Study On The Efficacy And Mechanism Of Four Traditional Chinese Medicines Against Hyperuricemi

Objective:Smilacis Glabrae Rhizoma(SRG),Alpinia officinarum Hance(AOH),Rhizoma Dioscoreae Septemlobae(RDS)and Plantago asiatica L.(PL)as traditional herbs of Chinese has been reported to have lowering uric acid and anti-inflammatory properties.Thus,they showing great potential for improving hyperuricemia(HUA).The aim of this study is to explore the potential pharmacological mechanisms of SRG,AOH,RDS and PL for improving HUA through perspectives of in vivo and in vitro experiments,gut microbiota and metabolites.Methods:1.Analysis of the main chemical constituents in SRG,AOH,RDS and PL using HPLC-Q-TOFMS.2.HUA mouse model established by intraperitoneal injection of PO(300 mg/kg).After 2 hour of PO injection,Benzbromarone(13 mg/kg),Febuxostat(10.4 mg/kg),SRG(1.560 g/kg),AOH(0.257 g/kg),AOH(0.378 g/kg)and PL(0.448 g/kg)were administered by gavage to each group except the control group.Once daily for 3 weeks.After the last dose,blood was collected from the inner canthus of the eye to isolate the serum for biochemical testing.Kidney,colorectal and liver were taken for WB and q RT-PCR assays,as well as for gut microbiota and metabolomics analysis.3.First,the effect of aqueous extracts of SRG,AOH,RDS and PL on the proliferative activity of HK-2 cells using the CCK8 assay.And the different concentrations of UA on the viability of HK-2 cells was also investigated.Finally,WB was used to detect the effect of the aqueous extracts of four Chinese herbal medicines on the expression level of uric acid transporter protein OAT1.4.Fecal samples from each group of mice were collected for 16 Sr RNA sequencing before the final dose.Then,based on diversity analysis,species composition analysis,functional prediction of intestinal flora and differential flora analysis,to explore the effects of aqueous extracts of four Chinese herbs on the intestinal microbiota of HUA.5.Identification of differential metabolite changes in mouse kidneys after treatment by ultra performance liquid chromatography coupled with high-definition mass spectrometry(UPLCHDMS).And exploring the effects of aqueous extracts of four Chinese medicines on the changes of metabolites in the kidney of HUA mice based on diversity analysis,differential metabolite screening and correlation analysis.Results:1.The aqueous extract of SRG is contains Neoastilbin,Astilbin,Neoisoastilbin,Isoastilbin and Engeletin.The aqueous extract of AOH is contains 2’-Hydroxyformononetin,Caffeic acid,Procyanidin B1,L-Epicatechin and Galangin.The aqueous extract of RDS is contains Palmitic acid,D-Mannose and Citric Acid.The aqueous extract of PL is contains 12-Hydroxyoctadecatrienoic acid,Luteolin,Eriodictyol-7-O-glucoside,Plumbagine B and(9Z,11 E,13S)-13-hydroxyoctadeca-9,11-dienoic acid.2.In vivo experiments have shown that four herbs can reduce the kidney and liver damage caused by HUA.SRG significantly reduced serum UA levels,ADA and XOD enzyme activities and significantly increased URAT1 protein expression in HUA mice.AOH significantly reduces serum UA levels and ADA enzyme activity in HUA mice.RDS can significantly reduce serum XOD enzyme activity and significantly increase URAT1 protein expression in HUA mice.PL significantly reduced serum BUN levels,ADA and XOD enzyme activities,and significantly increased URAT1 and OAT1 protein expression levels in HUA mice.In addition,in vitro experiments showed that the four herbs had different degrees of elevation on the reduced OAT1 protein expression level caused by UA.3.Four Chinese medicine can ameliorate abnormal changes in the abundance values of genus caused by HUA.such as Parasutterella,Dubosiella,Feacalibaculum,Turicibacter,Bifidobacterium and Parabacteroide.And the above gut microbiota was significantly correlated with the serum biochemical parameters of HUA mice.Thus,Four Chinese medicine may target the above genera to improve HUA.4.Four Chinese medicine reduced HUA-induced Xanthine,L-isoleucine,L-lysine,2-Hydroxycinnamic acid,Pantothenic acid,68-Dihydroxpurine,3-pyridinebutanoic,Piperidine Lyso PC(15:0/0:0)and Indoleacrylic acid differential metabolites were abnormally elevated.Furthermore,a proportion of the differential metabolites were significantly associated with gut microbiota.Thus,AOH,RDS and PL established a renal-intestinal axis to improve HUA.Conclusions:This study demonstrated that SRG,AOH,RDS and PL could improve PO-induced HUA.And reduce serum biochemical disorders,improve minor liver and kidney damage caused by HUA.Furthermore,SRG,AOH,RDS and PL also can regulate the HUA-induced disorders of gut microbiota and renal metabolites.Further correlation analysis speculated that AOH,RDS and PL may regulate the metabolic pathway of uric acid production through the intestinal-renal axis and improve HUA.