天麻素通过调节SREBP1c信号通路抑制非酒精性脂肪肝  

Gastrodin Inhibits Non-alcoholic Fatty Liver Disease via Mediating SREBP1c Signaling Pathway

作　　者：张孟莲 张耀文 唐林峰 巩仔鹏[2] 韩岚 王丹丹 ZHANG Menglian;ZHANG Yaowen;TANG Linfeng;GONG Zaipeng;HAN Lan;WANG Dandan(Anhui Province Key Laboratory of Traditional Chinese Medicine,School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China;State Key Laboratory of Efficacy and Utilization of Medicinal Plants,Guizhou Medical University,Guiyang 550014,China)

机构地区：[1]安徽中医药大学药学院现代中药安徽省重点实验室,合肥230012 [2]贵州医科大学省部共建药用植物功效与利用国家重点实验室,贵阳550014

出　　处：《中国实验方剂学杂志》2024年第11期70-77,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82300524);安徽省高等学校自然科学研究项目(KJ2021A0585);贵州医科大学药用植物功效利用国家重点实验室开放课题基金项目(FAMP202105K)。

摘　　要：目的:观察天麻素对高脂高胆固醇饲料(HFHC)诱导的小鼠在类固醇调节元件结合蛋白1c(SREBP1c)信号通路中的影响,研究天麻素治疗非酒精性脂肪肝(NAFLD)的作用机制。方法:体内采用8周龄雄性C57BL/6J小鼠,随机分为4组,每组6只,分别为正常组、天麻素组(50 mg·kg^(-1))、模型组、模型加天麻素组(50 mg·kg^(-1))。小鼠给予HFHC喂养4周建立NAFLD模型,4周后处死并取出小鼠肝脏组织。体外实验选用Huh7细胞,分为5组,游离脂肪酸(200μmol·L^(-1),油酸-棕榈酸2∶1)诱导NAFLD细胞模型;24 h后每组加入不同浓度的天麻素(0、5、10、20、40μmol·L^(-1))继续培养24 h。油红O染色检测小鼠肝脏和Huh7细胞内脂质蓄积情况,苏木素-伊红(HE)染色观察肝组织病理形态变化,全自动生化分析仪检测血清总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和甘油三脂(TG)的水平。相关试剂盒检测肝脏TC、TG和游离脂肪酸(FFA)的含量。实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)分别检测脂质合成关键蛋白脂肪酸合成酶(FASN)、乙酰辅酶A羧化酶1(ACC1)和硬脂酰辅酶A去饱和酶1(SCD1)的表达。结果:与正常组比较,模型组小鼠血清TC、LDL-C和TG含量显著升高(P<0.01),出现脂肪肝病变且肝内TC、TG和FFA含量显著升高(P<0.01),Huh7细胞脂质蓄积水平显著升高(P<0.01),小鼠和Huh7细胞脂质合成相关基因SREBP1c、FASN、ACC1和SCD1的表达水平显著升高(P<0.01)。与模型组比较,天麻素治疗后小鼠血清TC、LDL-C和TG水平明显降低(P<0.05,P<0.01),脂肪肝病变明显改善,肝脏TC、TG和FFA含量显著降低(P<0.05,P<0.01),Huh7细胞脂质蓄积水平明显降低(P<0.05,P<0.01),小鼠和Huh7细胞脂质合成相关基因SREBP1c、FASN、ACC1和SCD1的表达水平明显降低(P<0.05,P<0.01)。结论:天麻素能够降低肝内脂质积累与血脂水平,改善HFHC诱导的NAFLD,其作用机制可能与调节SREBP1c脂质合成相�Objective:To observe the effect of gastrodin on the steroid regulatory element-binding protein 1c(SREBP1c)signaling pathway in high-fat high-cholesterol diet(HFHC)-induced mice and explore the mechanism of gastrodin in the treatment of non-alcoholic fatty liver disease(NAFLD).Method:Eightweek-old male C57BL/6J mice were used in vivo and divided into the following four groups,with six mice in each group:normal group,gastrodin group(50 mg·kg^(-1)),model group,and model+gastrodin group(50 mg·kg^(-1)).NAFLD model was established by feeding mice with HFHC for four weeks,and the mice were euthanized and the liver tissues were collected after four weeks.In vitro experiments were performed using Huh7cells which were divided into five groups,and induced with free fatty acids(FFA,200μmol·L^(-1),oleic acidpalmitic acid 2∶1)to establish an NAFLD cell model.After 24 h,different concentrations of gastrodin(0,5,10,20,and 40μmol·L^(-1))were added to each group and cultured for another 24 h.Oil red O staining was used to detect lipid accumulation in mouse liver and Huh7 cells.Hematoxylin-eosin(HE)staining was used to observe pathological changes in liver tissue.Levels of serum total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and triglycerides(TG)were measured using an automatic biochemical analyzer.Relevant assay kits were used to detect liver TC,TG,and FFA levels.Real-time quantitative polymerase chain reaction(Real-time PCR)and Western blot were used to detect the expression of lipid synthesis-related proteins fatty acid synthase(FASN),acetyl-CoA carboxylase 1(ACC1),and stearoyl-CoA desaturase 1(SCD1).Result:Compared with the normal group,the model group showed significantly increased serum TC,LDL-C,and TG levels(P<0.01),liver TC,TG,and FFA levels(P<0.01),increased lipid accumulation in Huh7 cells(P<0.01),and significantly increased expression levels of lipid synthesis-related genes SREBP1c,FASN,ACC1,and SCD1 in mice and Huh7 cells(P<0.01).Compared with the model group,after gastrodin treatment,the serum

关 键 词：天麻素 非酒精性脂肪肝 固醇调节元件结合蛋白1C 血脂紊乱 脂肪变性 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33