隔药灸对克罗恩病肠纤维化大鼠结肠成纤维细胞CTGF、FN、Smad表达的影响

Effect of Medicinal Cake Moxibustion on the Expressions of Colonic Fibroblast CTGF, FN and Smad in Crohn Disease Rats with Intestinal Fibrosis

目的观察隔药灸对克罗恩病肠纤维化大鼠结肠成纤维细胞(conlonic fibroblast,CFB)CTGF、FN蛋白和Smad3、Smad7 mRNA表达的影响,探讨隔药灸治疗克罗恩病肠纤维化的作用机制。方法 SD大鼠随机分为正常组、模型组、隔药灸组和西药组。在成功制备克罗恩病肠纤维化大鼠模型的基础上,隔药灸组取天枢穴(双)、气海穴进行治疗,西药组以美沙拉嗪灌胃治疗。各组治疗干预结束后,留取结肠组织,分离并体外培养大鼠CFB。采用免疫细胞化学法观察各组大鼠CFB的CTGF、FN蛋白表达,应用实时荧光定量PCR技术检测各组大鼠CFB的Smad3、Smad7 mRNA表达。结果与正常组比较,模型组大鼠CFB的CTGF、FN蛋白表达升高(均P<0.01);与模型组比较,隔药灸组CFB的CTGF、FN蛋白表达降低(均P<0.01)。各组大鼠比较,CFB的Smad3和Smad7 mRNA表达变化均不显著(均P>0.05)。结论隔药灸能下调克罗恩病肠纤维化大鼠肠成纤维细胞CTGF、FN的蛋白表达。

Objective To investigate the effect of medicinal cake moxibustion on the expressions of colonic fibroblast （CFB） CTGF,fibronectin （FN）,and Smad3 and Smad7 mRNA in Crohn disease rats with intestinal fibrosis and explore the mechanism of medicinal cake moxibustion treatment for intestinal fibrosis in Crohn disease.Method SD rats were randomly allocated to normal,model,medicinal cake moxibustion and Western drug groups.After a rat model of Crohn＇s disease-associated intestinal fibrosis was made successfully,the medicinal cake moxibustion group was treated by selecting points Tianshu （ST25,bilateral） and Qihai （CV6） and the Westem drug group,by oral gavage of mesalazine.After treatment was finished in every group,colonic tissues were taken and rat CBF was separated and cultivated outside the body.The expressions of CTGF and FN in CFB were examined by immunocytochemistry and the expressions of Smad3 and Smad7 mRNA in CFB,by fluorescence-based quantitative real-time PCR in every group of rats.Result The expressions of CTGF and FN in CFB increased in the model group compared with the normal group （both P＜0.01） and decreased in the medicinal cake moxibustion group compared with the model group （both P＜0.01）.There were no statistically significant differences in the expressions of Smad3 and Smad7 mRNA in CFB between various groups of rats （all P＞0.05）.Conclusion Medicinal cake moxibustion can down-regulate the expressions of colonic fibroblast CTGF and FN in Crohn disease rats with intestinal fibrosis.