Decreasing of Trimethylamine N-Oxide by Cecal Microbiota and Choline-Trimethylamine Lyase are Associated with Sishen Pill on Diarrhea with Kidney-Yang Deficiency Syndrome

Mingmin Guo,1,2 Yi Wu,2,3 Maijiao Peng,1,2 Nenqun Xiao,1,2 Zhijun Lei,1,2 Zhoujin Tan2,3 1School of Pharmacy, Hunan University of Chinese Medicine, Changsha, People’s Republic of China; 2Hunan Key Laboratory of Traditional Chinese Medicine Prescription and Syndromes Translational Medicine, Changsha, People’s Republic of China; 3School of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha, People’s Republic of ChinaCorrespondence: Zhijun Lei, School of Pharmacy, Hunan University of Chinese Medicine, Changsha, 410208, People’s Republic of China, Email [email protected] Zhoujin Tan, School of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha, 410208, People’s Republic of China, Email [email protected]: Sishen Pill (SSP) is a traditional Chinese medicine prescription commonly used to treat diarrhea with kidney-yang deficiency syndrome. The aim was to investigate the underlying mechanisms of SSP’s therapeutic effects, providing experimental evidence for its mechanism of action.Methods: A mouse model of diarrhea with kidney-yang deficiency syndrome was induced using adenine combined with Folium sennae. After successful model replication, SSP decoction was administered. CutC activity, TMAO, IL-6, TNF-α levels, and cecal content microbiota were measured.Results: SSP significantly improved the general behavioral characteristics of diarrhea mice, and reduced CutC activity, TMAO and IL-6 levels. Sequencing results indicated significant changes at the phylum and genus levels. Correlation analysis revealed a positive correlation between CutC activity and Faecalibaculum (p< 0.05) and Chryseobacterium (p< 0.05), and a significant negative correlation with Prevotellaceae UCG− 001, Rikenella (p< 0.05), Acinetobacter (p< 0.05), Parasutterella (p< 0.05), and Lacticaseibacillus (p< 0.05). TNF-α levels showed a significant negative correlation with Lacticaseibacillus (p< 0.05), Prevotellaceae UCG− 001 (p< 0.01), Parasutterella (p< 0.05), and Candidatus Saccharimonas (p< 0.05). IL-6 levels exhibited a significant negative correlation with Rikenella (p< 0.05), Acinetobacter (p< 0.05), Prevotellaceae UCG− 001 (p< 0.05), Lacticaseibacillus (p< 0.01), and Parasutterella (p< 0.05), and a significant positive correlation with Faecalibaculum (p< 0.05), Chryseobacterium (p< 0.01), and A2. Serum TMAO levels showed a significant positive correlation with Faecalibaculum (p< 0.05) and Chryseobacterium (p< 0.01), and hepatic TMAO levels exhibited a significant positive correlation with Chryseobacterium (p< 0.05).Conclusion: SSP significantly alleviated the symptoms of diarrhea with kidney-yang deficiency syndrome by modulating the cecal microbiota, downregulating CutC activity, and reducing TMAO and inflammatory factor levels. The cecal microbiota-CutC-TMAO-inflammatory cytokine axis may be a key mechanism underlying the therapeutic effects of SSP.Keywords: Diarrhea, Kidney-Yang Deficiency Syndrome, Sishen Pill, CutC, Cecal microbiota, TMAO, Inflammation