL-10㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1的散点图3㊀讨㊀论HDP主要发生于妊娠期女性,大部分患者病情多轻微,早期症状不显著,少数患者可发生昏迷㊁抽搐等,危及母婴生命安全[8]㊂HDP发病机制非常复杂,多数学者认为和血管内皮受损㊁免疫失衡等因素有关[9-10]㊂因此,探寻HDP发病机制,寻找有效监测指标对其预防㊁诊断及治疗意义重大㊂miR属于内源性非编码RNA,于真核生物中存在广泛,可通过调节细胞基因表达对生物体生理活动进行干预㊂Hromadnikova等[11]指出,HDP发病可能和miR异常表达有关,miR对胎盘发育具有维持正常作用,可能成为HDP重要评估指标㊂miR-181b属于miR,当机体出现血管内皮损伤或炎症反应时其表达异常,可参与心血管疾病等发病过程㊂杨娜等[12]指出,miR-181b在HDP中呈高表达,可介导HDP发病过程㊂miR-210作为一种存在于人体11号染色体上的缺氧激活因子,当机体缺血缺氧时其呈高表达,可促使血管新生㊂Frazier等[13]报道,miR-210参与子痫前期发病过程,可通过介导机体炎症反应促进病情进展㊂miR-126属于与内皮细胞相关miR,对内皮祖细胞增殖分化具有促进作用,可使内皮细胞保持稳定,确保血管完整性㊂何晓焱等[14]指出,miR-126在HDP中表达下调,通过介导血管生成影响HDP发生发展㊂本研究中,观察组miR-181b㊁miR-210相对表达量比对照组高,miR-126相对表达量比对照组低,表明miR-181b㊁miR-210㊁miR-126参与HDP发病过程㊂炎症反应作为血管生成及损伤的重要调控因素,其与HDP发生发展存在一定关联㊂IL-1β㊁IL-6㊁IL-17属于促炎因子,其中IL-1β不仅对IL-6具有诱导作用,还能对血管内皮细胞及胎盘进行直接或间接作用,致使细胞损害;IL-6对Th1/Th2免疫平衡具有重要维持作用;IL-17由T细胞释放而成,可利用对内皮细胞等诱导作用促使IL-6等细胞因子产生,进而参与炎症反应㊂IL-10属于抑炎因子,对IL-6等促炎因子具有抑制作用,可调节机体免疫功能㊂TNF-α属于免疫调节因子,可参与机体炎症反应,诱导促炎因子释放㊂MCP-1属于趋化因子CC家族,不仅可结合血中单核细胞,并促使其转变为巨噬细胞,还可使血管内皮受损,降低一氧化氮功能,导致血管痉挛等㊂VCAM-1属于免疫球蛋白超家族,在细胞因子活化内皮细胞表面表达㊂当人体正常妊娠时,VCAM-1参与血管内皮与滋养细胞间黏附过程,有利于胎盘发育㊂本研究中,观察组IL-1β㊁IL-6㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1㊃151㊃比对照组高,IL-10比对照组低,表明炎症反应参与HDP发病过程㊂本研究中,GH组miR-181b㊁miR-210相对表达量及IL-1β㊁IL-6㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1均比MP组及SP组低,且MP组比SP组低;GH组miR-126相对表达量及IL-10均比MP组及SP组高,且MP组比SP组高,说明HDP患者病情愈重,miR-181b㊁miR-210表达及促炎因子水平愈高,miR-126表达及抑炎因子水平愈低㊂本研究进一步行相关性分析,发现HDP患者miR-181b㊁miR-210相对表达量与IL-1β㊁IL-6㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1均呈正相关,与IL-10呈负相关;HDP患者miR-126相对表达量与IL-1β㊁IL-6㊁IL-17㊁TNF-α㊁MCP-1㊁VCAM-1均呈负相关,与IL-10呈正相关,表明miR-181b㊁miR -210㊁miR-126可能通过与机体炎症反应相互作用而参与HDP发病过程㊂分析原因,HDP发病导致机体促炎因子大量释放,炎症反应被激活,血管内皮细胞受损,增加血管通透性,导致脏器缺血缺氧,加剧病情㊂miR-181b通过内皮细胞信号通路诱导促炎因子释放,使机体炎症反应加重,导致血管内皮受损,从而参与HDP发生过程㊂miR-210通过相应靶位将相应信号通路激活,引发机体氧化应激,致使机体免疫失衡,大量促炎因子释放,而抑炎因子释放受抑制,从而加剧机体炎症反应,参与HDP发生过程㊂miR-126通过靶基因PI3KR2㊁SPRED抑制PI3K㊁MAPK信号通路,阻止血管生成,致使母胎界面血供不足,胎盘结构出现异常,一些细胞碎片被释放而引起机体炎症反应,最终导致HDP发病㊂综上所述,miR-181b㊁miR-210在HDP中呈高表达,miR-126在HDP中呈低表达,且三者与HDP炎症反应密切相关㊂ʌ参考文献ɔ[1]㊀Lama S,Gurung P,Malla AP.Pregnancy induced hyperten-sive disorders among patients admitted to the department ofobstetric and gynecology in a tertiary care centre:a descrip-tive cross-sectional study[J].JNMA Nepal Med Assoc,2023,61(259):195-199.[2]㊀赵媛媛,王妙英,董有伟,等.妊娠期高血压疾病患者血清微小RNA-124-3p的表达及其临床意义研究[J].中国性科学,2022,31(3):116-120.[3]㊀Gu F,Lu D,Zhang L.MicroRNA-30a contributes to pre-ec-lampsia through regulating the proliferation,apoptosis,andangiogenesis modulation potential of mesenchymal stem cellsby targeting AVEN[J].Bioengineered,2022,13(4):8724-8734.[4]㊀Jin Y,Jia T,Wu X,et al.The predictive value of microRNAin early hypertensive disorder complicating pregnancy(HD-CP)[J].Am Transl Res,2021,13(6):7288-7293. [5]㊀任静华,季景环,孙志敏,等.妊娠期高血压患者凝血功能,炎性因子及血脂水平变化及临床意义[J].解放军医药杂志,2021,33(1):90-93.[6]㊀Feist H,Bajwa S,Pecks U.Hypertensive disease,pretermbirth,fetal growth restriction and chronic inflammatory disor-ders of the placenta:experiences in a single institution with astandardized protocol of investigation[J].Arch Gynecol Ob-stet,2022,306(2):337-347.[7]㊀谢幸,苟文丽.妇产科学[M].第8版.北京:人民卫生出版社,2013.64-71.[8]㊀Lin JH,Lyu X.Difficulties and confusion concerning themanagement of hypertensive disorders in pregnancy--inter-pretation of the guidelines for the diagnosis and treatment ofhypertensive disorders in pregnancy(2020)[J].SichuanDa Xue Xue Bao Yi Xue Ban,2022,53(6):1007-1011.[9]㊀Zhao L,Xiong M,Liu Y.Baicalin enhances the proliferationand invasion of trophoblasts and suppresses vascular endo-thelial damage by modulating long non-coding RNANEAT1/miRNA-205-5p in hypertensive disorder complica-ting pregnancy[J].Obstet Gynaecol Res,2021,47(9):3060-3070.[10]㊀Higaki A,Mogi M.Dendritic cells as potential initiators ofimmune-mediated hypertensive disorders[J].HypertensRes,2022,45(3):527-529.[11]㊀Hromadnikova I,Kotlabova K,Krofta L.Cardiovascular dis-ease-associated microRNA dysregulation during the firsttrimester of gestation in women with chronic hypertensionand normotensive women subsequently developing gestation-al hypertension or preeclampsia with or without fetal growthrestriction[J].Biomedicines,2022,10(2):256. [12]㊀杨娜,王微,张芳,等.nesfatin-1㊁miR-1233-3p㊁miR-181b在妊娠期高血压疾病中的表达及相关性[J].中国计划生育学杂志,2021,29(9):1916-1919. [13]㊀Frazier S,McBride MW,Mulvana H,et al.From animalmodels to patients:the role of placental microRNAs,miR-210,miR-126,and miR-148a/152in preeclampsia[J].Clin Sci(Lond),2020,134(8):1001-1025.[14]㊀何晓焱,彭诗寒,赵磊,等.血浆miRNA-126表达与妊娠期高血压疾病的相关性分析[J].中国医药导报,2019,16(8):76-79.㊃251㊃。

