青黛对溃疡性结肠炎体外炎症模型的抗炎作用及机制

Anti-inflammatory effects and mechanism of Qingdai on experimental ulcerative colitis in vitro

目的:研究青黛对溃疡性结肠炎体外炎症模型的抗炎作用,探讨青黛抗炎的作用机制。方法:人结肠上皮细胞接种于96孔板中,根据给药血清的不同分为正常组、模型组、柳氮磺吡啶(SASP)组、青黛组,除正常组外均用脂多糖诱导造模24h,分别加入相应的含药血清。干预24h后以ELISA法检测细胞中TNF-α、IL-12的含量;Western-blot法检测PI3K、AKT、p-AKT蛋白的表达;QPCR法检测PI3K、AKT、TNF-α、IL-12 mRNA的表达量。结果:模型组血清中TNF-α、IL-12含量显著高于正常组(P<0.01),SASP组和青黛组较模型组含量显著降低(P<0.01)。与模型组比较,青黛组PI3K、p-AKT蛋白的表达显著降低(P<0.01),且p-AKT蛋白表达在青黛组和SASP组间差异无统计学意义。与模型组比较,PI3K、TNF-α、IL-12 mRNA在SASP组和青黛组的表达显著下调,差异有统计学意义(P<0.01)。结论:青黛对溃疡性结肠炎体外炎症模型具有抗炎作用,其抗炎机制可能与其下调PI3K、p-AKT表达有关。

Objective:To study the anti-inflammatory effects of Qingdai on normal human colonic epithelial cells and explore the mechanism of Qingdai’s anti-inflammatory effects.Methods:HCoEpiC cells were inoculated into 96-well plates,and were divided into normal group,model group,SASP group,and Qingdai group according to different sera administered.Except the normal group,other groups were induced by LPS for 24 h.After 24 h-intervention,the content of TNF-αand IL-12 in cells serum were detected by ELISA.PI3 K,AKT and p-AKT protein were detected by Western-blot;The expression of PI3 K,AKT,TNF-αand IL-12 mRNA were detected by QPCR.Results:The serum concentration of TNF-αand IL-12 in the model group were significantly higher than those in the normal group(P<0.01),and the concentration in the SASP and Qingdai groups were significantly lower than those in the model group(P<0.01).Compared with the model group,the PI3 K and p-AKT protein expressions in the Qingdai group were significantly reduced(P<0.01),and the p-AKT protein expression was not statistically different between the Qingdai group and the SASP group.Compared with the model group,PI3 K,TNF-α,IL-12 mRNA expression of SASP group and Qingdai group was significantly down-regulated(P<0.01).Conclusion:Qingdai has anti-inflammatory effect on experimental ulcerative colitis in vitro,and the potential mechanism probably was related to its down-regulation of PI3 K and p-AKT expression.