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-现代医学

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

单位：沈阳市第四人民医院骨一科, 辽宁沈阳 110031

分类号：R684.3

出版年·卷·期（页码）：2020·39·第六期（723-727）

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+anti-miR-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-1286a 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-1286a 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-miR-NC), MIA+anti-miR-1286a (transfected with anti-miR-1286a), MIA+anti-miR-1286a+si-NC (co-transfected anti-miR-1286a and si-NC), MIA+anti-miR-1286a+si-XIAP (co-transfected anti-miR-1286a 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 ofCaspase-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-1286a 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-1286a showed the opposite effects. miR-1286a can inhibit the fluorescence activity of wild-type XIAP cells, and up-regulate XIAP after inhibiting miR-1286a.After inhibition of XIAP, the regulation of miR-1286a on MIA-induced osteoarthritis chondrocytes was reversed. Conclusion: Inhibition of miR-1286a can reduce apoptosis and oxidative stress of MIA-induced osteoarthritis chondrocyte, and the mechanism is related to targeting XIAP.

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