Interpreting The Property Of Traditional Chinese Medicine And The Lipid-lowering Mechanism Of Berberine By Gut Microbiota-drug Interaction

The property theory is a basic theory of traditional Chinese medicine(TCM)that generalize the properties,actions and application rules of TCMs,including four qi,five flavors,meridian,ascending and descending,floating and sinking,and toxicity.The property theory is not only the most characteristic theory of TCM,but also a core theoretical basis for guiding the clinical use of TCM.The interpretation of property theory by modern science and technology is an important basis and forerunner of the modern development of TCM,and can guide the research of all aspects of TCMs.Gut microbiota is a collection of microorganisms that inhabit host intestine.Numerous studies have demonstrated that the gut microbiota is an important biological indicator for human body syndrome,which can be used as the key medium for the transmission of TCM’s property in the body.On the one hand,the gut microbiota modulated by TCMs with different property showing the corresponding characteristics.On the other hand,the gut microbiota altered by TCMs further regulates the function of the human body,corrects the cold and hot changes of the body,so as to restore the balance of Qi movement to treat diseases.The gut microbiota reflects and transmits the properties of TCM,which provides a new idea and direction for the deep interpretation of the biological connotation and in vivo transmission mechanism of TCMs.Simultaneously,gut microbiota is also closely related to the efficacy of drugs.The gut microbes can metabolize herbal constituents into molecules with increased or decreased bioavailability and efficacy,thus alter the circulating intensity of the herb’s active components.On the other aspect,herbal medicines may change the composition of the gut microbiota,thereby improve the global functions of our body and remotely ameliorate diseases by enhancing beneficial germs and/or inhibiting bad germs.This study preliminarily explored the relationship between the TCM property and gut microbiota,thus proposed the corresponding relationship between the characteristics of four natures,five flavors,meridian and the alteration of gut microbiota.Then,we took bitter-flavored and cold-natured TCM Coptis chinensis Franch.as an example,and explored the regulation pattern of Coptis chinensis Franch.and its active component berberine(BBR)on gut microbiome,which provided a paradigm for exploring the mechanism of cold-natured drugs based on the alteration of gut microbiome.The dominant outcomes of present study are as follows:Section Ⅰ:Interpreting the property of traditional Chinese medicine by gut microbiota-drug interaction1.Traditional Chinese medicines differentially modulate the gut microbiota based on their nature(Yao-Xing)The in vivo study showed that antidiarrheal TCMs with different natures showed distinct impacts on the gut microbiota.Specifically,the alteration of gut microbial structure was gradually stronger with the herbal nature turned from hot to cold,among which hot-natured TCMs had no influence on the gut microbiota but cold-natured TCMs notably changed the structure of the gut microbial community.As for composition,pro-inflammatory bacteria Anaerotruncus,Tyzzerella and Ruminiclostridium were enriched by hot-natured TCMs,while that of anti-inflammatory bacteria Ruminococcaceae_UCG-010,Parasutterella and Bifidobacterium were increased by cold-natured TCMs,which consistent with their efficacy.The above results suggested that cold TCMs may exert their role by modulating the gut microbiota,while hot TCMs may through other pathways.2.The gut microbial co-abundance gene groups(CAGs)differentially respond to the flavor(Yao-Wei)of traditional Chinese medicineThe chemical component analysis showed that flavones,alkaloids,and saponins were the richest components for sweet-,bitter-,and pungent-flavored TCMs,respectively.Medicines with merged flavors(bitter-pungent and sweet-pungent)displayed mixed profiles of components.According to gut microbial analysis,modulation of TCMs belonging to the same flavor on the taxonomic classification was inconsistent to an extent,while the functional sets of gut microbiota,co-abundance gene groups(CAGs),strongly and differentially responded to distinct flavors.Moreover,these correlations were in line with their pharmacological actions.This indicated that the functional sets,CAGs,could be an appropriate indicator for TCM flavors.3.TCMs with varied meridians(Gui-Jing)differentially alleviate the adverse impact of Coptis chinensis on gut microbiotaCoptis chinensis administration dramatically disrupted the gut microbiota and decreased the alpha diversity.Co-administration of different TCMs alleviated the adverse impact of Coptis chinensis on gut microbiota in a meridian-dependent manner.TCMs belonging to Shaoyin meridian moderately shifted the gut microbiota,while TCMs belonging to Taiyin and especially Jueyin meridians remarkably recovered the gut microbial community to the normal status.Decreased Firmicutes(Clostridia and Bacilli)and Actinobacteria(Bifidobacteriales)and increased Proteobacteria(Enterobacteriaceae)were main features of Coptis chinensis-induced dysbiosis.TCMs belonging to Shaoyin,Taiyin and Jueyin meridians gradually reversed the changes of these bacteria to the normal condition.Simultaneously,the enriching effect of Coptis chinensis on functionally beneficial bacteria such as Akkermansia muciniphila and Blautia coccoides