淫羊藿总黄酮对衰老大鼠小肠干细胞及其微环境的调节作用

The Effect of Total Flavones of Epimedium on Small Intestinal Stem Cells and Microenvironment in Aging Rats

目的:探讨淫羊藿总黄酮对衰老大鼠小肠干细胞及其微环境变化的调节作用。方法:SD大鼠按其不同发育阶段分为6月龄成年组、24月龄衰老组及淫羊藿总黄酮低、中、高剂量组。采用常规HE染色,高碘酸-Schiff试剂(PAS)染色,免疫组化(荧光),Western bolt检测各组别小肠干细胞数量,增殖分化功能及其微环境相关信号的变化。结果:与成年组比较,衰老组小肠绒毛长度、隐窝深度、V/C值、杯状细胞数量及PCNA表达量降低,小肠干细胞数量显著减少,干细胞微环境Wnt/β-catenin和BMP信号均呈下调趋势。给予淫羊藿总黄酮后,与衰老组比较,小肠隐窝干细胞数量有所增加,增殖分化功能有所提升,表现为绒毛长度、隐窝深度、V/C值、杯状细胞数量及PCNA表达量增加,小肠干细胞微环境Wnt/β-catenin和BMP信号均呈上调趋势。结论:衰老过程中,小肠干细胞数量减少,增殖分化功能降低,干细胞微环境信号通路的蛋白表达减少;淫羊藿总黄酮可改善小肠干细胞的增殖分化功能,上调干细胞微环境Wnt/β-catenin和BMP信号通路相关蛋白表达。

Objective：To explore the effect of total flavones of Epimedium（TFE）on small intestinal stem cells and microenviron- ment in aging rats. Methods：Male SD rats were divided into 6 months old group （adult）, 24 months old group （aging）, three doses of TFE （ 10,20,60 mg/kg） groups. The conventional HE staining, periodic acid -Schiff＇ s （PAS） staining techniques, immunohistochemistry （IHC）, Immunofluorescence （IF）and Western blot were used for the detection of the number of intestinal stem cells （ISCs）, the prolif- eration and differentiation of intestinal stem cells and changes of related signals of intestinal stem cells microenvironment. Results ： Com- pared with adult group,the villus height, cyrpt depth, the ratio of villus height/crypt depth （V/C）, the number of globle cells, and the expresstion of PCNA, and the number of intestinal stem cells of aging group were decreased （P 〈 0. 05 ）. Wnt/β-catenin and BMP sig- nals were downregulated. Compared with aging group, the number of intestinal stem cells were increased, and the villus height, cyrpt depth, the ratio of V/C, the number of globle cells, and the expression of PCNA were raised in TFE treatment groups. TFE upregulated the expression of the Wnt/β-catenin and BMP signals. Conclusion：The proliferation and differentiation of intestinal stem ceils are dre- pressed and the Wnt/β-catenin signals were reduced in aging rats. TFE not only improves the proliferation and differentiation of intesti- nal stem cells, but also upregulates the positive effect of microenvironment to intestinal stem ceils.