天麻素对高脂和高胆固醇饮食致小鼠非酒精性脂肪性肝炎的治疗作用  

Therapeutic effect of gastrodin against high-fat and high-cholesterol diet-induced nonalcoholic steatohepatitis in mice

作　　者：李琳[1] 张营 赵晟珣[1] 肖方喜[3] 张林英[1] 刘菊[1] LI Lin;ZHANG Ying;ZHAO Sheng-xun;XIAO Fang-xi;ZHANG Lin-ying;LIU Ju(Department of General Medicine,Wuhan No.1 Hospital,Wuhan 430022,China;Department of Cardiology,General Hospital of the Yangtze River Shipping,Wuhan 430010,China;Department of Endocrinology,Wuhan No.1 Hospital,Wuhan 430022,China)

机构地区：[1]武汉市第一医院综合医疗科,湖北武汉430022 [2]长江航运总医院心内科,湖北武汉430010 [3]武汉市第一医院内分泌科,湖北武汉430022

出　　处：《中国药理学与毒理学杂志》2022年第11期819-829,共11页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(81700517);武汉市卫生健康委科研项目(WX20C02);武汉市卫生健康委科研项目(WZ22Q31)。

摘　　要：目的 探讨天麻素(GSTD)对高脂和高胆固醇(HFHC)饮食诱导的小鼠非酒精性脂肪性肝炎(NASH)的治疗作用及可能机制。方法 6周龄雄性C57BL/6小鼠按体重分为6组,每组8只:正常对照组、模型组、模型+GSTD 12.5,25.0和50.0 mg·kg^(-1)组及模型+二甲双胍(Met)100 mg·kg^(-1)阳性对照组。除正常对照组外,其余各组小鼠饲喂HFHC饮食24周建立NASH模型;第17周开始,治疗组同时ig给予相应分组药物,每天1次,连续8周。第24周末,测量小鼠肝湿重、体重和肝指数。HE染色观察肝组织形态,计算非酒精性脂肪性肝病活动积分(NAS),Masson染色观察肝组织胶原纤维增生,油红O染色观察肝组织脂质沉积,血糖仪测全血血糖,ELISA法检测血清胰岛素水平计算胰岛素抵抗指数;化学显色法检测血清谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性,肝组织中甘油三酯(TG)、胆固醇(TC)、游离脂肪酸(NEFA)、丙二醛(MDA)和谷胱甘肽(GSH)水平,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性;实时定量PCR(RT-qPCR)检测肝组织脂质合成基因[脂肪酸合成酶(FASN)、血小板反应蛋白受体(CD36)、硬脂酰辅酶A去饱和酶1(SCD1)、过氧化物酶增殖物活化受体γ(PPARγ)]脂质氧化基因PPARα、肉毒碱棕榈酰基转移酶1α(CPT1-α)、炎症因子基因[肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-1β、趋化因子配体2(CCL2)和CCL5]和纤维化标志物基因[Ⅰ型胶原α1(ColⅠα1)、肌动蛋白α2(Actα2)、结缔组织生长因子(CTGF)和转化生长因子β1(TGF-β1)]mRNA水平;Western印迹法检测肝组织胰岛素受体底物1(IRS^(-1))、蛋白激酶B(Akt)和NF-κB蛋白磷酸化水平。结果 与正常对照组相比,模型组肝湿重和体重增加(P<0.01),肝指数升高(P<0.05);肝组织出现严重的大泡性脂肪变性,并伴有大量炎症细胞浸润和胶原纤维增生,NAS评分升高(P<0.01);血糖和血清胰岛素水平、胰岛素抵抗指数以及血清GPT和GOT活性升高(P<0.01);OBJECTIVE To investigate the therapeutic effects of gastrodin(GSTD) against nonalcoholic steatohepatitis(NASH) in mice induced by a high-fat and high-cholesterol(HFHC) diet and to explore its underlying mechanism. METHODS Six-week-old male C57BL/6 mice were randomly divided into six groups by body weight: the normal control group, model group, model+GSTD 12.5 mg·kg^(-1),model+GSTD 25.0 mg·kg^(-1), model+GSTD 50.0 mg·kg^(-1), and model+Met 100 mg·kg^(-1). Except the normal control group, the mice were fed with HFHC for 24 weeks to establish a NASH model. Starting at 17th week, the treatment group was given GSTD or Met by ig once a day for eight weeks. At the end of the24thweek, the blood glucose and body weight of mice in each group were monitored. Histopathological changes, collagen fiber proliferation and lipid deposition in liver tissues were observed by using hematoxyline-eosin(HE), Masson′s trichrome and oil red O staining. The level of insulin was detected by ELISA and insulin resistance(HOMA-IR) was evaluated. The activities of serum glutamic pyruciv transaminase(GPT) and glutamic oxaloacetic transaminase(GOT), the levels of triglyceride(TG), cholesterol(TC), nonestesterified fatty acid(NEFA), malondialdehyde(MDA) and glutathione(GSH), and the activities of superoxide dismutase(SOD) and catalase(CAT) in liver tissues were measured. Real-time quantitative PCR(RT-qPCR) was used to detect the mRNA levels of lipid synthesis genes—fatty acid synthase(FASN), thrombospondin receptor(CD36), stearoyl-coenzyme A desaturase 1(SCD1), and peroxisome proliferator activated receptor γ(PPARγ), lipid oxidation genes—PPARα, carnitine palmitoyltransferase 1α(CPT1α);inflammatory factors gene: tumor necrosis factor α(TNF-α), interleukin-6(IL-6),IL-1β, C-C motif chemokine ligand 2(CCL2), and CCL5 and fibrotic marker genes—collagen type 1alpha 1(Col1α1), actin alpha2(Actα2), connective tissue growth factor(CTGF), and transforming growth factor beta 1(TGF-β1) in liver tissues. Western blotting was used to de

关 键 词：天麻素 非酒精性脂肪性肝炎 胰岛素抵抗 氧化应激 炎症 

分 类 号：R285.5[医药卫生—中药学]