降脂脉安方对非酒精性脂肪性肝病大鼠脂质代谢和炎症反应的影响

Effect of Jiangzhi Mai'an Fang(降脂脉安方)on Lipid Metabolism and Inflammation in NAFLD Rats

目的:探讨降脂脉安方(JZMA)对高脂饮食诱导的非酒精性脂肪性肝病(NAFLD)大鼠的影响及可能的作用机制。方法:53只大鼠中随机取8只作为对照组,其余大鼠为造模组。对照组大鼠予普通饲料喂养,造模组大鼠予高脂饲料喂养进行造模。造模成功后随机取40只NAFLD大鼠,随机分为模型组、易善复组(0.12 g/kg)、降脂脉安方低剂量组(4.95 g/kg)、降脂脉安方中剂量组(9.90 g/kg)、降脂脉安方高剂量组(19.80 g/kg),每组8只。末次给药后,HE染色观察各组肝脏形态学及病理学变化,检测血清甘油三酯(TG)、胆固醇(CHOL)、谷丙转氨酶(ALT)、谷草转氨酶(AST)水平及肝组织TG、CHOL水平,酶联免疫吸附法(ELISA)检测肝组织肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、游离脂肪酸(FFA)、C反应蛋白(CRP)水平。实时荧光定量PCR(qRT-PCR)技术检测肝组织SREBP-1c mRNA、FASN mRNA、SIRT1 mRNA、PTGS2/COX-2 mRNA表达水平,Western blotting法检测肝组织p-AMPK、FASN、SIRT1、PTGS2/COX-2及p-NF-κBp65蛋白表达水平。结果:降脂脉安方中、高剂量组能减轻NAFLD大鼠肝脏脂质沉积,降低血清TG、CHOL、AST、ALT水平,降低肝组织TNF-α、IL-6、CRP水平。降脂脉安方中、高剂量组能提高NAFLD大鼠肝脏p-AMPK的蛋白表达水平,降低SREBP-1c、FASN及p-NF-κB-p65蛋白表达水平。结论:降脂脉安方对NAFLD大鼠血脂水平具有一定的调节作用,对高脂血症引起的肝功能损伤具有改善作用,且可能通过激活AMPK通路调控肝脏脂质代谢及抑制NF-κB活性减轻肝细胞炎症反应,缓解NAFLD进程。

Objective:To explore the effect of Jiangzhi Mai'an Fang(JZMA)on non-alcoholic fatty liver disease(NAFLD)rats induced by high fat diet and its possible mechanism.Methods:Totally 8 rats were randomly selected from 53 rats as the control group,while the remaining rats were used as the modeling group.Rats were given regular fed in control group,while high fat feed for modeling in modeling group.After successful modeling,40 NAFLD rats were randomly selected and divided into model group,Yisanfu(易善复)group(0.12 g/kg),JZMA low dose(4.95 g/kg)group,JZMA medium dose(9.90 g/kg)group and JZMA high dose(19.80 g/kg)group,with 8 rats in each group.After the last administration,the morphological and pathological changes of liver were observed by HE staining,and the levels of serum triglyceride(TG),cholesterol(CHOL),glutamic pyruvic transaminase(ALT),glutamic oxaloacetic transaminase(AST)and the levels of TG and CHOL in liver tissue were detected.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),free fatty acid(FFA)and C-reactive protein(CRP)in liver tissue were measured by enzyme-linked immunosorbent assay(ELISA).The levels of SREBP-1c mRNA,FASN mRNA,SIRT1 mRNA and PTCS2/COX-2 mRNA in liver tissue were detected by real-time fluorescence quantitative PCR(qRT-PCR),and the protein levels of p-AMPK,FASN,SIRT1,PTGS2/COX-2 and p-NF-k Bp65 in liver tissue were detected by Western blotting.Results:The JZMA medium and high dose groups could reduce the lipid deposition in the liver of NAFLD rats,significantly decrease the levels of serum TG,CHOL,AST and ALT,and decrease the levels of TNF-α,IL-6 and CRP in liver tissue.In addition,the JZMA medium and high dose groups could significantly increase the protein expression level of p-AMPK in the liver of NAFLD rats,and reduce the expression levels of SREBP-1c,FASN and p-NF-kB-p65.Conclusion:Jiangzhi Mai'an Fang can regulate the blood lipid level of NAFLD rats and improve the liver function damage caused by hyperlipidemia,and it may reduce hepatocyte inflammation and alleviate the process of NAFLD by activating AMPK pathway to regulate liver lipid metabolism and inhibiting NF-k B activity.