脑胶质瘤中miR-181b的表达及临床意义

ＳｔａｔｅＫｅｙＬａｂｏｒｔｏａｒｙｆｏＯｎｃｏｌｏｇｙｉｎＳｏｕｔｈＣｈｉｎａ，ＤｅｐａｒｔｍｅｎｔｏｆＮｅｕｒｏｓｕｒｇｅｒｙ／Ｎｅｕｒｏ — ｏｎｃｏｌｏｇｙ，Ｓｕｎ
ｄｏｗｎｒｅｇｕｌａｔｅｄｉｎｓｅｖｅｒａｌｃａｎｃｅｒｔｙｐｅｓａｎｄｔｈｅａｂｅ￣ａｎｔｅｘｐｒｅｓｓｉｏｎｗａｓａｓｓｏｃｉａｔｅｄｗｉｔｈｄｒｕｇｒｅｓｉｓｔａｎｃｅ．ｍｉＲ一１８ｌｂｉｓｋｎｏｗｎｔｏｆｕｎｃｔｉｏｎａｓａｔｕｍｏｕｒｓｕｐｐｒｅｓｓｏｒｇｅｎｅａｎｄｓｅｌｖｅｓａｓａｐｒｏｇｎｏｓｔｉｃｍａｒｋｅｒｉｎｈｕｍａｎｇｌｉｏｍａ．ＴｈｅｐｕｒｐｏｓｅｏｆｔｈｉｓｓｔｕｄｙｉｓｔｏｅｖａｌｕａｔｅｍｉＲ一１８１ｂｅｘｐｒｅｓｓｉｏｎｉｎｇｌｉｏｍａｓ
瘤标本．以荧光定量ＰＣＲ法检测其中ｍｉＲ一１８１ｂ的表达．分析其与胶质瘤病理分级及患者预后的关系：利用ＣＣＫＩ８细胞毒性实验检测患者恶性胶质瘤细胞对替莫唑胺的敏感性．并分析其与ｍｉＲ１８１ｂ表达的关系。结果：ｍｉＲ．１８ｌｂ在胶质瘤组织中的表达显著低于正常脑组织＜０．０５１．ｍｉＲ一１８１ｂ的表达水平分别为：正常脑组织

