基于非靶代谢组学探讨荣肝复方对小鼠慢性肝纤维化的干预机制

Intervention Mechanism of Ronggan Compound Formula on Chronic Liver Fibrosis in Mice Based on Non-target Metabolomics

目的:基于非靶代谢组学探讨荣肝复方对CCL 4诱导的肝纤维化模型小鼠的干预机制,为荣肝复方治疗肝纤维化提供科学依据。方法:将30只小鼠随机分为3组,分别为对照组、模型组、荣肝复方组。首先建立CCL 4诱导的肝纤维化小鼠模型,在此基础上采用荣肝复方干预15 d后,取肝脏组织进行非靶向代谢组学及相关生物信息分析。运用UPLC-MS技术检测各组小鼠肝脏组织代谢组学变化,确定与荣肝复方作用机制密切相关的代谢途径。结果:在慢性肝损伤发病过程中,与对照组相比,模型组小鼠肝脏组织在阳离子模式和阴离子模式下分别检测出323、152个差异代谢物。与模型组相比,荣肝复方组小鼠肝脏组织在阳离子模式和阴离子模式下分别检测出235、130个差异代谢物。阳离子模式下,对3组小鼠肝脏组织样本进行趋势分析,有2个代谢物群具有显著趋势变化,分别表现为先上升后保持平稳和先下降后保持平稳。阴离子模式下,有2个代谢物群具有显著趋势变化,分别表现为先上升后下降和先下降后上升。代谢集富集到的相关通路说明,CCl 4诱导小鼠慢性肝损伤机制和荣肝复方干预机制与胰岛素抵抗、胰高血糖素信号通路、癌症中的中心碳代谢、代谢途径、不饱和脂肪酸的生物合成、ABC转运蛋白、精氨酸和脯氨酸代谢、脂肪细胞脂解的调控、蛋白质消化吸收等通路有关。尤其是ABC转运蛋白、脂肪酸代谢和胰岛素抵抗的调控尤其显著。结论:荣肝复方主要通过调控ABC转运蛋白、脂肪酸代谢和胰岛素抵抗等通路,缓解CCl 4导致的小鼠肝损伤。

Objective:To investigate the intervention mechanism of Ronggan Compound on CCl 4-induced liver fibrosis model mice based on non-target metabolomics,and to provide a scientific basis for the treatment of liver fibrosis by Ronggan Compound Formula.Methods:A total of 30 mice were randomly divided into three groups:control group,model group and Ronggan Compound Formula group.Firstly,a mouse model of CCl 4-induced liver fibrosis was established,and on this basis,15 days after the intervention of Ronggan Compound Formula,liver tissues were collected for non-targeted metabolomics and related bioinformatics analysis.UPLC-MS technology was used to detect the metabolomic changes in the liver tissue of mice in each group,and the metabolic pathways closely related to the mechanism of action of Ronggan Compound Formula were determined.Results:Compared with the control group,323 and 152 differential metabolites were detected in the liver tissues of mice with liver fibrosis after 7 weeks of CCl 4 modeling,respectively.Compared with the model group,235 and 130 differential metabolites were detected in the cation mode and anion mode,respectively.In cation mode,the trend analysis of liver tissue samples from three groups of mice showed that two metabolite groups had significant trend changes,which were first rising and then remaining stable,and first decreasing and then remaining stable,respectively.In the anion mode,there were two metabolite groups with significant trend changes,which were first rising and then decreasing,and first decreasing and then rising,respectively.The related pathways of metabolic enrichment showed that the mechanism of CCl 4-induced chronic liver injury in mice and the intervention mechanism of Ronggan Compound Formula were related to insulin resistance,glucagon signaling pathway,central carbon metabolism in cancer,metabolic pathways,biosynthesis of unsaturated fatty acids,ABC transporters,arginine and proline metabolism,regulation of adipocyte lipidosis,protein digestion and absorption,etc.In particular,the regulation of ABC transporters,fatty acid metabolism,and insulin resistance is particularly significant.Conclusion:Ronggan Compound Formula mainly alleviates the liver injury caused by CCl 4 in mice by regulating ABC transporter,fatty acid metabolism and insulin resistance.