摘要
目的 探讨不同配比丹参-三七(SM-PN)药对抗BV2细胞氧糖剥夺/再灌注(OGD/R)损伤的保护作用,并探索其可能的作用机制.方法 建立BV2细胞OGD/R模型,实验分为空白组、模型组及不同配比SM-PN+OGD/R组,采用CCK-8法筛选出细胞活力最好的SM-PN配比;Real-time PCR检测肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素-6(IL-6)、诱导型一氧化氮合酶(iNOS)、白细胞介素-10(IL-10)、转化生长因子-β(TGF-β)的mRNA表达;Western blot法检测核因子KB抑制蛋白α(IKBα)、磷酸化核因子κB抑制蛋白α(p-IκBα)、核因子-KB p65(NF-κB p65)、磷酸化核因子-κB p65(NF-κB p-p65)的磷酸化蛋白水平.结果 200 mg·L-1的SM-PN 5∶3组BV2细胞生存率较模型组显著升高(P＜0.05).与模型组比较,SM-PN组能显著下调IκBα和p65蛋白磷酸化水平;SM-PN组iNOS、TNF-α、IL-1β、IL-6 mRNA表达水平显著降低(P＜0.05);TGF-β、IL-10 mRNA表达水平显著增加(P＜0.05).结论 SM-PN能减轻OGD/R诱导的BV2细胞炎症损伤,这可能与抑制NF-KB信号通路,促进小胶质细胞从M1向M2型转化抑制炎症反应有关.
Abstract
Objective To determine the protective effect of salvia miltiorrhiza-panax notoginseng(SM-PN)with different proportions against BV2 cell injury induced by oxygen-glucose deprivation/reperfusion(OGD/R),and its possible mechanism.Methods BV2 cells were cultured in vitro and the models were established by OGD/R.The cells were randomly divided into a blank group,a model group,and an SM-PN+OGD/R group.The survival rate of cells in each group was screened by CCK-8,and the best SM-PN matching group was selected for subsequent experiments.The expression of TNF-α,IL-1β,IL-6,iNOS,TGF-β and IL-10 mRNA was analyzed by Real-time PCR.The protein levels of IκBα,p-IκBα,p65 and p-p65 were determined by Western blot.Results Compared with the model group,the cell viability of SM-PN 5∶3 groups at 200 mg·L-1 significantly increased,IκBα and p65 protein phosphorylation levels markedly decreased,the mRNA expresion level of iNOS,TNF-α,IL-1β,and IL-6 was significantly lower(P＜0.05),and that of TGF-β and IL-10 was significantly higher(P＜0.05).Conclusion SM-PN can inhibit OGD/R-induced neuroin flammatory responses in BV2 cells,whose mechanism may be related to the inhibition of NF-κB pathway,promoting the polarization of the microglia from M1 to M2 phenotype.
基金项目
河南省中医药科学研究专项(No.20-21ZY2211)
河南省中医药科学研究专项(No.20-21ZY3012)
NETL
NSTL
万方数据