MG53蛋白对脂多糖诱导的小鼠海马神经元细胞氧化损伤的影响
Effect of MG53 protein on lipopolysaccharide-induced oxidative damage in mice hippocampal neuron cells

目的:研究MG53蛋白对脂多糖(LPS)损伤的小鼠海马神经元细胞HT22的保护作用。方法:用不同质量浓度(25、50、100、250、500、1 000、1 500 mg/L)LPS分别处理HT22细胞 12、24、36、48、60 h后,CCK-8法检测细胞增殖,筛选LPS的作用时间及浓度。将HT22细胞分为正常对照组、MG53(30 mg/L)组、LPS组和MG53+LPS组,分别采用CCK-8法、Annexin V-FITC/PI双染法、流式细胞术和划痕实验检测各组细胞存活率、凋亡、细胞周期和迁移距离,ELISA法检测细胞内SOD活性和MDA含量,采用Western blot法检测Bax、Bcl-2 和Cyclin D1蛋白的表达。结果:经筛选,选定以500 mg/L的LPS作用48 h为细胞模型的实验条件。MG53可降低HT22细胞的凋亡率,促进G0/G1期细胞向S期转运,增加其迁移距离; 升高细胞内SOD活性,降低MDA含量,增加细胞内Bcl-2和Cyclin D1蛋白表达,降低Bax蛋白表达(P<0.05)。结论:MG53蛋白对LPS诱导的HT22细胞氧化损伤具有保护作用。
Aim: To investigate the improved effect of MG53 protein on lipopolysaccharide(LPS)-induced damage in mice hippocampal neuron HT22 cells.Methods: HT22 cells were treated with different concentrations(25, 50, 100, 250, 500, 1 000, 1 500 mg/L)of LPS for 12, 24, 36, 48 and 60 h, and CCK-8 assay was used to detect proliferation. HT22 cells were allocated into four groups: normal control group, MG53(30 mg/L)group, LPS group and MG53+LPS group. The proliferation, apoptosis, cell cycle and migration distance were detected by CCK-8 assay, Annexin V-FITC/PI, flow cytometry and Transwell assay, respectively, SOD and MDA were measured by ELISA kit, and the expressions of Bax, Bcl-2 and Cyclin D1 protein were detected by Western blot.Results: The cell proliferation inhibition rate of HT22 cells treated by 500 mg/L LPS for 48 h was close to 50%, so the experimental condition was determined. migration distance, cells in S phase of the HT22 cells treated by MG53 increased significantly, while apoptosis rate and cells in G0/G1 phase decreased significantly(P<0.05); SOD activity increased but MDA content decreased; meanwhile, the expression of Bcl-2 and Cyclin D1 protein increased, and Bax expression decreased significantly(P<0.05).Conclusion: MG53 protein could improve the LPS-induced oxidative damage in HT22 cells