化浊解毒抑溃煎对溃疡性结肠炎大鼠Th1/Th2平衡、应激反应及肠黏膜的影响

Effects of Huazhuo Jiedu Yikui decoction on Th1/Th2 balance,stress response and intestinal mucosa in rats with ulcerative colitis

目的探讨化浊解毒抑溃煎对溃疡性结肠炎大鼠Th1/Th2平衡、应激反应及肠黏膜的影响。方法SPF级别Wistar大鼠随机分为对照组、模型组、化浊解毒抑溃煎低、中、高剂量(1.25、2.5、5 kg·L^(-1))组及阳性药美沙拉嗪(0.05 kg·L^(-1))组,共6组。除对照组外,其余各组大鼠均建立溃疡性结肠炎动物模型,分别灌胃给予相应药物。ELISA法检测TNF-α、IL-8、IL-13、IL-10、IFN-γ及IL-4水平;相关试剂盒检测SOD、MDA及GSH-Px水平;HE染色观察肠道组织病理形态并计算评分;TUNEL法检测肠道上皮细胞凋亡率;免疫印迹检测肠组织NF-B、TLR4、MyD88、Bcl-2及Bax蛋白水平。结果与模型组相比,化浊解毒抑溃煎组TNF-α、IL-8、IL-13、MDA、IFN-γ、病理评分、肠上皮细胞凋亡率、NF-B、TLR4、MyD88及Bax水平降低(P<0.05),IL-10、IL-4、SOD、GSH-Px、Bcl-2水平升高(P<0.05)。结论化浊解毒抑溃煎可明显抑制溃疡性结肠炎大鼠氧化应激及炎症反应,促使Th1/Th2平衡,并发挥肠黏膜保护作用,其作用机制与抑制NF-B/TLR4/MyD88信号通路相关。

Aim To investigate the effect of Huazhuo Jiedu Yikui decoction(HJYD)on Th1/Th2 balance,stress response and intestinal mucosal in rats with ulcerative colitis.Methods Wistar rats of SPF grade were randomly divided into six groups:control group,model group,HJYD low,medium and high dose groups(1.25,2.5,5 kg·L^(-1)),and positive mesalazine group(0.05 kg·L^(-1)).Except for the control group,the rats in other groups were established into ulcerative colitis animal models and were intragastrically administered with drug.TNF-α,IL-8,IL-13,IL-10,IFN-γand IL-4 were detected by ELISA.The levels of SOD,MDA and GSH-Px were detected by related kits.The pathological morphology of intestinal tissue was observed using HE staining and the score was calculate.The apoptosis rate of intestinal epithelial cells was detected by TUNEL method.The protein levels of NF-κB,TLR4,MyD88,Bcl-2 and Bax in intestinal tissue were detected by Western blot.Results Compared with the model group,the levels of TNF-α,IL-8,IL-13,MDA,IFN-γ,pathological score,intestinal epithelial cell apoptosis rate,NF-κB,TLR4,MyD88 and Bax in the HJYD groups were reduced(P<0.05),and the levels of IL-10,IL-4,SOD,GSH-Px and Bcl-2 were raised(P<0.05).Conclusions HJYD can significantly inhibit oxidative stress and inflammatory response in rats with ulcerative colitis,promote Th1/Th2 balance,and play a protective role in intestinal mucosa.The mechanism is related to the inhibition of NF-κB/TLR4/MyD88 signaling pathway.