Toll样受体4对高糖诱导胰岛微血管内皮细胞株凋亡蛋白表达的影响  

Effect of Toll-like Receptor 4(TLR4) on the Expression of Apoptotic Proteins in High Glucose induced Islet Microvascular Endothelial Cells

作　　者：刘明明[1] 张晓艳[1] 武清斌[1] 尚飞[1] 李炳蔚[1] 李爱玲[1] 李宏伟[1] 修瑞娟[1] LIU Ming-ming ZHANG Xiao-yan Hong-wei XIU Rui-juan(Key Laboratory of Microcirculation, Academy of Medieal Seienees and WU Qing-bin, SHANG Fei, LI Bing-wei, LI Ai-ling , LI Ministry of Health, Institute of Peking Union Medical College, Microcirculation, Chinese Beijing 100005, China)

机构地区：[1]中国卫计委微循环重点实验室中国医学科学院北京协和医学院微循环研究所,北京100005

出　　处：《微循环学杂志》2017年第3期16-21,共6页Chinese Journal of Microcirculation

基　　金：协和青年基金;中央高校基本科研业务费专项资助(333201065;3332015200)

摘　　要：目的:观察小鼠胰岛微血管内皮细胞(MS1)Toll样受体4(TLR4)的表达及其对高糖诱导凋亡蛋白表达的影响。方法:离体小鼠MS1分为低浓度葡萄糖(5.6mM)对照组(C组)、高浓度葡萄糖(25.0mM和35.0mM)诱导组(HG组)、TLR4抑制剂干预组(35.0mM葡萄糖+1μM CLI-095,HG+CLI组)及TLR4抑制剂对照组(5.6mM葡萄糖+1μM CLI-095,CLI组)。平行培养24h或48h后,先用CCK-8法检测C组和HG组MS1细胞活力,选定高糖干预浓度(35.0mM)及时间(48h);再分别应用免疫组化和Western Blotting检测各组MS1细胞TLR4表达水平;应用Western Blotting检测各组MS1细胞凋亡蛋白Caspase-9、Caspase-3表达水平;应用明胶酶谱分析法检测基质金属蛋白酶2(MMP2)活性变化。结果:与C组比较,HG组MS1的TLR4蛋白表达显著增加(P<0.01);凋亡蛋白Caspase-9和Caspase-3表达均显著升高(P<0.01,P<0.05);MMP2活性明显增强(P<0.05);与HG组比较,HG+CLI组Caspase-9水平均明显降低(P<0.01),Caspase-3水平仅有降低趋势,差异无统计学意义(P>0.05);TLR4的表达和MMP2活性虽有下降,但差异亦无统计学意义(P>0.05)。结论:MS1高表达TLR4可能参与高糖诱导的MS1细胞凋亡。Objective：To investigate the expression of Toll-like receptor4（TLR4）on islet microvascular endothelial cells（MS1）and to assess its effect on the expression of apoptotic proteins in high glucose induced MS1.Method：MS1was divided into four groups according to the treatments by glucose and TLR4 signaling inhibitor CLI-095 as following：5.6mM glucose treated group（C group）,25.0mM and 35.0mM glucose treated groups（HG group）,TLR4 inhibitor treated group（35.0mM glucose plus 1μM CLI-095,HG＋CLI group）and TLR4 inhibitor control group（5.6mM glucose plus 1μM CLI-095,CLI group）.After cultured for 24 hor 48h,cell viability of C group and HG group was determined by CCK-8to identify the optimal concentration（35.0mM）and intervention time（48h）of high glucose.Then,the expression of TLR4 was determined by immunohistochemistry and Western Blotting and the expressions of apoptotic proteins（Caspase-9and Caspase-3）were determined by Western Blotting.MMP2 was evaluated by gelatin enzyme spectrum analysis.Results：Compared with C group,the expression of TLR4 in HG group increased significantly（P〈0.01）,the expressions of Caspase-9and Caspase-3were significantly higher in HG group than those in C group（P〈0.01 and P〈0.05,respectively）,meanwhile MMP2 activity increased significantly（P〈0.05）in HG group.Compared with HG group,the expressions of Caspase-9decreased significantly（P〈0.01）in HG＋CLI group,however no significant decreases were found on Caspase-3expression,TLR4 expression and MMP2 activity between HG group and HG＋CLI group（P〈0.05）although there were decreased tendencies.Conclusion：High expression of TLR4 in MS1associates with high glucose induced MS1 apoptosis.

关 键 词：胰岛微血管内皮细胞 TOLL样受体4 细胞凋亡 基质金属蛋白酶 糖尿病 

分 类 号：R363.2[医药卫生—病理学]