Research Of Synergistic Reaction Between Intestinal Microflora Quorum Sensing Inhibitor And Jiedu Quyu Ziyin Recipe On Treating MRL/lpr Mice

Objective To explore the effect of intestinal microbiota-responsive signal AI-2 inhibitors in the treatment of MRL/Lpr lupus mice by Jiedu Quyu Ziyin Recipe and to find new ways to treat SLE from the microorganism.Methods 28 SPF MRL/lpr female lupus mice were randomly divided into four groups,namely model group,Jiedu Quyu Ziyin Recipe group,brominated furanones group,Jiedu Quyu Ziyin Recipe and brominated furanones combination group.After 7 weeks,the blood,feces,kidneys and intestinal tissues of mice were collected under sterile conditions.The serum creatinine,urea nitrogen,lactate dehydrogenase and alanine transaminase concentration were detected by Blood Biochemical Automatic Detector.The concentration of anti ds-DNA antibody was detected by ELISA.The pathological changes of mice kidneys were observed by HE staining.The expression of IL-1β,IL-6 in kidneys,TLR-7,TLR-9 and IL-6 in intestinal were detected by RT-q PCR.Highthroughput sequencing of Illumina Mi Seq was used for structural analysis and bioinformatics analysis of intestinal microbiota in mice.Principal component analysis and statistical methods were used to analyze the difference intestinal microflora among each group.Results 1.Serological indicators: Compared with model group,the LDH in brominated furanones group decreased(p<0.05),the anti ds-DNA antibody in the medicine group and the combination group decreased(p<0.05).Compared with each group,SCr and LDH were significantly decreased(p<0.01)in combination group while BUN and ALT had no changes.2.Renal pathological detection: Kidneys of model group have mesangial cell and matrix myperplasia proliferation,inflammatory cell infiltration and renal tubular cytoplasmic red dye while medicine group with less inflammatory cell infiltration and mesangial cell proliferation.Brominated furanones group with focal mesangial cell and matrix hyperplasia proliferation.Combined group with little focal mesangial cell proliferation.3.Detection of cytokines: Compared with model group,the other three groups can down-regulate the expression of IL-1β in the kidney,TLR-7,TLR-9,IL-6 in intestinal and the effect of the combination group is the best.4.The variation of alpha diversity of intestinal flora: There were no significant changes among the groups in Shannon,Simpson,Chaol,PD whole tree indexes,which showed that there was no obvious change of species diversity of alpha in each group.5.LEfse on-line analysis: Based on comparison with model group,Propioniba cterium and Rubellimicrobium were reduced in three treatment groups.Lactobac illus and Rikenella were the common change bacterias of the brominated furan ones and combination group.Jeotgalicoccus,Pseudomonas were the common c hanges genus in brominated furanones and medicine group.Dehalobacterium,A naerostipes,Coprococcus,Ruminococcus,Helicobacter and Odoribacter were th e specific changes genus of the brominated furanones group.Parabacteroides,Mucispirillum,Blautia and Oscillospira were specific bacterias of combination group,Desulfovibrio,Anaerotruncus and Bilophila were specific change generas of the medicine group.Based on comparison with combination group,there was no common change genera in the other three groups;Mucispirillum was the common change genera of the brominated furanones and model group;Rikenella and Parabacteroides were the common changes bacterias of the model group and medicine group;Ruminococcus,Helicobacter,Bilophila,Prevotella,Halomonas,Coprobacillus were specially altered genera in the brominated furanones group;Propionibacterium,Lactobacillus,Rubellimicrobium,Blautia,Oscillospira were specially altered genera in the model group;Anaerostipes,Bacteroides,Allobaculum were specially altered genera in the medicine group.Conclusion 1.Jiedu Quyu Ziyin Recipe,brominated furanones and the combination of the two can release the condition of lupus mice and the effect of the combination group was best,which showed that intestinal microbiota-responsive signal AI-2 inhibitors of brominated furanones can cooperate with Jiedu Quyu Ziyin Recipe to treat SLE.2.The treatment groups can regulate the structure of intestinal microorganism and have a different impact on the intestinal microflora.3.In MRL/Lpr lupus mice,intestinal microorganism of Propionibacterium and Rubellimicrobium are related with disease;The intestinal microorganism of anaerostipes can specifically regulate by brominated furanones.Bilophila microo rganism can specifically regulate by Jiedu Quyu Ziyin Recipe.