miR-181基因家族在临床常见疾病中的研究进展

㊃综述㊃D O I:10.3969/j.i s s n.1672-9455.2023.16.026m i R-181基因家族在临床常见疾病中的研究进展*王春芳综述,柴富ә审校右江民族医学院附属医院检验科,广西百色533000摘要:微小R N A(m i R N A)是一类小的非编码R N A,广泛存在于真核生物体内,能够在转录后通过抑制目的基因信使R N A(m R N A)的翻译或促进m R N A降解来调节基因表达㊂m i R N A已成为多种生物过程的关键调控因子,各种调控机制不仅控制着它们的表达,还控制着它们的活性和生物利用度,这取决于m i R N A在人类相关蛋白质编码基因的潜在结合位点㊂m i R N A在免疫系统的发育和调控中也发挥着重要的作用,可以调控机体关键的细胞过程,包括细胞分化㊁血管生成和炎症反应的发生㊂现已发现很多生理过程和病理结果高度依赖于m i R N A,包括癌症㊁心血管疾病和代谢性疾病㊂其中,m i R-181基因家族在胚胎发育㊁细胞增殖㊁凋亡㊁自噬㊁线粒体功能和免疫反应等关键生物过程中发挥调节作用㊂该文综述了近年来m i R-181基因家族在临床疾病中的诊断和治疗方向及研究前沿,以期为m i R N A的临床疾病治疗提供新的思路㊂关键词:m i R-181;基因家族;生物标志物;靶标;疾病中图法分类号:R446.9文献标志码:A文章编号:1672-9455(2023)16-2416-05A d v a n c e s i n t h e s t u d y o f m i R-181g e n e f a m i l y i n c o m m o n c l i n i c a l d i s e a s e s*WA N G C h u n f a n g,C HA I F uәD e p a r t m e n t o f C l i n i c a l L a b o r a t o r y,t h e A f f i l i a t e d H o s p i t a l o f Y o u j i a n g M e d i c a l U n i v e r s i t yf o r N a t i o n a l i t i e s,B a i s e,G u a ng x i533000,Chi n aA b s t r a c t:m i c r o R N A(m i R N A)i s a k i n d o f s m a l l n o n-c o d i n g R N A w h i c h i s w i d e l y f o u n d i n e u k a r y o t i c o r-g a n i s m s a n d i s a b l e t o r e g u l a t e g e n e e x p r e s s i o n a f t e r t r a n s c r i p t i o n b y r e p r e s s i n g m e s s e n g e r R N A(m R N A) t r a n s l a t i o n o r p r o m o t i n g m R N A d e g r a d a t i o n.m i R N A h a v e e m e r g e d a s k e y r e g u l a t o r s o f a v a r i e t y o f b i o l o g i c a l p r o c e s s e s,a n d v a r i o u s r e g u l a t o r y m e c h a n i s m s c o n t r o l n o t o n l y t h e i r e x p r e s s i o n b u t a l s o t h e i r a c t i v i t y a n d b i o-a v a i l a b i l i t y,d e p e n d i n g o n t h e p o t e n t i a l b i n d i n g s i t e s o f m i R N A i n g e n e s e n c o d i n g r e l e v a n t h u m a n p r o t e i n s. m i R N A a l s o p l a y a n i m p o r t a n t r o l e i n t h e d e v e l o p m e n t a n d r e g u l a t i o n o f t h e i mm u n e s y s t e m,r e g u l a t i n g k e y c e l l u l a r p r o c e s s e s i n t h e b o d y,i n c l u d i n g c e l l d i f f e r e n t i a t i o n,a n g i o g e n e s i s a n d i n f l a mm a t i o n.M a n y p h y s i o l o g i c a l p r o c e s s e s a n d p a t h o l o g i c a l o u t c o m e s h a v e b e e n f o u n d t o b e h i g h l y d e p e n d e n t o n m i R N A,i n c l u d i n g c a n c e r,c a r-d i o v a s c u l a r a n d m e t a b o l i c d i s e a s e s.A m o n g t h e m,t h e m i R-181g e n e f a m i l y h a s i m p o r t a n t r o l e s i n t h e r e g u l a-t i o n o f k e y b i o l o g i c a l p r o c e s s e s s u c h a s e m b r y o n i c d e v e l o p m e n t,c e l l p r o l i f e r a t i o n,a p o p t o s i s,a u t o p h a g y,m i t o-c h o n d r i a l f u n c t i o n a n d i mm u n e r e s p o n s e.T h i s a r t i c l e r e v i e w s t h e d i a g n o s t i c a n d t h e r a p e u t i c d i r e c t i o n s a n d r e-s e a r c h f r o n t i e r s o f t h e m i R-181g e n e f a m i l y i n c l i n i c a l d i s e a s e s i n r e c e n t y e a r s,w i t h t h e a i m o f p r o v i d i n g n e w i-d e a s f o r t h e c l i n i c a l d i s e a s e t r e a t m e n t o f m i R N A.K e y w o r d s:m i R-181;g e n e f a m i l y;b i o m a r k e r;t a r g e t;d i s e a s e微小R N A(m i R N A)是一类长约22个核苷酸的内源性非编码小R N A分子,通过与目的基因信使R N A(m R N A)的3'非翻译区(3'U T R)不完全互补配对结合,导致靶m R N A发生降解或沉默,从而在转录后水平参与靶基因表达的调控[1]㊂m i R-181具有丰富的生物学功能,参与心脏㊁肺㊁骨骼肌和免疫系统的正常生长发育[2-3],其表达异常时可与多种人类疾病的发生㊁发展相关,在心血管㊁免疫系统疾病的发生中扮演着重要的角色,但具体机制仍有待进一步研究[4-6]㊂现将m i R-181在系统性红斑狼疮(S L E)㊁白血病㊁脑卒中㊁肝癌㊁肺癌等疾病中的表达调控作用及具体机制进行综述,以期为临床治疗和预后评估提供新的方向㊂1 m i R-181基因家族的基因结构m i R-181基因家族由m i R-181a㊁m i R-181b㊁m i R-181c和m i R-181d组成,表达出8个成熟序列,分别是m i R-181a-3p㊁m i R-181a-2-3p㊁m i R-181a-5p㊁m i R-181b-3p㊁m i R-181c-3p㊁m i R-181b-5p㊁m i R-181c-5p和m i R-㊃6142㊃检验医学与临床2023年8月第20卷第16期 L a b M e d C l i n,A u g u s t2023,V o l.20,N o.16*基金项目:国家自然科学基金地区科学基金项目(81960303);广西自然科学基金项目(2021J J A140882)㊂ә通信作者,E-m a i l:c f248248248@163.c o m㊂Copyright©博看网. All Rights Reserved.181d-5p,其中m i R-181c的成熟体有22个核苷酸, m i R-181a㊁m i R-181b和m i R-181d的成熟体有23个核苷酸㊂虽然m i R-181a1㊁m i R-181a2㊁m i R-181b1与m i R-181b2前体结构不同,但成熟体序列却是一致的㊂m i R-181所有成员含有共同的5'端种子序列A C A U U C A,造就它们成为在进化过程中基因组结构非常保守的关键序列,且常成簇地排列在染色体上,使其碱基序列相差甚微,功能相近,因此会协同表达及发挥功能[7]㊂2 m i R-181基因家族的生物学作用m i R-181基因家族是免疫系统调节的重要调控因子,可调控T细胞和B细胞的增殖㊁分化㊁免疫应答㊁免疫耐受等生理㊁病理过程[8-9]㊂大量研究表明, m i