was preserved when co-administration of Shaoyin,Taiyin and Jueyin meridians TCMs.4.Gut fungi differentially response to the antipyretic(heat-clearing)and diaphoretic(exterior-releasing)TCMs in Coptis chinensis-conditioned gut microbiotaOral administration of Coptis chinensis significantly changed the structure of gut bacteria but showed little influence on gut fungi.Co-treatment with antipyretic or diaphoretic TCMs alleviated the impact of Coptis chinensis on gut bacteria to a similar degree.However,combined with either antipyretic or diaphoretic TCMs significantly strengthened the influence of Coptis chinensis on gut fungi,with the latter superior to the former.The antipyretic TCMs enriched Penicillium spp.while diaphoretic TCMs promoted Fusarium spp.Further analysis revealed that the diaphoretic TCMs-enriched fungi Fusarium spp.were positively related to Akkermansia spp.,a beneficial bacteria that interacts with Toll-like receptor 4(TLR4)and regulates thermogenesis,thus providing a potential bacteria-related mechanism of diaphoretic TCMs.Section Ⅱ:Elucidating the lipid-lowering mechanism of berberine by modulating gut microbiota5.Dual modulation of gut bacteria and fungi manifests the gut-based anti-hyperlipidemic effect of Coptis chinensisPharmacological results showed that Coptis chinensis significantly decreased serum lipids and inhibited the hepatic lipid accumulation in the HFD-fed mice.Mechanistically,Coptis chinensis reduced the diversity of gut bacteria and fungi,as well as changed their composition.Fungus Aspergillus species(A.chevalieri,A.luteovirescens,A.oryzae,A.sp.F51)and Penicillium(P.expansum,P.janthinellum,P.sp.BAB-5649 and P.sp.GZU-BCECYN66-5)were decreased in Coptis chinensis-treated group,while Tilletia bornmuelleri,Tilletia bromi were increased.Furthermore,there are complex association between intestinal fungi and bacteria.For example,fungus Aspergillus(Aspergillus chevalieri,Aspergillus luteovirescens,Aspergillus oryzae)was negative associated with bacterium Blautia coccoides,but positive associated with Lactobacillus(L.johnsonii,L.sparagasseri,L.taiwanensis,L.amylovorus).These results suggested that Coptis chinensis may exert anti-hyperlipidemic effects by altering gut bacterial and fungal composition as well as modulating their interactions.6.Gut microbiota specifically mediates the anti-hyperlipidemic effect of BBRThrough different administration approach,antibiotics treatment and fecal microbiota transplantation,we found gut microbiota is necessary and sufficient for BBR’s hyperlipidemia-ameliorating effect.The enhancement of short chain fatty acids(SCFAs)-producing bacteria such as Clostridium Ⅺ,Anaerostipes,Blautia,Akkermansia and Coprobacillus is tightly associated with its anti-hyperlipidemia effect.Furthermore,BBR also decreased the relative abundance of Alistipes,Helicobacter,Enterorhabdus and Desulfovibrio.The above results indicate that the gut microbiota can specifically mediate the anti-hyperlipidemic effects of BBR.7.The BBR-enriched gut commensal Blautia producta ameliorates high-fat diet(HFD)-induced hyperlipidemia and stimulates liver LDLR expressionConsistent with previous study,orally administration of BBR(200 mg/kg)decreased serum and liver lipid levels in HFD-induced hyperlipidemic mice.Microbiome analysis indicated that Blautia was closely associated with BBR’s lipid-modulating activities.Further analysis revealed that BBR selectively promoted the growth of SCFA-producing bacteria Clostridium ⅩⅣa and Blautia producta.Orally treatment of HFD mice with live Blautia producta reduced obesity and alleviated hyperlipidemia.Notably,Blautia producta significantly increased LDLR expression in the liver,and its culture supernatant upregulated the LDLR level and promoted LDL uptake by HepG2 cells.This study not only identified the specific strains that mediate the lipid-lowering efficacy of BBR,but also linked the modulation effect of BBR on gut microbiota and the upregulation of hepatic LDLR expression,thus providing new insights into the anti-hyperlipidemic mechanism of BBR.8.Berberine,a potential prebiotic to indirectly promote Akkermansia growth through stimulating gut mucin secretionGavage of BBR dose-dependently increased the abundance of Akkermansia in mice.However,it did not stimulate Akkermansia growth in direct incubation,suggesting that BBR may promote Akkermansia in a host-dependent way.Oral administration of BBR significantly increased the transcription of mucin-producing genes and mucin secretion in colon.Untargeted metabolomics analysis showed that BBR increased polyamines production in feces which are known to stimulate goblet cell proliferation and differentiation,but treatment with eukaryotic polyamine synthase inhibitor DFMO did not abolish the stimulating effect of BBR on mucin secretion and Akkermansia growth,indicating that the gut bacteria-derived but not the host-derived polyamines may involve in the BBR-promoted Akkermansia growth.In conclusion,this study explored the inherent association between the gut microbiota and the property theory of TCM for the first time,which laid a foundation for the modern interpretation of the property theory of TCM.Meanwhile,taking Coptis chinensis and its active component BBR as an example,we explored the potential molecular mechanism of their anti-hyperlipidemia effect based on the gut microbiome.This work will promote the understanding of TCM’s classic theory and provide new research directions for the interpretation of the pharmacological mechanisms of some TCMs.