阻断RAS对HSkMCs细胞株细胞凋亡及Mfn2表达的影响

The Effects of Blocking Renin-angiotensin System on the Expression of Mfn2 mRNA and Apoptosis in HSkMCs

目的:观察氯沙坦对高糖培养人骨骼肌细胞(Human skeletal muscle cells,HSk MCs)中线粒体融合蛋白2(mitofusin2,Mfn2)的表达及其对细胞凋亡的影响。方法:1.使用不同浓度的葡萄糖养基(葡萄糖浓度分别为5.55 mmol/L,11.1 mmol/L,22.2 mmol/L)分别培养HSk MCs细胞株48小时,检测各组细胞中血管紧张素Ⅰ型受体(Angiotensin II type I receptor,AT1R)基因、基因Mfn2的表达,并用流式细胞术检测细胞凋亡。2.根据1中实验结果,选择对Mfn2影响最大的葡萄糖浓度(此组葡萄糖浓度为22.2mmol/L)作为后续实验的条件。加入血管紧张素受体Ⅱ拮抗剂(Angiotensin Receptor Blockers,ARB)氯沙坦(Losartan),处理人骨骼肌细胞(HSk MCs)48 h,以未加氯沙坦为对照组,观察其对线粒体融合蛋白2(Mfn2表达的影响,并行流式细胞术检测细胞凋亡。结果:氯沙坦干预组HSk MCs细胞中Mfn2表达上调,细胞凋亡减少。结论:阻断肾素血管紧张素系统(Renin-angiotensin System,RAS)能上调HSk MCs细胞株中的Mfn2表达,并减少细胞凋亡。

Objective： To observe the effect of losartan on the expression of Mfn2 m RNA and apoptosis in Human skeletal muscle cells（HSk MCs） cultured by high glucose medium. Methods： HSk MCs were cultivated in vitro and treated with glucose of different concentration（5.55 mmol/L group, 11.1 mmol/Lgroup, 22.2 mmol/Lgroup） for 48 hours. Then the expression of AT1 R and Mfn2 was detected by PCR, apoptosis was detected by flow cytometry.2. Select 22.mmol/Lcgroup which has the most influence on the expression of Mfn2 from the above experiments as the following experimental conditions. Added losartan（ ARB） to treat HSk MCs for 48 h, in compared with the group without losartan. To observe its effect on the expression of Mfn2 and apoptosis via flow cytometry. Results： The expression of Mfn2 in HSk MCs cultivated added losatan group was up- regulated, while apoptosis was reduced. Conclusions： Blocking RAS could up-regulate the expression of Mfn2 and reduce cell apoptosis in HSk MCs.