SP600125对1型糖尿病小鼠颈动脉内皮功能及单核细胞趋化蛋白1表达的影响

[文章编号] 1007 3949(2011)19 10 0795 07 实验研究 SP600125对1型糖尿病小鼠颈动脉内皮功能及单核细胞趋化蛋白1表达的影响王启章1,2,陈茂刚1,李达文1,刘朝来1,徐格林1,刘新峰1[1.南方医科大学南京临床学院(南京军区南京总医院)神经内科,江苏省南京市210002;2.广州医学院附属深圳沙井医院神经内科,广东省深圳市518104][关键词] 1型糖尿病; 颈动脉; JNK信号通路; 信号转导子和转录激活子1; 单核细胞趋化蛋白1[摘 要] 目的 探讨c Jun氨基末端激酶(JNK)特异性抑制剂SP600125对1型糖尿病小鼠颈动脉内皮功能及单核细胞趋化蛋白1表达的影响。

方法 本研究分5组。

第1组为C57BL/6野生型雄性小鼠,喂养8周;第2组为INS2AKITA雄性小鼠,每日腹腔注射生理盐水50 L,共8周;第3、4、5组为INS2AKITA雄性小鼠,每日各腹腔注射SP60012510、20和30mg/kg;连续8周。

8周后麻醉处死动物,取静脉血检测血清髓过氧化物酶、丙二醛、一氧化氮、一氧化氮合酶;取双侧颈总动脉分别行凝胶迁移或电泳迁移率实验检测颈动脉信号转导子和转录激活子1活性,行Western blot检测p JNK、JNK、单核细胞趋化蛋白1蛋白表达,行免疫组化检测颈动脉内皮单核细胞趋化蛋白1表达。

结果 与野生型雄性小鼠相比,1型糖尿病小鼠血清髓过氧化物酶、丙二醛明显增高,而血清一氧化氮合酶、一氧化氮明显减低(P<0 01)。

给予1型糖尿病小鼠SP600125不同剂量干预后,髓过氧化物酶浓度较1型糖尿病组明显减低,而一氧化氮、一氧化氮合酶浓度明显增高,且SP600125剂量越高,一氧化氮、一氧化氮合酶浓度增加越明显(P<0 05)。

同时,1型糖尿病小鼠p JNK/JNK、单核细胞趋化蛋白1蛋白表达、颈动脉信号转导子和转录激活子1活性以及颈动脉内皮单核细胞趋化蛋白1表达比正常小鼠明显增高(P<0 05)。

而经过3种剂量SP600125干预后,单核细胞趋化蛋白1蛋白表达分别下降21 82%、39 09%、68 18%;信号转导子和转录激活子1活性分别下降25 70%、33 20%、61 29%;颈动脉内皮单核细胞趋化蛋白1表达分别下降19 51%、39 14%、59 28%。

结论 JNK特异性抑制剂可通过抑制信号转导子和转录激活子1,从而升高糖尿病血清一氧化氮及一氧化氮合酶水平,改善血管舒张功能,并且降低单核细胞趋化蛋白1、髓过氧化物酶表达水平,并具有剂量依赖性。

进一步表明JNK通过信号转导子和转录激活子1参与了糖尿病颈动脉内皮功能紊乱及炎症因子表达,并为开发预防1型糖尿病大血管并发症的药物提供了一个新的理论依据。

[中图分类号] R363[文献标识码] AThe Effect of SP600125on Endothelial Function and Monocyte Chemotactic Protein 1Expression in Carotid Artery in Type1Diabetic MiceWANG Qi Zhang1,2,CHEN Mao Gang1,LI Da Wen1,LIU Chao Lai1,XU Ge Lin1,and LIU Xin Feng1(1.Department of Neurology,Jinling Hospital,Southern Medical University,Nanjing,Jiangsu210002,China;2.Department of Neurology,The Affiliated Shenzhen Shajing Hospital of Guangzhou Medical University(Shajing People s Hospital), Shenzhen,Guangdong518104,China)[KEY WORDS] Type1Diabetes; Carotid Artery; c Jun Amino terminal Kinase Signal Pathway; Signal Trans ducer and Activator of Transcription 1; Monocyte Chemotactic Protein 1[ABSTRACT] Aim The incidence of stroke in diabetes is increasing seriously,which is associated with endothelial dysfunction and increased inflammation in carotid artery. Our study is to investigate the effect of c Jun amino terminal ki[收稿日期] 2011 05 23[基金项目] 国家自然科学基金(青年基金)项目(81000502)及江苏省科技计划项目(BK2009319)资助[作者简介] 王启章,博士研究生,主治医师,研究方向为缺血性脑血管病介入及动脉粥样硬化的临床及基础研究,E mail为wqzflm@yahoo com cn。

陈茂刚,博士研究生,副主任医师,研究方向为动脉粥样硬化的基础及临床研究。

通讯作者刘新峰,主任医师,教授,博士研究生导师,研究方向为脑血管病介入及动脉粥样硬化的临床及基础研究,E mail为xfliu2@ya hoo com cn。

nase(JNK)specific inhibitor SP600125on carotid endothelial function and monocyte chemotactic protein 1(MCP 1)ex pression in type1diabetes. Methods Animals were divided into five groups in this study. The1st group was C57BL/6wild type male mice;the2nd group was INS2AKI TA male mice,intraperitoneal injection with0 9%normal saline (NS)each day for8weeks;the3rd,4th,5th group were INS2AKITA male mice,intraperitoneal injection with SP60012510 mg/kg,20mg/kg,30mg/kg respectively each day for consecutive8weeks. Then the mice were killed and sreum my eloperoxidase(MPO),malondialdehyde(MDA),nitric oxide(NO),total nitric oxide synthase(TNOS)were measured. HE staining was performed in carotid artery and immunohistochemistry was performed to investigate MCP 1protein ex pression in endothelial cells of carotid artery. P JNK,JNK,MCP 1protein expression in carotid artery was measured by Western blot,signal transducer and activator of transcription 1(STAT 1)DNA binding activity was assayed by electro phoretic mobility shift assay(EMSA). Results Compared with wild type mice,serum MPO and MDA expression in creased prominently(P<0 05),whereas TNOS,NO decreased and p JNK/JNK,MCP 1expression increased significant ly in INS2AKI TA mice(P<0 05);STAT 1DNA binding activity increased prominently in type1diabetic group compared with that in control group(P<0 05). Compared with type1diabetic mice,after treatment with SP600125,MPO de creased and NO,TNOS increased and p JNK/JNK,endothelial MCP 1expression decreased(P<0 05). The effects of SP600125on MPO,NO,TNOS,MCP 1were increased accordingly as the dose of SP600125increased. As the dose of SP600125increased,MCP 1protein expression decreased21 82%,39 09%,68 18%respectively;STAT 1DNA binding activity decreased25 70%,33 20%,61 29%respectively compared with type1diabetic mice. Conclusions JNK specific inhibitor SP600125could inhibit STAT 1DNA binding activity,thus increase serum NO and TNOS,and improve carotid diastolic function. SP600125could also decrease MCP1and MPO in a dose dependent manner. The results also showed JNK signal pathway is associated with carotid endothelial dysfunction and inflammatory expression via activation of STAT 1in type1diabetes. Our study provides a novel pharmacologic agent to prevent the promotion of macrovascular complication in type1diabetes.近年来基础和临床研究发现,糖尿病(diabetes mellitus,DM)患者卒中发病率较正常人明显增高,主要原因在于糖尿病患者易出现颈动脉粥样硬化及斑块。