Wuwei Qingzuo San (WWQZS), as a renowned traditional Mongolian patent medicine approved by Chinese State Food and Drug Administration, is used to treat hyperlipidemia, indigestion, and other ailments related to disorder of production of essence and phlegm, a typical abnormal metabolism of blood in traditional Mongolian medicine. A combination of network pharmacology and validation experiments in hyperlipidemia hamster is used to understand the potential mechanism of WWQZS for hypolipidemic effects, further for an integrated concept of traditional theory, bioactive constituents, and molecular mechanism for TMM. Through network pharmacology, we obtained 212 components, 219 predicted targets, and 349 known hyperlipidemia-related targets form public database and used Metascape to carry out enrichment analysis of 43 potential and 45 candidate targets to imply numerous BP concerned with metabolism of lipid, regulation of kinases and MF related to lipid binding, phosphatase binding, and receptor ligand activity that are involved in anti-hyperlipidemia. In addition, KEGG pathways that explicated hypolipidemic effect were involved in pathways including metabolism associated with kinase function according to MAPK signaling pathway, AMPK signaling pathway, and PI3K-Akt signaling pathway. Meanwhile, in HFD-induced hamster model, WWQZS could significantly reduce TC and ALT and help decrease TG, LDL-C as well; liver pathological section implied that WWQZS could relieve liver damage and lipid accumulation. Western blot indicated that WWQZS may upregulate CYP7A1 and activate AMPK to suppress the expression of HMGCR in livers. In conclusion, our results suggest that WWQSZS plays important dual hypolipidemic and liver-protective role in livers in HFD-induced hamster model. Through this research, a new reference is also provided to other researches in the study of ethnopharmacology.

Ethnopharmacological relevance: Total flavonoids of Rhizoma Drynariae (TFRD), an important component extracted from the dried rhizome of Drynaria roosii Nakaike [Polypodiaceae]. TFRD has been widely used in China to treat osteoporosis (OP). However, the specific molecular mechanism of TFRD against OP has not been fully clarified. Aim of the study: The aim of this study was to further elucidate molecular mechanism of TFRD for treating OP based on metagenomic and metabolomic analyses. Materials and methods: In this study, hematoxylin-eosin (H&E) staining, micro computed tomography (micro-CT) and bone mineral density (BMD) analysis were used to observe pharmacological effects of TFRD against ovariectomized (OVX)-induced OP. Subsequently, multiomics analysis including metagenomics, untargeted and short chain fatty acids (SCFAs) metabolomics were carried out to identify whether the anti-osteoporosis mechanism of TFRD correlated with gut microbiota and related metabolites. Results: TFRD could improve the microstructure and density of trabecular bone in OVX rats. 17 differential species, which mainly from Akkermansia, Bacteroides, and Phascolarctobacteriumgenus, 14 related differential metabolites and acetic acid in SCFAs were significantly altered by OVX and reversed by TFRD. Furthermore, according to results of untargeted metabolomics analysis, it was found several metabolic pathways such as phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis and so on might play an important role in TFRD against OP. In order to further study the relationship between gut microbiota and related metabolites, spearman correlation analysis was used, and showed that gut microbiota such as Akkermansia muciniphila might be closely related to several metabolites and metabolic pathways. Conclusions: TFRD treatment could reduce the effects of OVX-induced OP by altering community composition and abundance of gut microbiota, regulating metabolites and SCFAs. It was speculated that the gut microbiota especially Akkermansia muciniphila and related metabolites might play an important role in TFRD against OP, and deserve further study by follow-up experiment. This conclusion provides new theoretical support for mechanism research of TFRD against OP.