Bisantrene_COA_22927_MedChemExpress
Bisantrene_SDS_MedChemExpress
Inhibitors, Agonists, Screening LibrariesSafety Data Sheet Revision Date:Mar.-30-2017Print Date:Mar.-30-20171. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :BisantreneCatalog No. :HY-100875CAS No. :78186-34-21.2 Relevant identified uses of the substance or mixture and uses advised againstIdentified uses :Laboratory chemicals, manufacture of substances.1.3 Details of the supplier of the safety data sheetCompany:MedChemExpress USATel:609-228-6898Fax:609-228-5909E-mail:sales@1.4 Emergency telephone numberEmergency Phone #:609-228-68982. HAZARDS IDENTIFICATION2.1 Classification of the substance or mixtureNot a hazardous substance or mixture.2.2 GHS Label elements, including precautionary statementsNot a hazardous substance or mixture.2.3 Other hazardsNone.3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms:CL216942Formula:C22H22N8Molecular Weight:398.46CAS No. :78186-34-24. FIRST AID MEASURES4.1 Description of first aid measuresEye contactRemove any contact lenses, locate eye-wash station, and flush eyes immediately with large amounts of water. Separate eyelids with fingers to ensure adequate flushing. Promptly call a physician.Skin contactRinse skin thoroughly with large amounts of water. Remove contaminated clothing and shoes and call a physician.InhalationImmediately relocate self or casualty to fresh air. If breathing is difficult, give cardiopulmonary resuscitation (CPR). Avoid mouth-to-mouth resuscitation.IngestionWash out mouth with water; Do NOT induce vomiting; call a physician.4.2 Most important symptoms and effects, both acute and delayedThe most important known symptoms and effects are described in the labelling (see section 2.2).4.3 Indication of any immediate medical attention and special treatment neededTreat symptomatically.5. FIRE FIGHTING MEASURES5.1 Extinguishing mediaSuitable extinguishing mediaUse water spray, dry chemical, foam, and carbon dioxide fire extinguisher.5.2 Special hazards arising from the substance or mixtureDuring combustion, may emit irritant fumes.5.3 Advice for firefightersWear self-contained breathing apparatus and protective clothing.6. ACCIDENTAL RELEASE MEASURES6.1 Personal precautions, protective equipment and emergency proceduresUse full personal protective equipment. Avoid breathing vapors, mist, dust or gas. Ensure adequate ventilation. Evacuate personnel to safe areas.Refer to protective measures listed in sections 8.6.2 Environmental precautionsTry to prevent further leakage or spillage. Keep the product away from drains or water courses.6.3 Methods and materials for containment and cleaning upAbsorb solutions with finely-powdered liquid-binding material (diatomite, universal binders); Decontaminate surfaces and equipment by scrubbing with alcohol; Dispose of contaminated material according to Section 13.7. HANDLING AND STORAGE7.1 Precautions for safe handlingAvoid inhalation, contact with eyes and skin. Avoid dust and aerosol formation. Use only in areas with appropriate exhaust ventilation.7.2 Conditions for safe storage, including any incompatibilitiesKeep container tightly sealed in cool, well-ventilated area. Keep away from direct sunlight and sources of ignition.Recommended storage temperature:Powder-20°C 3 years4°C 2 yearsIn solvent-80°C 6 months-20°C 1 monthShipping at room temperature if less than 2 weeks.7.3 Specific end use(s)No data available.8. EXPOSURE CONTROLS/PERSONAL PROTECTION8.1 Control parametersComponents with workplace control parametersThis product contains no substances with occupational exposure limit values.8.2 Exposure controlsEngineering controlsEnsure adequate ventilation. Provide accessible safety shower and eye wash station.Personal protective equipmentEye protection Safety goggles with side-shields.Hand protection Protective gloves.Skin and body protection Impervious clothing.Respiratory protection Suitable respirator.Environmental exposure controls Keep the product away from drains, water courses or the soil. Cleanspillages in a safe way as soon as possible.9. PHYSICAL AND CHEMICAL PROPERTIES9.1 Information on basic physical and chemical propertiesAppearance Pink to red (Solid)Odor No data availableOdor threshold No data availablepH No data availableMelting/freezing