首页|Sestrin2通过调控mTORC1通路在骨性关节炎中的保护机制研究

Sestrin2通过调控mTORC1通路在骨性关节炎中的保护机制研究

扫码查看
原文链接
  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
目的 探索Sestrin2(SESN2)通过介导哺乳动物雷帕霉素靶蛋白复合体1(mTORC1)信号通路调控软骨细胞自噬活性的机制.方法 利用白细胞介素-1β(IL-1β)处理正常软骨细胞制备骨性关节炎(OA)软骨细胞模型,并分为对照组和IL-1β处理组,通过实时定量PCR(qPCR)及Western印迹检测对照组和IL-1β处理组细胞基质金属蛋白酶13(MMP13)、Ⅱ型胶原(COL2A1)及SESN2的表达水平.通过细胞转染技术获得软骨细胞过表达SESN2组与敲低SESN2组细胞模型,并用qPCR检测各组SESN2表达水平,用Western印迹检测各组SESN2、MMP13、COL2A1、mTORC1通路相关蛋白及自噬相关蛋白表达水平,用CCK-8和细胞划痕实验检测各组SESN2对细胞增殖和迁移的影响.结果 (1)经IL-1β处理后的软骨细胞MMP13表达水平显著上调,COL2A1及SESN2表达水平显著下降.(2)与对照组相比过表达SESN2组OA软骨细胞p-mTORC1、核糖体蛋白S6激酶1(S6K1)、MMP13蛋白表达显著下调,而5'-腺嘌呤核苷酸活化蛋白激酶(AMPK)和软骨保护基因COL2A1表达显著上升、苄氯素-1(Beclin-1)表达水平及微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)/(LC3-Ⅰ)比值增高,同时,过表达SESN2可上调软骨细胞增殖、迁移活性,敲低SESN2后结果则相反.结论 SESN2可以通过抑制mTORC1通路活性增强软骨细胞自噬、增殖及迁移活性,为揭示OA的发病机制及探寻新的治疗方法提供实验依据.
Sestrin2 protects against osteoarthritis by regulating the mTORC1 pathway
Objective To explore the mechanism by which Sestrin2(SESN2)regulates autophagy activity of chondrocytes by mediating mammalian rapamycin target protein complex 1(mTORC1)signaling pathway.Methods The normal chondrocytes were treated with interleukin-1 β(IL-1β)to establish an osteoarthritis(OA)chondrocyte model,which was divided into the control group and the IL-1 β-treated group.Real-time quantitative PCR(qPCR)and Western blot were used to detect the expression levels of matrix metalloproteinase 13(MMP13),type Ⅱ collagen(COL2A1)and SESN2 in the two groups.The cell models of the chondrocyte overexpression SESN2 group and knockdown SESN2 group were obtained via cell transfection technology,and the expression levels of SESN2 in each group were detected by qPCR while those of SESN2,MMP13,COL2A1,mTORC1 pathway-related proteins and autophagy-related proteins in each group were detected by Western blot.The effects of SESN2 on cell proliferation and migration were detected by CCK-8 and cell scratch assay.Results(1)The expression level of MMP13 in the IL-1 β-treated group was significantly up-regulated,while the expression levels of COL2A1 and SESN2 were significantly decreased.(2)Compared with the control group,the expressions of p-mTORC1,ribosomal protein S6 kinase 1(S6K1),and MMP13 protein in OA chondrocytes in the overexpression group were significantly down-regulated,while the expressions of adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)and chondroprotective gene COL2A1 were significantly increased,and the expression level of Beclin-1 and the ratio of microtubule associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)/(LC3-Ⅰ)were increased.Meanwhile,overexpression of SESN2 could up-regulate the proliferation and migration of chondrocytes,but the results were opposite after knockdown of SESN2.Conclusion SESN2 can enhance autophagy,proliferation and migration of chondrocytes by inhibiting mTORC1 pathway,which has provided data for revealing the pathogenesis of OA and exploring new therapeutic methods.

osteoarthritischondrocytesestrin2mTORC1autophagycell proliferationcell migration

刘泽众、李彩霞、刘晓光、付道通、刘长杰、张益民、赵世波

展开 >

山东第二医科大学,山东潍坊 261000

潍坊市人民医院牙科,山东潍坊 261000

潍坊市人民医院关节外科,山东潍坊 261000

骨性关节炎 软骨细胞 Sestrin2 mTORC1 自噬 细胞增殖 细胞迁移

山东省医药卫生科技发展计划项目山东省自然科学基金课题面上项目山东省自然科学基金青年基金项目潍坊市卫健委科研计划项目金天格中青年科研培养基金

202104070723ZR2020MH094ZR2019PH026WFWSJK-2022-057

2024

10.7644/j.issn.1674-9960.2024.08.004

军事医学
军事医学科学院

军事医学

CSTPCD
影响因子：0.586
ISSN：1674-9960
年,卷(期)：2024.48(8)