过表达THEM4对高糖诱导的人肾小管上皮细胞胶原分泌的影响

发布时间：2018-05-15 23:34

  本文选题：硫酯酶超家族成员 + PIK/Akt　； 参考：《中国药理学通报》2017年12期

【摘要】：目的观察过表达硫酯酶超家族成员4(thioesterase superfamily member 4,THEM4)对高糖诱导的人肾小管上皮细胞胶原分泌的影响。方法体外培养人肾小管上皮细胞(human renal proximal tubular epithelial cell,HKC),将细胞随机分为正常对照组(5.5 mmol·L~(-1)normal glucose,Control)、高糖组(30 mmol·L~(-1)high glucose,HG)、高糖+p Yrads-4-THEM4质粒转染组(HG+THEM4)、高糖+p Yr-adshuttle-4空质粒对照组(HG+V)。后3组经高糖刺激48 h后终止培养,Western blot检测THEM4、phospho-Akt(Ser473)、TGF-β1、α-SMA的表达;免疫细胞荧光检测THEM4蛋白表达和定位;ELISA方法检测细胞上清液中Ⅰ型胶原(collagenⅠ,ColⅠ)、Ⅲ型胶原(collagenⅢ,ColⅢ)分泌情况。结果高糖刺激可导致人肾小管上皮细胞内THEM4表达降低,phospho-Akt(Ser 473)、TGF-β1、α-SMA的表达增强及ColⅠ、ColⅢ分泌;过表达THEM4能够逆转高糖刺激所导致人肾小管上皮细胞phospho-Akt(Ser 473)蛋白的活化,下调α-SMA、TGF-β1表达,抑制ColⅠ、ColⅢ的分泌。结论过表达THEM4能够降低高糖诱导的胶原分泌,可能是通过抑制phospho-Akt(Ser 473)蛋白的活化,下调α-SMA、TGF-β1表达而实现的。
[Abstract]:Aim to investigate the effect of 4(thioesterase superfamily member 4 (THEM4), a member of the superfamily of thioesterase, on collagen secretion induced by high glucose in human renal tubular epithelial cells. Methods Human renal tubular epithelial cells were cultured in vitro. The cells were randomly divided into normal control group (n = 5.5 mmol L~(-1)normal), high glucose group (n = 30 mmol L~(-1)high), high glucose p Yrads-4-THEM4 plasmid transfection group (n = 30) and control group (n = 5). The expression of THEM4phospho-AktPhospho-Akttre-Ser473TGF- 尾 _ 1, 伪 -SMA in the supernatant was detected by blot, the expression of THEM4 protein was detected by immunofluorescence and the secretion of collagen 鈪，

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