目的 探讨化滞柔肝颗粒治疗非酒精性脂肪肝（nonalcoholic fatty liver disease，NAFLD）的作用机制。方法 高脂饮食饲养8周构建大鼠NAFLD模型，化滞柔肝颗粒ig给药4周。半自动生化分析仪测定大鼠血脂相关指标；油红O染色观察肝脏组织病理变化；Illumina Miseq测序平台对V3～V4可变区进行扩增和测序，超高效液相色谱串联质谱（UPLC-MS/MS）开展血浆代谢组学研究，并结合Spearman进行肠道菌群与代谢组学之间的关联分析。结果 与模型组相比，化滞柔肝颗粒中、高剂量组可显著降低大鼠体质量（P＜0.05、0.01）和血清总胆固醇（total cholesterol，TC）、三酰甘油（triacylglycerol，TG）水平（P＜0.01），升高高密度脂蛋白（high-density lipoprotein cholesterol，HDL-C）水平（P＜0.05、0.01）。此外，化滞柔肝颗粒高剂量组还可显著降低大鼠低密度脂蛋白（low-density lipoprotein cholesterol，LDL-C）水平（P＜0.05）。油红O染色显示各给药组脂滴分布较模型组减少。16S rDNA测序显示化滞柔肝颗粒可提高NAFLD模型大鼠杜氏乳杆菌属Dubosiella、脱硫弧菌属Desulfovibrio、Ruminococcaceae-UCG-010等菌群的相对丰度。代谢组学共筛选出18个差异代谢物，通路分析显示化滞柔肝颗粒抗NAFLD主要通过调控氨酰-tRNA生物合成、胆汁分泌、胆固醇代谢等通路。结论 化滞柔肝颗粒改善NAFLD的作用机制可能与调控肠道菌群组成和胆汁分泌、胆固醇代谢等相关代谢通路有关。

Objective To investigate the mechanism of Huazhi Rougan Granule (化滞柔肝颗粒) in treatment of non-alcoholic fatty liver disease (NAFLD). Methods Rats were fed a high-fat diet for eight weeks to establish a NAFLD model, and Huazhi Rougan Granules were administered intragastrically for four weeks. The semi-automatic biochemical analyzer was used to determine the blood lipid-related indicators in rats; Oil red O was used to observe the pathological changes of liver tissue; The Illumina Miseq sequencing platform was used to amplify and sequence the V3—V4 variable region, and UPLC-MS/MS was used to research plasma metabolomics. The correlation between intestinal flora and metabolomics was analyzed by Spearman. Results Compared with model group, the body weight (P < 0.05, 0.01) and serum level of total cholesterol (TC) and triacylglycerol (TG) (P < 0.01) of rats were significantly decreased and high density lipoprotein cholesterol (HDL-C) of rats was significantly increased (P < 0.05, 0.01) in Huazhi Rougan Granule medium and high dose groups. In addition, low density lipoprotein cholesterol (LDL-C) of rats was also significantly decreased in Huazhi Rougan Granule high dose group (P < 0.05). Oil red O staining showed that the fat distribution in each administration group was lower than that of the model group. 16S rDNA sequencing showed that Huazhi Rougan Granule could increase the relative abundance of Dubosiella, Desulfovibrio, Ruminococcaceae-UCG-010 bacteria in NAFLD model rats. A total of 18 differential metabolites were identified by metabolomics, pathway analysis showed that the anti-NAFLD effects of Huazhi Rougan Granules was mainly through the regulation of aminoacyl-tRNA biosynthesis, bile secretion, cholesterol metabolism and other pathways. Conclusion The mechanism of Huazhi Rougan Granules improving NAFLD may be related to the regulation of intestinal flora composition, cholesterol metabolism and other related metabolic pathways.

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河南省医学科技攻关计划联合共建项目（LHGJ20190273）；郑州大学第一附属医院横向课题（K2020-0004）