The Significance Of The ASICs Express In Human Bladder Epithelial Cells And Smooth Muscle Cells

Objective:To explore the expression of ASICs in human bladder epithelial cells and human bladder smooth muscle cell, and observe the effect of amiloride and pH value of medium change on the expression of ASICs and cell current changes.Methods:1.Culture human bladder urothelial cells SV-HUC-1and human bladder smooth muscle cell HBdSMC. And use fluorescence immunoassay to detect the expression protein of ASIC1a and ASIC3;2.Add the inhibitor amilorides of acid sensing ion channels into the medium, detect the expression of ASIC la and ASIC3protein in SV-HUC-1cell line&HBdSMC cell line.3.Change the concentration of extracellular H+and add the ASICs inhibitor amiloride into the medium, detect the current changes of human bladder urothelial cell by using the patch clamp technique..Results:1.Immunofluorescence indicated:containing ASICs in human bladder epithelial cells and smooth muscle cells, not obvious difference between the expression of ASIC1a and ASIC3in bladder smooth muscle cells, but in the bladder epithelial cells, the expression of ASIC3was higher than ASIC1a.2. After the addition of the ASICs inhibitor amiloride, the expression of ASIC1a in human bladder urothelial cell lines decrease (P<0.05), the expression of ASIC3was significantly decreased (P<0.01); the expression of ASIC1a&ASIC3protein in human bladder smooth muscle cell lines have no significant changes (P>0.05).3. Patch clamp tests showed:the corresponding current of ASICs in human bladder urothelial cell line grew with the increased of extracellular H+concentration, the more H+concentration, the higher strength of the current(R=0.832>0.06). Add amiloride the inhibitor of ASICs, the current intensity caused by ASICs in human bladder urothelial cell lines decreased obviously with the same pH.(P<0.05).Conclusions:1. Human bladder epithelial cells and bladder smooth muscle cells contain ASICs, not obvious difference between the expression of ASIC la and ASIC3in human bladder smooth muscle cells, but the expression of ASIC3protein in bladder urothelial cell lines was significantly higher than ASIC1a.2. Amiloride the inhibitor of acid sensing ion channels can decrease the protein expression of ASIC la and ASIC3in human bladder urothelial cells; no obvious effect on the ASICs in human bladder smooth muscle cell line;3. The expression of ASICs in human bladder urothelial cell lines Affected by H+, the over expression and current caused by the change of ASICs can be inhibited by amiloride.