香青兰总黄酮对H_2O_2诱导的U87细胞损伤的保护机制研究

Protective mechanism of total flavonoids of Dracocephalum Moldavica L. on H_2O_2-induced U87 cell injury

目的探讨香青兰总黄酮对过氧化氢(H_2O_2)诱导的U87细胞损伤的保护作用,以及ERK/MAPK信号通路在该过程中的作用机制。方法以正常培养的U87细胞为正常对照组,U87细胞加入0.31 mmol/L的H_2O_2损伤20min为模型组,采用不同浓度香青兰总黄酮(200.00、100.00、50.00、25.00、12.50、6.25、3.13、1.56μg/mL)预给药U87细胞2h,分别加入0.31mmol/L的H_2O_2刺激U87细胞20min作为实验组,采用CCK-8法检测U87的增殖活力,用ROS检测试剂盒法对活性氧(ROS)含量进行测定,Western blot法检测P-ERK1/2、ERK1/2蛋白的表达水平。结果与模型组比较,实验组细胞增殖活力提高,差异有统计学意义(P<0.05),与模型组相比,实验组细胞ROS含量降低,差异有统计学意义(P<0.05)。Western blot实验表明,与模型组比较,香青兰总黄酮不同浓度(100、50、25、5μg/mL)实验组P-ERK1/2蛋白含量均高于模型组,且呈现剂量依赖,差异有统计学意义(P<0.05)。结论香青兰总黄酮通过激活P-ERK1/2通路,抑制ROS过量产生,起到拮抗H_2O_2对U87细胞增殖的抑制作用。

Objective To investigate the protective effect of total flavonoids of Dracocephalum Moldavica L.on U87 cell injury induced by hydrogen peroxide（H_2O_2）and the role of ERK/MAPK signal pathway in this process.Methods U87 cell without treatment was used as normal control group,U87 cell treated with 0.31 mmol/L H_2O_2 for 20 min was used as model group,U87 cell in the test group was treated with different concentrations of total flavonoids（200.00,100.00,50.00,25.00,12.50,6.25,3.13,1.56μg/mL）for2.0 h,and then stimulated with 0.31 mmol/L H_2O_2 for 20 min to establish oxidative stress injury.The cell proliferation activity and the content of ROS of these groups were detected by CCK-8 and ROS detection kit,respectively.The expression of P-ERK1/2 and ERK1/2 was detected by western blot.Results Compared with the model group,the cell proliferation activity was significantly increased in the test group（P0.05）;the content of ROS in the test group at different concentrations of total flavonoids were lower than that in the model group（P 0.05）.The expression of P-ERK1/2 in the test group at different concentrations of total flavonoids（100.00,50.00,25.00,5.00μg/mL）was significantly higher than that in the model group（P 0.05）,and the change was dose dependent.Conclusion Total flavonoids of Dracocephalum Moldavica L.could alleviate the injury of U87 cells induced by H_2O_2 by activating the P-ERK1/2 pathway,and inhibiting the overproduction of ROS.