AMD3100 Targeting The CXCL12/CXCR4 Signaling To Selectively Suppress CXCR4+ T Cell Chemotaxis Alleviating Chronic Prostatitis

Research background and purpose:Chronic non-bacteria prostatitis（CNP）is a common disease in urology,and a considerable number of adult men will experience prostatitis-like symptoms.Its main manifestations are pain and discomfort in the lower abdomen and perineum,and abnormal urination,which can lead to male infertility and sexual dysfunction,and seriously affect the patient’s physical and mental health and quality of life.At present,the pathogenesis and pathophysiological changes of CNP are not very clear,but more and more evidences show that immune factors play a key role in the occurrence and development of CNP.Therefore,the purpose of this study is to find genes/pathways that play a key role in the development of CNP,reveal its mechanism,and provide some inspiration for the treatment of CNP patients.Materials and methods:The prostate was extracted from male Sprague-Dawley rats for antigen preparation,and healthy male NOD/Lt J mice were used for CNP model construction.After the model was successfully constructed,the mouse prostate was taken out and divided into two parts,one was used for immunohistochemistry（IHC）staining to assess the inflammatory infiltration state,and the other was saved for RNA sequencing.After acquiring the sequencing data,make scatter plots,volcano plots and cluster plots to display the information of m RNAs,lnc RNAs and circ RNAs,conduct pathway analysis,evaluate the biological functions or signal pathways of genes,drawing and enrichment analysis are performed through python Software execution.After obtaining CNP-related enrichment pathways,KEGG pathway enrichment analysis was used.Select classic enrichment pathways and treat CNP mice with pathway inhibitors to evaluate the clinical significance of these selected pathways.Result:Through RNA sequencing and immunohistochemistry methods,we found that CXCR4,CXCL12,CD44,and OFLM4 were highly expressed in infiltrating inflammatory T cells in the prostate tissue of CNP model mice,but less expressed in epithelial cells.According to the results of enriched pathways,we apply pathway inhibitors to inhibit these classic pathways.We found that AMD3100 targets the CXCL12/CXCR4 axis and significantly reduces the inflammatory infiltration of the CNP model.When we replaced AMD3100 with adeno-associated virus（AAV）-sh Cxcl12,we got similar results.To clarify the underlying mechanism of how the CXCL12/CXCR4 axis affects the pathogenesis of CNP,we tested the classic downstream pathway.The results suggest that p-Akt,p STAT3,and p-NF-κB are highly expressed in the CNP model,and are partially inhibited after the application of AMD3100 or aaf-shcxcl12,suggesting that the CXCL12/CXCR4 axis may signal Akt/NF-κB and STAT3 The pathway affects the pathogenesis of CNP.Conclusion:The m RNAs,lnc RNAs and circ RNAs expression differences between CNP model and negative control were verified by RNA sequencing technology,and CXCR4,CXCL12,CD44 and OLFM4 were determined as diagnostic markers.The CXCL12/CXCR4 axis may affect the pathogenesis of the disease through the AKT/NF-κB and STAT3 signaling pathways.In summary,these results indicate that targeting the newly discovered CXCL12/CXCR4 axis with small molecules AMD3100 or AAVsh Cxcl12 may help to develop new treatments to better inhibit CNP.Our research results provide a good foundation for future research.