岩黄连总碱对代谢相关脂肪性肝病小鼠的治疗作用及分子机制研究  

Study on effect and molecular mechanism of Corydalis saxicola total alkaloids on nonalcoholic fatty liver mice

作　　者：陈萍 鞠霖杰 成俊 刁和芳 赵开军 邱志霞[1] 黄芳 CHEN Ping;JU Linjie;CHENG Jun;DIAO Hefang;ZHAO Kaijun;QIU Zhixia;HUANG Fang(Department of Chinese Pharmacology and Traditional Chinese Medicine,China Pharmaceutical University,Nanjing 211198,China;Nanjing Zhongshan Pharmaceutical Limited Company,Nanjing 210046,China;Jiangsu Hongdian Institute of Traditional Chinese Medicine Industry,Nanjing 210042,China)

机构地区：[1]中国药科大学中药药理与中医药学系,江苏南京211198 [2]南京中山制药有限公司,江苏南京210046 [3]江苏弘典中药产业研究院,江苏南京210042

出　　处：《药物评价研究》2021年第3期468-477,共10页Drug Evaluation Research

基　　金：“重大新药创制”科技重大专项(2017ZX09301026)。

摘　　要：目的研究岩黄连总碱对高糖高脂饮食诱导的代谢相关脂肪性肝病(MAFLD)小鼠的治疗作用。方法随机取C57BL/6小鼠7只设置为对照组,喂以正常饲料;造模小鼠给予高脂饮食和高糖饮水(含20%果糖水),连续喂养10周;将造模小鼠按体质量随机分为模型组、盐酸二甲双胍(阳性药,200 mg/kg)组和岩黄连总碱低、高剂量(25、100 mg/kg)组,继续饲以高脂高糖饮食,连续ig给药5周。记录小鼠体质量,取肝脏并拍照,测定小鼠肝脏质量,计算肝脏指数;应用血糖仪测定小鼠空腹血糖(FBG)及口服糖耐量(OGTT);试剂盒法测定血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)及游离脂肪酸(NEFA)的水平;取肝组织进行HE染色、油红O染色和Masson染色;Western blotting检测肝组织中AMP依赖的蛋白激酶(AMPK)、p-AMPK、磷脂酰肌醇3-激酶(PI3K)、p-PI3K、蛋白激酶B(Akt)、p-Akt蛋白表达情况。结果与对照组比较,模型组小鼠体质量、肝脏指数显著升高;给药后小鼠体质量及肝脏指数显著下降(P<0.05、0.01)。与模型组比较,岩黄连总碱显著降低小鼠的FBG及OGTT水平(P<0.01);显著降低血清TC、TG、LDL-C及NEFA水平(P<0.01);显著改善小鼠肝组织脂肪变及纤维化;显著上调MAFLD小鼠肝组织p-AMPK、p-PI3K、p-Akt蛋白水平(P<0.01)。结论岩黄连总碱对MAFLD发挥显著治疗作用,其作用机制可能与通过激活AMPK/PI3K/Akt信号通路,减轻肝脏脂质沉积有关。Objective To study the effect and mechanism of Corydalis saxicola total alkaloids(CSTA)on the high-fat and high-sugar(HFHC)diet induced metabolic associated fatty liver disease(MAFLD)mice.Methods Seven C57 BL/6 mice were randomly selected as control group and fed with normal diet.Mice were fed high fat diet and high sugar drinking water(containing 20%fructose water)for 10 weeks.The model mice were randomly divided into model group,metformin hydrochloride(positive drug,200 mg/kg)group and CSTA low and high-dose(25,100 mg/kg)groups according to body weight.Record the body weight and liver coefficient,the fasting blood glucose(FBG)and oral glucose tolerance test(OGTT)of the mice.The total cholesterol(TC),triglycerides(TG),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)and non-esterified fatty acids(NEFA)were investigated to explore weather CSTA could alleviate dyslipidemia in MAFLD mice.HE staining,oil red O staining and Masson staining of liver sections were carried out according to the manufacture’s protocol.The expression of AMPK,p-AMPK,PI3K,p-PI3K,AKT,p-AKT in mouse livers were detected by Western blotting.Results The body weight and liver index of MAFLD mice were significantly increased,which were ameliorated by CSTA(P<0.05 and 0.01).Meanwhile,abnormal FBG and impaired OGTT in MAFLD mice were also normalized by CSTA(P<0.01).CSTA dramatically decreased the serum levels of TC,TG,LDL-C and NEFA in MAFLD mice(P<0.01).HFHC diet could exacerbate lipid accumulation in hepatic cells and hepatic fibrosis in mice,while these entities were relieved by CSTA.Western bloting furtherly indicated that the level of p-AMPK,p-PI3K,and p-Akt protein in the liver tissue of MAFLD mice could be upregulated by CSTA(P<0.01).Conclusion These data clarified CSTA ameliorated hepatic steatosis and dyslipidemia of MAFLD mice via a novel mechanism involved AMPK/PI3K/Akt pathway.

关 键 词：岩黄连总碱 代谢相关脂肪性肝病 高脂高糖饮食 AMPK/PI3K/Akt信号通路 

分 类 号：R285.5[医药卫生—中药学]