miR-145对TNF-α诱导人关节软骨细胞增殖、凋亡及炎症反应的影响  

Effect of miR-145 on the proliferation,apoptosis and inflammation of TNF-α-induced human articular chondrocytes

作　　者：阿布都艾尼·热吾提 肖伟[1] 周文正[1] 车立新[1] 徐江波[1] 孙俊刚[1] ABUDUAINI Re-wu-ti;XIAO Wei;ZHOU Wen-zheng;CHE Li-xin;XU Jiang-bo;SUN Jun-gang(Department of Orthopedic Trauma,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi,Xinjiang,830001,China)

机构地区：[1]新疆维吾尔自治区人民医院骨科中心创伤病区,乌鲁木齐830001

出　　处：《中国骨与关节杂志》2021年第7期521-526,共6页Chinese Journal of Bone and Joint

基　　金：新疆维吾尔自治区自然科学基金(2019D01C106)。

摘　　要：目的明确miR-145在肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)诱导人关节软骨细胞增殖、凋亡及炎症反应中的作用。方法利用0、5、10、15及30 ng/ml TNF-α诱导人C-20/A4和CH8关节软骨细胞48 h,提取总RNA并反转录,采用实时荧光定量聚合酶链反应检测miR-145的表达水平。瞬时转染miR-145模拟物(mimics)至30 ng/ml TNF-α诱导的人C-20/A4和CH8细胞,分别采用噻唑蓝比色法[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]、蛋白质印迹及酶联免疫吸附剂测定(enzyme-linked immunosorbnent assay,ELISA)观察细胞增殖、凋亡及炎症因子浓度的变化。结果随着TNF-α诱导浓度的提高,C-20/A4和CH8细胞中白介素-1β(interleukin-1β,IL-1β)、IL-6及IL-13的浓度呈剂量依赖性升高(P<0.05),而miR-145的表达水平呈剂量依赖性降低(P<0.05)。与对照组相比,miR-145 mimics显著提高C-20/A4和CH8细胞中miR-145的表达水平(P<0.05)。与对照组相比较,miR-145 mimics显著降低C-20/A4和CH8细胞增殖能力,并增加胱天蛋白酶3(caspase 3,CASP3)、cleaved CASP3、Bcl-2相关X蛋白(Bcl-2 related X protein,BAX)的表达水平及IL-1β、IL-6及IL-13的浓度(P<0.05)。结论miR-145在TNF-α诱导的人关节软骨细胞中呈剂量依赖性下调。体外过表达miR-145抑制TNF-α诱导的人关节软骨细胞增殖,促进凋亡及炎症反应。Objective To identify the role of miR-145 in the proliferation,apoptosis and inflammation of human articular chondrocytes induced by tumor necrosis factorα(TNF-α).Methods Human C-20/A4 and CH8 articular chondrocytes were treated with 0,5,10,15,and 30 ng/ml of TNF-αfor 48 h.Total RNA was extracted and reversely transcribed,and the expression levels of miR-145 were detected by real-time quantitative polymerase chain reaction.miR-145 mimics were transiently transfected into C-20/A4 and CH8 cells,and cell proliferation,apoptosis and inflammatory factors were evaluated by using 3-(4,5-dimethylth-iazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT),Western blotting and enzyme-linked immunosorbent assay(ELISA),respectively.Results With the increase of induced concentration of TNF-α,the concentrations of inflammatory factors including interleukin-1β(IL-1β),IL-6 and IL-13 in C-20/A4 and CH8 cells increased in a dose-dependent manner(P<0.05),whereas the expression level of miR-145 decreased in a dose-dependent manner(P<0.05).Compared with the control group,miR-145 mimics significantly increased the expression levels of miR-145 in C-20/A4 and CH8 cells(P<0.05).Compared with the control group,miR-145 mimics significantly reduced proliferation ability of C-20/A4 and CH8 cells,and increased the expression levels of caspase-3(CASP3),cleaved CASP3,Bcl-2 related X protein(BAX),and concentrations of IL-1β,IL-6 and IL-13(P<0.05).Conclusions miR-145 is dose-dependently down-regulated in human articular chondrocytes induced by TNF-α.Over-expression of miR-145 inhibits the proliferation,and promotes apoptosis and inflammation of human articular chondrocytes induced by TNF-αin vitro.

关 键 词：微RNAS 肿瘤坏死因子类 软骨细胞 细胞增殖 炎症 

分 类 号：R684[医药卫生—骨科学]