姜黄素通过Notch信号通路对H_2O_2诱导的脊髓星形胶质细胞损伤的影响研究

Effects of curcumin on spinal astrocyte damage induced by H_2O_2 through Notch signaling pathway

目的:探讨姜黄素对脊髓损伤(SCI)星形胶质细胞(AS)及Notch信号通路的影响。方法:分离培养脊髓AS,选择0. 1、0. 2、0. 4、0. 6、0. 8、1 mmol/L的过氧化氢(H_2O_2)处理AS,CCK8法检测细胞活力,建立AS损伤模型;实验分组为对照组、H_2O_2组和H_2O_2+姜黄素组,流式细胞仪检测各组细胞凋亡及活性氧簇(ROS)含量; ELISA试剂盒检测各组细胞中白介素-6(IL-6)、肿瘤坏死因子(TNF-α)的含量; Western blot检测各组细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、Bcl-2相关X蛋白(Bax)、Notch1和Hes1蛋白表达。结果:不同浓度的H_2O_2均可抑制AS活力,且随H_2O_2浓度升高细胞抑制率升高,选择IC_(50)为0. 4 mmol/L的H_2O_2作为损伤模型;与正常对照组比较,H_2O_2组细胞凋亡率、ROS含量、IL-6和TNF-α含量及Cleaved caspase3、Bax、Notch1和Hes1的蛋白表达均显著升高,与H_2O_2组比较,H_2O_2+姜黄素组细胞凋亡率、ROS含量、IL-6和TNF-α含量及Cleaved caspase3、Bax、Notch1和Hes1的蛋白表达均显著降低(P<0. 05)。结论:姜黄素可通过下调Notch信号通路降低脊髓AS凋亡、ROS含量及炎症因子IL-6和TNF-α含量,从而对SCI起保护作用。

Objective:To investigate the effect of curcumin on Asrocytes and Notch signaling pathway in spinal cord injury.Methods Spinal cord Asrocytes was isolated and cultured,and cells were treated with 0.1,0.2,0.4,0.6,0.8,1 mmol/L hydrogen peroxide(H2O2),the cell viability was measured by CCK8,AS damage model was established.Methods:The experimental grouping was control group,H2O2 group and H2O2+curcumin group,cell apoptosis and ROS content were detected by flow cytometry;the content of IL-6 and TNF-αwere detected by ELISA kit;the expression of Cleaved Caspase3,Bax,Notch1 and Hes1 protein were detected by Western blot.Results:The activity of AS was inhibited by different concentrations of H2O2,and the inhibition rate of cell increased with the increase of H2O2 concentration.IC 50 for 0.4 mmol/L′s H2O2 was selected to establish a damage model;compared with the normal control group,the apoptosis rate,the content of ROS,IL-6,TNF-αand expression of Cleaved Caspase3,Bax,Notch1 and Hes1 protein expression were increased significantly in H2O2 group,compared with H2O2 group,the apoptosis rate,the content of ROS,IL-6,TNF-αand expression of Cleaved Caspase3,Bax,Notch1 and Hes1 protein expression were lower significantly in H2O2+curcumin group(P<0.05).Conclusion:Curcumin can reduce the apoptosis of Asrocytes,the content of ROS and the contents of inflammatory factors IL-6 and TNF-αin spinal cord by down regulating the Notch signaling pathway,thereby protecting spinal cord injury.