OBJECTIVE To observe possible mechanism of the Qingchang Huashi Fang's therapeutic effect on Ulcerative Co-litis(UC), through observing the expression level of tumor necrosis factor-α(TNF-α) and claudin-1 of mucosal tissue in the experimental colitis rats. METHODS Rats successfully modeled with TNBS/anhydrous alcohol were divided into blank, model, Qichang Huashi Fang(12.8 g crude drug/kg/d) and Mesalazine group(0.67 g/kg/d). Single administration for 10 days successively , all groups of rats were executed to collect colon tissues cut out for related inspections. Using ELISA method to measure the TNF-α concentration of the colon tissues. The expression of claudin-1 protein in colonic epithelium was checked up with im-munohistochemical method and relative expression of claudin-1 mRNA with real-time PCR method. RESULTS The TNF-α (pg/mL) concentration of model group and Qichang Huashi Fang group are respectively 99.40±32.37 and 55.07±12.80(P＜ 0.01);And claudin-1 gene relative expression levels are respectively 2.18±0.78 and 3. 94±0.91 (P＜0.01); Qichang Huashi decoction can decrease TNF-aconcentration and increase claudin-1 mRNA relative expression significantly(P＜0.01), thus reducing claudin-1 protein damage(P＜0.05). The curative effect between Qichang Huashi Fang and Mesalazine group has no significant difference. CONCLUSION Qichang Huashi Fang can decrease TNF-α concentration and protect tight junction protein claudin-1. That is the one of the mechanisms of Qichang Huashi Fang to treat UC.%目的 观察清肠化湿方对实验性大鼠结肠炎结肠组织肿瘤坏死因子-α(TNF-α),紧密连接跨膜蛋白claudin-1表达水平的影响,以探讨其治疗溃疡性结肠炎(UC)的可能机制.方法 使用TNBS/无水乙醇造模成功后,分为空白组、模型组、清肠化湿方组12.8 g/(kg·d)、美沙拉嗪组(0.67 g/(kg·d).一次性ig给药,连续10d后,处死大鼠,留取结肠组织；ELISA法检测结肠组织TNF-a水平,免疫组化法观察结肠上皮claudin-1蛋白的表达情况,实时荧光定量PCR法检测claudin-1 mRNA相对表达水平.结果 清肠化湿方可以降低大鼠结肠炎模型结肠组织TNF-α水平[模型组,清肠化湿组分别为(99.40±32.37),(55.07士12.80)pg/mL],(P＜0.01),提高claudin-1 mRNA相对表达水平[模型组,清肠化湿组分别为(2.18±0.78),(3.94±0.91)](P＜0.01),减轻对claudin-1蛋白的损伤(P＜0.05),清肠化湿方组与美沙拉嗪组疗效无明显差异.结论 清肠化湿方降低结肠组织TNF-α水平,保护紧密连接蛋白claudin-1,是其治疗UC的机制之一.
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