Effects Of SCF On The Expression Of C-kit MRNA And Protein Of Interstitial Cells Of Cajal-like Cells In The Bladder Of Guinea Pig In The High Glucose Environment

Purpose:To examine SCF intervention to the interstitial cells of Cajal-like(ICCs) of c-kit mRNA and protein in the bladder of guinea pig in high glucose through reverse transcription polymerase chain reaction (RT-PCR) and laser scanning confocal microscopy(LSCM), and discover the possibility and feasibility which SCF intervention to the shape and function changed ICCs in the high sugar environment.Methods:The ICCs-like cells were isolated from the bladder of guinea pig by collagenase separation, extraction, purification and cultivated for 24h, then glouse medium of 15mmol/L, 30mmol/L concentration was added and continued for 72h (c-kit obviously decrease era).Then different SCF concentration (50ng/ml, 100ng/ml) medium were added for training 24 hours or 72 hours. Last mRNA expression of c-kit was detected by reverse transcription polymerase chain reaction (RT-PCR) and protein expression of c-kit was tested and analyzed by laser scanning confocal microscope.Results:Compared to control group, c - kit mRNA and protein expression levels of high glucose were significantly decreased (P<0.01). In 24h group, c-kit mRNA (P>0.05) and protein expression (P>0.05) of SCF 50ng/ml and 15mmol/L high glucose control group had no difference; c-kit mRNA (P< 0.05) and protein expression(P<0.05)of SCF50ng/ml increased than 15mmol/L high glucose control group; the c-kit mRNA content and protein expression of SCF 50ng/ml and SCF 100ng/ml intervention group has no disparity with 30mmol/L high sugar control group. In72h group, c-kit mRNA (P< 0.01) and protein expression (P<0.01) of SCF50ng/ml and SCF100ng/ml increased significantly than 15mmol/L high glucose control group; c-kit mRNA (P< 0.01) and protein expression (P<0.01) of SCF 50ng/ml increased than 30mmol/L high glucose control group. Conclusion:The microenvironment high sugar environment can reduce the c-kit expression in the guinea pig bladder ICCs, which disturb the SCF/c - kit signal pathway. High SCF concentrations can increase the expression of c-kit in interstitial cells of Cajal which have changed in shape and function, and promote the SCF/c - kit signal pathway to maintain a guinea pig bladder ICCs phenotype and function. From the level of molecular biology, the high SCF intervention in microenvironment had stimulative effect on the c-kit expression of shape and function changed ICCs, which were positively correlated with time and concentration of SCF.