受体相互作用蛋白质激酶3通过上调整合素β3促进骨关节炎相关病变  

Receptor-interacting Protein Kinase 3 Promotes Osteoarthritis-related Changes by Upregulating Integrin Subunit Beta 3

作　　者：程锦[1] 蒋艳芳[1] 段小宁[1] 傅欣[1] 张继英[1] 胡晓青[1] 敖英芳[1] CHENG Jin;JIANG Yan-Fang;DUAN Xiao-Ning;FU Xin;ZHANG Ji-Ying;HU Xiao-Qing;AO Ying-Fang(Department of Sports Medicine,Peking University Third Hospital,Institute of Sports Medicine of Peking University,Beijing Key Laboratory of Sports Injuries,Beijing 100191,China)

机构地区：[1]北京大学第三医院运动医学科,北京大学运动医学研究所,北京市运动医学关节伤病重点实验室,北京100191

出　　处：《中国生物化学与分子生物学报》2021年第3期391-400,共10页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.32000923,No.82072486);北京市自然科学基金(No.7214304,No.7171014)资助。

摘　　要：骨关节炎(osteoarthritis,OA)是最常见的慢性致残性关节疾患,目前尚无针对病因的有效治疗手段。程序性坏死在多种疾病中扮演关键角色,受体相互作用蛋白质激酶3(receptor-interacting protein kinase 3,RIP3)是程序性坏死进程的关键调控因子。有研究显示,RIP3在人与鼠骨关节炎退变软骨组织中表达水平显著上调,提示程序性坏死的发生,但RIP3在软骨中的具体病生理角色仍不明确。本研究拟对过表达RIP3前后的软骨细胞转录物组进行测序分析,探索RIP3在骨关节炎进程中发挥作用的具体机制。RNA测序结果显示,RIP3的过表达诱发软骨细胞中244个基因表达上调,277个表达下调。通过进一步构建基因间共表达作用网络,筛选出16个候选靶基因在mRNA水平进行验证,证实RIP3对磷脂酰肌醇3激酶调节亚单位5(phosphoinositide-3-kinase,regulatory subunit 5,Pik3r5)、整合素β3(integrin subunit beta 3,Itgb3)及成髓细胞瘤转录因子第2亚型(MYB proto-oncogene like 2,Mybl2)的表达上调作用最为显著。CCK-8以及乳酸脱氢酶活性检测结果表明,利用siRNA沉默Itgb3的表达可显著抑制RIP3诱发的软骨细胞活力下降及程序性坏死,同时也抑制了RIP3对软骨细胞中分解代谢相关基因Mmp1、Mmp13与Il6的表达上调作用,以及其对合成代谢相关基因Acan、Col2a1与Sox9的下调作用。本研究证实,RIP3通过上调软骨细胞中Itgb3的表达诱发软骨细胞坏死与软骨基质代谢紊乱,并最终导致软骨退变,为骨关节炎的临床治疗提供了新靶点,同时进一步明确了程序性坏死的病理生理学意义。Osteoarthritis(OA)is the most common chronic disabling joint disease,and currently there is no effective treatment for the cause.Necroptosis plays a key role in many diseases,and receptor-interacting protein kinase 3(RIP3)is a key regulator during necroptosis process.Studies have shown that the expression level of RIP3 was significantly upregulated in human and mouse OA degenerative cartilage tissues,suggesting the occurrence of necroptosis.However,the specific pathophysiological role of RIP3 in cartilage is still unclear.This study intends to sequence and analyze the transcriptome of chondrocytes before and after RIP3 overexpression,and explore the specific functional mechanism of RIP3 in OA pathogenesis.RNA sequencing results showed that overexpression of RIP3 induced upregulation of 244 genes and downregulation of 277 genes in chondrocytes.Sixteen candidate target genes were screened out by constructing gene co-expression network for further verification at mRNA level,and the results suggested that RIP3 had the most significant inductive effect on the expression of phosphoinositide-3-kinase,regulatory subunit 5(Pik3r5),integrin subunit beta 3(Itgb3)and MYB proto-oncogene like 2(Mybl2).Results from CCK-8 and lactate dehydrogenase activity analysis showed that silencing the expression of Itgb3 by siRNA significantly rescued chondrocyte viability decline and necroptosis induced by RIP3,and it also inhibited the upregulating effect of RIP3 on the expression of catabolism-related genes Mmp1,Mmp13 and Il6,as well as the downregulating effect of RIP3 on the expression of anabolism-related genes Acan,Col2a1 and Sox9.This study has demonstrated that RIP3 promotes chondrocyte necrosis and cartilage matrix metabolism disorders by upregulating the expression of Itgb3 in chondrocytes,and ultimately leads to cartilage degeneration.These findings provided potential novel targets for the clinical treatment of OA,and further clarified the pathophysiological significance of necroptosis.

关 键 词：骨关节炎 软骨细胞 程序性坏死 受体相互作用蛋白质激酶3 整合素Β3 

分 类 号：R684[医药卫生—骨科学]