point No data availableBoiling point/range No data availableFlash point No data availableEvaporation rate No data availableFlammability (solid, gas)No data availableUpper/lower flammability or explosive limits No data availableVapor pressure No data availableVapor density No data availableRelative density No data availableWater Solubility No data availablePartition coefficient No data availableAuto-ignition temperature No data availableDecomposition temperature No data availableViscosity No data availableExplosive properties No data availableOxidizing properties No data available9.2 Other safety informationNo data available.10. STABILITY AND REACTIVITY10.1 ReactivityNo data available.10.2 Chemical stabilityStable under recommended storage conditions.10.3 Possibility of hazardous reactionsNo data available.10.4 Conditions to avoidNo data available.10.5 Incompatible materialsStrong acids/alkalis, strong oxidising/reducing agents.10.6 Hazardous decomposition productsUnder fire conditions, may decompose and emit toxic fumes.Other decomposition products - no data available.11.TOXICOLOGICAL INFORMATION11.1 Information on toxicological effectsAcute toxicityClassified based on available data. For more details, see section 2Skin corrosion/irritationClassified based on available data. For more details, see section 2Serious eye damage/irritationClassified based on available data. For more details, see section 2Respiratory or skin sensitizationClassified based on available data. For more details, see section 2Germ cell mutagenicityClassified based on available data. For more details, see section 2CarcinogenicityIARC: No component of this product present at a level equal to or greater than 0.1% is identified as probable, possible or confirmed human carcinogen by IARC.ACGIH: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by ACGIH.NTP: No component of this product present at a level equal to or greater than 0.1% is identified as a anticipated or confirmed carcinogen by NTP.OSHA: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by OSHA.Reproductive toxicityClassified based on available data. For more details, see section 2Specific target organ toxicity - single exposureClassified based on available data. For more details, see section 2Specific target organ toxicity - repeated exposureClassified based on available data. For more details, see section 2Aspiration hazardClassified based on available data. For more details, see section 212. ECOLOGICAL INFORMATION12.1 ToxicityNo data available.12.2 Persistence and degradabilityNo data available.12.3 Bioaccumlative potentialNo data available.12.4 Mobility in soilNo data available.12.5 Results of PBT and vPvB assessmentPBT/vPvB assessment unavailable as chemical safety assessment not required or not conducted.12.6 Other adverse effectsNo data available.13. DISPOSAL CONSIDERATIONS13.1 Waste treatment methodsProductDispose substance in accordance with prevailing country, federal, state and local regulations.Contaminated packagingConduct recycling or disposal in accordance with prevailing country, federal, state and local regulations.14. TRANSPORT INFORMATIONDOT (US)This substance is considered to be non-hazardous for transport.IMDGThis substance is considered to be non-hazardous for transport.IATAThis substance is considered to be non-hazardous for transport.15. REGULATORY INFORMATIONSARA 302 Components:No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302.SARA 313 Components:This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title III, Section 313.SARA 311/312 Hazards:No SARA Hazards.Massachusetts Right To Know Components:No components are subject to the Massachusetts Right to Know Act.Pennsylvania Right To Know Components:No components are subject to the Pennsylvania Right to Know Act.New Jersey Right To Know Components:No components are subject to the New Jersey Right to Know Act.California Prop. 65 Components:This product does not contain any chemicals known to State of California to cause cancer, birth defects, or anyother reproductive harm.16. OTHER INFORMATIONCopyright 2017 MedChemExpress. The above information is correct to the best of our present knowledge but does not purport to be all inclusive and should be used only as a guide. The product is for research use only and for experienced personnel. It must only be handled by suitably qualified experienced scientists in appropriately equipped and authorized facilities. The burden of safe use of this material rests entirely with the user. MedChemExpress disclaims all liability for any damage resulting from handling or from contact with this product.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。
