Effect Of Estradiol On Disease Activity And Gut Microbiota In Systemic Lupus Erythematosus Mice

Objective:To explore the effect of estrogen on disease severity,gut microbiota and lymphocytes in B6.NZM2410sle1/2/3 mice(TC mice) with systemic lupus erythematosus.Methods:Treatment group:Ten female TC mice aged 8 weeks were implanted with 17β-estradiol release pellet(25ug/day,60 days) in the back of the ears.Control group:Ten female TC mice implanted with 100μL PBS.By observing and recording the symptoms of these two gloups start to appearing.Using ELISA method to detect the serum anti-dsDNA antibodies and B cell activation factor levels.Using hematoxylin and eosin staining method to detect the skin and kidney pathology change of the mice.By using immunofluorescence and immune histochemical methods to detect the immunoglobulin G deposition in the kidney.Using high-throughput sequencingin method to detect the gut microbiota changes of the feces.By using the flow cytometry technology to analysis the differents of intestinal lymphocyte.Resullts:Compared with the TC mice without treated,E2-treated TC mice developed symptoms such as patchy hair loss and anal skin ulceration.HE staining showed a marked atrophy of the skin epithelial layer and a decrease of collagen and the numberh of llicles in the dermis.Renal HE staining showed that glomerular atrophy,sclerosis,tubular atrophy and degeneration,renal interstitial fibrosis.Immunofluorescence and histochemical staining showed that the amount of renal IgG deposition were higher than the control group.Sequencing of feces showed that there were 611 OTUs that these two groups shared together and the number of OTUs in the E2-treated group was higher than the control group.On the class level,the relative abundance of Bacteroidesin the experimental group ware significantly lower than the control group,while Bacillus,difficile,unclassified Bacteroides were significantly elevated;Theα-diversity of gut microbiota showed that the chao1 and shannon index in experimental groups were lower than the control group but there was no significant difference.Theβ-divisity of gut microbiota showed that the LDA index of the treatment group was significantly higher than that of the control group in Firmicutes,Heliconbacteraceae,and Lachnospiraceae.The quantities of Bacteroidia,faecalibaculum and muribaculaceae in the E2 treated group were lower than the control group(P<0.05).Flow cytometry showed that the spleen plasma cells and plasmablasts in treatment group were significantly increased,and the intestinal lamina propria CD4~+ and CD8~+T lymphocytes were significantly higher than the control mice.Conclusion:Estrodiol aggravates the skin and kidney lesions of lupus mice,altering gut microbiota diversity and lymphocyte distribution.