清热活血方调控Rho激酶信号通路介导溃疡性结肠炎E-cadherin及TGF-β的影响  

The effect of Qingre Huoxue decoction mediated by Rho kinase signaling pathway on the expression of E-cadherin and TGF-β in ulcerative colitis

作　　者：李华燕[1] 陈玉[2] 李斌[1] 黄建斌[1] 张涛[2] 

机构地区：[1]江门市第二人民医院内科,广东江门529050 [2]广西中医药大学附属瑞康医院消化科,广西南宁530011

出　　处：《中国中西医结合消化杂志》2016年第6期414-417,421,共5页Chinese Journal of Integrated Traditional and Western Medicine on Digestion

基　　金：广西自然基金(No:2013GXNSFAA019116);江门市科技局项目(No:12A078)

摘　　要：[目的]基于检测Rho激酶信号通路介导溃疡性结肠炎(UC)E-cadherin及TGF-β中相关指标变化,探讨清热活血方介导上皮-间质转化,防治UC及其相关癌变的机制。[方法]100只Balb-c雄性小鼠按体重随机分为5组,即正常组、模型组、治疗组(低剂量、中剂量及高剂量)。采用3%DSS自由饮用2周法制备UC模型,自第3周起,分组给药,模型组予等剂量生理盐水灌胃,治疗组分别予不同剂量清热活血方灌胃,连续1周。末次给药后处死全部小鼠。应用光镜及透射电镜观察UC小鼠组织结构及超微结构变化、应用免疫应迹及荧光定量PCR技术检测Rho-GTP、E-cadherin、TGF-β等指标变化。[结果]基于结肠黏膜组织结构和超微结构分析:模型组无论光镜或电镜下均提示结肠黏膜上皮细胞脱落,细胞坏死明显,腺体分泌减少,黏膜层及黏膜下层大量炎性细胞细胞浸润,可见溃疡形成;而治疗组光镜下见肠壁黏膜及黏膜下层轻度充血水肿,少量炎细胞浸润,及成纤维母细胞增加,电镜下见上皮细胞体积明显变小,核染色质边集、核固缩,核膜断裂,核碎裂,细胞器明显扩张肿胀、碎裂,凋亡小体形成,并以凋亡为主。模型组结肠黏膜Rho激酶表达上升,E-cadherin及TGF-β表达下降,而治疗组结肠黏膜肠Rho激酶下降,而E-cadherin及TGF-β表达上升,与模型组比较,差异有统计学意义(P<0.05)。[结论]清热活血方可能通过下调Rho激酶表达,上调E-cadherin及TGF-β表达,诱导肠上皮细胞凋亡,改善肠道炎症,促进黏膜上皮细胞修复及溃疡愈合,介导上皮-间质转化,发挥缓解UC的效用。[Objective]We aim to investigate the mechanism of Qingre Huoxue decoction treating the ulcerative colitis associated carcinogenesis with epithelial mesenchymal by the observation of E-cadherin and TGF-βthrough the Rho kinase signaling pathway.[Methods]100Balb-c rats were randomly divided into normal group,blank group,low dosage group,medium dosage group and high dosage group.The ulcerative colitis models were prepared by 3% DSS for 2weeks.From the third week,the blank group was treated with equal dose of normal saline,and the intervention groups were treated with different doses of Qingre Huoxue decoction for 1week.After the last administration,all the rats were killed.The changes of tissue structure and ultrastructural in ulcerative colitis mucosa were observed by light microscope and transmission electron microscope.The changes of Rho-GTP,E-cadherin and TGF-βwere detected by Western blotting and PCR.[Results]The blank group whether light or electron microscopy showed that colonic mucosal epithelial cells were shedding with some necrotic cells and decreased glandular secretion in it.There was al-so a large number of inflammatory cell infiltrating in the colonic mucosa tissue with visible ulceration.In the intervention group,there was a little amount of inflammatory cell infiltrating the intestinal mucosa with hyperemia and edema under the light microscopy.we also can see the epithelial cells＇volume became small,the nuclear chromatin became marginated with nuclear pyknosis and nuclear membrane rupture which lead to apoptosis under the electron microscope.In the blank group,the expression of Rho kinase was increased,while the E-cadherin and TGF-βwere decreased.But in the intervention group,the Rho protein kinase decreased,while the expression of E-cadherin and TGF-βincreased significantly,compared with the blank group（P0.05）.[Conclusion]The Qingre Huoxue decoction may regulate the expression of the Rho protein kinase,E-cadherin and TGF-βin order to induce the apoptosis and inflammation of intestinal epi

关 键 词：清热活血方 Rho激酶信号通路 溃疡性结肠炎 E-钙黏附素 转化生长因子β 

分 类 号：R574.62[医药卫生—消化系统]