硕士论文--多奈哌齐透皮贴剂的药学研究
多奈哌齐透皮给药研究
Ｓｔｕｄｙ ｏｆ Ｄｏｎｅｐｅｚｉｌ Ｔｒａｎｓｄｅｒｍａｌ Ｐａｔｃｈ
Ａｂｓｔｒａｃｔ
Ｄｏｎｅｐｅｚｉｌ（ＤＰＢ）ｉｓ ａ ｐｉｐｅｒｉｄｉｎｅ—ｂａｓｅｄ ｒｅｖｅｒｓｉｂｌｅ ａｃｅｔｙ７ｌｃｈｏｌｉｎｅｓｔｅｒａｓｅ ｉｎｈｉｂｉｔｏｒ． Ｃｕｒｒｅｎｔｌｙ ＤＰＢ ｉｓ ｔｈｅ ｍｏｓｔ ｐｒｅｓｃｒｉｂｅｄ ｐｈａｒｍａｃｏｌｏｇｉｃａｌ ａｇｅｎｔ ｆｏｒ ｔｈｅ ｔｒｅａｔｍｅｎｔ ｏｆ ｍｉｌｄ ｔｏ ｍｏｄｅｒａｔｅ Ａｌｚｈｅｉｍｅｒ’Ｓ ｄｉｓｅａｓｅ．Ｉｔ ａｌｓｏ ｐｌａｙｓ ｔｈｅ ｉｍｐｏｒｔａｎｔ ｒｏｌｅ ｉｎ ｉｍｐｒｏｖｉｎｇ ｔｈｅ ｐａｔｉｅｎｔ’Ｓ ｍｅｎｔａｌ ｓｔａｔｅ ａｎｄ ｋｅｅｐｉｎｇ ｔｈｅ ｂｒａｉｎ ａｃｔｉｖｅ．Ｎｏｗ ｔｈｅ ｄｒｕｇ ｄｏｓａｇｅ ｆｏｒｍ ｏｆ ＤＰＢ ｈａｓ ｔａｂｌｅｔｓ ａｎｄ ｃａｐｓｕｌｅｓ．Ｒｅｓｅａｒｃｈｅｓ ｈａｖｅ ｓｈｏｗｅｄ ｔｈａｔ ａｉｚｈｅｉｍｅｒ’Ｓ ｄｉｓｅａｓｅ ｉｓ ｔｈｅ ｍｏｓｔ ｃｏｍｍｏｎ ｃａｕｓｅ ｏｆ
研究结果表明当使用混合丙烯酸酯压敏胶作为基质时（Ｂ／Ｃ＝３／Ｉ，ｗ／ｗ），多奈哌齐 经皮贴剂中药物浓度超过１５％ｗｔ＿ｉ产生结晶，透皮贴剂能够保证长时间的给药需求， 贴剂中有关物质的生成量较少，药物的稳定性较好，但药物的体外经皮渗透速率较低， 需要加入促渗剂提高药物的经皮渗透速率。使用脂肪酸酯１和有机酸作为混合促渗剂时， 能够起到协同促渗作用，与对照组相比，可将多奈哌齐累积透过量提高２．７倍。使用最 优处方进行大鼠体内药代动力学实验，结果表明，与口服给药相比，多奈哌齐经皮给药 后，血药浓度垒少可以维持４８小时以上，药物生物利用度显著提高，－、Ｉ‘均滞留时间和 半衰期也相应显著延长。
耐碳青霉烯类鲍曼不动杆菌耐药特性分析
临床医学研究与实践2021年2月第6卷第6期Analysis of drug-resistant properties of the carbonapenem-resistantAcinetobacter baumanniiCHEN Yihong,YANG Hancai,MO Jiajin(Clinical Laboratory Department,Luoding People's Hospital,Luoding 527200,China)ABSTRACT:Objective To analyze the drug -resistant properti es and distribution of the carbonapenem -resistant Acinetobacter baumannii (CRAB).Methods A total of 230patients with Acinetobacter baumannii (AB)infection detected in our hospital from January 2019to July 2020were selected as the research objects.According to whether the patients had CRAB infection,the patients were divided into two groups:76patients with CRAB infection were carbapenem-resistant group,and 154patients with non CRAB infection were non carbapenem-resistant group.The drug-resistant properties of the two groups were compared and the distribution and sources of CRAB were analyzed.Results The drug -resistantproperties of the drug -resistant of the carbapenem -resistant group was significantly higher than that of the non carbapenem-resistant group,and the difference was statistically significant (P <0.05);the drug-resistance rate of the drug-resistant of the carbapenem-resistant group to other 10antibiotics was higher than 92.25%,but the drug-resistance rate to amikacin was 26.92%;the antimicrobial resistance of the drug-resistant of the non carbapenem-resistant group in this experiment was lower than 23.51%.CRAB of 44.74%was distributed in ICU and 15.79%in neurology department;the main source of specimens was sputum,accounting for 81.58%,and other sources were less than 10.00%.Conclusion The drug-resistance properties of CRAB is relatively higher,and metallo-β-lactamases is the main reason for the drug resistance of CRAB.The hospital should pay more attention to the drug-resistance of AB,strengthen the disinfection in the hospital and the isolation between patients,so as to prevent the large-scale infection in the hospital and reduce thehospital infection rate and mortality of patients.KEYWORDS:carbonapenem -resistant Acinetobacter baumannii ;drug -resistant properties;metallo -β-lactamases;hospital infection调查分析DOI :10.19347/ki.2096-1413.202106001作者简介：陈毅红（1989-），女，汉族，广东罗定人，主管检验师，学士。
PHA-793887_DataSheet_MedChemExpress
Inhibitors, Agonists, Screening Libraries Data SheetBIOLOGICAL ACTIVITY:PHA–793887 is a novel pan–cdk inhibitor, including cdk1, cdk2, cdk4, cdk5, cdk7, and cdk9 with IC50 in the 5 to 140 nM range.IC50 Value: 8 nM (CDK2), 5 nM(CDK5), 10 nM (CDK7)Target: CDKsin vitro: PHA–793887 is cytotoxic for leukemic cell lines, including K562, KU812, KCL22, and TOM1, with IC50 of 0.3–7 μM, but it is not cytotoxic for normal unstimulated peripheral blood mononuclear cells or CD34+ hematopoietic stem cells. In colony assays,PHA–793887 shows very high activity against leukemia cell lines with IC50 less than 0.1 μM. PHA–793887 induces cell–cycle arrest,inhibits Rb and nucleophosmin phosphorylation, and modulates cyclin E and cdc6 expression at 0.2–1 μM and induces apoptosis at 5μM. PHA–793887 has low activity against