基于AMPK/ACC信号通路研究雷公菌活性成分对羟基苯甲醛对大鼠非酒精性脂肪肝的保护作用

Protective Effect of p-Hydroxybenzaldehyde, an Active Ingredient of Nostoc Commune on Non-alcoholic Fatty Liver in Rats Based on AMPK/ACC Signaling Pathway

关键词:  对羟基苯甲醛  非酒精性脂肪肝  AMPK/ACC  氧化应激  炎症反应

Key words:  p-hydroxybenzaldehyde  non-alcoholic fatty liver disease  AMPK/ACC  oxidative stress  inflammatory response

摘要:

目的 探究雷公菌活性成分对羟基苯甲醛(p-hydroxybenzaldehyde，HD)对高脂饮食诱导的非酒精性脂肪肝(non-alcoholic fatty liver disease，NAFLD)大鼠模型的保护作用及其对AMPK/ACC信号通路的影响。方法 将48只SD大鼠随机分为正常组、模型组、水飞蓟素(120 mg·kg^-1)组和HD高、中、低剂量组(200，100，50 mg·kg^-1)组。除正常组大鼠每天饲喂标准饲料外，其余各组大鼠连续喂养高脂饲料8周以建立模型。记录大鼠体质量、肝脏系数，检测肝功能(ALT、AST)、血脂(TC、TG、HDL-C、LDL-C)、肝脏抗氧化因子(GSH-Px、SOD、MDA)、肝脏炎性因子(TNF-α、IL-6、IL-1β)等指标的变化。采用HE染色观察肝组织的病理变化，Western blotting检测AMPK/ACC信号通路相关蛋白表达。结果 与模型组相比，HD各剂量组和水飞蓟素组大鼠体质量、肝脏系数显著降低。生化指标检测结果显示，中、高剂量组HD可显著降低模型大鼠血清TC、TG、AST、ALT、LDL-C和MDA水平，升高HDL-C含量；ELISA结果表明，中、高剂量HD能明显升高NAFLD大鼠肝组织GSH-Px和SOD活性，降低MDA含量，显著降低肝脏炎性细胞因子TNF-α、IL-6和IL-1β的水平。HE结果显示，HD可明显减轻肝组织脂肪变性和炎症细胞浸润。Western blotting结果表明，HD激活AMPK/ACC信号通路，显著增加模型大鼠肝脏p-AMPK和p-ACC蛋白表达。结论 HD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。

Abstract:

OBJECTIVE To investigate whether p-hydroxybenzaldehyde (HD),the active ingredient of Nostoc commune,has hepatoprotective efficacy in a high-fat diet-induced non-alcoholic fatty liver disease (NAFLD) rat model and its effect on AMPK/ACC signaling pathway. METHODS Forty-eight SD rats were randomly divided into normal group,model group,silymarin (120 mg·kg^-1) group and HD high,medium and low dose groups (200,100 and 50 mg·kg^-1).Except for the rats in the normal group,which were fed standard chow daily,the rats in all groups were continuously fed high-fat chow for 8 weeks to establish the model.The rats’ body weight,liver coefficients,changes in liver function (ALT,AST),blood lipids (TC,TG,HDL-C,LDL-C),liver antioxidant factors (GSH-Px,SOD,MDA),liver inflammatory factors (TNF-α,IL-6,IL-1β) and other indexes were recorded.HE staining was used to observe the pathological changes of liver tissues,and Western blotting was used to detect the protein expression related to AMPK/ACC signaling pathway.RESULTS Compared with the model group,the body mass and liver coefficients of rats in each dose group of HD and silymarin group were significantly reduced.The results of biochemical indexes showed that HD in the middle and high dose groups significantly reduced serum TC,TG,AST,ALT,LDL-C and MDA levels and increased HDL-C levels in the model rats.ELISA results showed that HD in the middle and high dose groups significantly increased GSH-Px and SOD activities and decreased MDA levels in the liver tissues of NAFLD mice,and significantly reduced the levels of liver inflammatory cytokines TNF-α,IL-6 and IL-1β.HE results showed that HD could significantly reduce hepatic steatosis and inflammatory cell infiltration.Western blotting showed that HD activated AMPK/ACC signaling pathway and significantly increased p-AMPK and p-ACC protein expression in the liver of NAFLD rats.CONCLUSION HD may ameliorate NAFLD by promoting AMPK/ACC signaling pathway to improve hepatic oxidative stress,inflammatory response,and reduce lipid synthesis.