附子理中丸对脾阳虚大鼠小肠上皮细胞CREB、Sirt1、UCP2表达和ATP合成的影响  

Effects of Fuzi Lizhong pills on CREB,Sirt1,UCP2 expression,and ATP synthesis in small intestinal epithelial cells of rats with spleen Yang deficiency

作　　者：孙大宇[1] 单德红[1] 刘旭东[1] 刘文俊[1] 王德山[1] SUN Dayu;SHAN Dehong;LIU Xudong;LIU Wenjun;WANG Deshan(Physiology Department of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]辽宁中医药大学生理教研室,沈阳110847

出　　处：《山东医药》2018年第41期26-29,共4页Shandong Medical Journal

基　　金：国家重点基础研究发展计划(973计划)项目(2013CB531702)

摘　　要：目的探讨附子理中丸对脾阳虚证大鼠小肠上皮细胞CREB、Sirt1、UCP2表达和ATP合成的影响。方法取SPF级SD雄性大鼠40只,按体质量分层随机分为4组,空白组、模型组、中药组、自愈组,每组10只。采用"饮食失节+劳倦"等复合因素方法复制脾气虚动物模型。在此基础上,采用"苦寒泻下"法复制脾阳虚动物模型。动物模型复制成功后,中药组给予附子理中丸1∶1水溶液灌胃治疗。在造模结束和中药治疗干预后,分别采用Western blotting法和RT-PCR法检测大鼠小肠黏膜上皮细胞中CREB、Sirt1及线粒体中UCP2蛋白和mRNA表达。采用免疫荧光法检测小肠上皮组织中ATP含量。结果与空白组相比,模型组大鼠小肠上皮组织中CREB与Sirt1蛋白和mRNA表达降低(P均<0. 01),UCP2蛋白和mRNA表达增加(P均<0. 01);与模型组相比,中药组大鼠小肠上皮组织中CREB与Sirt1蛋白和mRNA表达升高(P均<0. 05),UCP2蛋白和mRNA表达降低(P均<0. 01)。与空白组相比,模型组大鼠小肠组织中ATP含量降低(P <0. 01);与模型组相比,中药组大鼠小肠组织中ATP含量升高(P <0. 01)。结论附子理中丸可使脾阳虚大鼠小肠上皮细胞CREB、SIRT1表达升高,线粒体UCP2蛋白表达上调、肠上皮组织中ATP合成减少。Objective To investigate the effects of Fuzi Lizhong pills on CREB,Sirt1,mitochondrial UCP2 expression,and ATP synthesis in the small intestinal epithelial cells of rats with spleen Yang deficiency syndrome.Methods Forty male SD rats(SPF class)were randomly divided into four groups according to body mass:the blank group,model group,traditional Chinese medicine group,and self-recovery group,with 10 rats in each group.The animal model of spleen Qi deficiency was reproduced by diet incontinence in combination with fatigue.On this basis,the animal model of spleen Yang deficiency was reproduced by the method of diarrhea of bitter and cold.After the animal model was successfully established,the rats in the traditional Chinese medicine group were given a 1:1 aqueous solution of Fuzi Lizhong pills by gavage.Western blotting and PT-PCR were used to detect the protein and mRNA expression of CREB,Sirt1 in the intestinal epithelial cells and UCP2 in the mitochondria.The ATP production in the small intestinal epithelial cells was detected by immunofluorescence assay.Results Compared with the blank group,the protein and mRNA expression of CREB and Sirt1 in the small intestinal epithelial cells of rats decreased,while the expression of UCP2 protein and mRNA increased in the model group(all P<0.01);compared with the model group,the protein and mRNA expression levels of CREB and Sirt1 increased in the small intestinal epithelial cells of the traditional Chinese medicine group(P<0.05 or P<0.01),while the expression of UCP2 protein and mRNA significantly decreased(P<0.01).Compared with the blank group,the ATP production in the small intestinal epithelial cells of the rats in the model group significantly decreased(P<0.01).The ATP production in the traditional Chinese medicine group was significantly higher than that of the self-recovery group(P<0.01).Conclusion Fuzi Lizhong pills can increase the expression of CREB and Sirt1,up-regulate mitochondrial UCP2 protein expression and decrease the ATP synthesis in the small intestinal ep

关 键 词：脾阳虚证 附子理中丸 中药 小肠上皮细胞 信息沉默调节因子 环磷腺苷效应元件结合蛋白 解耦联蛋白2 三磷酸腺苷 大鼠 

分 类 号：R256.3[医药卫生—中医内科学]