替米沙坦对MIN6细胞的影响

目的:研究血管经张素Ⅱ受体拮抗剂(ARB)药物对胰岛素分泌细胞的影响及机制。方法:不同浓度替米沙坦及替米沙坦和甲苯磺丁脲共同干预MIN6细胞一定时间后,MTT法检测细胞活力变化,运用ROS(细胞内活性氧)捕获剂双氢溴乙啶(HE)、双氢-乙酰乙酸二氯荧光黄(DCFH-DA)和双氢罗丹明123(DHR123)孵育细胞,用荧光显微镜观察细胞内荧光、流式细胞仪检测MIN6细胞的MFI(平均荧光强度)而测得细胞内ROS水平。用Annexin-vFITC和PI双标记细胞凋亡检测法通过流式细胞仪测定细胞凋亡情况。结果:替米沙坦单独干预MIN6细胞后,细胞活力无明显抑制;细胞内的MFI无明显增加;细胞凋亡率亦无明显增加;而替米沙坦和甲苯磺丁脲共同孵育MIN6细胞后,细胞活力相对甲苯磺丁脲单独干预时增强(P>0.05);细胞内的MFI明显减少;细胞凋亡率亦明显减少(前期实验已经做过甲苯磺丁脲对MIN6细胞影响)。结论:替米沙坦对甲苯磺丁脲诱导的MIN6细胞氧化应激和凋亡有保护作用。说明ARB类药物对胰岛β细胞功能有保护作用,延缓β细胞衰竭。

Effects of telmisartan on MIN6 cells

Objective:To explore the effects of angiotensin II type 1 receptor blocker(ARB) on pancreatic beta-cells and the possible mechanisms.Methods:MIN6 cells were cultured with different concentration of telmisartan with or without tolbutamide,and cell viability was detected by MTT.Dihydroethidium(HE),2,7-dichlorofluorescein diacetate(DCFH-DA) or dihydrorhodamine 123(DHR123) was used as reactive oxygen species(ROS) capture.The mean fluorescent intensity of ethidium,2’,7’-dichlorofluorescein or rhodamine 123,which was the product of intracellular oxidation by the above probes,was detected by fluorescent microscopy and flow cytometry.Cell apoptosis was detected by Annexin-V-FITC/PI kit.Results:As compared with normal control,telmisartan treatment showed no change in cell viability,intracellular fluorescent intensity and cell apoptosis.Contrarily,combined treatment with telmisartan and tolbutamide suggested increased cell viability,yet decreased intracellular fluorescent intensity and cell apoptosis compared to the tolbutamide treated group.Conclusion:Telmisartan may protect MIN6 cells induced by tolbutamide from damage,suggesting that ARB may have beneficial effects on beta cell function and prevent the progression of beta cell failure.