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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 the effects of high glucose and PI3K / Akt pathway on the expression of type Ⅳ collagen (Col Ⅳ) in podocytes. The mouse podocytes were cultured in vitro. After being treated with high glucose (30 mmol / L), the cells were harvested at 0, 12, 24 and 48 hours respectively. The expression of Col Ⅳ was detected by immunocytochemical staining and Western blot. Activation of Akt and Inhibitory Effect of LY294002 on Col Ⅳ Expression. The results showed that the expression of Col Ⅳ protein in podocytes induced by high glucose increased significantly at 24h, and there was a significant difference at each time point compared with that before high glucose stimulation (P <0.05); high glucose activated Akt protein phosphorylation, p-Akt With the stimulation of prolonged expression increased. PI4K / Akt pathway inhibitor LY294002 could inhibit the expression of Col Ⅳ in podocytes induced by high glucose for 24h (P <0.05). Therefore, high glucose may up-regulate type IV collagen expression in podocytes by activating PI3K / Akt pathway.