摘要
目的:研究当归芍药散对非酒精性脂肪肝(NAFLD)大鼠的防治作用及其机制。方法:60只SD雄性大鼠随机分为正常对照组、模型对照组、易善复0.144 g/kg组以及当归芍药散2.44、4.88、9.76 g/kg组。通过喂饲高脂饲料复制NAFLD大鼠模型,造模同时给予相应药物治疗,1次/d,连续8 w,检测血清中胆固醇(TC)、三酰甘油(TG)、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)、丙二醛(MDA)以及肝组织中TC、TG、游离脂肪酸(FFA)、SOD、GSH和MDA的含量或活性变化;苏木素-伊红(HE)染色和油红O染色法观察肝脏病理形态学变化;荧光实时定量PCR(qRT-PCR)和蛋白免疫印迹法(Western blot)法检测肝脏中Kelch样环氧氯丙烷相关蛋白1(Keap1)、核因子E2相关因子(Nrf2)、血红素加氧酶1(HO-1)、谷胱氨酸连接酶催化亚基(GCLC)和谷胱氨酸连接酶调节亚基(GCLM)的mRNA和蛋白表达情况。结果:与正常对照组相比,模型对照组大鼠血清中TC、TG、ALT、AST、MDA和肝组织中TC、TG、FFA、MDA的含量或活性均显著升高(P<0.01),血清和肝组织中SOD、GSH的活性以及肝组织中Keap1、Nrf2、Ho1、Gclc和Gclm的mRNA和蛋白表达均显著降低(P<0.01),肝细胞内有大量的脂滴空泡积聚和较多的气球样病变;与模型对照组比,当归芍药散各剂量组均能明显降低血清中TC、TG、ALT、AST、MDA和肝组织中TC、TG、FFA、MDA的含量或活性(P<0.05或P<0.01),升高血清和肝组织中SOD、GSH的活性以及肝脏中Keap1、Nrf2、Ho1、Gclc和Gclm的mRNA和蛋白表达(P<0.05或P<0.01),减轻肝脏脂肪变性程度。结论:当归芍药散可能通过调节Keap1/Nrf2信号通路,抵抗氧化损伤来防治NAFLD。
Objective:To explore the efficacy and mechanism of Danggui Shaoyao San in the treatment of nonalcoholic fatty liver disease(NAFLD)in rats.Methods:Sixty SD male rats were randomly assigned into normal control group,model group,essentiale(polyene phosphatidylcholine,0.144 g/kg)group,and Danggui Shaoyao San(2.44,4.88,and 9.76 g/kg)groups.The rats were fed with high-fat diet to establish the NAFLD model,and corresponding drugs were administrated once a day for eight successive weeks.The levels of total cholesterol(TC),triglyceride(TG),alanine aminotransferase(ALT),aspartate aminotransferase(AST),superoxide dismutase(SOD),glutathione(GSH),and malondialdehyde(MDA)in serum,as well as the levels of TC,TG,free fatty acid(FFA),SOD,GSH,and MDA in liver tissue,were measured.Hematoxylin eosin(HE)staining and Oil red staining were conducted for observation of the pathological changes in liver.The qRT-PCR and Western blotting were employed to determine the mRNA and protein levels of Kelch-like ECH-associated protein1(Keap1),nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),glutamate-cysteine ligase catalytic subunit(GCLC),and glutamate-cysteine ligase modifier subunit(GCLM)in liver tissue.Results:Compared with the normal control group,the modeling raised the levels of TC,TG,ALT,AST,and MDA in serum and TC,TG,FFA,and MDA in liver tissue(P<0.01),decreased the activities of SOD and GSH in serum and liver,and down-regulated the mRNA and protein levels of Keap1,Nrf2,HO-1,GCLC,and GCLM in liver tissue(P<0.01).Meanwhile,the rats in the model group showed a large number of lipid droplet vacuoles and balloon-like lesions in the hepatocytes.Compared with the model group,Danggui Shaoyao San(all the doses)lowered the levels of TC,TG,ALT,AST,and MDA in serum and TC,TG,FFA,and MDA in liver tissue(P<0.01 or P<0.05),increased the activities of SOD and GSH in serum and liver,and up-regulated the mRNA and protein levels of Keap1,Nrf2,HO-1,GCLC,and GCLM in liver tissue(P<0.01 or P<0.05).Furthermore,it alleviated liver steatosis.Conclusion:Danggui Shaoyao San can prevent and treat NAFLD by regulating the Keap1/Nrf2 signaling pathway and resisting oxidative damage.
作者
彪雅宁
张睦清
韩雪
陈健
高晶淼
李子璇
宋竹青
张一昕
Biao Yaning;Zhang Muqing;Han Xue;Chen Jian;Gao Jingmiao;Li Zixuan;Song Zhuqing;Zhang Yixin(School of Pharmacy,Application Technology Research Center of Traditional Chinese Medicine(TCM)Prescription in University of Hebei Province,Hebei University of Chinese Medicine,Shijiazhuang 050200;Hebei Provincial Hospital of TCM,Shijiazhuang 050911;Cangzhou Rongfu Military Sanatorium,Cangzhou 061000)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第1期13-18,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
河北省自然科学基金面上项目(编号:H2020423028)
河北省高等学校科学研究计划重点项目(编号:ZD2021079)
中央财政公共卫生专项“2018年中药资源”普查项目(编号:Z135080000022)
国家级大学生创新创业训练计划项目(编号:202114432006)
关键词
当归芍药散
非酒精性脂肪肝
氧化应激
Kelch样环氧氯丙烷相关蛋白1
核因子E2相关因子
Danggui Shaoyao San
nonalcoholic fatty liver disease(NAFLD)
oxidative stress
Kelch-like ECH-associated protein1(Keap1)
nuclear factor E2-related factor 2(Nrf2)
作者简介
通信作者:张一昕,博士,教授,博士生导师,研究方向:代谢性疾病的中医药防治和机制研究,E-mail:hbzyx123@163.com;彪雅宁,博士,研究方向:中药防治代谢性疾病的机制研究,E-mail:694888650@qq.com。
