17β⁃雌二醇对血管内皮细胞自噬的影响  

Effect of 17β⁃estradiol on autophagy of vascular endothelial cells

作　　者：林朴卿 阮云军[2] 洪长江 Lin Puqing;Ruan Yunjun;Hong Changjiang(Department of Cardiology,Guangdong Clifford Hospital,Guangzhou 511495;Department of Geriatrics,Nanfang Hospital,Southern Medical University,Guangzhou 510010,China)

机构地区：[1]广东祈福医院心血管内科,广东广州511495 [2]南方医科大学南方医院老年病科,广东广州510010

出　　处：《广州医科大学学报》2021年第2期12-16,共5页Academic Journal of Guangzhou Medical University

基　　金：广东省自然科学基金(2016A030313755)。

摘　　要：目的:研究17β⁃雌二醇(17β⁃E_(2))对血管内皮细胞自噬的影响。方法:将人脐静脉内皮细胞分为6组,空白组、17β⁃E_(2)组、过氧化氢(H_(2)O_(2))组、17β⁃E_(2)+H_(2)O_(2)组、17β⁃E_(2)+H_(2)O_(2)+ICI182780(雌激素受体抑制剂)组和17β⁃E_(2)+H_(2)O_(2)+3⁃甲基腺嘌呤(3⁃MA,自噬抑制剂)组。Western Blot检测各组自噬蛋白LC3、Beclin⁃1和p62;免疫荧光检测各组自噬小体数量。结果:(1)与空白组相比,H_(2)O_(2)组和17β⁃E_(2)+H_(2)O_(2)组Beclin⁃1、LC3⁃Ⅱ蛋白表达明显增多,p62表达减少(均P<0.05);与H_(2)O_(2)组相比,17β⁃E_(2)+H_(2)O_(2)组Beclin⁃1、LC3⁃Ⅱ蛋白增多,p62表达减少(均P<0.05);与17β⁃E_(2)+H_(2)O_(2)组相比,加入雌激素抑制剂的17β⁃E_(2)+H_(2)O_(2)+ICI182780组或加入自噬抑制剂的17β⁃E_(2)+H_(2)O_(2)+3⁃MA组Beclin⁃1、LC3⁃Ⅱ蛋白表达明显减少,p62表达增多(均P<0.05);空白组与雌激素组相比,自噬相关蛋白表达无差异(P>0.05)。(2)与空白组相比,H_(2)O_(2)组和17β⁃E_(2)+H_(2)O_(2)组自噬小体明显增多,且17β⁃E_(2)+H_(2)O_(2)组较H_(2)O_(2)组增多(均P<0.05);与17β⁃E_(2)+H_(2)O_(2)组相比,17β⁃E_(2)+H_(2)O_(2)+ICI182780组和17β⁃E_(2)+H_(2)O_(2)+3⁃MA组自噬小体数量明显减少(均P<0􀆰05);空白组与雌激素组自噬小体数量无明显差异(P>0.05)。结论:17β⁃E_(2)可起到一定的促进血管内皮细胞自噬的作用,而这种作用只有在细胞应激情况下发生。Objective:To investigate the effect of 17β⁃estradiol(17β⁃E_(2))on autophagy of vascular endothelial cells.Methods:Human umbilical vascular endothelial cells were divided into 6 groups,namely the blank group,17β⁃E_(2) group,hydrogen peroxide(H_(2)O_(2))group,17β⁃E_(2)+H_(2)O_(2) group,17β⁃E_(2)+H_(2)O_(2)+ICI182780(an estrogen receptor inhibitor)group and 17β⁃E_(2)+H_(2)O_(2)+3⁃methyladenine(3⁃MA,an autophagy inhibitor)group.Western blot was used to detect the autophagy⁃related proteins LC3,Beclin⁃1 and p62 in each group;immunofluorescence assay was use to examine the number of autophagosomes in each group.Results:(1)Compared with the blank group,the Beclin⁃1 and LC3⁃Ⅱ protein expression was significantly increased,and p62 expression was decreased,in H_(2)O_(2) group and 17β⁃E_(2)+H_(2)O_(2) group(all P<0.05).Compared with the H_(2)O_(2) group,the Beclin⁃1 and LC3⁃Ⅱ protein expression was increased,and the p62 expression was lowered,in the 17β⁃E_(2)+H_(2)O_(2) group(all P<0.05).Compared with the 17β⁃E_(2)+H_(2)O_(2) group,the protein expression of Beclin⁃1 and LC3⁃Ⅱ in the 17β⁃E_(2)+H_(2)O_(2)+ICI182780 group with estrogen inhibitor or the 17β⁃E_(2)+H_(2)O_(2)+3⁃MA group with autophagy inhibitor was significantly decreased,and the p62 expression was increased(all P<0.05).Between the blank group and the 17β⁃E_(2) group,there was no difference in the expression of autophagy⁃related proteins(P>0􀆰05).(2)Compared with the blank group,autophagosomes were significantly increased in the H_(2)O_(2) group and the 17β⁃E_(2)+H_(2)O_(2) group,with more increase in the 17β⁃E_(2)+H_(2)O_(2) group than in the H_(2)O_(2) group(both P<0.05).Compared with the 17β⁃E_(2)+H_(2)O_(2) group,the number of autophagosomes in the 17β⁃E_(2)+H_(2)O_(2)+ICI182780 group and the 17β⁃E_(2)+H_(2)O_(2)+3⁃MA group was significantly reduced(all P<0.05).There was no significant difference in the number of autophagosomes between the blank group and the 17β⁃E_(2) gro

关 键 词：17β⁃雌二醇 自噬 血管内皮细胞 

分 类 号：Q25[生物学—细胞生物学]