槲皮素调节AMPK/SIRT1/NF-κB通路对乙型肝炎大鼠肝组织损伤的影响  

Impact of Quercetin on Liver Tissue Damage in Hepatitis B Rats by Regulating AMPK/SIRT1/NF-κB Pathway

作　　者：刘文晶 孙世杰[2] 韩冰[2] 高杰[2] 董雅楠 LIU Wen-jing;SUN Shi-jie;HAN Bing;GAO Jie;DONG Ya-nan(Clinical College of Weifang Medical University,Weifang,Shandong,261000,China;Second Department of Hepatobiliary Surgery,Yantai Yuhuangding Hospital,Yantai,Shandong,264000,China)

机构地区：[1]潍坊医学院临床学院,山东潍坊261000 [2]烟台毓璜顶医院肝胆外二科,山东烟台264000

出　　处：《现代生物医学进展》2023年第4期624-629,共6页Progress in Modern Biomedicine

基　　金：山东省医药卫生科技发展计划项目(2018WS065)。

摘　　要：目的:探究槲皮素调节AMPK/SIRT1/NF-κB通路对乙型肝炎(HB)大鼠肝组织损伤的影响。方法:采用随机数字表法将65只Wistar大鼠分为Ctrl组、HB组、槲皮素低剂量组(槲皮素L组,50 mg/kg)、槲皮素高剂量组(槲皮素H组,200 mg/kg)及槲皮素H+AMPK抑制剂组(200 mg/kg槲皮素+10 mg/kg Compound C),每组各13只,采用尾静脉注射携带1.3拷贝HBV基因组的重组8型腺相关病毒(r AAV8-1.3HBV)法建立HB大鼠模型(Ctrl组除外);酶联免疫吸附实验(Elisa)检测血清中HB表面抗原(HBs Ag)、HB e抗原(HBeAg)、肝功能指标[谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL)]水平及肝组织中炎性因子[白介素-1β(IL-1β)、肿瘤坏死因子α(TNF-α)]水平;实时荧光定量PCR(qRT-PCR)检测血清中HBV-DNA水平;苏木素-伊红(HE)染色、曼森氏(Masson)染色观察肝组织病理改变;免疫印迹法(WB)检测肝组织中AMPK/SIRT1/NF-κB通路蛋白表达水平。结果:与Ctrl组比较,HB组血清中HBs Ag、HBeAg、HBV-DNA、ALT、AST、TBIL水平均升高(P<0.05);肝组织可见肝静脉扩张、充血,肝细胞排列紊乱、水肿、坏死,同时发生明显纤维化;肝组织中AMPK磷酸化、SIRT1蛋白水平均降低,IL-1β、TNF-α水平及核NF-κB蛋白水平均升高;经槲皮素L、槲皮素H干预后上述情况均得到改善,且槲皮素H干预改善更明显(P<0.05);而增加AMPK抑制剂干预后,槲皮素H干预的改善作用被削弱(P<0.05)。结论:槲皮素能够减轻HB大鼠肝组织损伤,保护其肝功能,其机制可能与调节AMPK/SIRT1/NF-κB通路有关。Objective:To explore the impact of quercetin regulating AMPK/SIRT1/NF-kB pathway on liver tissue damage in hepatitis B(HB)rats.Methods:65 Wistar rats were divided into Ctrl group,HB group,Quercetin low-dose group(Quercetin L group,50 mg/kg),Quercetin high-dose group(Quercetin H group,200 mg/kg)and Quercetin H combined with AMPK inhibitor group(200 mg/kg Quercetin combined with 10 mg/kg Compound C)by random number table method,with 13 rats in each group,the HB rat model(except Ctrl group)was established by tail vein injection of recombinant adeno-associated virus type 8(rAAV8-1.3HBV)carrying 1.3 copies of HBV genome.Enzyme-linked immunosorbent assay(Elisa)was used to detect serum levels of HB surface antigen(HBsAg),HB e antigen(HBeAg)and liver function indicators[alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL)]and inflammatory factors[interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)]in liver tissue.Real-time quantitative PCR(qRT-PCR)was used to detect the level of serum HBV-DNA.Hematoxylin-eosin(HE)staining and Masson(Masson)staining were used to observe the pathological changes of liver tissue.The protein expression of AMPK/SIRT1/NF-κB pathway in liver tissue was detected by Western blotting.Results:Compared with the Ctrl group,the serum levels of HBsAg,HBeAg,HBV-DNA,ALT,AST and TBIL in the HB group were increased(P<0.05).The liver tissue showed hepatic vein dilatation,congestion,disordered arrangement of liver cells,edema,necrosis,and obvious fibrosis.The phosphorylation of AMPK and the level of SIRT1 protein in liver tissue were decreased,the levels of IL-1β,TNF-αand nuclear NF-κB protein were increased.After Quercetin L and Quercetin H intervention,the above conditions were improved,and Quercetin H intervention improved more obviously(P<0.05).However,after the intervention of AMPK inhibitor,the improvement effect of Quercetin H intervention was weakened(P<0.05).Conclusion:Quercetin can alleviate liver tissue damage in HB rats,and protect liver function,and the me

关 键 词：槲皮素 AMPK/SIRT1/NF-κB通路 乙型肝炎 肝组织损伤 炎症反应 

分 类 号：R-33[医药卫生] R512.62