香叶木素对非酒精性脂肪肝病幼鼠的保护作用

The protective effect of diosmetin on non-alcoholic fatty liver disease of young rats

目的探索香叶木素对高脂饮食诱导的非酒精性脂肪肝病幼鼠的影响。方法高脂饮食诱导非酒精性脂肪肝病大鼠模型。酶联免疫吸附实验（enzyme-linked immunosorbent assay, ELISA）检测血脂及炎症因子水平。苏木素伊红（hematoxylin-eosin, HE）染色和脱氧核糖核苷酸末端转移酶介导的缺口末端标记（terminal deoxynucleotidyl transferase-mediated dUTP nick labeling, TUNEL）染色分析肝脏组织病变及细胞凋亡。蛋白印记检测p-AMPKα1,AMPKα1,CPT-1,PPAR-α,SREBP-1c,FAS,p-P38,P38,p-AKT,AKT,p-AKT,AKT表达。结果与对照组相比,高脂饮食组大鼠TC, TG, LDL-c水平升高,HDL-c水平下降（P〈0.01）。与高脂饮食组相比,香叶木素（10, 20, 50 mg/kg）组大鼠TC, TG, LDL-c水平降低,HDL-c水平上升（P〈0.01）。香叶木素可缓解高脂饮食诱导的肝脏组织病变。高脂饮食组大鼠肝脏组织细胞凋亡高于对照组（P〈0.01）。香叶木素（10, 20, 50 mg/kg）组大鼠肝脏组织细胞凋亡低于高脂饮食组（P〈0.01）。高脂饮食组大鼠p-AMPKα1,CPT-1和PPAR-α表达低于对照组（P〈0.01）。与高脂饮食组相比,香叶木素（20, 50 mg/kg）组大鼠p-AMPKα1,CPT-1和PPAR-α表达升高（P〈0.01）。高脂饮食组大鼠SREBP-1c和FAS表达高于对照组（P〈0.01）。与高脂饮食组相比,香叶木素（10, 20, 50 mg/kg）组大鼠SREBP-1c和FAS表达下降（P〈0.01）。而且,高脂饮食组大鼠IL-6, IL-1β和TNF-α水平高于对照组（P〈0.01）。与高脂饮食组相比,香叶木素（20, 50 mg/kg）组大鼠IL-6, IL-1β和TNF-α水平下降（P〈0.01）。另外,香叶木素可逆转compound C对AMPK通路的抑制作用。结论香叶木素激活AMPK通路减轻高脂饮食诱导的非酒精性脂肪肝病幼鼠脂质代谢异常,肝脏组织病变,细胞凋亡及炎症反应。

Objective To explore the effect of diosmetin on non-alcoholic fatty liver disease of young rats induced by high-fat diet. Methods The rats models of non-alcoholic fatty liver disease were induced by high-fat diet. The levels of serum lipid and inflammatory factors were detected by enzyme-linked immunosorbent assay（ELISA）. The pathological changes and apoptosis of liver tissues were observed by hematoxylin-eosin（HE） staining and terminal deoxynucleotidyl transferase-mediated dUTP nick labeling（TUNEL）. The levels of p-AMPKα1, AMPKα1, CPT-1, PPAR-α, SREBP-1 c, FAS, p-P38, P38, p-AKT, AKT,p-AKT, AKT were tested by western blot. Results Compared with control group, the levels of TC, TG,LDL-c in high fat diet group were increased with declined levels of HDL-c（P〈0.01）. Compared with high fat diet group, the levels of TC, TG, LDL-c in diosmetin（10, 20, 50 mg/kg） groups were reduced with enhanced levels of HDL-c（P〈0.01）. The high fat diet-induced pathological changes of liver tissues were reliefed by diosmetin. The apoptosis of liver tissues in high fat diet group was higher than that of control group（P〈0.01）.The apoptosis of liver tissues in diosmetin（10, 20, 50 mg/kg） groups was lower than that of high fat diet group（P〈0.01）. The levels of p-AMPKα1, CPT-1 and PPAR-α in high fat diet group were lower than that of control group（P〈0.01）. Compared with high fat diet group, the levels of p-AMPKα1, CPT-1 and PPAR-α in diosmetin（20, 50 mg/kg） groups were elevated（P〈0.01）. The levels of SREBP-1 c and FAS in high fat diet group were higher than that of control group（P〈0.01）. Compared with high fat diet group, the levels of SREBP-1 c and FAS in diosmetin（10, 20, 50 mg/kg） groups were decreased（P〈0.01）. Moreover, the levels of IL-6, IL-1β and TNF-α in high fat diet group were higher than that of control group（P〈0.01）. Compared with high fat diet group, the levels of IL-6, IL-1β and TNF-α in diosmetin（20, 50 mg/kg） groups were reduced（P〈0.01）. In addition, the inhibitory effect of compound C on AMPK pathway was reversed by diosmetin.Conclusion Diosmetin alleviates the abnormal lipid metabolism,pathological changes of liver tissues,apoptosis and inflammatory response in high-fat diet-induced non-alcoholic fatty liver disease rats by activating AMPK pathway.