失笑散降低氧糖剥夺再灌注BV2细胞损伤及NLRP3的表达

Shixiao Powder Reduces the Damage of BV2 Cells induced by Hypoxia Glucose Deprivation Reperfusion and the Expression of NLRP3

目的:研究失笑散对BV2细胞氧糖剥夺再灌注(OGD/R)模型的保护作用及潜在机制。方法:将BV2细胞分为对照组,模型(OGD/R)组及失笑散组。失笑散组用不同浓度(1、10、20、50、100μg/mL)预处理BV2细胞2 h。对照组正常培养,不给药,不进行糖氧剥夺(OGD);模型组不给药,与失笑散组同时进行OGD,缺氧结束后再灌注24 h,收集细胞及其上清液。CCK-8法检测BV2细胞的活力;流式细胞术检测BV2细胞的凋亡;实时定量PCR和ELISA分别检测BV2细胞TNF-α、IL-6和NLRP3及相关蛋白Caspase-1、IL-1β、IL-18的mRNA和蛋白表达。结果:不同浓度失笑散并不影响BV2细胞活力。与对照组比较,模型组BV2细胞凋亡显著增多(P<0.01),而高浓度失笑散明显减少BV2细胞凋亡(P<0.05)。实时定量PCR和ELISA结果表明,与模型组比较,失笑散观察组BV2细胞TNF-α、IL-6和NLRP3炎性小体及相关蛋白Caspase-1、IL-1β、IL-18的mRNA和蛋白表达量明显减少(P<0.05)。结论:失笑散减轻OGD/R对BV2细胞的损伤及降低炎症介质的表达,机制可能与调控NLRP3炎性小体介导的炎症信号通路有关。

Objective:To study the protective effects of Shixiao Powder on hypoxia glucose deprivation reperfusion(OGD/R)of BV2 cells model and explore the potential mechanism.Methods:BV2 cells were divided into a control group,a model group and a Shixiao Powder group.In Shixiao Powder group,BV2 cells were pretreated with different concentrations(1,10,20,50,100μg/mL)for 2 h.BV2 cells in the control group were cultured normally without drug administration and OGD;BV2 cells in the model group were not treated with OGD at the same time as Shixiao Powder group.After hypoxia,reperfusion 24 h after hypoxia,and the cells and their supernatant were collected.The BV2 cell viability were examined by CCK-8;the apoptosis BV2 cells were observed by flow cytometry;the mRNA and protein expression of TNF-α,IL-6,NLRP3 inflammasome and related proteins(Caspase-1,IL-1β,IL-18)in BV2 cells were detected by real-time quantitative PCR and ELISA.Results:CCK-8 assay showed that there were no significant changes in BV2 cell viability with different concentrations of Shixiao Powder.Compared with the model group,the apoptosis ratio of BV2 cells increased significantly(P<0.01),while high concentration Shixiao Powder markedly reduced the apoptosis of BV2 cells(P<0.05).The results of qPCR and ELISA demonstrated that the mRNA and protein expression of TNF-α,IL-6,NLRP3 inflammasome and related proteins in BV2 cells evidently decreased in Shixiao Powder treatment group(P<0.05).Conclusion:Shixiao Powder can alleviate the damage of BV2 cells induced by hypoxia glucose deprivation reperfusion and the expression of NLRP3.The underlying mechanism may be related to the regulation of inflammatory signal pathway mediated by NLRP3 inflammasome.