基于血浆代谢组学探究三七总皂苷对2型糖尿病小鼠的降糖作用  

Mechanistic investigation on the hypoglycemic effect of Panax notoginseng saponins in type 2 diabetic mice based on plasma metabolomics

作　　者：张金花 刘汉湘 刘雨轩 吴敏 常晋霞[2] 刘文虎[1] ZHANG Jin-hua;LIU Han-xiang;LIU Yu-xuan;WU Min;CHANG Jin-xia;LIU Wen-hu(Department of Pharmacy,North Sichuan Medical College,Nanchong 637100,China;School of Basic Medical Sciences&Forensic Medical,North Sichuan Medical College,Nanchong 637100,China)

机构地区：[1]川北医学院,药学院,四川南充637100 [2]川北医学院,基础医学与法医学院,四川南充637100

出　　处：《药学学报》2024年第4期1028-1039,共12页Acta Pharmaceutica Sinica

基　　金：国家级大学生创新创业项目(202210634038);四川省应用基础科研项目(2019YJ0378);川北医学院重点发展项目(CBY22-ZDA01);南充市市校合作项目(18SXHZ0402).

摘　　要：基于血浆代谢组学结合实验验证研究三七总皂苷(Panax notoginseng saponins,PNS)对2型糖尿病(type 2 diabetes mellitus,T2DM)小鼠的降糖作用。将40只C57BL/6J小鼠随机分为对照组和实验组,对照组常规饲养,实验组高脂饲料喂养12周后,经腹腔注射链脲佐菌素(streptozotocin,STZ)建立T2DM模型,所有动物实验经川北医学院实验动物伦理委员会批准(批准号:NSMC2022023)。造模成功小鼠随机分为模型组(T2DM)、低剂量[200 mg·kg^(-1)·d^(-1)]和高剂量[300 mg·kg^(-1)·d^(-1)]三七总皂苷组,灌胃6周,每周监测小鼠体重、摄食量、进水量和空腹血糖值(fasting blood glucose,FBG),灌胃第5周进行口服糖耐量实验(oral glucose tolerance test,OGTT)。摘眼球采血检测肝脏功能及血脂;试剂盒检测血液肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白介素-6(interleukin-6,IL-6)的水平及肝脏中抗氧化系统谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)和超氧化物歧化酶(superoxide dismutase,SOD)的活性。基于代谢组学研究血浆中内源性代谢物变化;使用KEGG数据库通路富集分析;Western blot检测肝脏NF-κB通路及TNF-α和IL-6的表达。结果显示,T2DM小鼠构建成功;高剂量三七总皂苷(high Panax notoginseng saponins,HPNS)能够降低T2DM小鼠的空腹血糖,低剂量三七总皂苷(low Panax notoginseng saponins,LPNS)对空腹血糖无显著影响。HPNS能够改善T2DM小鼠肝脏功能,降低血脂及血液TNF-α和IL-6的水平,增加肝脏GSH-Px、CAT和SOD的活性。代谢组学结果显示,模型组血浆中45种代谢物变化显著,HPNS治疗后20种代谢物显著变化且向正常组转归。通路富集表明,花生四烯酸代谢、亚油酸代谢、谷胱甘肽代谢及肉碱合成在T2DM小鼠血液中改变,HPNS可改善T2DM小鼠血液中花生四烯酸和亚油酸异常代谢。Western blot表明,HPNS能够抑制T2DM小鼠NF-κB通路,降低TNF-α和IL-6的表达,提示PNS可�Plasma metabolomics combined experimental verification was employed for investigating of the hypoglycemic effect of Panax notoginseng saponins(PNS)on type 2 diabetes mellitus(T2DM)mice.Forty C57BL/6J mice were randomly divided into control and experimental groups after one week of adaptive feeding.The mice in control group were fed conventionally,and the T2DM model was established in mice of the experimental group by intraperitoneal injection of streptozotocin following twelve weeks of feeding on a high-fat diet(HFD).All experiments were approved by the Ethical Committee Experimental Animal Center of North Sichuan Medical College(NSMC2022023).After the failure cases during modeling were eliminated,the remaining mice were randomly divided into model group(T2DM),low dose[200 mg·kg^(-1)·d^(-1)]and high dose[300 mg·kg^(-1)·d^(-1)]PNS groups.Mice in normal and model groups were given equal amounts of normal saline by gavage.The mice were administered intragastrically with PNS for 6 weeks,and their body weight,food intake,water intake and fasting blood glucose(FBG)were measured weekly.Oral glucose tolerance test(OGTT)was performed at the 5th week of administration.The changes of liver functions and blood lipids were detected by collecting blood from eyeballs.Tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)levels were detected in the blood and the activity of glutathione peroxidase(GSH-Px),catalase(CAT)and superoxide dismutase(SOD)were analyzed in the liver by kit,respectively.Subsequently,the changes in plasma endogenous metabolites from each group were determined based on metabolomics,and the pathway enrichment analysis of differential metabolites was performed using KEGG database.NF-κB signaling pathway,TNF-αand IL-6 in liver were detected by western blot,respectively.The results showed that T2DM mice were successfully constructed.High dose of panax notoginseng saponins(HPNS)can reduce the FBG in T2DM mice while low dose of PNS(LPNS)has no significant effect on FBG.HPNS improves the liver function,reduce

关 键 词：三七总皂苷 降糖作用 NF-ΚB通路 花生四烯酸代谢 亚油酸代谢 

分 类 号：R917[医药卫生—药物分析学]