痛泻要方对肝郁脾虚型溃疡性结肠炎大鼠肠黏膜的保护作用

Protective effects of Tongxie Yaofang on intestinal mucosa in rats with ulcerative colitis due to Liver Depression and Spleen Deficiency Pattern

目的探讨痛泻要方对肝郁脾虚型溃疡性结肠炎大鼠肠黏膜的保护作用。方法大鼠随机分为空白组、模型组、美沙拉嗪组(0.4g/kg)和痛泻要方高、中、低剂量组(20.8、10.4、5.2g/kg),采取DNBS/乙醇溶液灌肠法+束缚应激法+饮食失节法建立动物模型。ELISA法检测血清TNF-α、IL-8水平,HE染色观察结肠病理变化,RT-qPCR、Westernblot和免疫组化法检测结肠组织TLR4、MyD88、NF-κB p65mRNA及蛋白表达。结果与模型组比较,美沙拉嗪组与痛泻要方各剂量组大鼠血清TNF-α、IL-8水平降低(P<0.05,P<0.01),结肠组织MyD88、NF-κB p65mRNA及蛋白表达降低(P<0.05,P<0.01),美沙拉嗪组与痛泻要方高、中剂量大鼠结肠组织TLR4 mRNA及蛋白表达降低(P<0.05,P<0.01),痛泻要方低剂量组大鼠结肠组织TLR4蛋白表达降低(P<0.05)。结论痛泻要方可能通过调控大鼠TLR4/MyD88/NF-κB信号通路,抑制机体炎症反应,修复肠黏膜屏障损伤。

AIM To explore the protective effects of Tongxie Yaofang on the intestinal mucosa of rats with ulcerative colitis(UC)due to Liver Depression and Spleen Deficiency.METHODS The rats were randomly divided into the blank group,the model group,the mesalazine group(0.4 g/kg),the high-dose,medium-dose and low-dose Tongxie Yaofang groups(20.8,10.4,5.2 g/kg).The rat models were induced by DNBS/ethanol solution enema,restraint stress and inappropriate diet.All rats had their serum levels of IL-8 and TNF-αdetected by ELISA;their colonic pathological changes observed by HE staining;and their colonic TLR4,MyD88 and NF-κB p65 mRNA and protein expressions detected by RT-qPCR,Western blot and immunohistochemistry method.RESULTS Compared with the model group,all theother groups intervened with the medical agents displayed lower serum levels of TNF-a and IL-8(P<0.05,P<0.01);decreased colonic expressions of MyD88 and NF-κB p65 mRNA and protein(P<0.05,P<0.01).The mesalazine group and the high-dose and medium-dose Tongxie Yaofang groups shared decreased colonic expression of TLR4 mRNA and protein(P<0.05,P<0.01);and the low-dose Tongxie Yaofang group demonstrated decreased colonic TLR4 protein expression(P<0.05).CONCLUSION Tongxie Yaofang may inhibit the inflammatory reaction of the rats and repair their intestinal mucosal barrier damage via TLR4/MyD88/NF-κB signal pathway.