基于TLRs通路探讨脾胃培源灌肠方对溃疡性结肠炎大鼠干预作用

Research of Piwei Peiyuan Guanchang Decoction in Rats with Ulcerative Colitis Based on the Interfering in TLRs Signal Pathway

目的：观察脾胃培源灌肠方对溃疡性结肠炎（Ulcerative colitis,UC）大鼠血清CRP含量的影响,通过研究TLR4蛋白在溃疡性结肠炎大鼠结肠组织中的表达,探讨TLRs信号传导通路在溃疡性结肠炎发病过程中的作用以及脾胃培源灌肠方的治疗机制。方法：选用SD健康雄性大鼠120只,将大鼠随机分为：空白组、模型组、美沙拉嗪组、脾胃培源灌肠方（高、中、低剂量）组,每组20只,采用TNBS/乙醇法复制UC大鼠模型诱导溃疡性结肠炎。造模成功后,空白组和模型组分别予生理盐水3 m L/d灌肠;美沙拉嗪组予美沙拉嗪悬浊液（0.36 g/L）3 m L/d灌肠;脾胃培源灌肠方（高、中、低剂量）组分别按含不同生药浓度（0.56 g/L、0.28 g/L、0.14 g/L）的中药汤剂3 m L/d灌肠,持续15 d,末次给药24 h后处死大鼠,取结肠组织。通过HE染色观察大鼠结肠黏膜组织病理学的改变,并采用Elisa法和Western bloting法检测血清CRP含量及TLR4的蛋白表达。结果：模型组血清CRP含量及TLR4蛋白表达较空白组显著升高（P〈0.05）;脾胃培源灌肠方（高、中、低）组和美沙拉嗪组血清CRP含量及TLR4蛋白表达较模型组显著降低（P〈0.05）,但美沙拉嗪组和脾胃培源灌肠方（高、中、低剂量）组4组间采取两两比较,两组间比较差异无统计学意义（P〉0.05）。结论：脾胃培源灌肠方能明显改善模型大鼠的结肠组织损伤及有效抑制结肠组织中TLR4蛋白的表达,说明TLRs信号转导通路参与了UC发病过程,脾胃培源灌肠方治疗UC可能是通过干预该通路的表达而实现。

Objective：To observe the effect of CRP content in rats with ulcerative colitis（UC）by Piwei Peiyuan Guanchang Decoction,and to explore the treatment machanism of Piwei Peiyuan Guanchang Decoction and the effection of TLRs signal pathway by the expression of TLR4 protein in colon tissue of the rats with ulcerative colitis. Methods：Selected 120 healthy SD male rats and dividing them into four group with 20 in each group：vacuity contrast group,model group,mesalazine group and Piwei Peiyuan Guanchang Decoction（with high,medium and low dose）group. Inducing the ulcerative colitis by replicating UC rats model with TNBS/ethanol method. After the modeling,the rats in vacuity contrast group and model group were given 3 m L/d saline water enema;mesalazine group was given（0.36 g/L）3 m L/d mesalazine suspension enema;Piwei Peiyuan Guanchang Decoction（with high,medium and low dose）group respectively were given traditional Chinese medicine enema with different concentrations（0.56 g/L,0.28 g/L,0.14 g/L）3 m L/d for 15 days. And in the 16 th day killed the rats and took the specimen of blood and colon from abdominal aorta. The pathological changes of colonic mucosa in rats were observed by HE staining,and the CRP content and the expression of TLR4 protein were detected by Elisa and Western bloting methods. Results：The CRP content and TLR4 protein levels of model group were elevated significantly compared with vacurity contrast group（P〈0.05）;The levels of CRP content and TLR4 protein of mesalazine group and Piwei Peiyuan Guanchang Decoction group were significantly lower than that in model group（P〈0.05）,but there was no significant difference between these two groups（P〉0.05）by comparing each other in 4 groups. Conclusion：Piwei Peiyuan GuanchangDecoction can improve the tissue damage of the colon mucosa in rats model and can effectively inhibit the expression of TLR4 protein in the colon tissue. And indicating that TLRs signal pathway is involved in the pathwayogenesis of UC,Piwei Peiyuan Guanchang Decoction can treat UC by interfering the expression in signal pathwayway.