miR-199a调控Sirt1对SH-SY5Y细胞氧化损伤的影响  

Effects of miR-199a in regulation of Sirt1 on oxidative damage of SH-SY5Y nerve cells

作　　者：万静枝 王婷 廖勇 Wan Jingzhi;Wang Ting;Liao Yong(Affiliated Renhe Hospital of China Three Gorges University,Yichang 443001,Hubei,China;School of Medine,Wuhan University of Science and Technology,Wuhan 430065,Hubei,China)

机构地区：[1]三峡大学附属仁和医院,湖北宜昌443001 [2]武汉科技大学医学院,湖北武汉430065

出　　处：《贵州医药》2025年第1期3-6,共4页Guizhou Medical Journal

摘　　要：目的探讨miR-199a调控Sirt1对人神经母细胞瘤细胞(SH-SY5Y)氧化损伤的影响。方法采用H_(2)O_(2)(1200μmol/L)分别刺激体外培养的SH-SY5Y细胞3、6、9、12、15 h,MTT法检测细胞活力,DCFA-DA探针法检测细胞内ROS水平;miR-199a模拟子和抑制物分别转染SH-SY5Y细胞48 h,Western blot检测Sirt1蛋白的表达;miR-199a模拟子和抑制物分别转染SH-SY5Y细胞48 h后采用H_(2)O_(2)(1200μmol/L)刺激12 h,生化法检测细胞超氧化物歧化酶活性和丙二醛含量。结果不同时间H_(2)O_(2)刺激SH-SY5Y细胞时细胞活力,细胞内超氧化物歧化酶含量升高,且呈现时间依赖性;miR-199a模拟子可抑制SH-SY5Y细胞Sirt1蛋白表达,miR-199a抑制物增加SH-SY5Y细胞Sirt1蛋白表达;当SH-SY5Y细胞处于氧化损伤时,miR-199a模拟子可降低细胞超氧化物歧化酶活性和升高丙二醛含量,miR-199a抑制物增加细胞活力、降低细胞内超氧化物歧化酶含量、升高超氧化物歧化酶活性和降低丙二醛含量。结论当SH-SY5Y细胞处于氧化损伤状态时,miR-199a调控Sirt1进一步加重SH-SY5Y细胞氧化损伤。Objective To investigate the effects of miR-199a in regulation of Sirt1 on oxidative damage of SH-SY5Y nerve cells.Methods Cultured SH-SY5Y nerve cells were stimulated with H_(2)O_(2)(1200μmol/L)for 3 h,6 h,9 h,12 h and 15 h respectively.Cell viability was detected by MTT assay and intracellular ROS levels were detected by DCF-DA probe.SH-SY5Y nerve cells were transfected with miR-199a mimics and inhibitors for 48 h,the expression of Sirt1 protein was detected by Western blot.SH-SY5Y nerve cells were transfected with miR-199a mimics and inhibitors for 48 h respectively and stimulated with H_(2)O_(2)(1200μmol/L)for 12 h.The activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)were detected by enzyme biochemical method.Result When SH-SY5Y nerve cells were stimulated by H_(2)O_(2) at different time,the cell viability and intracellular ROS content increased in a time-dependent manner.The miR-199a mimicon inhibited Sirt1 protein expression in SH-SY5Y neurons,the miR-199a inhibitor increased Sirt1 protein expression in SH-SY5Y neurons.When SH-SY5Y nerve cells were in a state of oxidative damage,miR-199a mimics decreased SOD content and increased MDA content,while miR-199a inhibitor increased cell vitality,decreased intracellular ROS content,increased SOD content and decreased MDA content.Conclusion When SH-SY5Y nerve cells are in the state of oxidative damage,the regulation of Sirt1 by miR-199a further aggravated the oxidative damage of SH-SY5Y nerve cells.

关 键 词：氧化损伤 人神经母细胞瘤细胞 miR-199a SIRT1 

分 类 号：R34[医药卫生—基础医学]