Therapeutic effects of Qingre jiedu formula on mice with gout based on immune inflammation and gut microbiota
Objective To explore the anti-inflammatory effect of Qingre Jiedu(QRJD)formula on mice with gout and its effect on gut microbiota.Methods Forty C57BL/6 mice weighing 20～22 g were divided into control(CON),model(MOD),allopurinol(ALLO),and QRJD formula(QRJD)groups,and i.g.10 g/0.1 mL carboxymethyl cellulose was administered to the CON every morning from 1 to 35 days.A hyperuricemia mouse model was prepared by intragastric injection of a potassium oxalic acid(500 mg/kg)and yeast extract(10 g/kg)suspension.On day 29,50 μL sterile carboxymethyl cellulose was injected into the right ankle of mice in the CON group under isoflurane-induced anesthesia,and a gouty arthritis model was prepared by injecting the same volume of sodium urate(50 mg/mL)into the right ankle of mice in the other groups.Each group was treated with corresponding drugs every day.On day 35,samples were collected from mice that had been fasted for 6 hours without water.Blood indexes,such as uric acid,creatinine,and urea nitrogen,were assessed.Hematoxylin-eosin and saffranine O-fast green staining was performed on ankle joints.Anti-inflammatory indexes of interleukin-10(IL-10)and transforming growth factor-β1(TGF-β1)were detected by ankle joints immunohistochemical assay.The cecum contents of mice were collected,and changes in gut microbiota were analyzed by high-throughput sequencing of 16S rDNA.Results(1)After 7 days of treatment,compared with the MOD group,QRJD formula effectively reduced the blood concentrations of uric acid(P＜0.001),creatinine(P＜0.01),and urea nitrogen(P＜0.05),and effectively protected renal functions.(2)Compared with the MOD group,HE staining showed that synovial hyperplasia and inflammatory cell infiltration were reduced in the QRJD formula group after treatment.The cartilage arrangement of the compound was more orderly than before,cartilage destruction was less than that in the MOD group,and no matrix loss was observed.(3)Immunohistochemical analysis of the ankle joint indicated that IL-10 and TGF-β1 were not significantly increased in CON and MOD groups.Compared with the MOD group,IL-10 and TGF-β1 expression in the QRJD formula group were increased(P＜0.05).(4)In terms of biodiversity,the number of MOD-specific OTUs increased by 75 compared to the CON group,while the QRJD was able to reduce the number of MOD-specific OTUs to more closely resemble the CON group;no significant difference was found in α-diversity among the four groups(P＜0.05),whereas β-diversity was more similar to the CON group(P=0.001).(5)Compared with the CON group,the MOD group exhibited increased abundances(P＜0.05)of Ruminococcaceae spp.,Dubosiella sp.,Tyzzerella sp.,Ileibacterium sp.,and Bacteroidales spp..In contrast to the MOD group,the QRJD formula group showed elevated abundances(P＜0.05)of Lactobacillus sp.,Ligilactobacillus sp.,and Bacteroides sp..Furthermore,an interaction network of gut microbiota indicated mutual interactions among these microorganisms.(6)In the correlation analysis between gut microbiota and renal functions as well as anti-inflammatory factors,the relative abundances of Dubosiella sp.,Tyzzerella sp.,and Bacteroidales spp.were significantly positively correlated to SUA and SCR(P＜0.05).However,Lactobacillus sp.,Ligilactobacillus sp.,and Mitochondria spp.exhibited a positive correlation to anti-inflammatory factors IL-10 and TGF-β1 with a more significant association observed for TGF-β1(P＜0.05).(7)COG function prediction suggested that the functions of the QRJD formula group were concentrated on inorganic ion transport and metabolism,and carbohydrate transport and metabolism.Conclusions QRJD effectively modulates immune inflammation and gut microbiota dysbiosis,thereby treating gout.Its mechanism of gout prevention and treatment may involve regulation of gut microbiota diversity and abundance,as well as the control of the abundance of differential bacterial species,such as Ruminococcaceae spp.,Dubosiella sp.,and Lactobacillus sp.,to achieve gout therapy.
万方数据
维普