R-181在多种疾病的病理㊁生理中发挥调控作用,其中包括炎症反应㊁自身免疫及细胞的增殖分化[8-10]㊂m i R-181a为m i R-181家族重要成员,可表达于人体组织细胞中㊂有研究表明,m i R-181a在胸腺T细胞发育和外周T细胞激活期间可以调节T细胞激活阈值,并且与老年人的抗病毒和疫苗反应密切相关[10]㊂m i R-181a和m i R-181b(m i R-181a/b)位于1号和9号染色体上,m i R-181a/b的调控机制复杂,与不同的靶基因直接相关,有研究表明,m i R-181a/b在肺癌患者的肿瘤组织和血浆中存在异常表达[11]㊂m i R-181c㊁m i R-181d2个m i R-181家族成员也能参与免疫炎症反应的发生㊂其中,m i R-181c靶向结合炎症因子白细胞介素(I L)-2的3'U T R,从而抑制C D4+T细胞的激活,参与免疫炎症反应[12]㊂目前,m i R N A在肿瘤的研究中有了很大进展,有研究表明,m i R-181a在肝癌和乳腺癌的肿瘤组织中表达异常,同时还调控了肿瘤细胞的凋亡㊁侵袭㊁增殖和分化,并参与了W n t/β-连环蛋白和转化生长因子(T G F)-β信号通路的调控[13]㊂有研究发现,上调m i R-181b可通过靶向结合炎症细胞因子C X C L1和C X C L2来抑制乳腺癌细胞的迁移[14],也可通过靶向结合S o x6来抑制肺癌细胞的增殖和分化[15]㊂以上研究表明,m i R-181家族在疾病的发生㊁发展中发挥着重要的作用㊂3 m i R-181基因家族与疾病的相关性3.1 m i R-181与S L E 多种m i R N A目前被证实与S L E的发生㊁发展密切相关[16-18],m i R-181在其发病过程中扮演重要角色㊂S L E是一种多器官受累㊁临床异质性明显㊁免疫耐受失调的自身免疫性疾病,自身抗体的产生及免疫复合物的不同脏器累积是其标志性特征,其中狼疮性肾炎(L N)是最常见的并发症㊂S L E在西方国家㊁日本的发病率和病死率有下降趋势,但在我国的形势依然严峻㊂有研究发现,m i R-181a在调节T细胞和B细胞分化及免疫应答中起着关键作用,并在S L E患者血清中存在差异表达[19]㊂A L E X A N D E R等[20]研究显示,m i R-181a通过靶向调控I L-8㊁I L-1α㊁I L-1β㊁I L-6等免疫炎症因子的表达参与S L E的病理进程㊂有研究表明,在缺血/再灌注(I/R)暴露的肾脏中m i R-181d-5p的表达下调,可以改善肾脏功能,减少细胞凋亡和炎症反应[21],然而有关m i R-181d在S L E的发生㊁发展中的作用机制仍未被充分研究㊂上述研究表明,m i R-181可能成为评估S L E病情发展的关键性标志物㊂3.2 m i R-181与白血病白血病是一种造血系统的恶性肿瘤,由于发病机制复杂,导致治疗难度较大㊂有充分的证据表明,m i R N A在白血病患者中异常表达,参与了疾病的发生㊁发展[22]㊂目前,临床药物治疗白血病不良反应明显,很多患者都存在耐药现象,所以急需发现白血病新的潜在作用机制,寻求新的治疗方式㊂V E R MA等[23]研究表明,m i R-181家族成员与I L-1β㊁T o l l样受体(T L R)介导的白血病T C L1原癌基因及免疫相关基因表达呈负相关,m i R-181a的表达水平在M1/M2型患者较M3/M4型患者细胞中明显升高㊂有研究发现,m i R-181b表达水平在慢性淋巴细胞白血病(C L L)患者体内表达下调,这可能与C L L 患者预后不良相关[24]㊂多年来,化学免疫疗法氟达拉滨-环磷酰胺-利妥昔单抗(F C R)是一线治疗症状性B-C L L患者的金标准,有研究发现,血液m i R-181c是F C R疗效的有效预测生物标志物[25]㊂此外,Z HU 等[26]研究表明,m i R-181a/b在慢性粒细胞白血病(C M L)中表达下调,且其下调与不良预后有关㊂体外动物研究发现,m i R-181a/b可以调控靶基因B C L-2㊁M C L-1和X I A P,同时B C L-2㊁M C L-1和X I A P的表达下调与白血病细胞对化疗药物的耐药有关,提示m i R-181a/b与白血病细胞对化疗药物的耐药密切相关㊂有研究发现,m i R-181d在C M L中呈过表达,通过靶向结合组蛋白去甲基化酶(R B P2)促进白血病细胞增殖[27]㊂以上研究表明,m i R-181家族参与了白血病的多个生物学过程,可以在疾病的诊断和治疗中发挥重要作用,有望成为白血病的预后评估指标㊂3.3 m i R-181与脑缺血损伤脑卒中是一种严重威胁人类健康的脑血管疾病,由于该疾病的发病因素较多,发病率高,预后差,早期预防成为疾病治疗的关键㊂目前,研究已经发现m i R-181基因家族可通过调控N F-κB信号通路参与炎症反应,而炎症反应是脑卒中的重要并发症,可见m i R-181与脑卒中的病理进程密切相关㊂目前研究发现,能量衰竭㊁兴奋性氨基酸细胞毒性作用㊁氧化应激㊁免疫反应㊁凋亡是缺血性脑卒中主要的发病机制,其缺血性炎症反应过程可能通过激活T L R来发挥炎症作用㊂研究发现,m i R-181c 通过其3'U T R末端负向调控T L R4信号的表达,抑制N F-κB的激活,进一步抑制炎症因子T N F-α㊁I L-1α和I L-1β的表达㊂m i R-181c可减少神经胶质细胞释放T N F-α,进而也减少了神经细胞的凋亡[28];m i R-㊃7142㊃检验医学与临床2023年8月第20卷第16期 L a b M e d C l i n,A u g u s t2023,V o l.20,N o.16Copyright©博看网. All Rights Reserved.181a可通过直接下调单核细胞和巨噬细胞释放的I L-1α,从而达到抗感染作用,m i R-181a从整体上能引起星状胶质细胞功能障碍㊂上述研究表明,m i R-181家族分子在脑卒中发展过程中发挥着重要的作用㊂有趣的是,目前鲜有m i R-181d与脑卒中的相关报道,可以作为研究的新方向㊂但是一种m i R N A往往存在多个作用靶点,盲目地选用m i R N A反而会不利于疾病的治疗,所以临床要依据m i R N A的功能基础实施治疗㊂3.4 m i R-181与肝癌肝癌是我国最常见的实性肿瘤,分为原发性肝癌和继发性肝癌两大类,原发性肝癌是我国常见的恶性肿瘤之一,高发于东南沿海及广西地区㊂最新研究表明,肝细胞癌可以释放大量的m i R N A,m i R N A可以直接或间接地通过靶向有关蛋白来参与肿瘤调控,同时m i R N A可以改变肿瘤的微环境㊂研究发现,m i R-181基因家族m i R-181a㊁m i R-181b㊁m i R-181c和m i R-181d在肝细胞癌中的表达均明显上调,并且通过直接靶向调控细胞分化相关的转录因子C D X2㊁G A T A6及W n t信号通路抑制因子N L K的表达,达到维持肝癌细胞的特性[8],提示m i R-181有望成为治疗肝癌的分子靶标与生物标志物㊂Z H A O等[29]研究证实,高转移性结直肠癌细胞释放的富含m i R-181a-5p的细胞外囊泡通过激活肝星状细胞和重塑肿瘤微环境促进肝转移,为肝转移机制提供新的见解㊂WA N G等[30]研究发现,m i R-181b和m i R-181d高表达于被喂食无胆碱与限定氨基酸诱发的肝癌小鼠肝组织中,调控金属蛋白酶组织抑制因子-3(癌症抑制因子)的表达明显降低,同时发现m i R-181b在肝癌细胞中呈高表达,其表达水平受到T G F-β的调控,但是其作用机制尚未明确㊂W E I等[31]研究发现,肝癌细胞来源的外泌体通过m i R-181d-5p的传递和S O C S3/F A K/S r c通路在骨髓间质干细胞分化和肝癌转移中发挥作用㊂上述研究提示,m i R-181在肝癌的发生㊁发展过程中扮演重要的促癌基因角色㊂3.5 m i R-181与肺癌肺癌是全球发病率最高的癌症,大多数患者为非小细胞肺癌(N S C L C)㊂肺癌的治疗仍存在很多困难,如患者后期易出现多重耐药和癌细胞的转移㊂有研究指出,m i R N A可以作为肺癌诊断和预后的重要标志物,其中m i R-181与肺癌调控作用密切相关㊂C H E N等[32]通过微阵列表达谱检测筛选发现15种m i R N A在耐放疗N S C L C患者及非耐放疗N S C L C患者肺癌组织中存在明显差异,且发现m i R-181a的靶基因在磷脂酰肌醇3-羟基激酶(P I3K)/蛋白激酶B(A K T)信号通路㊁N S C L C信号通路等癌症信号通路中明显富集,靶向结合会对N S C L C的放射敏感性产生影响㊂有研究表明,小细胞肺癌(S C L C)患者体内m i R-181d的表达水平升高,可以靶向结合B C L-2,抑制肿瘤细胞的增殖[33]㊂Z HU等[34]进行了人肺癌细胞多药耐药性研究,发现与亲本细胞比较,耐药细胞株中m i R-181b的表达水平明显下调,且通过外源性转染过表达m i R-181b,使耐药细胞可恢复相应药物敏感性,此外,m i R-181b主要通过对B c l-2的调控而发挥作用㊂疾病的早期诊断至关重要,目前m i R N A可以对多种疾病患者进行生存预测,起到辅助治疗的作用,但m i R-181在肺癌组织中的作用机制及靶标调控仍有待进一步研究㊂3.6 m i R-181与妊娠期高血压疾病(H D C P) H D-C P是一种产科常见的综合征,是临床上发病率较高的多器官受累性疾病,由于缺乏有效的预防及治疗手段,HD C P严重危害着孕妇及胎儿的健康㊂H D C P 以高血压㊁蛋白尿㊁免疫系统失调为主要的临床特征㊂由于m i R N A在血液中表达水平较高且容易获得,所以血液中差异表达的m i R N A有望成为H D C P诊断的标志物㊂有研究发现,m i R-181b可能在H D C P诱导的心脏病中起到保护心肌的作用,有望成为新的标志物[35]㊂研究指出,m i R-181b在H D C P患者胎盘中高表达,主要作用机制是m i R-181b的3'U T R末端能与纤溶酶原激活物抑制剂1(P A I-1)m R N A结合并调控P A I-1的表达,P A I-1在血管内皮细胞及血管平滑肌细胞上表达丰富,在血管平滑肌细胞的肥大㊁增殖㊁重塑中发挥至关重要的作用,表明m i R-181b通过影响血管重塑可提高胎盘细胞的侵袭力,在H D C P的发生㊁发展中起重要作用[36]㊂有研究显示,m