【摘要】 目的:研究微小RNA-422a （miR-422a）靶向调控人性别决定区Y框6（SOX6）对过氧化氢（H₂O₂）刺激下大鼠肾上腺嗜铬细胞瘤细胞株PC12（具有神经细胞特性）损伤的作用。方法:采用real-time PCR检测H₂O₂处理后PC12细胞中miR-422a表达的变化,在PC12细胞中转染miR-422a mimics,MTT法测定细胞活力,试剂盒检测乳酸脱氢酶（LDH）漏出率,流式细胞术测定细胞凋亡水平。靶基因预测软件预测SOX6可能是miR-422a的靶基因,萤光素酶报告基因实验鉴定靶向关系。在PC12细胞中共转染miR-422a mimics和SOX6过表达载体,检测过表达SOX6对miR-422a mimics调控H₂O₂条件下PC12细胞活力、LDH漏出率和凋亡的影响。结果:H₂O₂处理后PC12细胞中miR-422a表达水平降低（P<0.05）。H₂O₂处理后的PC12细胞的活力降低,LDH漏出率和细胞凋亡率升高（P<0.05）。miR-422a mimics能够提高H₂O₂条件下PC12细胞活力,降低LDH漏出率和凋亡率（P<0.05）。miR-422a靶向负调控SOX6表达。SOX6过表达载体可以逆转miR-422a对PC12细胞活力提高和LDH漏出率、细胞凋亡率降低的作用（P<0.05）。结论:miR-422a通过靶向抑制SOX6可降低H₂O₂对PC12细胞的损伤作用。更多还原

【Abstract】 AIM: To investigate the effects of microRNA-422 a（miR-422 a） on the damage of rat adrenal gland pheochromocytoma PC12 cells induced by hydrogen peroxide（H₂O₂）. METHODS: The expression of miR-422 a in the PC12 cells treated with H₂O₂ was detected by real-time PCR. After miR-422 a mimics were transfected into PC12 cells, the cell viability was measured by MTT assay, the lactate dehydrogenase（LDH） leakage rate was detected, and the apoptosis was analyzed by flow cytometry. Target gene prediction software was used to predict that sex-determining region Y box 6（SOX6） may be the target gene of miR-422 a. Luciferase reporter assay was used to identify the targeting relationship. miR-422 a mimics and SOX6 over-expression vector were co-transfected into the PC12 cells. The effects of SOX6 over-expression on the viability, LDH leakage rate and apoptosis of PC12 cells treated with H₂O₂ and transfected with miR-422 a mimics were evaluated. RESULTS: The expression of miR-422 a in the PC12 cells was decreased after treatment with H₂O₂（P<0.05）. The viability of PC12 cells treated with H₂O₂ was decreased, and the LDH leakage rate and apoptotic rate were increased. Transfection with miR-422 a mimics enhanced the viability of PC12 cells treated with H₂O₂, and the leakage rate of LDH and apoptotic rate of the PC12 cells were reduced. The expression of SOX6 was negatively regulated by miR-422 a. SOX6 over-expression reversed the effects of miR-422 a on PC12 cell viability, LDH leakage and apoptosis. CONCLUSION: miR-422 a reduces the damage of PC12 cells induced by H₂O₂ via targeting SOX6.更多还原