清热活血方对溃疡性结肠炎模型小鼠β-catenin、Cdx1、Cdx2表达影响的研究

Study on the effect of Qingre Huoxue decoction on the expression of β-catenin,Cdx1 and Cdx2 in ulcerative colitis rat models

目的观察清热活血方对溃疡性结肠炎模型小鼠结肠黏膜组织中β-catenin、Cdx1、Cdx2表达的影响,探讨该方防治溃疡性结肠炎的可能机制及其效应靶点。方法将100只雄性Balb/c小鼠随机分为正常组、模型组、清热活血方低剂量组、清热活血方中剂量组及清热活血方高剂量组,除正常组外,其余组采用自由饮用3%DSS 2周法制备溃疡性结肠炎模型。自第3周起,正常组、模型组给予等剂量生理盐水,各给药组分别给予清热活血方3.3g/kg、6.6 g/kg、13.2 g/kg灌胃,均连续1周。末次给药后处死全部小鼠,应用HE染色法检测结肠黏膜组织病理学变化;应用免疫组化及荧光定量PCR技术检测结肠黏膜组织中β-catenin、Cdx1、Cdx2表达情况;应用免疫荧光染色法检测结肠黏膜组织中β-catenin核内转位变化。结果光镜下可见模型组结肠黏膜上皮脱落,大量炎性细胞浸润及溃疡形成;而各给药组结肠黏膜充血、肿胀、上皮细胞脱落及成纤维母细胞表达增加,黏膜固有层可见少许炎性细胞浸润,未见明显溃疡。与正常组比较,模型组小鼠结肠黏膜中β-catenin、Cdx1、Cdx2表达明显增加(P均<0.05);与模型组比较,各给药组β-catenin、Cdx1、Cdx2阳性表达明显减少(P均<0.05)。模型组小鼠结肠黏膜中β-catenin存在膜表达缺失、核内异常转位现象,而各给药组小鼠结肠黏膜中β-catenin表达以细胞膜为主,核内表达减少,部分可见核内异常转位。结论清热活血方可能通过促进β-catenin降解,下调Cdx1、Cdx2表达,调控肠上皮细胞增殖,修复肠黏膜屏障而发挥缓解溃疡性结肠炎的效用。

Objective It is to explore observe the effect of Qingre Huoxue decoction on the expression of β- catenin,Cdx2 and Cdx1 in the treatment of ulcerative colitis rat models,thus to explore the possible mechanism and effect target point of this treatment. Methods 100 Balb / c rats were randomly divided into normal group,blank group,low,middle and high dosage groups of Qingre Huoxue decoction. All the rats in every group except normal group were used to establish ulcerative colitis models by drinking 3% DSS for 2 weeks. Since the 3th week,the normal group and blank group was given the same dose of normal saline by intragastric administration,and every treatment group was treated with different dosage of Qingre Huoxue decoction（ 3. 3 g / kg,6. 6 g / kg,13. 2 g / kg respectively） for one week. All rats were killed at the end of the last administration. The pathological changes of colonic mucosa were detected by HE staining. The expression of Cdx1,β- catenin and Cdx2 were detected by immunohistochemical staining and PCR. The changes of β- catenin in the nucleus were detected by immunofluorescence staining. Results In the blank group,there was a large number of inflammatory cell infiltration and ulcers under the light microscope. While in the treatment group,there was a little inflammatory cell infiltration with no ulcer in it. In the blank group,the β- catenin expression on cell membrane of colon mucosa was absent with abnormal nuclear translocation.And the expression of β- catenin,Cdx2 and Cdx1 increased in the colon mucosa. In treatment group,the expression of β-catenin was mainly in the cell membrane,and the expression of β- catenin,Cdx2 and Cdx1 in the colon mucosa was decreased. The differences were statistically significant（ P〈0. 05）. Conclusion Qingre Huoxue decoction may regulate the expression of Cdx1 and Cdx2 by the degradation of β- catenin in order to promote the proliferation of intestinal epithelial cells,with repairing the intestinal mucosal barrier and relieving the symptoms of ulcerative colitis.