CDK1, CDK4, CDK9 and GSK3β with IC50 of 60 nM, 62 nM, 138 nM and 79 nM, respectively.PHA–793887 inhibits cell proliferation of many tumor cell lines, including A2780, HCT–116, COLO–205, C–433, DU–145, A375, PC3,MCF–7, and BX–PC3, with IC50 of 88 nM–3.4 μM. PHA–793887 (1 μM) shows a decrease in the S phase, a subsequent increase of the G1 phase and a slight accumulation of G2/M phase in A2780 cells. PHA–793887 (3 μM) significantly increases G2/M phase and reduces DNA synthsis.in vivo: PHA–793887 (20 mg/kg) is effective in xenograft models of K562 and HL60 cells, primary leukemic disseminated model, and a high–burden disseminated ALL–2 model derived from a relapsed Philadelphia–positive acute lymphoid leukemia patient. PHA–793887(10–30 mg/kg) shows good efficacy in the human ovarian A2780, colon HCT–116, and pancreatic BX–PC3 carcinoma xenograft models.PROTOCOL (Extracted from published papers and Only for reference)Animal administration [1]A2780 human ovarian adenocarcinoma–bearing CD–1 nude mice (Charles River Breeding Laboratories) were treated with 15 or 30mg/kg of PHA–793887 once a day by i.v. route for 2 days. Animals were sacrificed at 1.5, 6, and 24 hours after the last treatment, and tumor slices from mice treated with compound or vehicle (n = 9 animals per group) were collected and immediately soaked inRNAlater solution (Qiagen) for gene expression analysis. For pharmacokinetic analysis, plasma samples were collected from six animals at each time interval and analyzed by liquid chromatography–tandem mass spectrometry. Data were evaluated by nonlinear mixed effect modeling. Plasma levels of the compound were fitted using a two–compartment model with bolus input and first–order output mean, and random parameters were considered in the following equation: Pi = P.exp (η). The random effects were considered normal distributed with zero mean and variance ω2.References:[1]. Locatelli G et al Transcriptional analysis of an E2F gene signature as a biomarker of activity of the cyclin–dependent kinase inhibitor PHA–793887 inProduct Name:PHA–793887Cat. No.:HY-11001CAS No.:718630-59-2Molecular Formula:C 19H 31N 5O 2Molecular Weight:361.48Target:CDK Pathway:Cell Cycle/DNA Damage Solubility:10 mM in DMSOtumor and skin biopsies from a phase I clinical study. Mol Cancer Ther. 2010 May;9(5):1265–73.[2]. Massard C, Soria JC, Anthoney DA, Proctor A, Scaburri A, Pacciarini MA, Laffranchi B, Pellizzoni C, Kroemer G, Armand JP, Balheda R, Twelves CJ.A first in man, phase I dose–escalation study of PHA–793887, an inhibitor of multiple cyclin–dependent kinases (CDK2, 1 and 4) reveals unexpected hepatotoxicity in patients with solid tumors.Cell Cycle. 2011 Mar 15;10(6):963–70. Epub 2011 Mar 15.[3]. Alzani R, Pedrini O, Albanese C, Ceruti R, Casolaro A, Patton V, Colotta F, Rambaldi A, Introna M, Pesenti E, Ciomei M, Golay J.Therapeutic efficacy of the pan–cdk inhibitor PHA–793887 in vitro and in vivo in engraftment and high–burden leukemia models.Exp Hematol. 2010 Apr;38(4):259–269.e2. Epub 2010 Feb 16.[4]. Brasca MG, Albanese C, Alzani R, Amici R, Avanzi N, Ballinari D, Bischoff J, Borghi D, Casale E, Croci V, Fiorentini F, Isacchi A, Mercurio C, Nesi M, Orsini P, Pastori W, Pesenti E, Pevarello P, Roussel P, Varasi M, Volpi D, Vulpetti A, Ciomei M.Optimization of 6,6–dimethyl pyrrolo[3,4–c]pyrazoles: Identification of PHA–793887, a potent CDK inhibitor suitable for intravenous dosing.Bioorg Med Chem. 2010 Mar 1;18(5):1844–53. Epub 2010 Jan 25.[5]. Zoubir M, Flament C, Gdoura A, Bahleda R, Litvinova E, Soumelis V, Conforti R, Viaud S, Soria JC, Kroemer G, Zitvogel L, Chaput N.An inhibitor of cyclin–dependent kinases suppresses TLR signaling and increases the susceptibility of cancer patients to herpesviridae.Cell Cycle. 2011 Jan 1;10(1):118–26. Epub 2011 Jan 1.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。
碱基切除修复抑制剂甲氧胺联合β-榄香烯治疗恶性脑胶质瘤的实验研究
序言β-榄香烯属国家二类非细胞毒性抗肿瘤新药，临床研究证实其对包括脑胶质瘤在内的多种肿瘤疗效确切，且无其他传统化疗药常有的骨髓抑制、肝肾功能损害等毒副作用。
但目前临床应用的榄香烯乳注射液因其存在静脉炎发生率很高、剂型性质不稳定等缺点，其进一步的应用受到了较大的限制。
碱基切除修复抑制剂甲氧胺（Methoxyamine），可通过裂解核酸内切酶破坏DNA碱基切除修复过程，从而抑制肿瘤细胞对损伤作用的修复反应。
据此，可认为抑制DNA 碱基切除修复可能是增强肿瘤细胞化疗敏感性的潜在靶点，目前多项实验报道也已证实了甲氧胺可增强烷化剂和放疗的抗肿瘤效果。
近年来，通过纳米技术构建的纳米脂质体在提高药物溶解度、增加药物稳定性、降低药物副作用、缓控释给药等方面较普通的脂质体有了显著的提高。
研究表明，纳米脂质体对正常细胞和组织无损伤作用，并可长时间吸附于靶细胞周围，因此使药物能充分向靶组织渗透，也可以通过静电吸附效应与细胞膜接触而融合而进入细胞内。