i R-181a 在孕妇体内表达水平升高,尤其在子痫前期孕妇体内表达水平升高更明显,可见m i R-181a的异常表达在H D C P的发生㊁发展中具有重要的参考意义㊂3.7 m i R-181与其他疾病很多前瞻性研究报道了m i R-181家族成员可能与心脑血管疾病㊁炎症反应㊁肿瘤㊁病毒感染和神经系统疾病密切相关㊂J I N等[37]研究发现,m i R-181a-2-3p在预测胃癌患者顺铂治疗获益方面有着重要的价值,靶向下调m i R-181a-2-3p 可抑制肿瘤生长,抑制顺铂介导的耐药㊂此外,顺铂联合m i R-181a-2-3p下调可能是未来顺铂耐药胃癌患者的一种有前途的治疗选择㊂有研究发现,在人表皮生长因子受体2(H E R2)阳性乳腺癌中观察到高去甲基化酶(F T O)表达,则提示着乳腺癌晚期进展,通过F T O/m i R-181b-3p/A R L5B信号通路促进乳腺癌细胞的侵袭和迁移的致癌活性[38]㊂L I等[39]报道表明, G S K I P基因是m i R-181c-5p在宫颈鳞状细胞癌(S C C)中的靶基因,而G S K I P是W n t/β-c a t e n i n信号的激活剂,通过负向调节β-c a t e n i n抑制剂G S K3β, m i R-181c-5p通过抑制G S K I P的表达来减弱S i H a细胞的干细胞特性㊂m i R-181d是一种对肿瘤具有调控作用的m i c r o R N A,有研究发现m i R-181d可以通过调节胞内P I3K/A K T途径抑制胃癌细胞的增殖和转移[40]㊂以上研究均表明m i R-18的表达与多种疾病㊃8142㊃检验医学与临床2023年8月第20卷第16期 L a b M e d C l i n,A u g u s t2023,V o l.20,N o.16Copyright©博看网. All Rights Reserved.相关㊂4展望m i R N A作为一类内源性非蛋白编码基因,通过与目的基因m R N A的3'U T R不完全互补配对结合,从而精准地调控靶蛋白编码基因的表达水平,已成为后基因组时代医学及生物学领域的研究热点之一㊂尽管m i R-181在调控肿瘤㊁免疫系统及心血管等疾病发生㊁发展方面的研究已取得一定的进展,其主要研究着重于m i R-181的表达调控作用,但m i R-181如何导致其靶m R N A s抑制或沉默的机制仍未阐明,因此,有必要进行m i R-181对特定靶基因具体调控机制的研究㊂另外,m i R-181家族成员在相关疾病的动物模型或人体中发生了异常变化,但是其在人类疾病中的具体作用尚无确切的定论㊂然而,随着分子遗传学㊁分子生物学㊁人类基因组学研究的逐步深入,发现m i R-181家族在肝细胞癌㊁口腔鳞状细胞癌㊁胶质瘤㊁S L E等疾病的病理进程中扮演着重要的角色,有望成为一类新的生物标志物或治疗靶标,为临床诊断和治疗及预后提供有利的方向㊂参考文献[1]L U O B,Z HO U K,L I U F U Y,e t a l.N o v e l i n s i g h t i n t om i R N A b i o l o g y a n d i t s r o l e i n t h e p a t h o g e n e s i s o f s y s-t e m i c l u p u s e r y t h e m a t o s u s[J].F r o n t I mm u n o l,2022,13: 1059887.[2]I N D R I E R I A,C A R R E L L A S,C A R O T E N U T O P,e t a l.T h e p e r v a s i v e r o l e o f t h e m i R-181f a m i l y i n d e v e l o p m e n t, n e u r o d e g e n e r a t i o n,a n d c a n c e r[J].I n t J M o l S c i,2020,21(6):2092.[3]X I A O C,R A J E W S K Y K.m i c r o R N A c o n t r o l i n t h e i m-m u n e s y s t e m:b a s i c p r i n c i p l e s[J].C e l l,2009,136(1): 26-36.[4]P O P-B I C A C,P I N T E A S,C O J O C N E A N U-P E T R I C R,e t a l.m i R-181f a m i l y-s p e c i f i c b e h a v i o r i n d i f f e r e n t c a n c-e r s:a m e t a-a n a l y s i s v i e w[J].C a n c e r M e t a s t a s i s R e v,2018,37(1):17-32.[5]P R I C E N L,R O T L L A N N,C A N F RÁN-D U Q U E A,e ta l.G e n e t i c D i s s e c t i o n o f t h e I m p a c t o f m i R-33a a n d m i R-33b d u r i n g t h e P r o g r e s s i o n o f A t h e r o s c l e r o s i s[J].C e l lR e p o r t s,2017,21(5):1317-1330.[6]K R A U S S S,N A L A V A D E R,W E B E R S,e t a l.U p r e g u l a-t i o n o f m i R-25a n d m i R-181f a m i l y m e m b e r s c o r r e l a t e sw i t h r e d u c e d e x p r e s s i o n o f a t x n3i n l y m p h o c y t e s f r o mS C A3p a t i e n t s[J].M i c r o r n a,2019,8(1):76-85. 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[22]L E E Y G,K I M I,OH S,e t a l.S m a l l R N A s e q u e n c i n g㊃9142㊃检验医学与临床2023年8月第20卷第16期 L a b M e d C l i n,A u g u s t2023,V o l.20,N o.16Copyright©博看网. All Rights Reserved.p r o f i l e s o f m i R-181a n d m i R-221,t h e m o s t r e l e v a n t m i-c r o R N A s i n a c u t e m y e l o id le u k e m i a[J].K o r e a n J I n t e r nM e d,2019,34(1):178-183.[23]V E R MA P,P A N D E Y R K,P R A J A P A T I P,e t a l.C i r c u-l a t i n g m i c r o R N A s:p o t e n t i a l a n d e m e r g i n g b i o m a r k e r s f o rd i a g n o s i s o f h u m a n i n fe c t i o u s d i s e a s e s[J].F r o n t M i c r o b i-o l,2016,7:1274.[24]K O U Z,L I U H,WA N G Y C,e t a l.E x p r e s s i o n l e v e l a n d t a r g e t g e n e p r e d i c t i o n o f m i R-181b i n p a t i e n t s w i t h c h r o-n i c l y m p h o c y t i c l e u k e m i a[J].Z h o n g g u o S h i Y a n X u e Y e X u e Z a Z h i,2020,28(3):808-814.[25]D U R O U X-R I C HA R D I,G A G E Z A L,A L A T E R R E E,e ta l.m i R N A p r o f i l e a t d i a g n o s i s p r e d i c t s t r e a t m e n t o u t-c o m e i n p a t i e n t s w i t h B-c h r o n i c l y m p h o c y t i c l e u k e m i a:aF I L O s t u d y[J].F r o n t I mm u n o l,2022,13:983771.[26]Z HU D X,Z HU W,F A N G C,e t a l.m i R-181a/b s i g n i f i-c a n t l y e n h a n c e sd r u g se n s i t i v i t y i n c h r o n i c l y m p h o c y t i c l e u k e m i a c e l l s v i a t a r g e t i n g m u l t i p l e a n t i-a p o p t o s i s g e n e s [J].C a r c i n o g e n e s i s,2012,33(7):1294-1301. [27]Z HO U M,Y I N X,Z H E N G L,e t a l.m i R-181d/R B P2/N F-κB p65f e e d b a c k r e g u l a t i o n p r o m o t e s c h r o n i c m y e l o i d l e u k e m i a b l a s t c r i s i s[J].F r o n t O n c o l,2021,11:654411.