叶下红对实验性肝脂肪变大鼠的保护作用及其分子机制  

Protective effect of emilia sonchifolia on rats with experimental hepatic steatosis and its molecular mechanism

作　　者：郑永平[1] 肖亮生[1] 李庆南[1] 胡建凡 郑灿滨[1] 林奕芬[1] 

机构地区：[1]汕头市中心医院消化科,广东汕头515031

出　　处：《中国中西医结合急救杂志》2015年第6期591-595,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：广东省中医药强省科研课题（20131053）

摘　　要：目的：探讨叶下红对实验性肝脂肪变大鼠的保护作用及其分子机制。方法将70只SD大鼠按随机数字表法分为正常对照组、模型组、高或低剂量叶下红组及高剂量叶下红＋磷酸化细胞外调节蛋白激酶1/2（pERK1/2）抑制剂（PD98059）组（PD组）。正常对照组予以普通饲料；其余各组予以高脂低蛋白饲料联合30%四氯化碳（CCl4）花生油2 mL/kg每3 d皮下注射1次，连续3周制备动物模型。制模后叶下红组以高、低剂量叶下红提取物（5.0 g/kg和2.5 g/kg）灌胃，每日1次；PD组每日1次灌胃5.0 g/kg叶下红提取物后加用0.3 mg/kg PD98059每周1次尾静脉注射。3周后均改为普通饲料，第5周末取肝组织进行病理学观察；用全自动生化分析仪检测血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、总胆固醇（TC）、甘油三酯（TG）水平，采用免疫组化染色、蛋白质免疫印迹试验（Western Blot）及流式细胞术检测固醇调节元件结合蛋白1（SREBP-1）、pERK1/2、Toll样受体4（TLR4）、高迁移率族蛋白B1（HMGB1）阳性细胞计数及蛋白表达；用羟胺法及硫代巴比妥酸（TBA）法检测肝组织超氧化物歧化酶（SOD）、丙二醛（MDA）水平。结果高、低剂量叶下红组及PD组肝组织小叶炎症较模型组减轻（分：1.50±0.53、1.80±0.43、1.20±0.42比2.30±0.48），ALT、AST、TC、TG、SREBP-1、MDA均较模型组减少，以高剂量叶下红组最为明显〔ALT（U/L）为51.91±6.95比66.50±12.15，AST （U/L）为125.70±5.62比147.10±10.52，TC（mmol/L）为1.79±1.04比2.81±1.08，TG（mmol/L）为0.87±0.55比1.17±0.67，SREBP-1为（30.60±5.56）%比（53.10±5.02）%，MDA（nmol/mg）为5.20±0.87比10.61±5.45， P＜0.05或P＜0.01〕；pERK1/2、TLR4、HMGB1相对表达量与模型组比较差异无统计学意义〔pERK1/2为（43.77±4.93）%比（46.83±5.27）%，TLR4（相对荧光强度）为69.12±24.64比69.08±24.32，HMGB1（相对�Objective To investigate the preventive effects of emilia sonchifolia on experimental hepatic steatosis in rats and its molecular mechanism.Methods Seventy Sprague-Dawley （SD） rats were randomly divided into five groups： normal control, model, high dose emilia sonchifolia, low dose emilia sonchifolia groups and high dose emilia sonchifolia ＋ phosphorylated extracellular signal regulated protein kinase 1/2 （pERK1/2） inhibitor （PD98059） group （PD group）. In normal control group, the rats were fed with normal diet, and in the other four groups, the rats were fed with high fat and low protein diet combined with 30% carbon tetrachloride （CCl4） peanut oil 2 mL/kg subcutaneous injection, once every 3 days for consecutive 3 weeks to establish animal models with hepatic steatosis. In emilia sonchifolia high and low dose groups, 5.0 g/kg and 2.5 g/kg doses of emilia sonchifolia were given respectively by gavage, once a day. In PD group, after administration of emilia sonchifolia high dose by gavage once a day, additionally PD98059 0.3 mg/kg was injected through a tail vein, once a week. After 3 weeks, all rats were switched to normal diet and treatment continued as before. At the end of the 5th week, liver tissues were taken for pathological analyses. The serum levels of alanine transaminase （ALT）, aspartate transaminase （AST）, total cholesterol （TC）, and triglyceride （TG） were determinated by automatic biochenical analyzer. The positive cell count and protein expressions of sterol-regulatory element binding protein 1 （SREBP-1）, pERK1/2, toll like receptor 4 （TLR4） and high mobility group box-1 protein （HMGB1） were tested by immunohistochemistry, Western Blot and flow cytometry. The levels of superoxide dismutase （SOD） and malonaldehyde （MDA） in liver cell homogenate were detected by hydroxylamine and TBA method.Results Compared with the model group, the lobular inflammation in high and low dose emilia sonchifolia groups and PD group was attenuated （1.50±0.53,

关 键 词：叶下红 肝脂肪变性 固醇调节元件结合蛋白1 磷酸化细胞外调节蛋白激酶1/2 TOLL样受体4 高迁移率族蛋白B1 超氧化物歧化酶 丙二醛 

分 类 号：R459.3[医药卫生—治疗学]