目的 探究毛菊苣乙醇提取物对高脂饮食诱导的肥胖小鼠粪便胆汁酸谱的影响.方法 选取6周龄C57BL/6雄性小鼠24只随机分为正常组、模型组、给药组和二甲双胍组.正常组给予常规饮食,其余3组给予高脂饲料,同时给药组每日灌胃10 mL/kg毛菊苣乙醇提取物溶液,二甲双胍组每日灌胃10 mL/kg二甲双胍溶液.10周后收集小鼠肝检测肝甘油三酯(triglycerides,TG)、总胆固醇(total cholesterol,TC)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)和高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C);收集小鼠粪便通过超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)检测粪便胆汁酸含量.结果 与正常组相比,模型组小鼠体重(P＜0.0001)、血清TG、TC、LDL-C水平(P＜0.0001)和肝TG水平(P＜0.05)显著升高,肝HDL-C水平显著降低(P＜0.001),表现出增重和脂质代谢异常.毛菊苣乙醇提取物能显著降低小鼠体重(P＜0.0001)、血清TG(P＜0.05)、TC(P＜0.01)、LDL-C(P＜0.01)水平及肝TG(P＜0.0001)、LDL-C(P＜0.05)水平.方法学验证结果表明本研究建立的方法能准确定量粪便中的52种胆汁酸.分析各类胆汁酸浓度发现,毛菊苣乙醇提取物能显著增大次级胆汁酸/初级胆汁酸比例(P＜0.05).多元统计分析结果显示,各组的胆汁酸代谢模式发生明显改变.以第一主成分的变量投影重要度(variable importance in the projection,VIP)＞1,P＜0.05为条件筛选出给药组相对模型组的8种差异胆汁酸.通过检索京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)数据库,发现差异胆汁酸主要参与次级胆汁酸生物合成途径.相关分析表明,粪便中脱氧胆酸(rs=0.6445,P＜0.001)、异石胆酸(rs=0.5879,P＜0.01)、3β-脱氧胆酸(rs=0.6649,P＜0.001)和ω-鼠胆酸(rs=0.5387,P＜0.01)4种差异胆汁酸与体重具有较强的正相关.结论 毛菊苣乙醇提取物可能通过调控次级胆汁酸生物合成,改变粪便胆汁酸代谢轮廓从而发挥减重和改善脂质异常的作用.
Effect of Cichorium glandulosum Boiss.et Huet.on fecal bile acid profile in obese mice based on targeted metabolomics technique
Objective To investigate the effect of ethanolic extract of Cichorium glandulosum Boiss.et Huet.on fecal bile acid profiles in high-fat-diet-induced obese mice.Methods Twenty-four 6-week-old C57 BL/6 male mice were divided randomly into normal,model,drug-administration,and metformin groups.Mice in the normal group were fed a regular diet and mice in the other three groups were given high-fat diets.The drug-administration group was gavaged with 10 mL/kg ethanol extract of Cichorium glandulosum Boiss.et Huet.daily,and the metformin group was gavaged with 10 mL/kg metformin daily.After 10 weeks,livers were collected to measure hepatic total triglycerides(TG),total cholesterol(TC),low-density lipoprotein-cholesterol(LDL-C),and high-density lipoprotein(HDL)-C.Feces were collected and analyzed.Results Body weight(P＜0.0001),liver TG(P＜0.05),and TC(P＞0.05)were all significantly higher in model mice compared with normal mice,while LDL-C(P＞0.05)and HDL-C(P＜0.001)were significantly lower,indicating abnormal weight gain and lipid metabolism.Alcoholic extract of Cichorium glandulosum Boiss.et Huet.significantly reduced body weight(P＜0.0001),liver TG(P＜0.0001),serum TG(P＜0.05),TC(P＜0.01),and LDL-C(P＜0.05)in mice.Methodsological validation showed that the current method could accurately quantify 52 bile acids in feces.Analysis of the concentration of each type of bile acid revealed that alcoholic extract of Cichorium glandulosum Boiss.et Huet.significantly increased the secondary/primary bile acid ratio(P＜0.05).Multivariate analysis showed that the bile acid metabolic pattern was significantly altered in all groups.Eight differential bile acids were screened in the drug-administration group relative to the model group using variable importance of projection＞1 and P＜0.05.A search of the Kyoto Encyclopedia of Genes and Genomes database revealed that the differential bile acids were mainly involved in the secondary bile acid biosynthesis pathway.Correlation analysis showed that four differential bile acids,deoxycholic acid(r,=0.6445,P＜0.001),isolithocholic acid(r,=0.5879,P＜0.01),3β-deoxycholic acid(r,=0.6649,P＜0.001),and ω-rhamnoglutaric acid(rs=0.5387,P＜0.01),in feces were strongly positively correlated with body weight.Conclusions Cichorium glandulosum Boiss.et Huet.alcoholic extract may play a role in weight reduction and amelioration of dyslipidemia by modulating secondary bile acid biosynthesis and altering fecal bile acid metabolic profiles.
万方数据
维普
