首乌丸对衰老大鼠肝肾组织的作用及机制研究

Effect and mechanism of Shouwuwan on liver and kidney of aging rats

目的观察首乌丸对D-半乳糖致衰大鼠肝肾组织中SIRT1、p53、p21、FoxO4、DNMT1的影响,以探讨首乌丸延缓衰老的作用及可能机制。方法大鼠随机分为正常组、模型组、首乌丸组、维E组。模型组、首乌丸组、维E组用D-半乳糖造成衰老模型,造模同时首乌丸组以首乌丸混悬液灌胃,维E组以维生素E混悬液灌胃,正常组、模型组予等量生理盐水灌胃,连续6周。HE染色观察肝肾组织病理改变,三标免疫荧光染色观测大鼠肝肾组织p53、p21、SIRT1的表达,WB检测大鼠肝肾组织FoxO4的蛋白表达量,qPCR检测大鼠肝肾组织DNMT1 mRNA的表达。结果首乌丸可明显改善衰老大鼠肝细胞的脂肪变性以及肾脏结缔组织增生、肾小管扩张等病理改变。同时提高衰老大鼠肝肾组织中SIRT1阳性率,降低p53阳性率及p21阳性率,降低FoxO4蛋白表达量,促进DNMT1mRNA的表达。结论首乌丸可能通过上调肝肾组织中SIRT1的水平,抑制p53、p21及FoxO4的表达,促进衰老细胞凋亡,增强细胞的活性,同时上调DNMT1表达以增强肝肾组织的DNA甲基化水平,从而延缓衰老。

Objective To observe the effect of Shouwuwan on SIRT1,p53,p21,FoxO4 and DNMT1 in liver and kidney tissues of D-galactose-induced aging rats,so as to explore the anti-aging effect and possible mechanism of Shouwuwan.Methods Rats were randomly divided into normal group,model group,Shouwuwan group and vitamin E group.Model group,Shouwuwan group and vitamin E group were made into aging model with D-galactose.At the same time as the model was made,Shouwuwan group was gavaged with Shouwuwan pill suspension,vitamin E group was gavaged with vitamin E suspension,the normal group and the model group were gavaged with the same amount of normal saline for 6 weeks.HE staining was used to observe the pathological changes of liver and kidney.The expression of p53,p21 and SIRT1 in rat liver and kidney was observed by three label immunofluo⁃rescence staining.Detection of FoxO4 expression in rat liver and kidney tissues by WB.Detection of DNMT1 mRNA expression in rat liver and kidney tissues by qPCR.Results Shouwuwan can obviously improve the fatty degeneration of hepatocytes,the proliferation of connective tissue and the expansion of renal tubules in aging rats.At the same time,it can increase the positive rate of SIRT1,reduce the positive rate of p53 and p21,reduce the content of FoxO4 protein,and promote the expression of DNMT1 mRNA in liver and kidney tissues of aging rats.Conclusion Shouwuwan may increase the level of SIRT1 in liver and kidney tissue,inhibit the expression of p53,p21 and FoxO4,promote the apoptosis of aging cells and enhance the activity of cells.At the same time,the expression of DNMT1 may be up-regulated to enhance the DNA methylation level of liver and kidney tissues,so as to delay aging.