羟基红花黄色素A对糖氧剥夺/复糖复氧后星形胶质细胞保护作用及其机制研究

Protective effect and mechanism of hydroxysafflor yellow A on astrocytes after OGD/R

目的:探讨糖氧剥夺/复糖复氧(OGD/R)状态下羟基红花黄色素A(HSYA)对星形胶质细胞(Ast)的保护作用及其机制。方法:分离小鼠原代Ast,实验设置为正常对照组、模型组和HSYA处理组。MTT、LDH检测Ast活力;免疫荧光检测GFAP蛋白表达;RT-PCR和Western blot检测C3、S100A10、PTX3极化指标;RT-PCR和ELISA法检测TNF-α、IL-1β、IL-6、IL-10炎症因子表达;氧化检测试剂盒检测CAT、SOD、GSH-Px、RNS、ROS、MDA含量;Western blot检测GFAP、p-NF-κB(p65)、p-STAT3、Nrf2、HO-1蛋白表达。结果:Ast经OGD/R处理后被激活且形态发生改变;同时A1型Ast增加,TNF-α、IL-1β、IL-6等炎症因子及RNS、ROS、MDA表达升高;A2型Ast降低,CAT、SOD、GSH-Px降低。HSYA可减轻OGD/R状态下Ast的形态异常,减少A1型Ast,同时减少炎症因子分泌以及RNS、ROS和MDA含量,降低p-NF-κB(p65)和p-STAT3表达;增加IL-10分泌和CAT、SOD、GSH-Px活性,增加Nrf2和HO-1蛋白表达。结论:HSYA可能通过p-NF-κB(p65)/p-STAT3通路降低OGD/R后Ast的炎症反应和激活Nrf2/HO-1通路来减轻Ast的氧化应激发挥保护作用。

Objective:To study the protective effect and mechanism of hydroxysafflor yellow A(HSYA)on astrocytes(Ast)in the state of oxygen deprivation/reoxygenation(OGD/R).Methods:Primary Ast of mice were isolated and cultured.The cells were devided into 3 groups as follows:normal group,OGD/R group and OGD/R plus HSYA(OGD/R+HSYA)group.Cytotoxicity and cell viability of HSYA on Ast were detected by MTT assay and LDH,the expression of GFAP was examined by immunocytochemistry.RT-PCR and Western blot was used to determine the expressions of C 3,S 100 A 10,PTX 3.The levels of TNF-α,IL-1β,IL-6,IL-10 were detected with ELISA and RT-PCR.CAT,SOD,GSH-Px,RNS,ROS,MDA were detected by oxidative damage detection kits.The expressions of GFAP,p-NF-κB(p 65),p-STAT 3,Nrf 2 and HO-1 protein was examined by Western blot.Results:Ast was activated and changed in shape after OGD/R treatment.Type A 1 Asts were increased and the expressions of TNF-α,IL-1βand IL-6,and the content of RNS,ROS and MDA increased.While A 2 type Ast decreased,the content of CAT,SOD,GSH-Px decreased.After HSYA treatment,the expression of GFAP and type A 1 Ast as well as the secretions of TNF-α,IL-1βand IL-6,and the content of RNS,ROS and MDA were reduced,and p-NF-κB/p 65,p-STAT 3 were decreased,but the expressions of type A 2 Asts,the secretions of IL-10 and the activities of CAT,SOD,GSH-Px were increased.Increased expression of Nrf 2 and HO-1 protein.Conclusion:HSYA can regulate the release of inflammatory factors,which may be related to inhibit the p-NF-κB(p 65)/p-STAT 3 pathways.In addition,HSYA also reduced the oxidative response of Ast through the activation of Nrf 2/HO-1 pathway.