miR-1286a对MIA诱导的骨关节炎软骨细胞凋亡和氧化应激的影响  

Effects of miR-1286a on MIA-induced osteoarthritis chondrocyte apoptosis and oxidative stress

作　　者：董宝铁[1] 李泓[1] 高伟[1] 周振东[1] DONG Baotie;LI Hong;GAO Wei;ZHOU Zhendong(the First Department of Orthopedics,the 4th People's Hospital of Shenyang,Shengyang 110031,China)

机构地区：[1]沈阳市第四人民医院骨一科,辽宁沈阳110031

出　　处：《现代医学》2020年第6期723-727,共5页Modern Medical Journal

摘　　要：目的:探讨miR-1286a对碘乙酸钠(MIA)诱导的骨关节炎软骨细胞凋亡、氧化应激的调控机制。方法:用实时荧光定量反转录聚合酶链反应(qRT-PCR)实验检测正常软骨组织、骨关节炎软骨组织和不同处理方式的骨关节炎软骨细胞中miR-1286a、连锁凋亡抑制蛋白(XIAP)的表达;用脂质体法将MIA+antimiR-NC(转染anti-miR-NC)、MIA+anti-miR-1286a(转染anti-miR-1286a)、MIA+anti-miR-1286a+si-NC(共转染anti-miR-1286a和si-NC)、MIA+anti-miR-1286a+si-XIAP(共转染anti-miR-1286a和si-XIAP)转染至MIA诱导的骨关节炎软骨细胞;流式细胞术、蛋白质免疫印迹法检测细胞的凋亡和半胱氨酸天冬氨酸蛋白酶(Caspase-3)、XIAP蛋白的表达;酶联免疫吸附试验(ELISA)检测细胞中超氧化物歧化酶(SOD)、丙二醛(MDA)的含量;双荧光素酶报告实验检测细胞的荧光强度。结果:miR-1286a在骨关节炎软骨组织和MIA诱导的骨关节炎软骨细胞中的表达显著升高(P<0.05)。MIA诱导的骨关节炎软骨细胞的凋亡率明显升高,Caspase-3上调,SOD含量降低,MDA含量升高,抑制miR-1286a后则具有相反的作用。miR-1286a可抑制野生型XIAP细胞的荧光活性,并且抑制miR-1286a后上调XIAP。抑制XIAP还可逆转抑制miR-1286a对MIA诱导的骨关节炎软骨细胞的调控作用。结论:抑制miR-1286a可减轻MIA诱导的骨关节炎软骨细胞凋亡和氧化应激,其机制与靶向XIAP相关。Objective:To investigate the regulatory mechanism of miR-1286 a on the apoptosis and oxidative stress of osteoarthritis chondrocytes induced by sodium iodoacetate(MIA).Methods:The quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-1286 a and X-linked inhibitor of apoptosis protein(XIAP)in normal cartilage tissue,osteoarthritis cartilage tissue,and osteoarthritis chondrocytes with different treatment methods.Using liposome method,MIA+anti-miR-NC group(transfected with anti-miRNC),MIA+anti-miR-1286 a(transfected with anti-miR-1286 a),MIA+anti-miR-1286 a+si-NC(co-transfected anti-miR-1286 a and si-NC),MIA+anti-miR-1286 a+si-XIAP(co-transfected anti-miR-1286 a and si-XIAP)to MIA-induced osteoarthritis chondrocytes.Flow cytometry was used to detect osteoarthritis chondrocyte apoptosis induced by MIA.Western bloting was used to detect the protein expression of Caspase-3 and XIAP.Enzyme-linked immunosorbent assay(ELISA)was performed to evaluate the content of superoxide dismutase(SOD)and malondialdehyde(MDA)in the cells.The double luciferase report experiment was applied to determine the fluorescence intensity of the cells.Results:The expression of miR-1286 a was significantly increased in osteoarthritis cartilage tissue and MIA-induced osteoarthritis chondrocytes(P<0.05).MIA-induced osteoarthritis chondrocyte apoptosis rate enhanced greatly,Caspase-3 was up-regulated,SOD content decreased,MDA content increased,and inhibition of miR-1286 a showed the opposite effects.miR-1286 a can inhibit the fluorescence activity of wild-type XIAP cells,and up-regulate XIAP after inhibiting miR-1286 a.After inhibition of XIAP,the regulation of miR-1286 a on MIA-induced osteoarthritis chondrocytes was reversed.Conclusion:Inhibition of miR-1286 a can reduce apoptosis and oxidative stress of MIA-induced osteoarthritis chondrocyte,and the mechanism is related to targeting XIAP.

关 键 词：miR-1286a 连锁凋亡抑制蛋白 骨关节炎软骨细胞 凋亡 氧化应激 

分 类 号：R684.3[医药卫生—骨科学]