Study Of The Effects Of Hyaluronan And Melatonin On The Occurrence And Development Of Chronic Nonbacterial Prostatits

Part ? HA/CD44 regulates the Th1 cells differentiation by activating ANX A1/Akt/m TOR signaling to drive the pathogenesis of CNPBackground:Chronic nonbacterial prostatitis(CNP)is a common genitourinary disease in young and middle-aged men.However,current treatment strategies have not yet achieved satisfactory clinical efficacy due to its low cure rate,high recurrence rate and unclear pathogenesis.Numerous evidences indicate that immune dysregulation might play a considerable role in the occurrence and development of CNP.CD44,expressed on the membrane of lymphoid and non-lymphoid cells,takes part in multiple physiological reactions,including homing,activation,and proliferation of lymphocyte cells.Numerous evidences showed that CD44 plays an important role in the occurrence and development of various inflammatory diseases,but the role of CD44 in the pathogenesis of CNP is rarely reported.Materials and Methods:Firstly,the experimental autoimmune prostatitis(EAP)mouse model was established by subcutaneously injecting a mixture of prostate antigen and complete Freund's adjuvant.Lymphocytes from the peripheral blood of CNP patients and prostate tissues from EAP mice were collected for RNA-sequencing to find potential key molecules of CNP.Secondly,Based on EAP mice models,the effects of CD44 and hyaluronan(HA)on EAP were detected by interfering mice with sh CD44-adeno-associated virus and 4-methylumbelliferone.Finally,the potential regulatory molecular pathways of HA/CD44 affecting the occurrence and development of EAP was disclosed via co-immunoprecipitation,silver staining,mass spectrometry and in vitro experiments.Results:We found that CD44 was a key molecule of CNP and it mainly expressed on the infiltrated lymphocytes instead of the local prostate'epithelial cells.Inhibiting the expression of CD44 or HA could significantly alleviate prostatic inflammation of EAP mice.In vitro assay found that HA/CD44 was able to promote the differentiation of Th1 cells.An in-depth mechanism study found that HA can promote the protein interaction of CD44 and ANX A1,following by the activation of Akt/m TOR pathway,consequently bringing out the enhanced differentiation of Th1 cells.After inhibiting the expression of ANX A1 in na(?)ve CD4⁺T cells,the differentiation ratio of Th1 cells and expression of p-Akt and p-m TOR could be significantly reduced.These results indicated that the HA/CD44 axis could promote Th1 cells differentiation by promoting the expression of ANX A1 and activating Akt/m TOR pathway.Conclusions:This study systematically explores the biological role of HA/CD44 in CNP and identifies novel mechanisms that HA/CD44 promotes Th1 cell differentiation.Targeting the HA/CD44/ANX A1/Akt/m TOR signaling represents novel potential therapeutic strategies for patients with CNP.Part ? Melatonin attenuates prostatic inflammation and pelvic pain via Sirt1-dependent inhibition of the NLRP3 inflammasomeBackground: Chronic nonbacterial prostatitis(CNP)is a common male genitourinary system disease.As a neuroendocrine hormone,melatonin possesses a variety of biological functions,among which its anti-inflammatory effects through Sirt1 and NLRPS inflammasome have recently drawn substantial attention.The purpose of the current research was to study the effects of melatonin on CNP and the underlying mechanisms using the mouse model of experimental autoimmune prostatitis(EAP).Methods: We explored the anti-inflammatory effects of melatonin on CNP by measuring the expression of silent information regulator 1(Sirt1)and NLRP3inflammasome-related proteins in EAP mice via Western blotting and immunohistochemical staining.Results: The EAP model mice exhibited severe diffuse leukocyte infiltration and significantly increased pelvic pain compared to the control mice.In melatonin treatment group,the histological appearance of prostate tissues,pelvic pain development,and the levels of pro-inflammatory cytokines were significantly alleviated.Furthermore,we found that protective effects of melatonin were achieved through activation of the Sirt1 pathway and downregulation of the NLRP3 inflammasome.Conclusions: The results indicated that melatonin could attenuate prostate inflammation and pelvic pain by inhibiting the NLRP3 inflammasomes signaling pathway through the activation of Sirt1 in mice of EAP,and these efforts could provide a promising therapeutic strategy for CNP.