Tianhuang formula enhancing nonalcoholic fatty liver disease in mice through regulating gut microbiota

LUO Zhi-zhong; YANG Ling; HAN Yi; GUO Jiao; LUO Duo-sheng

doi:10.20251/j.cnki.1003-3734.2025.05.011

Abstract：Objective:To investigate the effect and mechanism of the Tianhuang formula(TH)in improving nonalcoholic fatty liver disease(NAFLD)in mice by regulating gut microbiota.Methods:High-fat diet was used to induce a NAFLD model.After 8 weeks of oral administration of atorvastatin or TH,fresh feces from mice in the high-dose TH group and the model group were collected to prepare microbiota suspensions for fecal microbiota transplantation experiments.At the end of the experiment,blood lipid and liver function indicators in mice were examined;hematoxylin and eosin staining and oil red O staining were used to analyze pathological changes in mice liver tissue;PCR technology was utilized to detect the expressions of mRNA of inflammatory factors,fatty acid synthesis,and fatty acid β-oxidation-related genes in liver tissues;16S rRNA sequencing was conducted to analyze changes in the gut microbiota of mice.Results:Compared with the model group,liver steatosis and liver injury were alleviated,and the body weight and serum lipid levels were significantly reduced(P<0.01)in the mice of the atorvastatin and low and high dose TH groups.Compared with the transplantation control group,significant improvement in liver steatosis,reduced levels of serum triglycerides,aspartate aminotransferase,liver inflammatory factors,and the expression of fatty acid synthesis-related gene mRNA were observed in the TH microbiota transplantation group(P<0.05),along with a significant increase in the expression of fatty acid β-oxidation-related gene mRNA(P<0.05).The 16S rRNA sequencing results showed that the gut microbiota structure changed in the TH microbiota transplantation group compared with the transplantation control group.At the genus level,the abundance of harmful bacteria such as Bacteroides decreased significantly(P<0.05),while the abundance of beneficial bacteria such as Lactobacillus increased significantly(P<0.05)in the TH microbiota transplantation group.Correlation analysis indicated significant associations of Lactobacillus,Bacteroides,etc.,with liver function,blood lipids,inflammatory factors,fatty acid synthesis,and fatty acid β-oxidation.Conclusion:This study reveals that TH may exert its therapeutic effects on NAFLD by ameliorating dysbiosis of the gut microbiota and affecting liver inflammatory factors,fatty acid synthesis,and fatty acid β-oxidation in mice.