D159687减轻神经元氧化应激损伤的作用机制研究  

Study on the mechanism of D159687 in reducing oxidative stress injury of neurons

作　　者：李俊 王伟杰[1] 丁涟沭[1] LI Jun;WANG Weijie;DING Lianshu(Huai’an First Hospital Affiliated to Nanjing Medical University, Huai’an 223300, Jiangsu, China)

机构地区：[1]南京医科大学附属淮安第一医院,江苏淮安223300

出　　处：《现代中西医结合杂志》2020年第16期1747-1752,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：淮安市科技局立项课题(HAB201726)。

摘　　要：目的研究D159687对过氧化氢(H2O2)所致的小鼠海马神经元HT22细胞损伤的保护作用并探讨其作用机制。方法实验分为正常对照组、模型组、D1596870.5μmol/L组、D1596871μmol/L组、D1596872μmol/L组、D1596874μmol/L组。将HT22细胞株接种于96孔板,对照组采用10%胎牛血清的DMEM高糖培养基培养,其他组加入不同浓度的D159687进行干预培养,后经H2O2损伤。CCK-8法测定各组细胞存活率,Western blot法检测Caspase-3和NADPH氧化酶亚基gp91phox、p47phox表达情况,TUNEL法检测细胞凋亡情况,并测定各组细活性氧(ROS)水平、丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性。结果模型组细胞存活率、SOD活性均显著低于正常对照组(P均<0.05),ROS表达水平、MDA含量及Caspase-3、p47phox、gp91phox蛋白表达水平均显著高于正常对照组(P均<0.05);D1596870.5μmol/L组、D1596871μmol/L组、D1596872μmol/L组、D1596874μmol/L组细胞存活率、SOD活性显著高于模型组(P均<0.05),ROS表达水平、MDA含量及Caspase-3、p47phox、gp91phox蛋白表达水平均显著低于模型组(P均<0.05)。结论在0.5~4μmol/L浓度范围内,D159687浓度依赖性地拮抗H2O2所致HT22细胞损伤,同时抑制HT22细胞凋亡,这种作用可能与其抑制NADPH氧化酶关键亚基gp91phox和p47phox蛋白表达有关。Objective It is to study the protective effect of D159687 on hydrogen peroxide(H2O2)-induced HT22 cell injury in mouse hippocampal neurons and to explore its mechanism.Methods The experiment was divided into 6 groups:normal control group,model group,D1596870.5μM group,D1596871μM group,D1596872μM group and D1596874μM group.The HT22 cell line was inoculated on 96-well plate,DMEM medium with 10%fetal bovine serum was used in control group,and D159687 of different concentrations was added in other groups for intervention culture,and then it was injured by H2O2.The cell survival rate was measured by CCK-8 method,the expression of Caspase-3 and NADPH oxidase subunits gp91phox and p47phox were detected by Western blot,the apoptosis was detected by TUNEL method,the level of reactive oxygen species(ROS),content of malondialdehyde(MDA)and the activity of superoxide dismutase(SOD)were measured.Results The cell survival rate and SOD activity of the model group were significantly lower than those of the normal control group(P<0.05),and the expression level of ROS,the content of MDA and the expression level of Caspase-3,p47phox,gp91phox protein were significantly higher than those of the normal control group(P<0.05);The cell survival rate and SOD activity of the D1596870.5μM group,D1596871μM group,D1596872μM group,D1596874μM group were significantly higher than those of the model group(P<0.05),and the expression level of ROS,the content of MDA,and the expression levels of Caspase-3,p47phox and gp91phox were significantly lower than those of the model group(P<0.05).Conclusion In the concentration range of 0.5-4μM,D159687 can inhibit the injury of HT22 cells induced by H2O2 and the apoptosis of HT22 cells in concentration-dependence,which may be related to the inhibition of the expression of gp91phox and p47phox,the key subunits of NADPH oxidase.

关 键 词：D159687 HT22细胞 氧化应激 

分 类 号：R-33[医药卫生]