因此将药物包封于纳米脂质体被认为可以改变被包封药物的体内分布，提高药物治疗指数，降低药物毒性。
基于增强β-榄香烯的疗效，减少毒副作用的目的，本课题研究内容分两部分：（一）联合碱基切除修复抑制剂甲氧胺，探讨是否在体内外抗瘤活性上具有协同作用，以期减少榄香烯用量，降低毒副反应，为其在临床的应用提供实验和理论依据。
（二）、利用纳米脂质体技术构建新型的β-榄香烯-纳米脂质体药物传递系统，初步探讨其体外抗瘤活性。
II碱基切除修复抑制剂甲氧胺联合β-榄香烯治疗恶性脑胶质瘤的实验研究中文摘要胶质瘤是成人神经系统最常见的原发性肿瘤，手术全切除率很低，复发率高，当前多种治疗效果仍不理想。
榄香烯属国家二类非细胞毒性抗肿瘤新药，临床研究发现其对多种肿瘤疗效确切，而且还具有提高和改善机体免疫功能，与放化疗协同作用等独特效果。
但是肿瘤细胞具有强大的DNA损伤修复机制，会对化疗药物产生抗性。
因此抑制这种内在的DNA修复过程，如碱基切除修复抑制剂甲氧胺的联合应用有利于提高化疗药物的抗瘤效果。
乙酰胆碱酯酶抑制剂
上海应用技术学院研究生课程《高等天然产物化学》试卷2014 / 2015 学年第1 学期课程代码：NX0702013论文题目：乙酰胆碱酯酶抑制剂的研究进展姓名：芮银146061414康满满146061409专业：制药工程学院：化工学院乙酰胆碱酯酶抑制剂的研究进展芮银，陈祎桐，康满满摘要：本文阐述了乙酰胆碱酯酶抑制剂(AChEI)的研究进展，介绍了用于药物治疗的乙酰胆碱酯酶抑制剂的各种来源如植物、微生物等，及其抑制乙酰胆碱的活性物质。
在此基础上，总结了几种现代分析技术，对AChEIs进行筛选，大大加快AD药物资源的开发利用进程。
这些方法主要有基于比色法的Ellman's法及相关的改进方法、薄层显色法、荧光显色法、电喷雾质谱法等。
但是，到目前为止，现代分析技术在AD药物资源中的应用还处在起步阶段。
关键词：乙酰胆碱酯酶抑制剂，筛选方法，薄层显色法，荧光显色法The progress of acetylcholinesteraseinhibitorsRui Yin, Chen Yitong, Kang ManmanAbstract：In this artical, the research elaborates progress of acetylcholinesterase inhibitors (AChEI), and introduces a variety of sources for drug treatment acetylcholinesterase inhibitors such as plants, microorganisms, and its active ingredients. On this basis, the review summarizes several modern analytic techniques such as Ellman's method which based on the colorimetric method, TLC chromogenic method, fluorescent color method, Electrospray ionization mass spectrometry and so on. However, at present, the application of modern analytic techniques in AD drug resources is still in infancy.Key word: Acetylcholinesterase inhibitors, Screening Methods, TLC chromogenic method, Fluorescent color method目录摘要.................................................................................................错误!未定义书签。
杯苋甾酮通过上调miR－204 表达抑制IL－1β 诱导的软骨细胞凋亡的实验研究
ʌ文章编号ɔ１００６－６２３３(２０２１)０３－０３６８－０７杯苋甾酮通过上调ｍｉＲ－２０４表达抑制ＩＬ－１β诱导的软骨细胞凋亡的实验研究杨少辉ꎬ㊀许红霞ꎬ㊀孙卫强ꎬ㊀崔慧君ꎬ㊀唐㊀丽(山东省文登整骨医院临床药学科ꎬ㊀山东㊀威海㊀２６４４００)ʌ摘㊀要ɔ目的:探讨杯苋甾酮对微小ＲＮＡ(ｍｉＲ)－２０４表达及白细胞介素(ＩＬ)－１β诱导的软骨细胞凋亡的影响ꎮ方法:体外培养人正常软骨细胞Ｃ２８/Ｉ２ꎬ设置①对照组(不做处理)㊁ＩＬ－１β组(５０ｍｇ/ＬＩＬ－１β)㊁低剂量杯苋甾酮组(２.５ｍｇ/Ｌ杯苋甾酮＋５０ｍｇ/ＬＩＬ－１β)㊁中剂量杯苋甾酮组(５ｍｇ/Ｌ杯苋甾酮＋５０ｍｇ/ＬＩＬ－１β)和高剂量杯苋甾酮组(１０ｍｇ/Ｌ杯苋甾酮＋５０ｍｇ/ＬＩＬ－１β)ꎻ②正常对照组(不转染)㊁阴性对照组(转染ＮＣ－ｓｉＲＮＡ)和ｍｉＲ－２０４ｓｉＲＮＡ组(转染ｍｉＲ－２０４ｓｉＲＮＡ)ꎬ三组均加入５０ｍｇ/ＬＩＬ－１β㊁１０ｍｇ/Ｌ杯苋甾酮处理ꎻ流式细胞仪检测各组Ｃ２８/Ｉ２细胞凋亡情况ꎻ酶联免疫(ＥＬＩＳＡ)法检测各组Ｃ２８/Ｉ２细胞上清液中炎症因子ＩＬ－６㊁ＩＬ－８㊁肿瘤坏死因子α(ＴＮＦ－α)水平ꎻ实时荧光定量ＰＣＲ(ｑＲＴ－ＰＣＲ)法检测各组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达情况ꎻ蛋白印迹(ＷＢ)法检测各组Ｃ２８/Ｉ２细胞中细胞色素Ｃ(ＣｙｔｏｃｈｒｏｍｅＣ)㊁Ｂｃｌ－２相关Ｘ蛋白(Ｂａｘ)㊁半胱天冬酶－３(Ｃａｓｐａｓｅ－３)蛋白表达情况ꎮ结果:与对照组相比ꎬＩＬ－１β组Ｃ２８/Ｉ２细胞活力㊁ｍｉＲ－２０４表达水平显著降低(Ｐ<０.０５)ꎬ细胞凋亡率㊁ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α表达水平及ＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白表达水平显著升高(Ｐ<０.０５)ꎻ与ＩＬ－１β组相比ꎬ随着杯苋甾酮剂量的增加ꎬＣ２８/Ｉ２细胞活力㊁ｍｉＲ－２０４表达水平逐次升高(Ｐ<０.０５)ꎬ细胞凋亡率㊁ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α表达水平及ＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白表达水平逐次降低(Ｐ<０.０５)ꎻ与阴性对照组相比ꎬｍｉＲ－２０４ｓｉＲＮＡ组相比凋亡率显著升高(Ｐ<０.０５)ꎮ结论:杯苋甾酮可能通过上调ｍｉＲ－２０４表达对ＩＬ－１β诱导的软骨细胞凋亡具有抑制作用ꎮʌ关键词ɔ㊀杯苋甾酮ꎻ㊀微小ＲＮＡ－２０４ꎻ㊀白细胞介素－１βꎻ㊀骨关节炎ꎻ㊀软骨细胞ʌ文献标识码ɔ㊀Ａ㊀㊀㊀㊀㊀ʌｄｏｉɔ１０.３９６９/ｊ.ｉｓｓｎ.１００６－６２３３.２０２１.０３.００４ＣｙａｓｔｅｒｏｎｅＩｎｈｉｂｉｔｓＩＬ－１βＩｎｄｕｃｅｄＣｈｏｎｄｒｏｃｙｔｅＡｐｏｐｔｏｓｉｓｂｙＵｐ－ｒｅｇｕｌａｔｉｎｇＭｉＲ－２０４ＥｘｐｒｅｓｓｉｏｎＹＡＮＧＳｈａｏｈｕｉꎬＸＵＨｏｎｇｘｉａꎬＳＵＮＷｅｉｑｉａｎｇꎬｅｔａｌ(ＷｅｎｄｅｎｇＯｒｔｈｏｐｅｄｉｃＨｏｓｐｉｔａｌꎬＳｈａｎｄｏｎｇＷｅｉｈａｉ２６４４００ꎬＣｈｉｎａ)ʌＡｂｓｔｒａｃｔɔＯｂｊｅｃｔｉｖｅ:ＴｏｉｎｖｅｓｔｉｇａｔｅｔｈｅｅｆｆｅｃｔｓｏｆＣｙａｓｔｅｒｏｎｅｏｎｔｈｅｅｘｐｒｅｓｓｉｏｎｏｆｍｉｃｒｏＲＮＡ(ｍｉＲ)－２０４ａｎｄｔｈｅａｐｏｐｔｏｓｉｓｏｆｃｈｏｎｄｒｏｃｙｔｅｓｉｎｄｕｃｅｄｂｙｉｎｔｅｒｌｅｕｋｉｎ(ＩＬ)－１β.Ｍｅｔｈｏｄｓ:Ｈｕｍａｎｎｏｒｍａｌｃｈｏｎｄｒｏ￣ｃｙｔｅｓＣ２８/Ｉ２ｗｅｒｅｃｕｌｔｕｒｅｄｉｎｖｉｔｒｏꎬａｎｄｓｅｔｕｐ①ｃｏｎｔｒｏｌｇｒｏｕｐ(ｎｏｔｒｅａｔｍｅｎｔ)ꎬＩＬ－１βｇｒｏｕｐ(５０ｍｇ/ＬＩＬ－１β)ꎬｌｏｗ－ｄｏｓｅＣｙａｓｔｅｒｏｎｅｇｒｏｕｐ(２.５ｍｇ/ＬＣｙａｓｔｅｒｏｎｅ＋５０ｍｇ/ＬＩＬ－１β)ꎬｍｅｄｉｕｍｄｏｓｅＣｙａｓｔｅｒｏｎｅｇｒｏｕｐ(５ｍｇ/ＬＣｙａｓｔｅｒｏｎｅ＋５０ｍｇ/ＬＩＬ－１β)ａｎｄｈｉｇｈｄｏｓｅＣｙａｓｔｅｒｏｎｅｇｒｏｕｐ(１０ｍｇ/ＬＣｙａｓｔｅｒｏｎｅ＋５０ｍｇ/ＬＩＬ－１β)ꎻ②ｎｏｒｍａｌｃｏｎｔｒｏｌｇｒｏｕｐ(ｎｏｔｒａｎｓｆｅｃｔｉｏｎ)ꎬｎｅｇａｔｉｖｅｃｏｎｔｒｏｌｇｒｏｕｐ(ｔｒａｎｓｆｅｃｔＮＣ－ｓｉＲＮＡ)ａｎｄｍｉＲ－２０４ｓｉＲＮＡｇｒｏｕｐ(ｔｒａｎｓｆｅｃｔｍｉＲ－２０４ｓｉＲＮＡ)ꎬｔｈｒｅｅｇｒｏｕｐｓｗｅｒｅａｄｍｉｎｓｔｅｒｅｄ５０ｍｇ/ＬＩＬ－１βａｎｄ１０ｍｇ/ＬＣｙａｓｔｅｒｏｎｅ.ＴｈｅｐｒｏｌｉｆｅｒａｔｉｏｎｏｆＣ２８/Ｉ２ｃｅｌｌｓｗａｓｄｅｔｅｃｔｅｄｂｙＣＣＫ－８ｍｅｔｈｏｄꎻｔｈｅａｐｏｐｔｏｓｉｓｏｆＣ２８/Ｉ２ｃｅｌｌｓｗａｓｄｅｔｅｃｔｅｄｂｙｆｌｏｗｃｙｔｏｍｅｔｒｙꎻｔｈｅｌｅｖｅｌｓｏｆＩＬ－６ꎬＩＬ－８ａｎｄｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ－α(ＴＮＦ－α)ｉｎｔｈｅｓｕｐｅＲ￣ＮＡｔａｎｔｏｆＣ２８/Ｉ２ｃｅｌｌｓｗｅｒｅｄｅｔｅｃｔｅｄｂｙｅｎｚｙｍｅ－ｌｉｎｋｅｄｉｍｍｕｎｏｓｏｒｂｅｎｔａｓｓａｙ(ＥＬＩＳＡ)ꎻｔｈｅｅｘｐｒｅｓｓｉｏｎｏｆｍｉＲ－２０４ｉｎＣ２８/Ｉ２ｃｅｌｌｓｗａｓｄｅｔｅｃｔｅｄｂｙｒｅａｌ－ｔｉｍｅｆｌｕｏｒｅｓｃｅｎｔｑｕａｎｔｉｔａｔｉｖｅＰＣＲ(ｑＲＴ－ＰＣＲ)ꎻｉｎａｄｄｉｔｉｏｎꎬｔｈｅｐｒｏｔｅｉｎｅｘｐｒｅｓｓｉｏｎｓｏｆＣｙｔｏｃｈｒｏｍｅＣꎬＢｃｌ－２－ｒｅｌａｔｅｄＸｐｒｏｔｅｉｎ(Ｂａｘ)ａｎｄＣａｓｐａｓｅ－３ｉｎＣ２８/Ｉ２ｃｅｌｌｓｗｅｒｅｄｅｔｅｃｔｅｄｂｙＷｅｓｔｅｒｎｂｌｏｔ(ＷＢ).