[28]L I U X,L I F,Z HA O S,e t a l.m i c r o R N A-124-m e d i a t e dr e g u l a t i o n o f i n h i b i t o r y m e m b e r o f a p o p t o s i s-s t i m u l a t i n g p r o t e i n o f p53f a m i l y i n e x p e r i m e n t a l s t r o k e[J].S t r o k e, 2013,44(7):1973-1980.[29]Z HA O S,M I Y,Z H E N G B,e t a l.H i g h l y-m e t a s t a t i c c o l o-r e c t a l c a n c e r c e l l r e l e a s e d m i R-181a-5p-r i c h e x t r a c e l l u l a r v e s i c l e s p r o m o t e l i v e r m e t a s t a s i s b y a c t i v a t i n g h e p a t i c s t e l l a t e c e l l s a n d r e m o d e l l i n g t h e t u m o u r m i c r o e n v i r o n-m e n t[J].J E x t r a c e l l V e s i c l e s,2022,11(1):e12186.[30]W A N G B,H S U S H,M A J UM D E R S,e t a l.T G F b e t a m e d i-a t e d u p r e g u l a t i o n o f h e p a t i c m i R-181b p r o m o t e s h e p a t oc a r c i-n o g e n e s i s b y t a r g e t i n g T I M P3[J].O n c o g e n e,2010,29(12): 1787-1797.[31]W E I H,WA N G J,X U Z,e t a l.H e p a t o m a c e l l-d e r i v e d e x-t r a c e l l u l a r v e s i c l e s p r o m o t e l i v e r c a n c e r m e t a s t a s i s b y i n-d u c i n g t he d if f e r e n t i a t i o n o f b o n e m a r r o w s t e m c e l l st h r o u g h m i c r o R N A-181d-5p a n d t h e F A K/S r c P a t h w a y [J].F r o n t C e l l D e v B i o l,2021,9:607001. [32]C H E N Y,L I A O W,Y U A N A,e t a l.m i R-181a r e d u c e sr a d i o s e n s i t i v i t y o f n o n-s m a l l-c e l l l u n g c a n c e r v i a i n h i b i-t i n g P T E N[J].P a n m i n e r v a M e d,2022,64(3):374-383.[33]A N J,P A N Y,Y A N Z,e t a l.m i R-23a i n a m p l i f i e d19p13. 13l o c i t a r g e t s m e t a l l o t h i o n e i n2A a n d p r o m o t e s g r o w t h i n g a s t r i c c a n c e r c e l l s[J].J C e l l B i o c h e m,2013,114(9): 2160-2169.[34]Z HU W,S HA N X,WA N G T,e t a l.m i R-181b m o d u l a t e sm u l t i d r u g r e s i s t a n c e b y t a r g e t i n g B C L2i n h u m a n c a n c e rc e l l l i n e s[J].I n t J C a n c e r,2010,127(11):2520-2529.[35]G A O Z,WA N G L,WA N G J,e t a l.M o l e c u l a r m e c h a n i s m o f m i R-181b i n h e a r t d i s e a s e d u e t o p r e g n a n c y-i n d u c e dh y p e r t e n s i o n s y n d r o m e[J].E x p T h e r M e d,2017,14(4): 2953-2959.[36]C H E N Y S,S H E N L,MA I R Q,e t a l.L e v e l s o f m i c r o R-N A-181b a n d p l a s m i n o g e n a c t i v a t o r i n h i b i t o r-1a r e a s s o-c i a t e d w i t h h y p e r t e n s i v e d i s o r d e r s c o m p l i c a t i n g p r e g n a n-c y[J].E x p T h e r M e d,2014,8(5):1523-1527.[37]J I N L,MA X,Z HA N G N,e t a l.T a r g e t i n g o n c o g e n i cm i R-181a-2-3p i n h i b i t s g r o w t h a n d s u p p r e s s e s c i s p l a t i n r e s i s t a n c e o f g a s t r i c c a n c e r[J].C a n c e r M a n a g R e s,2021, 13:8599-8609.[38]X U Y,Y E S,Z HA N G N,e t a l.T h e F T O/m i R-181b-3p/A R L5B s i g n a l i n g p a t h w a y r e g u l a t e s c e l l m i g r a t i o n a n d i n-v a s i o n i n b r e a s t c a n c e r[J].C a n c e r C o mm u n(L o n d), 2020,40(10):484-500.[39]L I N,C H E N G C,WA N G T.m i R-181c-5p m i t i g a t e s t u-m o r i g e n e s i s i n c e r v i c a l s q u a m o u s c e l l c a r c i n o m a v i a t a r-g e t i n g g l y c o g e n s y n t h a s e k i n a s e3βi n t e r a c t i o n p r o t e i n(G S K I P)[J].O n c o T a r g e t s T h e r,2020,13:4495-4505.[40]J I A N G K,X I E L F,X I A O T Z,e t a l.m i R-181d i n h i b i t sc e l l p r o l i f e r a t i o n a nd me t a s t a s i s t h r o u g h P I3K/A K Tp a t h w a y i n g a s t r i c c a n c e r[J].E u r R e v M e d P h a r m a c o l S c i,2019,23(20):8861-8869.(收稿日期:2022-09-20修回日期:2023-02-15)㊃综述㊃D O I:10.3969/j.i s s n.1672-9455.2023.16.027聚集诱导发光材料在细菌成像检测和感染治疗中的应用*谢岭平1,2,赖丽莎1,彭兰芬1综述,付文金1ә审校1.广东医科大学附属厚街医院检验科,广东东莞523000;2.广东医科大学基础医学院,广东东莞523808摘要:细菌感染已成为人类健康最大威胁之一,抗菌药物的过度使用导致了耐药菌的出现和传播㊂具有聚集诱导发光(A I E)特性的荧光材料已成为研究细菌检测和细菌感染治疗的热点,展现出巨大的应用潜力㊂该文将概述A I E的发光机制和性能特点,并对其相关材料在细菌成像检测和细菌感染治疗等方面的研究进展进行综述㊂关键词:细菌;聚集诱导发光;荧光;发展趋势中图法分类号:R446.5文献标志码:A文章编号:1672-9455(2023)16-2420-05㊃0242㊃检验医学与临床2023年8月第20卷第16期 L a b M e d C l i n,A u g u s t2023,V o l.20,N o.16*基金项目:广东省基础与应用基础研究基金项目重点项目(2019B151520004);广东省东莞市社会科技发展重点项目(202050715023181)㊂ә通信作者,E-m a i l:2804787909@q q.c o m㊂Copyright©博看网. All Rights Reserved.。

