Sirolimus inhibits the expression of type Ⅰ collagen and fibronectin in cultured renal cortical myofibroblasts / 北京大学学报(医学版)

ABSTRACT

Objective:To investigate the anti-fibrotic effect of sirolimus(rapamycin)at the cell level.Methods:The primary cultured rat renal cortical myofibroblasts were divided into two groups,control group and sirolimus 40 mg/L group at each time point.The protein levels of ?-SMA,Col-Ⅰ,fibronectin(FN)were analyzed by Western blot in both the whole cell lysates and supernatant culture media 12 h,24 h and 48 h after incubation,respectively.Real-time quantitative PCR was carried out to measure the levels of procollagen-ⅠmRNA 1 h,2 h,4 h,and 6 h after cell incubation.The activities of gelatinase MMP-2 and MMP-9 in the supernatant from the cultured cell media were assayed by gelatin zymography.Results:(1)Sirolimus had no effect on the expression of ?-SMA of myofibroblasts at differnet time points.(2)The expression of Col-Ⅰin the whole cell lysates both reduced at the end of 24 h and 48 h in sirolimus group significantly [(0.58?0.05)and(0.63?0.18),P