瓜蒌薤白半夏汤化裁降低高脂饮食模型小鼠血脂的肠道非靶向代谢组研究

Study on the Mechanism of Modified Gualou Xiebai Banxia Decoction(瓜蒌薤白半夏汤化裁)on Reducing Blood Lipid in Mice with High Fat Diet Based on Intestinal Non-target Metabonomics

目的考察瓜蒌薤白半夏汤化裁降低高脂饮食模型小鼠血脂过程中小鼠的肠道代谢物变化情况,揭示其中的肠道代谢机制。方法分别选择24只ApoE-/-小鼠及6只C57BL/6J小鼠,以高脂饲料及普通饲料饲食;ApoE-/-小鼠分为高、中、低3个治疗组,分别依据瓜蒌薤白半夏汤化裁14、11、7 g/(kg·d)的剂量进行灌胃;模型组则灌胃等体积0.9%氯化钠溶液。末次给药24 h后处死小鼠,并获得其动脉血及肠道内容物样本,分别检测各组小鼠的血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)及高密度脂蛋白胆固醇(HDL-C),同时以液相色谱-质谱联用仪(LC-MS)测定其肠道内容物的代谢组学变化,对获得的代谢组数据进行主成分分析(PCA),并以R程序获得其中的差异代谢物,再利用MetaboAnalyst等在线数据库对获得的差异代谢物进行功能估计分析。结果瓜蒌薤白半夏汤化裁高剂量组可以显著降低小鼠体内TG、TC及LDL-C水平,同时提升HDL-C的水平(P<0.05)。代谢组学的分析结果显示瓜蒌薤白半夏汤化裁高、中、低剂量组小鼠肠道内容物代谢组差异较大。进一步对正常组、模型组及瓜蒌薤白半夏汤化裁高剂量组3组的肠道代谢物分析后,共识别出13个下调和9个上调的阴离子差异代谢物;15个下调和6个上调的阳离子差异代谢物,功能富集的结果显示上述差异代谢物涉及到的最核心的通路为原胆汁酸生物合成路径(primary bile acid biosynthesis)。结论瓜蒌薤白半夏汤化裁可有效改善高脂血症模型小鼠的血脂异常症状,其可通过调节模型小鼠肠道代谢物的形式降低血脂,其中最重要的代谢通路为原胆汁酸生物合成路径。

Objective To investigate the changes of intestinal metabonomics in mice with high-fat diet and explore the intestinal metabolic mechanism.Methods 24 ApoE-/-mice and 6 C57BL/6J mice were fed with high-fat diet and common diet respectively;ApoE-/-mice in high,medium and low treatment groups were respectively given 14 g(/kg·d),11 g(/kg·d)and 7 g(/kg·d)of Modified Gualou Xiebai Banxia Decoction(瓜蒌薤白半夏汤化裁),while the model group was given the same volume of normal saline.The serum total cholesterol,triglyceride,low density lipoprotein cholesterol and high density lipoprotein cholesterol were detected respectively.The metabonomic changes of intestinal contents were detected byliquid chro-matograph mass spectrometer.The metabonomics data were analyzed by principal component analysis.The differential metabolites were obtained by R program.The results showed that the serum TG,TC,LD-C and HDL-C were significantly higher than those of the control group functional estimation analysis was performed for different metabolites.Results The high dose group of Modified Gualou Xiebai Banxia Decoction could significantly reduce the levels of TG,TC and LDL-C in mice,and increase the level of HDL-C(P<0.05).The results of metabonomics analysis showed that the metabonomics of intestinal contents in high,medium and low treatment groups were significantly different.After further analysis of intestinal metabolites in normal group,model group and high-dose group,13 down-regulated and 9-up-regulated anionic differential metabolites were identified;15 down-regulated and 6-up-regulated cationic differential metabolites were identified.The results of functional enrichment showed that the core pathway involved in the above differential metabolites was primary bile acid biosynthesis of protobile acids.Conclusion Modified Gualou Xiebai Banxia Decoction can effectively improve the dyslipidemia symptoms of hyperlipidemia model mice,which can reduce blood lipid by regulating intestinal metabolites of model mice,and the most important metabolic pathway is protobile acid biosynthesis.