四逆汤对溃疡性结肠炎大鼠TLR4/NF-κB信号通路相关因子及肠道菌群的影响

Effects of Sini Decoction(四逆汤)on TLR4/NF-κB Signaling Pathway Related Factors and Intestinal Flora in Rats with Ulcerative Colitis

目的 探讨四逆汤对溃疡性结肠炎大鼠Toll样受体4(Toll-like receptor 4,TLR4)/核因子-κB(nuclear factor-κB,NF-κB)信号通路相关因子及肠道菌群的影响。方法 选取健康雄性SD大鼠28只,常规饲料喂养,自由进食进水,实验开始前适应性饲养7 d。按照随机数字表法将28只大鼠分为正常组、模型组、四逆汤组、美沙拉嗪组,各7只。正常组大鼠注入氯化钠溶液,模型组、四逆汤组、美沙拉嗪组大鼠通过结肠内注入三硝基苯磺酸(trinitrobenzenesulfonic acid, TNBS)建立溃疡性结肠炎模型。造模完成后,四逆汤组大鼠采用四逆汤3.8 g/kg灌胃,美沙拉嗪组采用美沙拉嗪10 g/kg灌胃,1次/d。在给药前1 d及给药7 d后秤测量大鼠的体质量、疾病指数活动评分(disease activity index, DAI)评分,检测大鼠给药7 d后的TLR4、NF-κB、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)、白细胞介素-16(interleukin-16,IL-16)、黏蛋白2(mucin 2,MUC2)、三叶因子3(trefoil factor 3,TFF3)表达及肠道菌群。结果 给药前,正常组大鼠体质量显著高于模型组、四逆汤组、美沙拉嗪组(P<0.05);给药7 d后,四逆汤组、美沙拉嗪组大鼠体质量显著升高(P<0.05),正常组、四逆汤组、美沙拉嗪组大鼠体质量显著高于模型组(P<0.05)。给药前,正常组大鼠DAI评分显著低于模型组、四逆汤组、美沙拉嗪组(P<0.05);给药7 d后,四逆汤组、美沙拉嗪组大鼠DAI评分显著降低(P<0.05),模型组、四逆汤组、美沙拉嗪组大鼠DAI评分显著高于正常组(P<0.05),四逆汤组、美沙拉嗪组大鼠DAI评分显著低于模型组(P<0.05)。给药7 d后,正常组、四逆汤组、美沙拉嗪组大鼠TLR4、NF-κB表达均显著低于模型组(P<0.05)。给药7 d后,正常组、四逆汤组、美沙拉嗪组大鼠TNF-α、IL-10、IL-16水平均显著低于模型组(P<0.05)。给药7 d后,正常组、四逆汤组、美沙拉嗪组大鼠肠球菌、肠杆菌显著低于模型组,双歧杆菌、乳酸杆菌显著高于模型组(P<0.05)。给药7 d后,正常组、四逆汤组、美沙拉嗪组大鼠TFF3、MUC2表达显著高于模型组(P<0.05)。结论 四逆汤可用于溃疡性结肠炎的治疗,对于TLR4/NF-κB信号通路相关因子及肠道菌群均有一定调节作用。

Objective To investigate the effects of Sini Decoction(四逆汤)on Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway related factors and intestinal flora in rats with ulcerative colitis.s Methods Twenty-eight healthy male Sprague-Dawley(SD)rats were randomly fed with conventional diet and water intake.The rats were fed adaptively for 7 days before the experiment.According to the random number table method,28 rats were divided into normal group,model group,Sini Decoction group and mesalazine group,7 rats in each group.The rats in the normal group were injected with sodium chloride solution,and the rats in the model group,the Sini Decoction group and the mesalazine group were injected with trinitrobenzenesul-fonic acid(TNBS)into the colon to establish the ulcerative colitis model.After the modeling,the rats in Sini Decoction group were given Sini Decoction 3.8 g/kg by gavage,and the rats in mesalazine group were given mesalazine 10 g/kg by gavage,once a day.The body weight and disease activity index(DAI)of rats were measured 1 day before and 7 days after administration.The expressions of TLR4,NF-κB,tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),interleukin-16(IL-16),mucin 2(MUC2)and trefoil factor 3(TFF3)and intestinal flora of rats were detected 7 days after administration.s Results Before admin-istration,the body weight of the rats in the normal group was significantly higher than that in the model group,the Sini Decoction group and the mesalazine group(P<0.05).After 7 days of administration,the body weights of rats in the Sini decoction group and the mesalazine group were significantly increased(P<0.05),and the body weights of the rats in the normal group,the Sini Decoction group and the mesalazine group were significantly higher than those in the model group(P<0.05).Before administra-tion,the DAI score of the normal group was significantly lower than that of the model group,the Sini Decoction group and the me-salazine group(P<0.05).After 7 days of administration,the DAI scores of rats in the Sini Decoction group and the mesalazine group were significantly decreased(P<0.05).The DAI scores of the rats in the model group,the Sini decoction group and the mesalazine group were significantly higher than those in the normal group(P<0.05),and the DAI scores of rats in the Sini De-coction group and mesalazine group were significantly lower than those in the model group(P<0.05).After 7 days of adminis-tration,the expressions of TLR4 and NF-κB in the normal group,the Sini Decoction group and the mesalazine group were signifi-cantly lower than those in the model group(P<0.05).After 7 days of administration,the levels of TNF-α,IL-10 and IL-16 in the normal group,the Sini Decoction group and the mesalazine group were significantly lower than those in the model group(P<0.05).After 7 days of administration,Enterococcus and Enterobacter in the normal group,the Sini decoction group and the mesalazine group were significantly lower than those in the model group,and Bifidobacterium and Lactobacillus were significantly higher than those in the model group(P<0.05).After 7 days of administration,the expressions of TFF3 and MUC2 in the nor-mal group,the Sini Decoction group and the mesalazine group were significantly higher than those in the model group(P<0.05).n Conclusion Sini Decoction can be used in the treatment of ulcerative colon and has a certain regulatory effect on TLR4/NF-κB signaling pathway related factors and intestinal flora.