1,25二羟维生素D3通过PI3K/Akt信号通路调控自噬抑制足细胞凋亡  

1,25(OH)2 D3 regulates autophagy and inhibits podocyte apoptosis through PI3K/ Akt signaling pathway

作　　者：师朗 张雅飞 方旭 宋志霞[1,2] SHI Lang;ZHANG Yafei;FANG Xu;SONG Zhixia(The First College of Clinical Medical Science,China Three Gorges University,Yichang 443003,China;Department of Nephrology,Yichang Central People􀆳s Hospital,Yichang 443003,China)

机构地区：[1]三峡大学第一临床医学院,宜昌443003 [2]宜昌市中心人民医院肾内科

出　　处：《肾脏病与透析肾移植杂志》2020年第4期339-345,共7页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：国家自然科学基金项目(81600567);宜昌市科学技术局项目(A1630102)。

摘　　要：目的:探讨1,25二羟维生素D3[1,25(OH)2D3]对高糖诱导足细胞损伤的影响及作用机制。方法:将MPC⁃5细胞(小鼠永生系足细胞)分为6组:正常对照组(NG)、甘露醇渗透压对照组(NG+M)、高糖组(HG)、高糖+1,25(OH)2D3干预组(HG+VD)、高糖+1,25(OH)2D3+3⁃甲基腺嘌呤干预组(HG+VD+3⁃MA)、高糖+1,25(OH)2D3+PI3K抑制剂LY294002组(HG+VD+LY294002)。采用CCK⁃8法分析细胞存活率;通过细胞免疫荧光技术观察自噬相关蛋白LC3⁃Ⅱ的表达;Western印迹检测自噬相关蛋白Beclin⁃1、P62,通路相关蛋白PI3K、Akt,抗凋亡蛋白Bcl⁃2及凋亡蛋白Bax和caspase⁃3的表达。结果:(1)高糖可诱导足细胞凋亡,活性维生素D3可减少高糖诱导的足细胞凋亡;(2)活性维生素D3可逆转高糖诱导的足细胞自噬的下调,自噬抑制剂3⁃甲基腺嘌呤(3⁃MA)可阻断活性维生素D3对高糖诱导的足细胞的损伤的保护作用;(3)活性维生素D3激活PI3K/Akt信号,LY294002阻断了对足细胞的保护作用及上调自噬的作用。结论:活性维生素D3可通过PI3K/Akt信号通路调控自噬,减轻高糖诱导的足细胞凋亡和损伤。Objective:To investigate effect of 1,25(OH)2D3 on hyperglycosis⁃induced podocyte injury and its mechanism.Methodology:MPC⁃5 cells were divided into six groups:normal glucose concentration group(NG),high glucose group(HG),Mannitol osmotic pressure control(NG+M),high glucose+1,25(OH)2D3 intervention group(HG+VD),high glucose+1,25(OH)2D3+3⁃methyladenine intervention group(HG+VD+3⁃MA),high glucose+1,25(OH)2D3+PI3K inhibitor LY294002 group(HG+VD+LY294002)􀆰Cell viability was analyzed by CCK⁃8 method􀆰By immunofluorescence technique to observe autophagy related protein expression of LC3⁃Ⅱ;Western blotting was used to detect the expression of autophagy⁃related proteins Beclin⁃1,P62,pathway related proteins PI3K,Akt,anti⁃apoptotic proteins Bcl⁃2 and apoptotic proteins Bax and caspase⁃3.Results:1􀆰High glucose can induce apoptosis of podocytes,and active vitamin D3 can reduce the apoptosis of podocytes induced by high glucose.2􀆰Active vitamin D3 can reverse the down⁃regulation of autophagy in high⁃glycosylated podocytes,and autophagy inhibitor 3⁃methyladenine(3⁃MA)can block the protective effect of active vitamin D on high⁃glycosylated podocytes.3􀆰Active vitamin D3 activates PI3K/Akt signal,and LY294002 blocks the protective effect on podocytes and up⁃regulates autophagy.Conclusion:Active vitamin D3 can regulate autophagy through PI3K/Akt signaling pathway to alleviate high⁃glycose⁃induced podocyte injury and apoptosis.

关 键 词：1 25 二羟维生素D3 自噬 凋亡 足细胞 PI3K/AKT信号通路 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]