黄芩苷对IL-1β诱导大鼠软骨细胞凋亡和炎症反应的抑制作用及相关机制研究

Inhibitory effect and related mechanisms of baicalin on IL-1β-induced chondrocyte apoptosis and inflammatory response in rats

目的:探究黄芩苷(BA)对IL-1β诱导大鼠软骨细胞凋亡和炎症反应的影响及潜在的作用机制。方法:分离大鼠膝关节软骨细胞,CCK-8检测不同浓度BA对大鼠软骨细胞活性的影响;将细胞分为Control组、IL-1β组、BA 5μmol/L组、BA 10μmol/L组;流式细胞术检测细胞凋亡;ELISA检测细胞中NO和炎症因子含量;Western blot检测细胞中MMP-1、MMP-3、MMP-13、iNOS、环COX-2、ADAMTS-5及PI3K/AKT通路相关蛋白的表达水平。结果:BA浓度低于10μmol/L时对大鼠软骨细胞的存活率无显著影响,半数抑制浓度为73μmol/L。与Control组相比,IL-1β组细胞凋亡率显著提高;软骨细胞中NO、IL-6、PEG2和TNF-α含量均显著升高;细胞中MMP-1、MMP-3、MMP-13、iNOS、COX-2、ADAMTS-5、PI3K、p-AKT、p-p65表达水平均显著升高。与IL-1β组相比,BA 5、10μmol/L组细胞凋亡率显著降低;软骨细胞中NO、IL-6、PEG2和TNF-α含量均显著降低;细胞中MMP-1、MMP-3、MMP-13、iNOS、COX-2、ADAMTS-5、PI3K、p-AKT、p-p65表达水平均显著降低。结论:BA抑制IL-1β诱导的大鼠软骨细胞凋亡和炎症反应,其作用机制可能与抑制PI3K/AKT通路的激活相关。

Objective:To investigate effect and potential mechanism of baicalin(BA)on apoptosis and inflammatory response of rat chondrocyte induced by IL-1β.Methods:Chondrocyte of knee joint of rats were isolated,and effects of BA at different concentrations on chondrocyte activity were detected by CCK-8.Cells were divided into control group,IL-1βgroup,BA 5μmol/L group and BA 10μmol/L group.Flow cytometry was used to detect apoptosis,and ELISA was used to detect concentration of NO and inflammatory factors in cells.Expression levels of MMP-1,MMP-3,MMP-13,iNOS,COX-2,ADAMTS-5 and PI3K/AKT pathway related proteins were detected by Western blot.Results:Survival rate of rat chondrocytes was not significantly affected when BA concentration was lower than 10μmol/L,and half inhibitory concentration was 73μmol/L.Compared with control group,apoptotic rate of IL-1βgroup was significantly increased;contents of NO,IL-6,PEG2 and TNF-αin chondrocyte were significantly increased;expressions of MMP-1,MMP-3,MMP-13,iNOS,COX-2,ADAMTS-5,PI3K,p-AKT and p-p65 in chondrocyte were significantly increased.Compared with IL-1βgroup,apoptotic rate of BA 5 and 10μmol/L group was significantly decreased;contents of NO,IL-6,PEG2 and TNF-αin chondrocyte were significantly decreased;expressions of MMP-1,MMP-3,MMP-13,iNOS,COX-2,ADAMTS-5,PI3K,p-AKT and p-p65 in chondrocyte were significantly decreased.Conclusion:BA inhibits IL-1β-induced chondrocyte apoptosis and inflammation in rats,and its mechanism may be related to the inhibition of PI3K/AKT pathway activation.