The Barrier Functions Research Of Prostate Capillary Endothelial Cells And Glandular Epithelium Cells

Backgrounds: Prostatitis is a common and frequently-occurring disease of urology department,men of 35~50 years at some point in his life will be affected by the prostate syndrome.Currently,in clinical practice the presence medication effect of prostatitis is very poor.And the symptoms of chronic prostatitis patients are easy to recur.Cause great damage in patients’ psychological and physiology.Its exact etiology and pathogenesis is still unclear.In prostate disease,prostate stromal exist inflammation,but hemameba are not present in the prostatic fluid,at the same time many kinds of antibacterial drugs in the prostatic fluid concentration was lower than plasma.So some scholars suggest that there is a barrier mechanism between the blood circulation and prostate tissue.Currently,because of the blood prostate barrier makes antimicrobial agents rarely accumulating in the prostate,which leads to insufficient therapeutic concentrations.So the research of blood prostate barrier has important significance for the treatment of prostate disease.In blood brain barrier,capillary endothelial cells play an important role in the blood brain barriers’ barrier properties.In the process of brain development,the capillaries form the blood-brain barrier by differentiation,maturation,and whose permeability is generally determined by the biochemical features of brain capillary endothelial cells,and the permeability will be increased with the increase of the blood vessel diameter.For blood retinal barrier and blood labyrinth barrier,capillary endothelial cells formation of a wide range of tight junction with tissue surround.thereby limiting the biological macromolecules into the brain,for the blood prostate barrier the capillaries of the prostate gland are nonporous is similar with blood brain barrier,blood testis barrier,blood epididymis barrier and blood eye barrier,so capillary endothelial cells may also be involved in the prostate barrier.About the epithelial cells of the prostate gland,some studies of barrier antigen EBA continuity distribution in the reproductive system of rats found that the barrier antigen EBA in rat dorsolateral prostate are continuity distributed.Wenqiang Ma through the experiment also confirmed that the EBA antigen continuously expressed in rat dorsolateral prostate,Therefore the prostate gland epithelial cells may also be involved in the blood prostate barrier,and may play an important role in the blood prostate barrier.Objective: To culture rat prostate capillary endothelial cells and glandular epithelium cells and study their barrier functions in vitro.Methods: Rat prostate capillary endothelial cells were isolated and cultured by cell growth feature selection methods and a gland epithelial cell line was also grown.Prostate capillary endothelial cells were identified by immunofluorescence staining for Factor VIII-related antigen.A co-culture model was then established with both cell types.The tight junction protein claudin-1 was visualized by immunohistochemistry and tight junctions confirmed by Transmission Electron Microscopy(TEM).Transepithelial electrical resistances(TEERs)were monitored with a Millicell system.A phenol red leakage test was also used to assess permeability.And use the rat pulmonary capillary endothelial cells and brain capillary endothelial cells as negative and positive controls.Results:1.We successful isolated rat prostate capillary endothelial cells by growth feature selection methods,which were positive for the endothelial cell marker Factor VIII.2.Immunohistochemistry showed that the tight junction protein claudin-1 was expressed between adjacent capillary endothelial cells and glandular epithelial cells.TEM confirmed formation of tight junctions between the adjacent capillary endothelial and glandular epithelial cells.3.Cultures of rat prostate capillary endothelial cells,glandular epithelial cells,co-cultures model,brain capillary endothelial cells and pulmonary capillary endothelial cells.The peak TEERs of about 142.2±3.1?/cm2,201.3±3.5?/cm2,223±2.3?/cm2,248.20±6.2?/cm2,43.3±3.5?/cm2,respectively and these values remained stable for 2–3 days.As with TEER increased,capillary endothelial cells,glandular epithelial cells,co-cultures model and brain capillary endothelial cells layer permeability to phenol red was decreased.However,in negative control,pulmonary capillary endothelial cells,the change trend of phenol red in basal lateral is not obvious.Their have statistically difference between all cell types(P<0.05).Conclusion:1.Use growth characteristic selection method can successful isolation and culture rat prostate capillary endothelial cells.This method has the advantage of simple operation,no mechanical and chemical damage.2.Rat prostate capillary endothelial and glandular epithelial cells have similar structure and function with brain capillary endothelial cells.Capillary endothelial and glandular epithelial cells cultured in vitro have barrier function.Suggest that prostate capillary endothelial cells and glandular epithelium cells may play a very important role in the blood prostate barrier.3.The co-culture model was closer to the physiological state and could be useful as a research model of the blood–prostate barrier in vitro.