Traditional Mongolian Medicine Qiqirigan-8 Alleviates Non-alcoholic Fatty Liver Disease Via Restoring Gut Microbiota and Metabolism

dandan yang ¹ Ta Na ¹ Wuyun siqin ^1* Yan Niu ^2* Ha Shentuya ¹ An na ^1* Mingxing Ma ^1* Wenhui Zhao ^1* Menggen duxi ^1* Minjie Wang ^2*

• ¹ School of Traditional Mongolian Medicine, Inner Mongolia Medical University,, Hohhot, China
• ² School of Basic Medicine, Inner Mongolia Medical University, Hohhot, Inner Mongolia Autonomous Region, China

Background: Mongolian Medicine Qiqirigan-8 (MMQ-8) is a traditional Mongolian medicine formula used to treat fatty liver disease. However, the material basis and in vivo metabolic process of the therapeutic effect of MMQ-8 on non-alcoholic fatty liver disease (NAFLD) remain unclear. Methods: The chemical composition of MMQ-8 was determined using UHPLC-QE-MS. C57BL/6J mice were fed a choline-deficient diet for 12 weeks to induce a NAFLD model. HE staining, combined with serum biochemical indexes, was used to observe liver appearance and characterize the pathological changes and functions of the liver. HE staining and AB-PAS staining of the colon, along with ZO-1 immunofluorescence expression in the colon. The expression of intestinal tight junction genes was analyzed by qRT-PCR. Fecal metagenomics and serum non-targeted metabolomics were used to reveal the effects of MMQ-8 on the gut microbiota and metabolism in mice with NAFLD. Finally, we emphasize the interaction between gut microbiota and metabolites through Spearman correlation coefficient analysis. Results: MMQ-8 contains 17 active ingredients, which can reduce hepatic steatosis and lobular inflammation in mice with NAFLD, and have protective effects against liver injury. MMQ-8 reduced the infiltration of inflammatory cells in the colon epithelium of model mice while restoring the number of goblet cells. MMQ-8 significantly enhanced ZO-1 protein expression in the colon, as well as the mRNA expression of both ZO-1 and Occludin. Fecal metagenomics results showed that MMQ-8 reduced the Bacillota/Bacteroidota ratio in NAFLD mice. Increased the abundance of beneficial bacteria such as Porphyromonadaceae, Prevotella, and Bacteroidota. and suppressed the abundance of dysfunctional bacteria, such as Bacillota, Acetatifactor, and Erysipelotrichaceae. Furthermore, metabolomics studies revealed that MMQ-8 intervention significantly regulated the expression of metabolites related to glutathione metabolism, butyric acid metabolism, sphingolipid metabolism, and glycerophospholipid metabolism in NAFLD mice compared to the model group. Conclusion: In summary, these findings indicate that MMQ-8 has a synergistic anti-NAFLD effect through its multi-component, multi-target, gut microbiota-modulating and multi metabolic pathway characteristics. The host's regulation of specific gut microbiota and involvement in multiple metabolic pathways may be one of the important mechanisms by which MMQ-8 exerts its therapeutic effects on NAFLD.