高糖和LY294002干预影响足细胞内Ⅳ型胶原表达

Effects of High Glucose and LY294002 on the Expression of Collagen Ⅳ in Mouse Podocyte

探讨高糖和PI3K/Akt通路对足细胞内Ⅳ型胶原(Col Ⅳ)表达的影响。体外培养小鼠足细胞,给予高糖(30mmol/L)处理后,分别于0,12,24,48h收集细胞,采用免疫细胞化学染色法和Western blot技术检测Col Ⅳ的表达;Western blot技术检测Akt的活化及LY294002对Col Ⅳ表达的抑制效应。结果表明,高糖诱导足细胞内Col Ⅳ蛋白表达增多,24h明显,各时间点与高糖刺激前相比均有统计学差异(P<0.05);高糖激活Akt蛋白磷酸化,p-Akt随刺激时间延长表达增多。PI3K/Akt通路抑制剂LY294002孵育细胞24h后,可减弱高糖诱导的足细胞内Col Ⅳ的表达(P<0.05)。因此,高糖可能通过激活PI3K/Akt通路上调足细胞内Ⅳ型胶原表达。

To investigate effects of high glucose and phosphoinositide 3 kinese/protein kinase B （P13K/ Akt） pathway on the expression of collagen Ⅳ （Col Ⅳ） in mouse podocyte, we divided cultured mouse podocytes into high glucose （30 mmol/L, HG） group and normal glucose （5 mmol/L, NG） group. Ceils were collected respectively at 0, 12, 24, 48 h after stimulation. The expression of collogen Ⅳ was detected by immunocytochemistry and Western blot analysis. The expression of phospho-Akt and the inhibition of LY294002 on collogen Ⅳ were analized by Western blot. Compared with control group, the level of Col Ⅳ in mouse podocytes treated by high glucose was significantly increased （P〈0.05）, with reaching the peak at 24 h. The phosphorylation of Akt was observed in mouse podocytes induced by high glucose. The level of phospho-Akt was increased in a time-dependent manner. However, inhibition of activation of Akt with LY294002, a specific PI3K/Akt pathway inhibitor, attenuated the high glucoseinduced expression of Col IV at 24 h after intervention. We conclude that high glucose maybe up-regulate the expression of Col Ⅳ in mouse podocytes by activating PI3K/Akt pathway.