Ｒｅｓｕｌｔｓ:ＣｏｍｐａｒｅｄｗｉｔｈｔｈｏｓｅｉｎｃｏｎｔｒｏｌｇｒｏｕｐꎬｔｈｅｃｅｌｌｖｉａｂｉｌｉｔｙａｎｄｍｉＲ－２０４ｅｘｐｒｅｓｓｉｏｎｌｅｖｅｌｏｆＣ２８/Ｉ２ｃｅｌｌｓｉｎＩＬ－１βｇｒｏｕｐｗｅｒｅｓｉｇｎｉｆｉｃａｎｔｌｙｌｏｗｅｒ(Ｐ<０.０５)ꎬａｎｄｔｈｅａｐｏｐｔｏｓｉｓʌ基金项目ɔ山东省中医药科技发展计划编号ꎬ(编号:２０１３ＺＤＺＫ－１４０)ｒａｔｅꎬｔｈｅｅｘｐｒｅｓｓｉｏｎｌｅｖｅｌｓｏｆＩＬ－６ꎬＩＬ－８ａｎｄＴＮＦ－αꎬｔｈｅｐｒｏｔｅｉｎｅｘｐｒｅｓｓｉｏｎｌｅｖｅｌｓｏｆＣｙｔｏｃｈｒｏｍｅＣꎬＢａｘａｎｄＣａｓｐａｓｅ－３ｗｅｒｅｓｉｇｎｉｆｉｃａｎｔｌｙｈｉｇｈｅｒ(Ｐ<０.０５)ꎻｍｏｒｅｏｖｅｒꎬｃｏｍｐａｒｅｄｗｉｔｈｔｈｏｓｅｉｎｔｈｅＩＬ－１βｇｒｏｕｐꎬｗｉｔｈｔｈｅａｄｄｉｔｉｏｎｏｆＣｙａｓｔｅｒｏｎｅａｎｄｔｈｅｉｎｃｒｅａｓｅｏｆｉｔｓｄｏｓａｇｅꎬｔｈｅｃｅｌｌｖｉａｂｉｌｉｔｙａｎｄｔｈｅｅｘｐｒｅｓｓｉｏｎｌｅｖｅｌｏｆｍｉＲ－２０４ｉｎＣ２８/Ｉ２ｃｅｌｌｓｉｎｃｒｅａｓｅｄｇｒａｄｕａｌｌｙ(Ｐ<０.０５)ꎬａｎｄｔｈｅａｐｏｐｔｏｓｉｓｒａｔｅꎬｔｈｅｅｘｐｒｅｓｓｉｏｎｌｅｖｅｌｓｏｆＩＬ－６ꎬＩＬ－８ａｎｄＴＮＦ－αꎬｔｈｅｐｒｏｔｅｉｎｅｘｐｒｅｓｓｉｏｎｌｅｖｅｌｓｏｆＣｙｔｏｃｈｒｏｍｅＣꎬＢａｘａｎｄＣａｓｐａｓｅ－３ｄｅｃｒｅａｓｅｄｇｒａｄｕａｌｌｙ(Ｐ<０.０５).ＣｏｍｐａｒｅｄｗｉｔｈｎｅｇａｔｉｖｅｃｏｎｔｒｏｌｇｒｏｕｐꎬｔｈｅａｐｏｐｔｏｓｉｓｒａｔｅｏｆｍｉＲ－２０４ｓｉＲＮＡｇｒｏｕｐｗｅｒｅｓｉｇｎｉｆｉｃａｎｔｌｙｉｎｃｒｅａｓｅｄ(Ｐ<０.０５).Ｃｏｎｃｌｕｓｉｏｎ:ＣｙａｓｔｅｒｏｎｅｍａｙｉｎｈｉｂｉｔｔｈｅＩＬ－１β－ｉｎｄｕｃｅｄａｐｏｐｔｏｓｉｓｏｆｃｈｏｎｄｒｏｃｙｔｅｓｂｙｕｐ－ｒｅｇｕｌａ￣ｔｉｎｇｔｈｅｅｘｐｒｅｓｓｉｏｎｏｆｍｉＲ－２０４.ʌＫｅｙｗｏｒｄｓɔ㊀Ｃｙａｓｔｅｒｏｎｅꎻ㊀ＭｉｃｒｏＲＮＡ－２０４ꎻ㊀Ｉｎｔｅｒｌｅｕｋｉｎ－１βꎻ㊀Ｏｓｔｅｏａｒｔｈｒｉｔｉｓꎻ㊀Ｃｈｏｎｄｒｏｃｙｔｅ㊀㊀骨关节炎(ｏｓｔｅｏａｒｔｈｒｉｔｉｓꎬＯＡ)是一种常见的慢性㊁进行性骨关节疾病ꎬ多见于中老年人ꎬ关节软骨破坏及继发的软骨细胞凋亡是其典型特征[１]ꎮ目前对于ＯＡ尚无满意的治疗方法ꎬ通常针对细胞外基质降解酶ꎬ寻找相应的酶活性抑制剂ꎬ但会出现不良反应[２]ꎮ虽然ＯＡ进展缓慢ꎬ但患者运动过程中负担很大ꎮ长期以来ꎬ大型负重关节的疼痛㊁僵硬会导致严重的残疾ꎬ需要进行外科手术治疗ꎬ影响生活质量的同时ꎬ还往往伴随着巨大的医疗保健成本ꎮ研究发现ꎬ软骨细胞凋亡是导致关节软骨退变并发展为ＯＡ的重要环节ꎬ抑制软骨细胞凋亡可能是治疗ＯＡ的有效途径[３]ꎮ白细胞介素(ｉｎｔｅｒｌｅｕｋｉｎꎬＩＬ)－１β作为一种致炎细胞因子ꎬ可诱导软骨细胞凋亡ꎬ在ＯＡ发生发展中起重要作用[４]ꎮ微小ＲＮＡ(ｍｉｃｒｏＲＮＡꎬｍｉＲＮＡ)是一种小分子非编码ＲＮＡꎬ能够调节软骨细胞增殖㊁分化㊁凋亡等生物学行为ꎬ在ＯＡ发生发展过程中起重要作用[５]ꎮ有研究发现ꎬｍｉＲ－２０４可通过抑制ＩＬ－１β的产生参与ＯＡ进展[６]ꎮ杯苋甾酮是苋科植物川牛膝的主要化学成分ꎬ可双向调节破骨和成骨ꎬ是一种潜在的骨质疏松治疗药物[７]ꎮ本研究将通过探讨杯苋甾酮通过ｍｉＲ－２０４途径对ＩＬ－１β诱导的软骨细胞凋亡的影响ꎬ以期为ＯＡ治疗提供参考ꎮ１㊀材料与方法１.１㊀试剂与仪器:人正常软骨细胞Ｃ２８/Ｉ２购自美国ＡＴＣＣ细胞库ꎻ胎牛血清购自澳大利亚ＡｕｓＧｅｎｅＸ公司ꎻＤＭＥＭ培养基购自德国ｍｅｒｃｋ/ｓｉｇｍａ公司ꎻ重组人ＩＬ－１β购自美国ｓａｎｔａ公司ꎻ杯苋甾酮(批号:Ｄ００７３ꎬＨＰＬＣȡ９８％)购自上海宝曼生物科技有限公司ꎻＣＣＫ－８试剂购自日本同仁化学研究所ꎻＡｎｎｅｘｉｎＶ－ＦＩＴＣ/ＰＩ细胞凋亡检测试剂盒购自哈尔滨新海基因检测有限公司ꎻ实时荧光定量ＰＣＲ(ｑｕａｎｔｉｔａｔｉｖｅｒｅａｌ－ｔｉｍｅＰＣＲꎬｑＲＴ－ＰＣＲ)试剂盒购自美国Ｐｒｏｍｅｇａ公司ꎻＬｉｐｏ￣ｆｅｃｔａｍｉｎｅ２０００转染试剂(货号:１１６６８０３０)购自美国ＬｉｆｅＴｅｃｈｎｏｌｏｇｉｅｓꎻＮＣ－ｓｉＲＮＡ㊁ｍｉＲ－２０４ｓｉＲＮＡ及ｍｉＲ－２０４㊁Ｕ６引物均由生工生物工程(上海)股份有限公司合成ꎻＩＬ－６㊁ＩＬ－８㊁肿瘤坏死因子α(ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃ￣ｔｏｒ－ａｌｐｈａꎬＴＮＦ－α)㊁酶联免疫(ｅｎｚｙｍｅ－ｌｉｎｋｅｄｉｍｍｕ￣ｎｏｓｏｒｂｅｎｔａｓｓａｙꎬＥＬＩＳＡ)试剂盒㊁细胞色素Ｃ(Ｃｙｔｏ￣ｃｈｒｏｍｅＣ)抗体㊁Ｂｃｌ－２相关Ｘ蛋白(Ｂｃｌ－２ａｓｓｏｃｉａｔｅｄＸｐｒｏｔｅｉｎꎬＢａｘ)抗体㊁半胱天冬酶－３(Ｃａｓｐａｓｅ－３)抗体㊁ＧＡＰＤＨ抗体购自ａｂｃａｍ公司ꎻ辣根过氧化物酶标记的羊抗兔(批号:０２９５Ｇ－ＨＲＰ)购自美国ｓａｎｔａ公司ꎻ恒温培养箱(型号:ＭＩＲ－１６２－ＰＣ/ＭＩＲ－２６２－ＰＣ)ꎬ日本松下公司ꎻ流式细胞仪(型号:ＢＤＦＡＣＳＣａｎｔｏＩＩ)ꎬ美国ＢＤ公司ꎻ荧光定量ＰＣＲ仪(型号:ＡＢＩ７５００)为美国ＡｐｐｌｉｅｄＢｉｏｓｙｓｔｅｍｓ公司产品ꎻ酶标仪㊁化学发光成像系统(型号:ＭＯＤＥＬ５５０㊁ＣｈｅｍｉＤｏｃＸＲＳ)ꎬ美国Ｂｉｏ－Ｒａｄ公司ꎮ１.２㊀实验方法１.２.１㊀细胞培养:将Ｃ２８/Ｉ２细胞接种于含１０％胎牛血清㊁１００Ｕ/ｍＬ青霉素－链霉素的ＤＭＥＭ培养液中ꎬ置于３７ħ㊁５％ＣＯ２培养箱中培养ꎬ待细胞融合率达到８０％左右时ꎬ用胰蛋白酶消化㊁传代培养ꎮ取对数期Ｃ２８/Ｉ２细胞以１ˑ１０４个/孔的密度接种于９６孔培养板ꎬ设置①对照组㊁ＩＬ－１β组㊁低剂量杯苋甾酮组㊁中剂量杯苋甾酮组和高剂量杯苋甾酮组ꎬ对照组不进行处理ꎬＩＬ－１β组加５０ｍｇ/ＬＩＬ－１β处理[８]ꎬ低剂量杯苋甾酮组㊁中剂量杯苋甾酮组㊁高剂量杯苋甾酮组在加５０ｍｇ/ＬＩＬ－１β处理的基础上ꎬ分别加入杯苋甾酮ꎬ终浓度分别为２.５ｍｇ/Ｌ㊁５ｍｇ/Ｌ㊁１０ｍｇ/Ｌ[７]ꎻ②正常对照组㊁阴性对照组和ｍｉＲ－２０４ｓｉＲＮＡ组ꎮ三组均加入５０ｍｇ/ＬＩＬ－１β㊁１０ｍｇ/Ｌ杯苋甾酮处理ꎬ其中阴性对照组和ｍｉＲ－２０４ｓｉＲＮＡ组Ｃ２８/Ｉ２细胞使用Ｌｉｐｏ￣ｆｅｃｔａｍｉｎｅ２０００转染试剂分别转染ＮＣ－ｓｉＲＮＡ㊁ｍｉＲ－２０４ｓｉＲＮＡꎮ１.２.２㊀ＣＣＫ－８法检测Ｃ２８/Ｉ２细胞增殖情况:将Ｃ２８/Ｉ２细胞培养２４ｈ㊁４８ｈ㊁７２ｈꎬ同时设置仅含培养基的空白组ꎬ加入ＣＣＫ－８试剂后继续培养２ｈꎬ使用全自动酶标仪于波长４５０ｎｍ处检测各孔吸光度(ｏｐｔｉｃａｌｄｅｎｓｉｔｙꎬＯＤ)值ꎬ计算细胞活力ꎬ细胞活力＝(ＯＤ处理组－ＯＤ空白组)/(ＯＤ对照组－ＯＤ空白组)ˑ１００％ꎮ１.２.３㊀流式细胞仪检测Ｃ２８/Ｉ２细胞凋亡情况:将Ｃ２８/Ｉ２细胞培养４８ｈꎬ收集细胞ꎬ胰蛋白酶消化ꎬＰＢＳ洗涤ꎬ使用４００μＬ１ˑ结合缓冲液调整成细胞浓度为１ˑ１０６个/ｍＬ的细胞悬浮液ꎬ按照ＡｎｎｅｘｉｎＶ－ＦＩＴＣ/ＰＩ凋亡检测试剂盒操作说明书步骤ꎬ分别加入ＡｎｎｅｘｉｎＶ－ＦＩＴＣ㊁ＰＩ各５μＬꎬ避光孵育１ｈꎬ使用流式细胞仪检测细胞凋亡率ꎮ１.２.４㊀ＥＬＩＳＡ法检测Ｃ２８/Ｉ２细胞上清液中炎症因子水平:将Ｃ２８/Ｉ２细胞培养４８ｈꎬ收集各组Ｃ２８/Ｉ２细胞上清液ꎬ严格按照ＥＬＩＳＡ试剂盒说明书操作ꎬ酶标仪于４５０ｎｍ波长下检测各孔ＯＤ值ꎬ计算ＩＬ－６㊁ＩＬ－８和ＴＮＦ－α水平ꎮ１.２.５㊀ｑＲＴ－ＰＣＲ法检测Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达情况:将Ｃ２８/Ｉ２细胞培养４８ｈꎬ收集细胞ꎬ使用Ｔｒ￣ｉｚｏｌ试剂提取总ＲＮＡꎬ逆转录合成ｃＤＮＡꎬ采用ｑＲＴ－ＰＣＲ法检测ｍｉＲ－２０４表达水平ꎬ内参基因为Ｕ６ꎮ反应程序:９５ħ预变性１０ｍｉｎꎻ９５ħ变性１０ｓꎬ５５ħ退火３５ｓꎬ７２ħ延伸１ｍｉｎꎬ循环４０次ꎮｍｉＲ－２０４上游引物:５'－ＡＣＧＣＧＴＧＴＡＧＴＡＧＣＴＧＣＴＧＡＧ－３'ꎬ下游引物:５'－ＧＣＧＧＵＡＵＣＧＵＡＵＣＧＵＡＵＧＣＵＡＧ－３'ꎻ内参Ｕ６上游引物:５'－ＣＴＣＧＣＴＴＣＧＧＣＡＧＣＡＣＡ－３'ꎬ下游引物:５'－ＡＡＣＧＣＴＴＣＡＣＧＡＡＴＴＴＧＣＧＴ－３'ꎮＣｔ值为扩增产物的荧光信号达到临界阈值时对应的循环数ꎬ相对表达量以２－ΔΔＣｔ法表示ꎮ１.２.６㊀蛋白印迹(ｗｅｓｔｅｒｎｂｌｏｔꎬＷＢ)法检测Ｃ２８/Ｉ２细胞中ＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白表达情况:将Ｃ２８/Ｉ２细胞培养４８ｈꎬ收集细胞ꎬ使用蛋白提取试剂盒提取各组细胞总蛋白ꎬ使用ＢＣＡ蛋白测定试剂盒测定蛋白浓度并进行定量ꎮ电泳分离等量蛋白质并转至ＰＶＤＦ膜上ꎬ用含５％脱脂奶粉的ＴＢＳＴ封闭１ｈꎬ分别加入ＣｙｔｏｃｈｒｏｍｅＣ抗体㊁Ｂａｘ抗体㊁Ｃａｓｐａｓｅ－３抗体㊁ＧＡＰＤＨ抗体(１:５００)ꎬ４ħ孵育过夜ꎬＴＢＳＴ洗涤后加辣根过氧化物酶标记的羊抗兔二抗(１:５０００)ꎬ室温孵育１ｈꎬ免疫反应化学发光法显色ꎬＴａｎｏｎ６００图像分析系统拍摄图像并分析条带灰度ꎬ目的蛋白相对表达量＝目的蛋白灰度值/内参ＧＡＰＤＨ灰度值ꎮ１.３㊀统计学分析:数据处理采用ＳＰＳＳ２４.０统计学软件ꎮ计量资料采用均数ʃ标准差( ｘʃｓ)表示ꎬ多组间比较采用单因素方差分析ꎬ进一步两两比较采用ＳＮＫ－ｑ检验ꎮ当Ｐ<０.０５时ꎬ差异有统计学意义ꎮ２㊀结㊀果２.１㊀各组Ｃ２８/Ｉ２细胞增殖情况:各个时间点ꎬ与对照组相比ꎬＩＬ－１β组Ｃ２８/Ｉ２细胞活力显著降低(Ｐ<０.０５)ꎻ与ＩＬ－１β组相比ꎬ随着杯苋甾酮剂量的升高ꎬＣ２８/Ｉ２细胞活力逐次升高ꎮ见表１ꎮ表１㊀各组Ｃ２８/Ｉ２细胞活力比较( ｘʃｓꎬｎ＝６)组别细胞活力(％)２４ｈ㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀４８ｈ㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀７２ｈ对照组１００ʃ０.