益肾排毒丸调节AMPK/ACC信号通路对db/db小鼠肝损伤的保护作用研究

目的：探究益肾排毒丸（YSPDW）对db/db小鼠肝损伤的保护作用及其对脂代谢通路的影响机制。方法：C57BL/6小鼠作为空白对照组、8周龄的db/db小鼠分为非酒精性脂肪肝（non-alcoholic fatty liver disease，NAFLD）模型组、益肾排毒丸治疗组和二甲双胍阳性对照组。给药8周后检测小鼠肝脏系数、随机血糖、肝功能（丙氨转氨酶ALT、谷草转氨酶AST）、血脂（总胆固醇TC、三酰甘油TG、高密度脂蛋白胆固醇HDL-C、低密度脂蛋白胆固醇LDL-C）、肝脏脂质（TC、TG）、肝脏抗氧化因子（谷胱甘肽过氧化物酶GSH-Px、谷胱甘肽GSH、超氧化物歧化酶SOD、丙二醛MDA、过氧化氢酶CAT）、肝脏炎性因子（TNF-α、IL-6、IL-1β、MCP-1）等指标的变化。HE和PAS染色评估小鼠肝脏形态变化、脂肪变性和糖原沉积。Western blot检测脂代谢AMPK/ACC信号通路相关蛋白表达。结果：与模型组相比，益肾排毒丸给药组和二甲双胍组小鼠随机血糖和肝脏系数显著降低。生化指标检测结果显示益肾排毒丸可显著降低NAFLD模型小鼠血清AST、ALT、TC、TG水平和肝组织TC、TG、MDA水平，升高HDL-C含量，发挥肝保护作用；ELISA结果表明益肾排毒丸能明显升高NAFLD小鼠肝组织GSH-Px、GSH和SOD活性，显著降低肝脏炎性细胞因子TNF-α、IL-6、IL-1β和MCP-1的水平，表明益肾排毒丸可增加机体抗氧化能力，抑制炎症因子释放。HE和PAS结果显示益肾排毒丸可明显减轻肝组织脂肪变性和炎症细胞浸润，改善肝细胞的结构和形态完整。Western blot结果表明，益肾排毒丸能激活AMPK/ACC信号通路，显著增加模型小鼠肝脏p-AMPK和p-ACC蛋白表达。结论：益肾排毒丸可能通过促进AMPK/ACC信号通路来改善肝脏氧化应激、炎性反应，减少脂质合成，从而改善NAFLD的肝损伤。

Protective effect of Yishen Paidu Pill on liver injury by regulating AMPK/ACC signaling pathway in db/db mice

OBJECTIVE To explore the protective effect of Yishen Paidu Pill(YSPDW)on liver injury and elucidate its potential mechanism of lipid metabolism in mice.METHODS C57BL/6 mice were selected as normal control group.And 8-week-old db/db mice were divided into three groups of NAFLD model,YSPDW treatment and metformin treatment.After 8-week treatment,liver coefficient,random blood glucose level,serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),hepatic TC/TG,hepatic antioxidant factors(GSH-Px,GSH,SOD,MDA&CAT)and inflammatory factors(TNF-α,IL-6,IL-1β&MCP-1)of each group were measured.Hematoxylin-eosin(HE)and periodic acid-Schiff(PAS)stains were employed for evaluating liver morphology,steatosis and glycogen deposition.The relative expressions of AMPK/ACC pathway proteins were measured by Western blot.RESULTS As compared with model group,random blood glucose and liver coefficient of YSPDW and metformin treatment groups markedly decreased.The results of biochemical test showed that YSPDW could significantly reduce the serum levels of AST/ALT/TC/TG,lower the levels of hepatic TC/TG/MDA and boost the level of HDL-C.Enzyme-linked immunosorbent assay(ELISA)revealed that YSPDW could enhance the activities of GSH-Px/GSH/SOD and lower hepatic inflammatory cytokines of TNF-α,IL-6,IL-1βand MCP-1.It further indicated that YSPDW could enhance the antioxidant capacity and inhibit the release of inflammatory cytokines.Results of HE/PAS staining showed that YSPDW could significantly reduce liver fatty degeneration and inflammatory cell infiltration and improve the structure and morphology of liver cells.Western blot indicated that YSPDW could activate AMPK/ACC signaling pathway and significantly up-regulated the expressions of p-AMPK and p-ACC in murine liver.CONCLUSION YSPDW may improve liver oxidative stress and inflammatory response,reduce lipid synthesis through AMPK/ACC pathway and ameliorate liver damage of NAFLD.