消脂方调控AMPK/ACC信号通路对DIO小鼠减重降脂作用研究

Study on the Effect of Xiaozhi Decoction(消脂方)Regulating AMPK/ACC Signal Pathway on Weight and Lipid Reduction in DIO Mice

目的通过对高脂饮食诱导肥胖(Diet-induced obesity,DIO)模型小鼠的干预,探讨消脂方减重降脂的作用机制。方法 C57BL/6J雄性小鼠高脂饮食造模成功后的DIO模型小鼠,分为肥胖模型组(蒸馏水,n=6)、低剂量实验组(消脂方汤剂,药物浓度0.458 3 g/mL,n=6)、中剂量(等效剂量)实验组(消脂方汤剂,药物浓度0.916 5 g/mL,n=6),高剂量实验组(消脂方汤剂,药物浓度1.833 0 g/mL,n=6)、阳性对照组(荷丹片,药物浓度0.028 5 g/mL,n=6),并设未经高脂造模的正常对照组(蒸馏水,n=6),干预4周。观察各组干预后Lee’s指数、BF、肝脏指数、血脂(FFA、LDL、HDL、TG、TC)、肝、脂肪组织病理学、脂肪细胞横截面面积、相关蛋白(ACC、AMPK)及其磷酸化水平的差异。结果与模型组比较,消脂方呈剂量依赖性降低小鼠Lee’s指数、BF、肝脏指数、脂肪细胞面积、FFA、LDL、TC、TG;升高HDL、p-ACC蛋白、p-AMPK蛋白表达水平及两者磷酸化比值。与荷丹片组比较,消脂方中剂量组Lee’s指数、横截面面积、FFA更低;高剂量组Lee’s指数、BF、脂肪细胞面积、FFA、LDL、TG更低,HDL更高。消脂方各剂量组之间以高剂量组作用最佳,低剂量组最次。差异均有统计学意义(P<0.05或P<0.01)。肝HE、油红O及脂肪HE染色结果显示:各干预组小鼠与模型组比较均有改善,消脂方高剂量组与正常组最为趋近。结论消脂方对DIO模型小鼠的减重降脂作用可能通过上调ACC、AMPK磷酸化水平调节AMPK/ACC信号通路起效。

Objective Through the intervention of DIO model mice,to explore the mechanism of Xiaozhi Decoction (消脂方) in reducing weight and fat. Methods C57BL/6J male mice were feeded with high fat diet to induce DIO model mice. The DIO model mice were divided into obesity model group(distilled water,n=6),low dose experimental group(Xiaozhi Decoction,drug concentration 0.458 3 g/mL,n=6),medium dose experimental group(Xiaozhi Decoction,equivalent dose,drug concentration 0.916 5 g/mL,n=6),high dose experimental group(Xiaozhi Decoction,drug concentration 1.833 0 g/mL,n=6),positive control group [Hedan Tablet(荷丹片),drug concentration 0.028 5 g/mL,n=6] and normal group without high fat modeling(distilled water,n=6). They were all treated for 4 weeks. The differences of Lee’s index,body fat ratio,liver index,blood lipid(FFA,LDL,HDL,TG,TC),liver,fatty histopathology,fatty cell area,related proteins(ACC,AMPK)and their phosphorylation levels were observed. Results Compared with the model group,Xiaozhi Decoction decreased Lee’s index,BF,liver index,fat cell cross-sectional area,FFA,LDL,TC,TG,increased the expression level of HDL,p-ACC protein,p-AMPK protein and their phosphorylation ratio in a dose-dependent manner. Compared with Hedan tablet group,Lee’s index,crosssectional area and FFA were lower in the medium dose group;Lee’s index,BF,cross-sectional area,FFA,LDL,TG were lower and HDL were higher in high dose group. The high dose group had the best effect,the low dose group the worst. The difference was statistically significant(P<0.05 or P<0.01). The results of liver HE,oil red O and fatty HE staining showed that:compared with the model group,the mice in each intervention group were improved,and the mice in the high dose group of Xiaozhi Decoction were the most similar to those in the normal group. Conclusion The effect of Xiaozhi Decoction on weight loss and lipid reduction in DIO model mice may be through the regulation of AMPK/ACC signal pathway by up regulating the levels of ACC and AMPK phosphorylation.