硒介导PI3K/Akt/mTOR信号通路对子痫前期滋养细胞自噬的调控作用及机制  

Regulatory effect of selenium on autophagy in preeclampsia trophoblast cells through the PI3K/Akt/mTOR signaling pathway

作　　者：刘婉儿 张乐乐 王婷[1] 郭伟[1] LIU Wan'er;ZHANG Lele;WANG Ting;GUO Wei(Department of Obstetrics and Gynecology,The First Affiliated Hospital of Shandong First Medical University(Shandong Provincial Qianfoshan Hospital),Jinan 250014,China)

机构地区：[1]山东第一医科大学第一附属医院(山东省千佛山医院)妇产科,济南250014

出　　处：《山东医药》2024年第34期34-37,共4页Shandong Medical Journal

摘　　要：目的探讨硒通过PI3K/Akt/mTOR信号通路对子痫前期(PE)人绒毛膜滋养细胞(HTR-8-Svneo)自噬及增殖迁移的影响及其机制。方法体外培养HTR-8-Svneo,随机分为正常对照组、乏氧组以及乏氧加硒组;对照组HTR-8-Svneo细胞置于37℃CO_(2)培养箱,乏氧组HTR-8-Svneo细胞置于37℃乏氧培养箱,乏氧加硒组HTR-8-Svneo细胞置于37℃乏氧加硒(硒浓度为10μmol/L)培养箱,三组均培养24 h。采用CCK-8试剂盒测量HTR-8-Svneo细胞活性,细胞划痕试验检测HTR-8-Svneo细胞增殖迁移能力,Western blotting法检测HTR-8-Svneo细胞PI3K/Akt/mTOR信号通路蛋白(p-PI3K、p-AKT、p-mTOR)以及自噬相关蛋白(LC3、Beclin-1、p62)表达,免疫荧光观察HTR-8-Svneo细胞LC3表达量。结果对照组、乏氧组、乏氧加硒组细胞活力OD值分别为1.5±0.21、1.04±0.05、1.48±0.27;乏氧组细胞活力较对照组降低、乏氧加硒组细胞活力较乏氧组增强(P均<0.05)。对照组、乏氧组、乏氧加硒组细胞划痕愈合率分别为17.25%、6.08%、18.13%;乏氧组细胞较对照组划痕愈合率降低、乏氧加硒组较乏氧组细胞划痕愈合率升高(P均<0.05)。乏氧组与对照组、乏氧加硒组与乏氧组比较,HTR-8-Svneo细胞通路蛋白以及自噬相关蛋白表达差异有统计学意义(P均<0.05)。对照组、乏氧组、乏氧加硒组HTR-8-Svneo细胞荧光表达量分别为73.26±4.18、134.83±3.03、74.7±12.37;乏氧组细胞荧光表达量较对照组升高,乏氧加硒组细胞荧光表达量较乏氧组降低,差异均有统计学意义(P均<0.05)。结论HTR-8-Svneo细胞加硒培养可提高细胞活力和增殖迁移能力,其机制可能与硒通过PI3K/AKT/mTOR通路调控PE滋养细胞自噬相关。Objective To explore the effects of selenium on autophagy and proliferation and migration of human trophoblast cells(HTR-8-Svneo)in preeclampsia(PE)through the PI3K/Akt/mTOR signaling pathway and the underlying mechanisms.Methods HTR-8-Svneo cells were cultured in vitro and randomly divided into the normal control group,hypoxia group,and hypoxia plus selenium group.HTR-8-Svneo cells in the control group were placed in a 37℃CO2 incubator,HTR-8-Svneo cells in the hypoxia group were placed in a 37℃ hypoxia incubator,and HTR-8-Svneo cells in the hypoxia plus selenium group were placed in a 37℃ hypoxia incubator with selenium supplementation(at the concentration of 10μmol/L),and all groups were cultured for 24 h.CCK-8 assay kit was used to measure the viability of HTR-8-Svneo cells,the cell scratch test was used to detect the proliferation and migration abilities of HTR-8-Svneo cells,and Western blotting was used to detect the expression of PI3K/Akt/mTOR signaling pathway proteins(p-PI3K,p-Akt,p-mTOR)and autophagy-related proteins(LC3,Beclin-1,p62)in HTR-8-Svneo cells,and immunofluorescence was used to observe the expression of LC3 in HTR-8-Svneo cells.Results The OD values of cell viability in the control,hypoxia,and hypoxia plus selenium groups were 1.5±0.21,1.04±0.05,and 1.48±0.27,respectively;the cell viability was lower in the hypoxia group than in the control group,and the cell viability was higher in the hypoxia plus selenium group than in the hypoxia group(both P<0.05).The cell scratch healing rates in the control,hypoxia,and hypoxia plus selenium groups were 17.25%,6.08%,and 18.13%,respectively;the scratch healing rate in the hypoxia group was lower than that in the control group,and the scratch healing rate in the hypoxia plus selenium group was higher than that in the hypoxia group(both P<0.05).There were statistically significant differences in the expression of pathway proteins and autophagy-related proteins in HTR-8-Svneo cells between the hypoxia group and the control group,and between the hyp

关 键 词：子痫前期 硒 人绒毛膜滋养细胞 PI3K/Akt/mTOR通路 细胞自噬 

分 类 号：R714.24[医药卫生—妇产科学]