骨形态发生蛋白-7对高糖刺激足细胞snaill表达及转分化的影响

发布时间：2018-04-19 04:09

本文选题：足细胞 + 上皮细胞-间充质细胞转分化　；参考：《青岛大学》2014年硕士论文

【摘要】：目的通过观察高糖环境下足细胞snaill表达变化及骨形态发生蛋白-7(BMP-7)干预对高糖刺激足细胞p-smad2/3、snaill表达和足细胞EMT的影响,探讨snaill在足细胞EMT中的作用及BMP-7抑制高糖诱导足细胞EMT的可能机制。方法以体外培养的永生化小鼠足细胞(MPC5)为研究对象,分别给予不同时间(6h、12h、24h、48h)高糖刺激；将足细胞随机分为以下4组：正常糖组(NG组)、高糖刺激组(HG组)、高糖+BMP-7干预组(BMP-7组)和甘露醇对照组(MG组),各组培养时间均为48h；采用倒置相差显微镜观察各组足细胞形态,采用实时荧光定量RT-PCR检测足细胞snaill、nephrin、α-平滑肌肌动蛋白(a-SMA) mRNA表达量,采用western(?)迹法检测足细胞snail、p-smad2/3、nephrin、α-SMA蛋白表达量。结果与NG组相比,高糖刺激可以使足细胞形态发生明显改变,在转录水平和蛋白水平诱导snail、α-SMA的表达、减少nephrin的表达(P0.05),且随着刺激时间的延长,作用更加明显；与HG组相比,给予BMP-7干预在一定程度上可以逆转高糖诱导的足细胞形态改变及p-smad2/3、snail、α-SMA的表达异常,增加足细胞nephrin表达(P0.05)；NG组与MG组足细胞各因子表达水平的差异无统计学意义(P0.05)。结论高糖刺激可以呈时间依赖性上调足细胞snaill表达,诱导足细胞转分化；BMP-7可能通过拮抗TGFβ/smad通路调节高糖诱导足细胞snaill的异常表达,抑制足细胞转分化。
[Abstract]:Objective to observe the changes of snaill expression in podocytes and the effects of bone morphogenetic protein-7 BMP-7 on the expression of p-smad2 / 3nsnaill and podocyte EMT in podocytes stimulated by high glucose.To investigate the role of snaill in podocyte EMT and the possible mechanism of BMP-7 inhibiting high glucose induced podocyte EMT.Methods the immortalized mouse podocytes (MPC5) were cultured in vitro and were stimulated with high glucose at different time (6 h, 12 h, 24 h, 48 h).Podocytes were randomly divided into the following four groups: normal glucose group (NG group), high glucose stimulation group (HG group), high glucose BMP-7 intervention group (BMP-7 group) and mannitol control group (MG group) for 48 h.The expression of snaillonnephrin (伪 -smooth muscle actin) mRNA in podocytes was detected by real-time fluorescence quantitative RT-PCR.The expression of snaila p-smad 2 / 3 nephrin, 伪 -SMA protein in podocytes was detected by trace method.Results compared with NG group, high glucose stimulation could significantly change the morphology of podocyte, induce Snail, 伪 -SMA expression at transcription and protein level, and decrease the expression of nephrin P0.05, and the effect was more obvious with the prolongation of stimulation time, compared with HG group.The intervention of BMP-7 could reverse the morphological changes of podocytes induced by high glucose and the abnormal expression of p-smad2 / 3 snailand 伪 -SMA in podocytes to a certain extent, but there was no significant difference in the expression of podocytes between P0.05 and MG groups.Conclusion High glucose stimulation can up-regulate the expression of snaill in podocyte in a time-dependent manner. BMP-7 may regulate the abnormal expression of snaill in podocyte induced by high glucose by antagonizing the TGF 尾 -smad pathway and inhibiting the transdifferentiation of podocyte.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692

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