miR181b影响ApoE——小鼠动脉粥样硬化形成的自噬调节机制

miR181b影响ApoE—/—小鼠动脉粥样硬化形成的自噬调节机制作者：董轶杨绍南潘旭东马爱军李舒裴浩天来源：《青岛大学学报（医学版）》2018年第02期[摘要]目的探讨微小RNA 181b（miR181b）是否通过调节自噬相关靶蛋白参与动脉粥样硬化发展。

方法通过颈总动脉套环诱导ApoE-/-小鼠颈动脉粥样硬化斑块形成。

通过尾静脉注射miR181b 慢病毒载体构建其过表达（miR181bOE）及低表达（miR181bKD）模型。

检测血浆中脂质水平，苏木精伊红染色对动脉粥样硬化斑块大小和体积进行定量，Western Blot检测自噬相关蛋白的表达。

结果各组ApoE-/-小鼠血浆中总胆固醇、三酰甘油、低密度脂蛋白胆固醇水平均无显著差异（P>0.05）。

MiR181bKD和miR181bOE分别显著减少和增加颈动脉斑块面积和体积（F=118.95～322.03，P结论抑制miR181b可能通过下调p70S6K表达增加自噬活性，延缓颈动脉粥样硬化斑块的形成。

[关键词]动脉粥样硬化；微RNAs；自噬体；核糖体蛋白质S6激酶类，70kDa[中图分类号]R743.1；R543.5[文献标志码]A[文章编号] 20965532（2018）02014406动脉粥样硬化已经成为威胁人类健康的最危险的疾病之一，颈动脉是多发部位[12]。

颈动脉狭窄是缺血性卒中的危险因素，可导致高达10%～20%的卒中或短暂性脑缺血发作[3]。

研究认为，动脉粥样硬化是一种炎癥性疾病，其起始和发展与大动脉和中动脉内膜中的胆固醇积累有关[4]。

但是，目前动脉粥样硬化的病理生理机制还不完全清楚，新的治疗方法和生物标志物都需进一步阐明。

自噬是一种高度保守过程，细胞质内的物质均可通过溶酶体依赖途径降解[56]。

最近的证据表明，动脉粥样硬化斑块中的自噬标志物的表达可在低密度脂蛋白、炎症和代谢应激等内在刺激下被相应上调。

值得注意的是，前期研究表明自噬也会被其他信号因子调控。

抑制miR181b通过上调PKG—1减轻大鼠心肌细胞肥大

抑制miR181b通过上调PKG—1减轻大鼠心肌细胞肥大目的：通过检测miR181b在心肌肥厚患者外周血中的表达，研究miR181b 与PRKG-1在心肌肥大中的作用机制，为心肌肥厚临床诊断和治疗提供新的靶点。

方法：采用实时定量PCR方法检测50例临床诊断为心肌肥厚患者及25例正常健康人的外周血miR181b的水平，分析其表达水平与临床病理特征的关系。

结果：miR181b在心肌肥厚患者外周血中表达（3.35±0.22）升高，差异有统计学意义（P＜0.05）；体外大鼠原代心肌细胞在镜下呈多角形，采用α-横纹肌肌动蛋白抗体和DAPI进行免疫荧光染色，镜下90%以上细胞发出绿色荧光，表明原代培养心肌细胞纯度较高；采用去氧肾上腺素处理心肌细胞72 h后，心肌细胞总蛋白表达显著升高（P＜0.05）；流式细胞术结果显示PE组心肌细胞大小明显增加（P＜0.05）；实时定量PCR检测结果表明，PE组中心肌肥大相关基因β-MHC、α-SA、ANP显著升高（P＜0.05），证明体外构建心肌肥大模型成功。