００１００.００ʃ０.００１００.００ʃ０.００ＩＬ－１β组５６.７４ʃ４.０２ａ４１.６７ʃ４.８６ａ３６.５１ʃ４.６２ａ低剂量杯苋甾酮组６３.７３ʃ４.４３５２.０８ʃ４.９３ｂ４７.０２ʃ４.４９ｂ中剂量杯苋甾酮组７４.６５ʃ５.２８ｂｃ６４.５８ʃ６.５７ｂｃ５５.９３ʃ４.５３ｂｃ高剂量杯苋甾酮组８５.２６ʃ５.７４ｂｃｄ８１.２５ʃ６.４４ｂｃｄ７３.６６ʃ５.０４ｂｃｄＦ８８.４１０１２２.０８９２１３.５２０Ｐ０.００００.００００.０００㊀㊀注:与对照组相比ꎬａＰ<０.０５ꎻ与ＩＬ－１β组相比ꎬｂＰ<０.０５ꎻ与低剂量杯苋甾酮组相比ꎬｃＰ<０.０５ꎻ与中剂量杯苋甾酮组相比ꎬｄＰ<０.０５２.２㊀各组Ｃ２８/Ｉ２细胞凋亡情况:与对照组相比ꎬＩＬ－１β组Ｃ２８/Ｉ２细胞凋亡率显著升高(Ｐ<０.０５)ꎻ与ＩＬ－１β组相比ꎬ低㊁中㊁高剂量杯苋甾酮组Ｃ２８/Ｉ２细胞凋亡率显著降低(Ｐ<０.０５)ꎻ随着杯苋甾酮剂量的升高ꎬＣ２８/Ｉ２细凋亡率逐次降低(Ｐ<０.０５)ꎮ见图１㊁表２ꎮ表２㊀各组Ｃ２８/Ｉ２细胞凋亡率比较( ｘʃｓꎬｎ＝６)组别凋亡率(％)对照组５.６４ʃ０.８２ＩＬ－１β组３８.９２ʃ２.３５ａ低剂量杯苋甾酮组３０.０４ʃ２.０２ｂ中剂量杯苋甾酮组２４.１１ʃ２.１８ｂｃ高剂量杯苋甾酮组１７.２５ʃ１.５７ｂｃｄＦ２７３.７５３Ｐ０.０００㊀㊀注:与对照组相比ꎬａＰ<０.０５ꎻ与ＩＬ－１β组相比ꎬｂＰ<０.０５ꎻ与低剂量杯苋甾酮组相比ꎬｃＰ<０.０５ꎻ与中剂量杯苋甾酮组相比ꎬｄＰ<０.０５２.３㊀各组Ｃ２８/Ｉ２细胞上清液中炎症因子表达水平:与对照组相比ꎬＩＬ－１β组Ｃ２８/Ｉ２细胞上清液中ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α表达水平显著升高(Ｐ<０.０５)ꎻ与ＩＬ－１β组相比ꎬ低㊁中㊁高剂量杯苋甾酮组Ｃ２８/Ｉ２细胞上清液中ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α表达水平显著降低(Ｐ<０.０５)ꎻ随着杯苋甾酮剂量的升高ꎬＣ２８/Ｉ２细胞上清液中ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α表达水平逐次降低(Ｐ<０.０５)ꎮ见表３ꎮ２.４㊀各组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达情况:与对照组相比ꎬＩＬ－１β组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达水平显著降低(Ｐ<０.０５)ꎻ与ＩＬ－１β组相比ꎬ低㊁中㊁高剂量杯苋甾酮组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达水平显著升高(Ｐ<０.０５)ꎻ随着杯苋甾酮剂量的升高ꎬＣ２８/Ｉ２细胞中ｍｉＲ－２０４表达水平逐次升高(Ｐ<０.０５)ꎮ见图２㊁表４ꎮ表３㊀各组Ｃ２８/Ｉ２细胞上清液中炎症因子表达水平比较( ｘʃｓꎬｎ＝６)组别ＩＬ－６(ｐｇ/ｍＬ)ＩＬ－８(ｐｇ/ｍＬ)ＴＮＦ－α(ｐｇ/ｍＬ)对照组４４.２８ʃ６.８２３６.９４ʃ５.０６２４.５５ʃ４.５４ＩＬ－１β组１５２.６６ʃ２２.３５ａ１１９.２６ʃ１１.２３ａ８８.２６ʃ１０.３６ａ低剂量杯苋甾酮组１２２.４７ʃ２０.１６ｂ９８.５５ʃ１１.５４ｂ６９.５４ʃ９.６７ｂ中剂量杯苋甾酮组９６.８４ʃ１７.６５ｂｃ７４.８８ʃ８.６３ｂｃ５０.１７ʃ７.５８ｂｃ高剂量杯苋甾酮组７０.０４ʃ１３.０３ｂｃｄ５９.７１ʃ６.７５ｂｃｄ３８.９４ʃ７.０３ｂｃｄＦ３７.９３９７６.９５４５７.６４６Ｐ０.００００.００００.０００㊀㊀注:与对照组相比ꎬａＰ<０.０５ꎻ与ＩＬ－１β组相比ꎬｂＰ<０.０５ꎻ与低剂量杯苋甾酮组相比ꎬｃＰ<０.０５ꎻ与中剂量杯苋甾酮组相比ꎬｄＰ<０.０５图１㊀各组Ｃ２８/Ｉ２细胞凋亡情况２.５㊀各组Ｃ２８/Ｉ２细胞中ＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白表达情况:与对照组相比ꎬＩＬ－１β组Ｃ２８/Ｉ２细胞中ＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白表达水平显著升高(Ｐ<０.０５)ꎻ与ＩＬ－１β组相比ꎬ低㊁中㊁高剂量杯苋甾酮组Ｃ２８/Ｉ２细胞中ＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白表达水平显著降低(Ｐ<０.０５)ꎻ随着杯苋甾酮剂量的升高ꎬＣ２８/Ｉ２细胞中ＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白表达水平逐次降低(Ｐ<０.０５)ꎮ见图３㊁表５ꎮ图２㊀五组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达电泳图２.６㊀下调ｍｉＲ－２０４表达对杯苋甾酮抑制Ｃ２８/Ｉ２细胞凋亡的影响:正常对照组与阴性对照组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达水平㊁凋亡率的差异无统计学意义(Ｐ<０.０５)ꎮ与阴性对照组相比ꎬｍｉＲ－２０４ｓｉＲＮＡ组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达水平显著降低ꎬ凋亡率显著升高(Ｐ<０.０５)ꎮ见图４㊁图５㊁表６ꎮ表４㊀各组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达水平比较( ｘʃｓꎬｎ＝６)组别ｍｉＲ－２０４对照组１.０２ʃ０.０９ＩＬ－１β组０.５１ʃ０.０７ａ低剂量杯苋甾酮组０.６２ʃ０.０６ｂ中剂量杯苋甾酮组０.７５ʃ０.０９ｂｃ高剂量杯苋甾酮组０.８９ʃ０.１１ｂｃｄＦ３３.９７０Ｐ０.０００㊀㊀注:与对照组相比ꎬａＰ<０.０５ꎻ与ＩＬ－１β组相比ꎬｂＰ<０.０５ꎻ与低剂量杯苋甾酮组相比ꎬｃＰ<０.０５ꎻ与中剂量杯苋甾酮组相比ꎬｄＰ<０.０５图３㊀各组Ｃ２８/Ｉ２细胞中ＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白表达情况图４㊀下调ｍｉＲ－２０４表达对杯苋甾酮抑制Ｃ２８/Ｉ２细胞凋亡的影响表５㊀各组Ｃ２８/Ｉ２细胞中ＣｙｔｏｃｈｒｏｍｅＣＢａｘＣａｓｐａｓｅ－３蛋白表达水平比较( ｘʃｓꎬｎ＝６)组别ＣｙｔｏｃｈｒｏｍｅＣ/ＧＡＰＤＨＢａｘ/ＧＡＰＤＨＣａｓｐａｓｅ－３/ＧＡＰＤＨ对照组０.１７ʃ０.０４０.２３ʃ０.０５０.１５ʃ０.０３ＩＬ－１β组０.９５ʃ０.１１ａ０.８７ʃ０.１１ａ０.８６ʃ０.１０ａ低剂量杯苋甾酮组０.７４ʃ０.０９ｂ０.６２ʃ０.０８ｂ０.５９ʃ０.０８ｂ中剂量杯苋甾酮组０.４８ʃ０.０６ｂｃ０.５０ʃ０.０６ｂｃ０.４８ʃ０.０８ｂｃ高剂量杯苋甾酮组０.３６ʃ０.０６ｂｃｄ０.３９ʃ０.０６ｂｃｄ０.３９ʃ０.０５ｂｃｄＦ９８.５３４６２.２０２７８.０１１Ｐ０.００００.００００.０００㊀㊀注:与对照组相比ꎬａＰ<０.０５ꎻ与ＩＬ－１β组相比ꎬｂＰ<０.０５ꎻ与低剂量杯苋甾酮组相比ꎬｃＰ<０.０５ꎻ与中剂量杯苋甾酮组相比ꎬｄＰ<０.０５表６㊀各组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达水平及细胞凋亡率比较( ｘʃｓꎬｎ＝６)组别ｍｉＲ－２０４细胞凋亡率(％)正常对照组１.０２ʃ０.０８１８.６７ʃ１.７４阴性对照组１.０１ʃ０.０６１８.１３ʃ１.５９ｍｉＲ－２０４ｓｉＲＮＡ组０.６８ʃ０.０６ａ３３.１５ʃ１.８２ａＦ４９.５４４１４７.３４７Ｐ０.００００.０００㊀㊀注:与阴性对照组相比ꎬａＰ<０.０５图５㊀三组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达电泳图３㊀讨㊀论现有研究已对ＯＡ发病机制有一定程度的了解ꎬ大量的候选药物也在为用于ＯＡ治疗开展相关实验ꎮ然而ꎬ目前进入临床实验的试验药物都是高特异性的ꎬ属于针对某种细胞外基质降解酶的抑制剂ꎬ这种治疗手段并不能改善ＯＡ的整个疾病进程ꎬ治疗效果有限ꎬ且部分药物具有副作用[２]ꎮＩＬ是一类调节细胞生长及分化的因子ꎬ主要由白细胞分泌ꎬ目前已知与软骨损伤有关的ＩＬ因子有ＩＬ－１β㊁ＩＬ－６㊁ＩＬ－８等[９]ꎮ其中ꎬＩＬ－１β是破坏关节软骨最直接的ＩＬ细胞因子ꎬ其水平与关节软骨退变程度呈正比ꎬ能够促进软骨基质的降解ꎬ并诱导软骨细胞的凋亡ꎬ常用于诱导关节软骨细胞凋亡建立关节炎软骨细胞模型ꎮＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α是参与炎症反应的常见因子ꎬ均参与ＯＡ的发生发展过程[１０]ꎮ本研究结果显示ꎬ与对照组相比ꎬＩＬ－１β组Ｃ２８/Ｉ２细胞ＯＤ值显著降低ꎬ凋亡率显著升高ꎬ上清液中ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α表达水平显著升高ꎬ提示ＩＬ－１β可以抑制Ｃ２８/Ｉ２细胞增殖ꎬ并诱导其凋亡ꎬ这可能是通过促进ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α等炎症因子表达ꎬ增强炎症反应发挥作用的ꎮｍｉＲＮＡ是一种单链小分子非编码ＲＮＡꎬ广泛存在于真核生物中ꎬ参与对炎症反应的调控ꎮ李静等[１１]研究表明ꎬｍｉＲ－２０４能通过下调ＮＯＴＣＨ２减轻心肌缺血再灌注大鼠的心肌炎症反应及氧化应激反应ꎮ徐琰瑛等[１２]研究表明ꎬｍｉＲ－２０４在角膜损伤愈合过程中ꎬ不仅与小梁网细胞的凋亡及生存能力有关ꎬ还与炎症介质的表达有着重要联系ꎮＳｏｎｇ等[６]研究发现ꎬＯＡ软骨组织中ｍｉＲ－２０４水平明显降低ꎬ使用ＩＬ－１β处理软骨细胞可降低ｍｉＲ－２０４水平ꎬ而过表达ｍｉＲ－２０４可抑制软骨细胞中ＩＬ－１β蛋白水平ꎬ并消除ＩＬ－１β对软骨细胞中ＣＯＸ－２㊁ＩＬ－６表达的促进作用ꎮ本研究结果亦显示ꎬＩＬ－１β组Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达水平较对照组显著降低ꎬ提示ＩＬ－１β可抑制Ｃ２８/Ｉ２细胞中ｍｉＲ－２０４表达ꎮ川牛膝是一种苋科植物ꎬ其根干燥后具有祛瘀止痛㊁活血通经㊁强筋健骨等功效ꎬ常用来治疗骨折及其炎症反应[１３]ꎮ杯苋甾酮是川牛膝的主要化学成分ꎬ在治疗骨质疏松症时具有抑制破骨分化和促进成骨分化的双向作用ꎬ在骨折修复中有助于加快骨折愈合并缩短骨折愈合时间[１４]ꎮ本研究发现ꎬ与ＩＬ－１β组相比ꎬ杯苋甾酮组Ｃ２８/Ｉ２细胞ＯＤ值及ｍｉＲ－２０４表达水平显著升高ꎬ细胞凋亡率及上清液中ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α表达水平显著降低ꎬ提示使用杯苋甾酮可以促进ｍｉＲ－２０４表达ꎬ降低炎症因子ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α的表达ꎬ促进Ｃ２８/Ｉ２细胞增殖并减少其凋亡ꎮ进一步研究显示ꎬ在使用ＩＬ－１β㊁杯苋甾酮处理Ｃ２８/Ｉ２细胞基础上ꎬ下调ｍｉＲ－２０４表达可导致细胞凋亡率升高ꎬ提示ｍｉＲ－２０４表达水平与Ｃ２８/Ｉ２细胞凋亡过程有关ꎬ杯苋甾酮减少Ｃ２８/Ｉ２细胞凋亡可能是通过上调ｍｉＲ－２０４表达发挥作用的ꎬ并进一步推测杯苋甾酮通过上调ｍｉＲ－２０４表达ꎬ消除ＩＬ－１β引起的炎症因子ＩＬ－６㊁ＩＬ－８㊁ＴＮＦ－α水平变化ꎬ进而影响Ｃ２８/Ｉ２细胞的增殖㊁凋亡ꎬ但ｍｉＲ－２０４影响炎症因子变化的途径还有待深入研究[６]ꎮＣｙｔｏｃｈｒｏｍｅＣ是重要的凋亡激活因子ꎬ当细胞受到病理因素刺激后ꎬ从线粒体释放至细胞质中ꎬ激活细胞质中的凋亡反应ꎬ诱导细胞凋亡发生ꎮ有研究发现ꎬＩＬ－１β可以促进ＣｙｔｏｃｈｒｏｍｅＣ释放ꎬ经由线粒体途径诱导关节软骨细胞的凋亡[１５]ꎮＢａｘ㊁Ｃａｓｐａｓｅ－３分别为Ｂｃｌ－２家族㊁ｃａｓｐａｓｅ家族的重要成员ꎬ参与细胞凋亡ꎮ本研究结果显示ꎬＩＬ－１β促进Ｃ２８/Ｉ２细胞凋亡及杯苋甾酮通过促进ｍｉＲ－２０４表达发挥抑制ＩＬ－１β对Ｃ２８/Ｉ２细胞的促凋亡作用过程中ꎬＣｙｔｏｃｈｒｏｍｅＣ㊁Ｂａｘ㊁Ｃａｓｐａｓｅ－３蛋白水平均有显著变化ꎬ表明杯苋甾酮促进ｍｉＲ－２０４表达后ꎬ减轻ＩＬ－１β引起的炎症反应ꎬ并可能通过调控ＣｙｔｏｃｈｒｏｍｅＣ释放ꎬ进而减少促凋亡蛋白Ｂａｘ㊁Ｃａｓｐａｓｅ－３表达ꎬ从而抑制Ｃ２８/Ｉ２细胞凋亡ꎮ综上所述ꎬ杯苋甾酮可能通过上调ｍｉＲ－２０４表达ꎬ进而抑制ＩＬ－１β诱导的软骨细胞Ｃ２８/Ｉ２凋亡ꎮ这可能为骨关节炎症的治疗提供了一定参考ꎬ但ｍｉＲＮＡ调控机制复杂ꎬ下一步研究将更深入明确其具体调控机制ꎮʌ参考文献ɔ[１]㊀ＧｅＱꎬＷａｎｇＨꎬＸｕＸꎬｅｔａｌ.ＰＤＫ１ｐｒｏｍｏｔｅｓａｐｏｐｔｏｓｉｓｏｆｃｈｏｎｄｒｏｃｙｔｅｓｖｉａｍｏｄｕｌａｔｉｎｇＭＡＰＫｐａｔｈｗａｙｉｎｏｓｔｅｏａｒｔｈｒｉｔｉｓ[Ｊ].ＴｉｓｓｕｅＣｅｌｌꎬ２０１７ꎬ４９(６):７１９~７２５.[２]㊀ＲｏｏｓＥＭꎬＡｒｄｅｎＮＫ.Ｓｔｒａｔｅｇｉｅｓｆｏｒｔｈｅｐｒｅｖｅｎｔｉｏｎｏｆｋｎｅｅｏｓ￣ｔｅｏａｒｔｈｒｉｔｉｓ[Ｊ].ＮａｔＲｅｖＲｈｅｕｍａｔｏｌꎬ２０１６ꎬ１２(２):９２~１０１. [３]㊀ＧｕＲꎬＬｉｕＮꎬＬｕｏＳꎬｅｔａｌ.ＭｉｃｒｏＲＮＡ－９ｒｅｇｕｌａｔｅｓｔｈｅｄｅｖｅｌ￣ｏｐｍｅｎｔｏｆｋｎｅｅｏｓｔｅｏａｒｔｈｒｉｔｉｓｔｈｒｏｕｇｈｔｈｅＮＦ－ｋａｐｐａＢ１ｐａｔｈ￣ｗａｙｉｎｃｈｏｎｄｒｏｃｙｔｅｓ[Ｊ].Ｍｅｄｉｃｉｎｅ(Ｂａｌｔｉｍｏｒｅ)ꎬ２０１６ꎬ９５(３６):１~７.[４]㊀武豪杰ꎬ王晓ꎬ张明辉ꎬ等.白细胞介素－１β和基质金属蛋白酶－９在兔膝关节骨性关节炎模型滑膜中的表达及意义[Ｊ].中华实验外科杂志ꎬ２０１６ꎬ３３(３):７４１~７４３. [５]㊀ＷａｎｇＪꎬＣｈｅｎＬꎬＪｉｎＳꎬｅｔａｌ.ＡｌｔｅｒｅｄｅｘｐｒｅｓｓｉｏｎｏｆｍｉｃｒｏＲＮＡ－９８ｉｎＩＬ－１β－ｉｎｄｕｃｅｄｃａｒｔｉｌａｇｅｄｅｇｒａｄａｔｉｏｎａｎｄｉｔｓｒｏｌｅｉｎｃｈｏｎｄｒｏｃｙｔｅａｐｏｐｔｏｓｉｓ[Ｊ].ＭｏｌＭｅｄＲｅｐꎬ２０１７ꎬ１６(３):３２０８~３２１６.[６]㊀ＳｏｎｇＸꎬＺｈｕＭꎬＳｕｎＹꎬｅｔａｌ.ＭｉＲ－２０４ｅｎｈａｎｃｅｓｔｈｅｐｒｏｇｒｅｓ￣ｓｉｏｎｏｆｏｓｔｅｏａｒｔｈｒｉｔｉｓｂｙｓｕｐｐｒｅｓｓｉｎｇｔｈｅｐｒｏｄｕｃｔｉｏｎｏｆＩＬ－１β[Ｊ].Ｐｈａｒｍａｚｉｅꎬ２０１７ꎬ７２(１０):５８７~５９２.[７]㊀纪亲龙ꎬ孔祥东ꎬ戚珊红ꎬ等.杯苋甾酮抑制破骨分化和促进成骨分化的双向作用治疗骨质疏松症[Ｊ].昆明医科大学学报ꎬ２０１８ꎬ３９(５):２１~２８.[８]㊀章国银ꎬ章礼炜ꎬ范少华ꎬ等.槲皮苷对ＩＬ－１β诱导的软骨细胞凋亡的影响及其机制[Ｊ].武汉大学学报(医学版)ꎬ２０１８ꎬ３９(５):７４５~７７４.[９]㊀ＹｕＣＤꎬＭｉａｏＷＨꎬＺｈａｎｇＹＹꎬｅｔａｌ.ＩｎｈｉｂｉｔｉｏｎｏｆｍｉＲ－１２６ｐｒｏｔｅｃｔｓｃｈｏｎｄｒｏｃｙｔｅｓｆｒｏｍＩＬ－１βｉｎｄｕｃｅｄｉｎｆｌａｍｍａｔｉｏｎｖｉａｕｐｒｅｇｕｌａｔｉｏｎｏｆＢｃｌ－２[Ｊ].ＢｏｎｅＪｏｉｎｔＲｅｓꎬ２０１８ꎬ７(６):４１４~４２１.[１０]㊀杜津莉ꎬ樊炜骏ꎬ李建ꎬ等.独活寄生汤含药血清对膝关节ＯＡ大鼠滑膜细胞ＴＮＦ－α㊁ＩＬ－６㊁ＩＬ－８㊁ＩＬ－１０表达的影响及其机制[Ｊ].山东医药ꎬ２０１７ꎬ５７(４４):１４~１７. [１１]㊀李静ꎬ武敏ꎬ朱海慧ꎬ等.ｍｉＲ－２０４改善心肌缺血再灌注大鼠炎症及氧化应激[Ｊ].医学分子生物学杂志ꎬ２０１９ꎬ１６(５):４１４~４２１.[１２]㊀徐琰瑛.ｍｉＲ－２０４调控眼科疾病的研究进展[Ｊ].中华实验眼科杂志ꎬ２０１７ꎬ３５(８):７６１~７６３.[１３]㊀周巧ꎬ刘健ꎬ忻凌ꎬ等.基于关联规则挖掘中药内外合治骨关节炎对免疫炎症指标的影响[Ｊ].中华中医药学刊ꎬ２０１７ꎬ３５(８):２０７０~２０７３.[１４]㊀陈鸿泰ꎬ罗毅文ꎬ陈东风ꎬ等.牛膝杯苋甾酮激发大鼠骨髓间充质干细胞体外迁移及ＣＸＣＲ４表达的研究[Ｊ].中国骨质疏松杂志ꎬ２０１９ꎬ２５(１１):１５５０~１５５５.[１５]㊀范忠诚ꎬ王琮仁ꎬ李杨.ＳＩＲＴ２对白细胞介素－１β诱导的兔关节软骨细胞凋亡的影响[Ｊ].中国老年学杂志ꎬ２０１８ꎬ３８(１８):４４８８~４４９２.ʌ文章编号ɔ１００６－６２３３(２０２１)０３－０３７４－０６复方芪参提取物对糖尿病足溃疡模型大鼠创面愈合及ＨＩＦ－１α/ＶＥＧＦ/ＶＥＧＦＲ２通路的影响于泽洋ꎬ㊀李天博ꎬ㊀王江宁(首都医科大学附属北京世纪坛医院矫形外科ꎬ㊀北京㊀１０００３８)ʌ摘㊀要ɔ目的:探讨复方芪参提取物对糖尿病足溃疡模型大鼠创面愈合及缺氧诱导因子－１α(ＨＩＦ－１α)/血管内皮生长因子(ＶＥＧＦ)/血管内皮生长因子受体２(ＶＥＧＦＲ２)通路的影响ꎮ方法:建立糖尿病足溃疡大鼠模型ꎬ将造模成功４８只大鼠随机分为模型组㊁二甲双胍组(０.２ｇ/ｋｇ)㊁复方芪参提取物低剂量组(４ｇ/ｋｇ)㊁复方芪参提取物高剂量组(８ｇ/ｋｇ)ꎬ每组１２只ꎻ１２只正常大鼠以相同方法建立溃疡创面伤口作为对照组ꎮ造模成功后第１天ꎬ二甲双胍组㊁复方芪参提取物低剂量组㊁复方芪参提取物高剂量组分别灌胃０.２ｇ/ｋｇ二甲双胍㊁４ｇ/ｋｇ㊁８ｇ/ｋｇ复方芪参提取物ꎬ对照组和模型组灌胃等体积生理盐水ꎬ每天１次ꎬ连续灌胃４周ꎮ给药周期结束后ꎬ测定大鼠溃疡创面愈合率ꎬ观察糖尿病足大鼠和正常大鼠溃疡创面组织病理改变ꎬ测定糖尿病足大鼠和正常大鼠溃疡创面组织中ＨＩＦ－１α㊁ＶＥＧＦ㊁ＶＥＧＦＲ２ｍＲＮＡ和蛋白水平ꎮ结果:对照组创面组织结构正常ꎬ可见大量成纤维细胞及毛细血管ꎻ模型组创面组织大量炎症细胞浸润ꎬ成纤维细胞及毛细血管数目明显减少ꎻ二甲双胍和复方芪参提取物干预后ꎬ创面组织成纤维细胞及毛细血管增加ꎬ炎症细胞浸润减少ꎮ与对照组比较ꎬ模型组大鼠创面愈合率㊁溃疡创面组织ＨＩＦ－１α㊁ＶＥＧＦ㊁ＶＥＧＦＲ２ｍＲＮＡ和蛋白水平降低(Ｐ<０.０５)ꎻ与模型组比较ꎬ二甲双胍组㊁复方芪参提取物低㊁高剂量组大鼠创面愈合率㊁溃疡创面组织ＨＩＦ－１α㊁ＶＥＧＦ㊁ＶＥＧＦＲ２ｍＲＮＡ和蛋白水平升高(Ｐ<０.０５)ꎻ复方芪参提取物高剂量组大鼠创面愈合率㊁溃疡创面组织ＨＩＦ－１α㊁ＶＥＧＦ㊁ＶＥＧＦＲ２ｍＲＮＡ和蛋白水平高于复方芪参提取物低剂量组(Ｐ<０.０５)ꎻ与二甲双胍组比较ꎬ复方芪参提取物低剂量组大鼠上述指标降低(Ｐ<０.０５)ꎻ复方芪参提取物高剂量组与二甲双胍组大鼠上述指标差异无统计学意义(Ｐ>０.０５)ꎮ结论:复方芪参提取物可促进糖尿病足溃疡模型大鼠创面愈合ꎬ其机制可能与复方芪参提取物促进大鼠糖尿病足溃疡创面组织中ＨＩＦ－１α㊁ＶＥＧＦ㊁ＶＥＧＦＲ２ｍＲＮＡ和蛋白表达ꎬ进而激活ＨＩＦ－１α/ＶＥＧＦ/ＶＥＧＦＲ２通路有关ꎮʌ关键词ɔ㊀复方芪参提取物ꎻ㊀糖尿病足溃疡ꎻ㊀ＨＩＦ－１α/ＶＥＧＦ/ＶＥＧＦＲ２通路ʌ文献标识码ɔ㊀Ａ㊀㊀㊀㊀㊀ʌｄｏｉɔ１０.３９６９/ｊ.ｉｓｓｎ.１００６－６２３３.２０２１.０３.００５ＥｆｆｅｃｔｓｏｆＣｏｍｐｏｕｎｄＱｉｓｈｅｎＥｘｔｒａｃｔｏｎＷｏｕｎｄＨｅａｌｉｎｇａｎｄＨＩＦ－１α/ＶＥＧＦ/ＶＥＧＦＲ２ＰａｔｈｗａｙｉｎＤｉａｂｅｔｉｃＦｏｏｔＵｌｃｅｒＭｏｄｅｌＲａｔｓʌ基金项目ɔ北京市科技计划课题ꎬ(编号:Ｚ１７１１００００１０１７０７０)ʌ通讯作者ɔ王江宁。
降胆固醇药物的新作用
降胆固醇药物的新作用
麦田
【期刊名称】《心血管病防治知识》
【年(卷),期】2009(0)3
【摘要】@@ 早在2008年初,就提出一个医学难题:为什么降低胆固醇药物,不能减少动脉的粥样斑块?
【总页数】1页(P53)
【作者】麦田
【作者单位】(Missing)
【正文语种】中文
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