Mechanism Of Microbiota-Mediated NR3C1 Gene 1₇ Promoter Methylation Regulating The Comorbidity Of NIH-Ⅲ Prostatitis And Depression

BackgroundChronic prostatitis is a common and refractory disease in adult male patients.The proportion of comorbidity with depression is high.But the specific mechanism is not clear.Gut microbiota,prostate microbiota and their metabolites play important roles in the comorbidity of chronic pain and depression,but their roles and molecular mechanisms in the occurrence and development of comorbidity have not been fully elucidated.Objective1.To explore the alteration of prostate microbiota in EAP rat complicated with depression.2.To explore the alteration of gut microbiota in EAP rat complicated with depression.3.To verify the effects of prostate and gut microbiota diversity on the methylation level of NR3C1 gene 17 promoter methylation(DNAm)level after NIH-Ⅲ prostatitis secondary to depression,and to analyze the correlation of peripheral blood mononuclear cells(PBMC)with hippocampal tissue and prostate tissue DNAm level.MethodsA rat model of experimental autoimmune prostatitis(EAP)was constructed by prostate injection with purified prostatitis protein solution.A rat model of prostatitis and depression(EAP+CUMS)was constructed by chronic unpredictability mild stress(CUMS)inducing depression-like behavior.Behavioral,pathological and hematological tests were used to verify the successful modeling.16SrRNA method was used to detect the diversity of prostatic and intestinal flora.The methylation level of CpG island in the 17 promoter region of NR3C1 gene in prostate tissue,peripheral blood mononuclear cells(PBMC)and hippocampus was quantitatively determined by high-throughput heavy sulfite sequencing.Results1.A rat model of EAP complicated with depression was established and confirmed by increases in IL-1β,IL-6,and TNF-α as well as the occurrence of depressive-like behaviors.EAP+CUMS significantly altered the richness,evenness,and composition of prostate microbiota.The alpha diversity and beta diversity of prostatic microbiota changed significantly.The relative abundance of Proteobacteria and Bacteroidetes increased significantly in EAP+CUMS rats,while that of Arthrobacter decreased significantly.Metabolic phenotypic analysis showed that the metabolic pathway of protein N-glycosylation(bacteria)in EAP+CUMS model prostatic microbiota was significantly up-regulated,which is regulated by DCE29,Nocardioes,Helicobacter and Dorea.2.16S rRNA sequence analysis proved the abnormal abundance,uniformity and composition of gut microbiota in EAP+CUMS rats.In EAP CUMS rats.There was no significant difference in the alpha diversity of gut microbiota,but there was a significant change in beta diversity.The abundance of Proteobacteria increased significantly,the abundance of Firmicutes decreased significantly,the abundance of Lactobacillus decreased significantly,and the abundance of Shigella,Blautia,Bacteroides,Dorea and Isobacteria increased significantly in EAP+CUMS rats.In addition,metabolomics profiling revealed the significant difference among three groups,including superpathway of demethylmenaquinol-6 biosynthesis Ⅱ,3-phenylpropanoate and 3(hydroxyphenyl)propanoate degradation to 2-oxopent-4-enoate,4hydroxyphenylacetate degradation,which is regulated by helicobacter,flexispira,and shigella.3.The secretion of proinflammatory cytokines(IL-1β,IL-6,TNF-α)in prostate tissue of rats with EAP/depression increased,which led to prostate tissue edema and glandular duct blockage.At the same time,proinflammatory cytokines(IL-6,TNF-α)could enter the central nervous system through the blood-brain barrier,inducing hypomethylation at key sites such as CpG-10 of NR3C1 gene 17 promoter in hippocampal tissue,resulting in up-regulation of NR3C1 gene expression.At the same time,activate IDO in glial cells,induce its overexpression and induce depressive symptoms.Conclusion1.The disorder of prostatic microbiota and the difference of metabolic phenotype of protein N-glycosylation(bacteria)are the main pathological basis of EAP complicated with depression.2.The disorder of gut microbiota and the difference of short-chain fatty acid metabolism phenotype are one of the pathological bases of EAP complicated with depression.3.The methylation level of NR3C1 gene 17 promoter was decreased due to prostate microbiota disorder and gut microbiota disorder after NIH-Ⅲ prostatitis and depression comorbidities.The CpG-10 methylation level of PBMC was positively correlated with hippocampal tissue,which had certain predictive value.