目的 观察血管紧张素Ⅱ(AngⅡ)刺激及氯沙坦干预对足细胞小窝蛋白1(caveolin-1)表达的影响,探讨caveolin-1在足细胞损伤中的作用.方法 体外培养永生化小鼠足细胞(MPC),AngⅡ(10-6mol/L)刺激不同时间(3 h、6h、12 h和24 h);Losartan(10-6 mol/L)提前预处理3h后与AngⅡ(10-6 mol/L)共孵育6h,Hoechst-33342检测足细胞凋亡率,Western-blot法检测各组细胞caveolin-1蛋白表达,免疫荧光检测caveolin-1及其磷酸化水平.结果 ①AngⅡ刺激3h,足细胞即开始发生凋亡,随着刺激时间的延长,足细胞凋亡明显增多(P<0.05).②AngⅡ刺激不同时间caveolin-1表达总量无明显改变(P>0.05),从3h开始,caveolin-1磷酸化水平明显增高(P<0.05).③Losartan干预后caveolin-1磷酸化水平明显降低(P<0.05).结论 caveolin-1可能在AngⅡ诱导的足细胞损伤中发挥着重要的作用.
Objective To evaluate the possible role of caveolin-1 in angiotensin Ⅱ (Ang Ⅱ )-induced podocyte damage. Methods Immortalized mouse podocytes were cultured in vitro. Podocytes were exposed to AngⅡ(10-6 mol/L) for different time lengths (3, 6, 12, and 24 h) with or without pretreatment of Losartan (10 ^-6 mol/L). Apoptosis was evaluated by cell nucleus staining. Total proteins were prepared and evaluated for the expression of caveolin-1 by Western-blot analysis; Caveohn-1 and p-caveolin-1 expression was detected by using immunofluorescence. Results (1) Ang H induced podocyte apoptosis in a time-dependent manner (P〈0. 05) ; (2) The expression of caveolin-1 protein had no significant change after AngⅡ stimulation (P〉0. 05), but the phosphorylation level of caveolind was enhanced greatly after exposure to Ang Ⅱ for 3 h, which was decreased significantly by pretreatment with Losartan (P〈0. 05). Conclusions Caveolin-1 may play an important role in AngⅡ-induced podocyte damage.