The Role Of Exosomes In Type ?A Chronic Prostatitis

Background:Category III prostatitis(CP/CPPS)is the most common type of chronic prostatitis.The typical clinical symptoms include genital or pelvic discomfort,with urinary symptoms and sexual dysfunction,lasting for at least 3 months.CP/CPPS is subclassified as an inflammatory type(IIIA)and a noninflammatory type(IIIB)according to the presence of leukocytes in EPS or EPS-urine.For some patients,the therapeutic effort of conventional treatments is poor,clinical symptoms relapse frequently.The etiology of CP/CPPS is unknown,many factors are involved.Recent researches show that inflammation and immunity have important roles in the disease.Exosomes are extracellular membraneous nanovesicles with a diameter of 30-150 nm,derived by cells,can be found in different body fluids such as plasma?urine?saliva?cerebrospinal fluid.The membrane of exosomes is rich in an array of lipids,such as cholesterol,sphingomyelin,ceramide.Exosomes bear proteins,lipids,and RNAs,mediating intercellular communication between different cell types in the body,and thus affecting normal and pathological conditions.The release of exosomes is markedly increased in inflammation and caner.Emerging evidences have indicated that exosomes play important roles in inflammation and immunological diseases,and as messager s between cells,exosomes have pro-inflammatory effect.They induce the production of inflammation-associated cytokines,promote inflammatory response in diseases.So we speculate that by delivering inflammatory information,exosomes may promote inflammatory response and contribute to CP/CPPS.In this study,we want to isolate and characterize exosomes from IIIA CP/CPPS patients and healthy controls,explore weather EPS-derived exosomes have pro-inflammatory effect on prostate stroma cell(WPMY-1)and the role of exosomes in CP/CPPS.Methods:1?Exosomes were isolated from the EPS and EPS-urine of IIIA CP/CPPS patients and healthy controls by ultracentrifugation.Transmission electron microscopy,Western-blot and particulate size description analyser were used to characterize exosomes.2?Proteins were quantified by BCA assay.Exosomes were quantified using Exosome Quantification Assay Kit(ExoELISA-ULTRA CD63).3?Labeled by lipophilic dye(PKH-26)exosomes were incubated with WPMY-1 cells.After 12 h,WPMY-1 cells were stained and analyzed using confocal laser scanning microscopy.4?WPMY-1 cells were incubated with EPS-derived exosomes for 48 h.Production of IL-1?,IL-8 and TNF-a were measured.Results:1?We isolated exosomes from the EPS and EPS-urine of IIIA CP/CPPS patients and healthy controls by ultracentrifugation.Exosomes were identified by transmission electron microscopy?Western-blot and particulate size description analyser.2? The levels of EPS-derived exosomes and EPS-urine derived exosomes were significantly higher in IIIA CP/CPPS patients than in healthy controls.3?After incubated for 12 h,EPS-derived exosomes were internalized by WPMY-1 cells.4?WPMY-1 cells were co-cultured with EPS-derived exosomes for 48 h.EPS-derived exosomes from IIIA CP/CPPS patients induced a higher production of IL-8 compared to healthy exosomes.IL-1? and TNF-a have no difference.Conclusion:The levels of EPS-derived exosomes and EPS-urine derived exosomes were increased in IIIA CP/CPPS.The EPS-derived exosomes from IIIA CP/CPPS patients have pro-inflammatory effect,induced WPMY-1 cells to produce significantly more IL-8.Exosomes may play a role in IIIA CP/CPPS.