六味地黄丸基于SIRT6/NF-κB信号通路对糖尿病伴肝损伤的保护作用

Protective Effect of Liuwei Dihuangwan on Diabetes Mellitus with Liver Injury Based on SIRT6/NF-κB Signaling Pathway

目的:从炎症角度探讨六味地黄丸对糖尿病伴肝损伤的保护机制。方法:选取自发性2型糖尿病动物模型db/db雄性小鼠18只,按空腹血糖(FBG)随机分模型组、六味地黄丸组(9. 75 g·kg-1·d-1,1次/d)、白藜芦醇组(42. 6 mg·kg-1·d-1,1次/d); 12只同窝野生型db/m小鼠,按FBG随机分正常组和六味地黄丸对照组(9. 75 g·kg-1·d-1,1次/d),正常组和模型组等量蒸馏水灌胃,各组给药16周后内眦静脉取血后处死小鼠,检测FBG,甘油三酯(TG)和丙氨酸氨基转移酶(ALT)等变化,苏木素-伊红(HE)染色观察肝组织病理改变,蛋白免疫印迹法(Western blot)检测肝组织沉默信息调节因子6(SIRT6),核转录因子-κB p65(NF-κB p65),单核细胞趋化蛋白-1(MCP-1),血管细胞黏附因子-1(VCAM-1)和细胞间黏附因子-1(ICAM-1)蛋白表达。结果:与正常组比较,模型组小鼠FBG,TG和ALT显著升高(P <0. 01),NF-κB p65,MCP-1,VCAM-1和ICAM-1蛋白表达显著升高(P <0. 01),SIRT6蛋白表达显著降低(P <0. 01);与模型组比较,六味地黄丸组和白藜芦醇组血FBG,TG显著降低(P <0. 01),NF-κB p65,MCP-1和VCAM-1蛋白表达明显降低(P <0. 05,P <0. 01),SIRT6蛋白表达明显升高(P <0. 05),改善小鼠肝组织的肝细胞脂肪变性及炎细胞浸润。结论:六味地黄丸对糖尿病小鼠伴肝脏损伤有保护作用,其机制与调节血脂代谢,抑制炎症反应有关。

Objective: To investigate the protective mechanism of Liuwei Dihuangwan on diabetes mellitus with liver injury in terms of inflammation. Method: The 18 db/db mice were selected from animal model of spontaneous type 2 diabetes, mice were randomly divided into model group, Liuwei Dihuangwan group( 9. 75 g·kg^-1·d^-1,once a day),resveratrol group( 42. 6 mg·kg^-1·d^-1,once a day) based on fasting bloodglucose( FBG). The 12 litter wild db/m mice were randomly divided into normal group,Liuwei Dihuangwan group( 9. 75 g·kg^-1·d^-1,once a day). The normal group and the model group were given the same amount of distilled water by gavage. The mice in each group were treated for 16 weeks. FBG,triglyceride( TG) and alanine aminotransferase( ALT) were examined. Histopathological changes were observed by liver biopsy hematoxylin-eosin( HE) staining. Silent information regulator 6( SIRT6),nuclear factor-kappaB p65( NF-κB p65),monocyte chemoattractant protein-1( MCP-1),vascular cell adhesion molecule-1( VCAM-1) and intercellular adhesion molecule-1( ICAM-1) in the liver tissue were detected by Western blot. Result: Compared with normal group,FBG,TG and ALT in model group increased significantly( P < 0. 01),protein expression levels of NF-κB p65,MCP-1,VCAM-1 and ICAM-1 in model group were significantly up-regulated( P < 0. 01),the protein expression level of SIRT6 in model group was significantly down-regulated( P < 0. 01). Compared with model group,Liuwei Dihuangwan and resveratrol could significantly reduce the levels of serum FBG and TG( P < 0. 01),the protein expression of SIRT6 was significantly increased( P < 0. 01),NF-κB p65,MCP-1 and VCAM-1 were significantly decreased( P < 0. 05, P < 0. 01), and liver steatosis and inflammatory cells infiltration were alleviated.Conclusion: Liuwei Dihuangwan can protect the diabetes mice from liver injury,and its mechanism is related to the improvement of lipid metabolism and the inhibition of inflammatory response.