17β-雌二醇通过PI3K/Akt/mTOR通路抑制白介素1β诱导的大鼠椎间盘髓核细胞的凋亡  

17β-estradiol inhibits interleukin-1β-induced rat nucleus pulposus cell apoptosis through the PI3K/Akt/mTOR signal pathway

作　　者：郭洪涛 郭尔斐[1] 徐建杰[1] 张斌[1] 高雁冰 武建忠[1] GUO Hong-Tao;GUO Er-Fei;XU Jian-Jie;ZHANG Bin;GAO Yan-Bing;WU Jian-Zhong(Department of Spinal Surgery,Shijiazhuang first hospital,Shijiazhuang 050011,China)

机构地区：[1]石家庄市第一医院脊柱外科,石家庄050011

出　　处：《生理学报》2021年第1期62-68,共7页Acta Physiologica Sinica

基　　金：supported by the Fund Project of Health Commission of Hebei Province,China(No.20180999)。

摘　　要：髓核细胞(nucleus pulposus cells,NPCs)的异常凋亡是导致椎间盘退变(intervertebral disc degeneration,IVDD)的主要原因。本研究组前期研究显示,17β-雌二醇(17β-estradiol,E2)能够通过PI3K/Akt信号通路抑制白介素1β(interleukin-1β,IL-1β)诱导的大鼠椎间盘NPCs凋亡。本研究旨在探讨PI3K/Akt途径的下游蛋白是否参与E2对NPCs凋亡的抑制作用。用胰蛋白酶消化法分离原代大鼠NPCs,采用E2和PI3K/Akt信号通路下游蛋白的不同抑制剂预处理后用IL-1β处理,用Annexin V/PI染色法检测凋亡率,用CCK-8法检测细胞活力,用细胞黏附试验检测NPCs与Ⅱ型胶原的黏附能力,用Western blot检测哺乳动物雷帕霉素靶蛋白(mammalian target of Rapamycin,mTOR)、糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)和核因子κB(nuclear factor kappaB,NF-κB)磷酸化水平。结果显示,E2显著抑制IL-1β诱导的NPCs凋亡,逆转由IL-1β引起的细胞活力和黏附能力的降低,抑制IL-1β对mTOR磷酸化水平的下调作用,而雷帕霉素可以阻断E2的这些保护作用。以上结果提示,E2可能通过PI3K/Akt/mTOR信号通路抑制IL-1β诱导的NPCs凋亡。The apoptosis of nucleus pulposus cells(NPCs)is the main cellular process of intervertebral disc degeneration(IVDD).Our previous studies showed that 17β-estradiol(E2)protects rat NPCs from interleukin-1β(IL-1β)-induced apoptosis via the PI3K/Akt signaling pathway.This study was aimed to investigate whether downstream proteins of PI3K/Akt pathway were involved in inhibition of E2 on NPCs’apoptosis.Primary culture of rat NPCs was isolated by trypsin digestion.Being pretreated with E2 and different inhibitors of downstream proteins of PI3K/Akt pathway,the NPCs were treated with IL-1β.Cellular apoptosis was detected by Annexin V/PI staining.Cell viability was detected by CCK-8.Cell adhesion was evaluated by cell-collagen binding assay.Phosphorylation levels of mammalian target of Rapamycin(mTOR),glycogen synthase kinase-3β(GSK-3β)and nuclear factorκB(NF-κB)were detected by Western blot.The results showed that E2 significantly inhibited the IL-1β-induced apoptosis of NPCs,reversed the decrease of cell viability and adhesion induced by IL-1β,and inhibited the down-regulation of mTOR phosphorylation level induced by IL-1β.Rapamycin could block these protective effects of E2.These results suggest that E2 may inhibit IL-1β-induced NPCs’apoptosis through the PI3K/Akt/mTOR signaling pathway.

关 键 词：17Β-雌二醇 髓核细胞 mTOR 白介素1Β 凋亡 

分 类 号：R34[医药卫生—基础医学]