茱萸丸通过抑制肠道菌群产生氧化三甲胺改善金黄地鼠皮脂溢出作用与机制研究OA

Objective:This paper aims to observe the effect of Zhuyu(茱萸)Pill on sebum secretion induced by androgen in golden hamsters and its related mechanism.Methods:An animal model of sebum secretion was established by using dihydrotestosterone to induce the auricular tissue of golden hamsters.The total modeling period was 18 days.50 successfully modeling golden hamsters were randomly divided into a model control group,Zhuyu Pill groups at doses of 106,212,and 425 mg/kg,and an isotretinoin group at a dose of 3.7 mg/kg,with 10 hamsters in each group.Another 10 hamsters were selected as the normal control group.After modeling,the normal control group and the model control group were given the same volume of sterile distilled water by gavage for four weeks.The pathological changes of auricu-lar morphology and hematoxylin-eosin(HE)staining were evaluated.The contents of non-esterified fatty acid(NEFA)and triacylglycerol(TG)in the auricular tissue were detected by biochemical methods.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of testosterone(T),estradiol(E2),and luteinizing hormone(LH)in the serum.16S rDNA technology was used to detect the differences in Alpha and Beta diversity and species composition of the gut microbiota in the feces of golden hamsters.UHPLC-MS/MS was used to detect the contents of trimethylamine ox-ide(TMAO)and trimethylamine(TMA)in the plasma of golden hamsters.Pearson was used to measure and analyze the correlation between gut microbiota and TMAO and TMA.Immunohistochemistry was used to detect the protein ex-pression levels of NOD-like receptor family pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein containing CARD(ASC),and cysteinyl aspartate specific protease 1(Caspase-1)in the auricular tissue of golden ham-sters.Western blot and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)were used to de-tect the protein and mRNA expression levels of myeloid differentiation primary response protein 88(MyD88),interleu-kin-1 receptor-associated kinase 1(IRAK1),tumor necrosis factor receptor-associated factor 6(TRAF6),and nuclear factor kappa-B p65(NF-κB p65).Results:Compared with the normal control group,the model control group showed thickened auricular skin,excessive keratinization,elevated serum T and LH levels,decreased E2 levels,decreased Alpha diversity of gut microbiota,increased Firmicutes and Dubosiella,decreased Bacteroidota,Actinobacteria,Ligilactobacillus,Faecaculum,and Blautia,as well as increased expression levels of TMAO,TMA,NLRP 3 inflammasome,and MyD88/I-RAK1/TRAF6/NF-κB pathways in the plasma(P<0.01).Compared with those in the model control group,the auricu-lar lesions in groups with different doses of Zhuyu Pill were improved,the levels of T and LH were decreased in the ser-um,the E2 level was increased.Alpha diversity of gut microbiota was increased,the Verrucomicrobiota,Bacteroidota,Ligilactobacillus,Blautia,and Akkermansia were increased,while the Firmicutes and Dubosiella were decreased.The lev-els of TMAO and TMA in the plasma were decreased,and the protein and mRNA expressions of NLRP3 inflammasome and MyD88/IRAK1/TRAF6/NF-κB pathways were decreased(P<0.05 or P<0.01).Conclusion:Zhuyu Pill can regu-late the levels of sex hormones and inhibit the pathological changes of sebum excretion.Its mechanism may be related to correcting the imbalance of the microbiota,reducing the production of TMAO and TMA,regulating the MyD88/IRAK1/TRAF6/NF-κB signaling pathways and the expression of its downstream NLRP3 inflammasome.