含WW结构域的转录因子1对软骨细胞合成和分解代谢的调控作用  

Regulation of chondrocyte synthesis and catabolism by WW structural domain-containing transcription factor1

作　　者：王远兴 张飞 王涛 谢志鸿 李豪 任超 彭吾训 Wang Yuanxing;Zhang Fei;Wang Tao;Xie Zhihong;Li Hao;Ren Chao;Peng Wuxun(Clinical School of Medicine,Guizhou Medical University,Guizhou 550004,China;Department of Emergency Orthopedics,Affiliated Hospital of Guizhou Medical University,Guizhou 550004,China)

机构地区：[1]贵州医科大学临床医学院,贵州550004 [2]贵州医科大学附属医院急诊骨科,贵州550004

出　　处：《中华实验外科杂志》2024年第7期1409-1412,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(82260429、82260434、82060397);贵州省自然科学基金(黔科合基础-ZK[2022]一般399);贵州医科大学附属医院博士科研启动基金(gyfybsky-2022-14);贵州医科大学附属医院“学科优秀后备人才”项目。

摘　　要：目的探讨含WW结构域的转录因子1(WWTR1或TAZ)在创伤后骨性关节炎(PTOA)中对软骨细胞的调控作用。方法由贵州医科大学动物实验中心提供20只c57bl/6J小鼠简单随机法分为假手术组(Sham)组、内侧半月板(DMM)不稳定组,每组6例,3个月后取材。苏木精-伊红(HE)染色和番红O固绿(SafraninO)染色验证模型是否构建成功和根据国际骨关节炎研究学会(OARSI)对骨关节软骨损伤进行分级,免疫组织化学法检测TAZ的表达。分离培养膝关节原代软骨细胞,将其分为对照组(Control)和TAZ抑制组(TAZinhibitor-1)进行体外实验。通过免疫荧光检测TAZinhibitor-1对TAZ表达的影响。蛋白质印迹法(Westernblot)和实时荧光定量聚合酶链反应(qPCR)检测TAZ inhibitor-1对软骨细胞合成和分解代谢相关因子表达的影响。组间比较采用t检验。结果HE结果显示,DMM组滑膜比对照组有增厚和有炎性细胞浸润。SafraninO染色显示,根据OARSI评分DMM组关节软骨浅层比对照组有降解和软骨下骨增厚[(4.333±1.731)分比(1.000±0.167)分,t=21.240,P<0.01]。免疫组织化学结果显示,DMM组TAZ表达低于对照组(63.333±2.667比44.500±1.500,t=19.210,P<0.01)。在体外实验中,免疫荧光结果显示,TAZ inhibitor-1组软骨细胞内TAZ的表达低于对照组(44.406±1.731比54.772±5.419,t=15.860,P<0.01)。Westernblot结果显示,TAZinhibitor-1组二型胶原酶I(Col2al)、糖胺聚糖(ACAN)表达低于对照组(0.470±0.038比0.937±0.330,0.589±0.015比0.859±0.037,t=20.830、17.160,P<0.01),TAZinhibitor-1组基质金属蛋白酶-13(MMP-13)、含TSP样基序的去整合素金属蛋白酶(ADAMTS)-7表达高于对照组(0.897±0.004比0.540±0.018、0.956±0.007比0.605±0.013,t=4.920、8.056,P<0.01)。qPCR结果显示,TAZinhibitor-1组MMP-13、ADAMTS-7、SOX5等基因表达高于对照组(1.697±0.030比1.000±0.007、5.242±0.343比1.000±0.031、0.769±0.018比1.000±0.030,t=22.820、23.820、12.270,P<0.01),TAZinhibitor-1组CO12al、ACAN、Objective To investigate the role of WW structural domain-containing transcription factor 1(WWTR1 or TAZ)in the regulation of chondrocytes in post-traumatic osteoarthritis(PTOA).Methods Totally,20 c57bl/6 mice provided by the Animal Experiment Center of Guizhou Medical University were divided into the sham-operated(Sham)group and the medial meniscus(DMM)instability group by a simple randomization method,6 cases per group,and the knee joints were extracted 3 months later.The model was successfully constructed by employing hematoxylin and eosin(HE)staining and Safranin O staining and grading of osteoarthritic cartilage damage according to the Osteoarthritis Research Society International(OARSI),and the expression of TAZ was detected by immunohistochemistry.Primary chondrocytes of the knee joint were isolated,cultured,and divided into control group and TAZ inhibitor group(TAZ inhibitor-1)for in vitro experiments.Then the effect of TAZ inhibitor-1 on TAZ expression in chondrocytes was detected by immunofluorescence.Western blotting assays and real-time quantitative polymerase chain reaction(qPCR)were performed to detect the effect of TAZ inhibitor-1 on the expression of factors related to chondrocyte synthesis and catabolism.Comparisons between groups were made using the ttest.Results HE staining showed that the synovium was thickened and infiltrated with inflammatory cells in the DMM group compared to the control group.Safranin O staining showed that superficial articular cartilage in the DMM had more degradation and thickening of the subchondral bone than the control group according to the the OARSI score(4.333±1.731 vs.1.000±0.167,t=21.240,P<0.01).Immunohistochemical results showed that the TAZ expression was lower in the DMM group than in the control group(63.333±2.667 vs.44.500±1.500,t=19.210,P<0.01).In vitro experiments,Immunofluorescence results showed that the expression of TAZ in chondrocytes in the TAZ inhibitor-1 group was lower than that in the control group(44.406±1.731 vs.54.772±5.419,t=15.860,P<0

关 键 词：创伤 骨性关节炎 软骨细胞 转录因子1 代谢 

分 类 号：R684.3[医药卫生—骨科学]