结论：miR181b 在心肌肥厚患者外周血中表达显著上调；体外成功培养乳鼠原代心肌细胞，通过PE处理法成功构建体外心肌肥大模型；在肥大心肌细胞中，miR181b与prkg-1 mRNA及蛋白的表达有显著相关性；miR181b可能通过调控prkg-1 mRNA及蛋白水平的表达参与心肌肥大，有望成为心肌肥厚临床诊断和治疗的新的靶点。

标签：心肌肥大；心室重构；miR181b；PRKG1心肌肥厚是很多慢性疾病的一个共同的临床病理性改变，主要发生在长期压力负荷过重的情况下，作为一种代偿方式。

但这种代偿功能也有其不利之处，主要因为肥大的心肌需氧增加，而冠状动脉的供血量往往不能给予满足，造成心肌缺血，这将最后导致心肌收缩力的减退，是导致患者心衰进而引起死亡的重要原因[1]。

1 材料与方法1.1 材料1.1.1 心肌肥大患者外周血采集收集2012年1月-2014年3月在本院治疗的50例经心脏超声诊断有心肌肥厚的患者，男40例，女10例，年龄64～72岁，平均68.5岁，中位年龄67岁。

miR-181b通过下调CYLD蛋白影响甲状腺乳头状癌细胞的增殖和凋亡

［摘要］目的探讨ｍｉＲ１８１ｂ通过下调ＣＹＬＤ蛋白影响甲状腺乳头状癌细胞的增殖和凋亡行为及其机制。方法ｑＰＣＲ检测ｍｉＲ１８１ｂ在甲状腺乳头状癌组织和正常甲状腺组织中的表达情况及差异；Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ实验检测ｍｉＲ１８１ｂ与ＣＹＬＤ蛋白之间的调控作用；克隆形成实验检测抑制ｍｉＲ１８１ｂ后甲状腺癌细胞增殖能力的变化情况；流式细胞术实验检测抑制ｍｉＲ１８１ｂ后甲状腺癌细胞凋亡行为的变化情况。结果与正常甲状腺组织相比，甲状腺乳头状癌组织中ｍｉＲ１８１ｂ的表达水平相对上调，差异具有统计学意义（Ｐ＜０．０５）；在ＦＴＣ１３３细胞株中ｍｉＲ１８１ｂ的表达水平相对较高；Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ实验证实ｍｉＲ１８１ｂ能直接调控ＣＹＬＤ蛋白的表达水平；抑制ｍｉＲ１８１ｂ的表达后可以抑制甲状腺癌细胞的增殖行为，并在一定程度上促进其凋亡。结论ｍｉＲ１８１ｂ可以调控ＣＬＹＤ的表达影响甲状腺癌细胞的增殖和凋亡行为。
目前研究指出参与甲状腺乳头状癌发展的最常见的遗传改变包括ＲＥＴＲＡＳＢＲＡＦ基因激、ＲＥＴ基因重排、ＢＲＡＦＶ６００Ｅ点突变及ＲＡＳ突变等［５］。虽然这些遗传改变会导致相同的信号通路的激活，但针对其具体机制作用的发挥目前不甚清楚。在人类基因组中，
［基金项目］陕西省科学技术研究发展计划基金（２０１２ＳＦ２０２０５）
Ｋｅｙｗｏｒｄｓ：ｍｉＲ１８１ｂ；ｔｈｙｒｏｉｄｐａｐｉｌｌａｒｙｃａｒｃｉｎｏｍａ；ＣＹＬＤ；ａｐｏｐｔｏｓｉｓ
甲状腺癌是最常见的内分泌异常的恶性肿瘤，占女性所有类型恶性肿瘤的５％左右，在中国，每年约有２００００的恶性甲状腺癌患者死亡。［１２］甲状腺乳头状癌（ＰＴＣ）是甲状腺恶性肿瘤中最常见的肿瘤类型，约占８０％［３］。针对甲状腺乳头状癌的治疗目前主要采取手术联合放疗，但是治愈率不高［４］。探讨新型肿瘤分子标记物是目前急需解决的问题。

MicroRNA在骨性关节炎中的最新研究进展

MicroRNA在骨性关节炎中的最新研究进展李小林;陈洁琳;王大平【摘要】微RNA（ miRNA）是一类内源性基因编码的单链RNA小分子，通过参与靶基因的表达调控对骨性关节炎（ OA）的病理进展发挥促进、抑制或双重作用。

在OA患者外周血中异常表达的miRNA，以稳定的复合物形式存在，在未来可能成为诊断OA并判断其预后的重要生物学标志物。

通过研究OA的miRNA治疗靶点来干预miRNA对OA的病理进展，为临床疾病的治疗提供了新思路。

%MicroRNA(miRNA),a class of single-stranded RNA molecules encoded by endogenous genes ,plays posi-tive,negative or dual roles in pathological progression of osteoarthritis ( OA) by participating in the regulation of target gene expression .The abnormal expressed miRNAs in peripheral blood of OA patients ,existing as a stable compound ,may become important biological markers in the diagnosis and prognosis of OA .Searching for target genes of miRNAs in OA to intervene OA pathological progression may provide a novel way for the clinical treatment .【期刊名称】《医学综述》【年(卷),期】2017(023)001【总页数】5页(P12-15,20)【关键词】微RNA;骨性关节炎;软骨细胞;靶基因【作者】李小林;陈洁琳;王大平【作者单位】安徽医科大学附属深圳市第二人民医院临床医学院，广东深圳518000; 深圳市组织工程重点实验室，广东深圳518000;深圳市组织工程重点实验室，广东深圳518000;安徽医科大学附属深圳市第二人民医院临床医学院，广东深圳 518000; 深圳市组织工程重点实验室，广东深圳518000【正文语种】中文【中图分类】R336骨性关节炎(osteoarthritis，OA)多见于中老年人，女性发病率高于男性［1］。

检测血清miR-181b、miR-210诊断妊娠期高血压疾病价值

定量 PCR 检测 miR
１８１b、
miR
２１０水平,观察分析二者与临床指标关系,采用受试者工作特征 (
ROC)曲线评估 miR
１８１b、
miR
２１０诊断效能. 结果:与对照组相比,观察组血清 miR１８１b、miR２１０水平均升高 (
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检测血清 miR
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