The Role Of Ketamine In Human Uroepithelial Cells Apoptosis

Objectives:To evaluate the effects of ketamine-induced apoptosis of human uroepithelial cells (SV-HUC-1), to provide a theoretical basis for the preliminary pathogenesis of ketamine-induced urinary system injury.Methods:The SV-HUC-1cells were cultured under varying concentrations of ketamine (Ommol/L,0.5mmol/L,1mmol/L,2mmol/L) for24hours. Annexin V-FITC/PI double staining flow cytometry was used to detect the ratio of cell apoptosis, and the relative protein levels of Bax, Bcl-2, pro-caspase-3, cleaved Caspase-3in SV-HUC-1cells were analyzed by western blotting.Then, select an appropriate concentration as the optimal working concentration for next experiment, culture SV-HUC-1cells at different duration of times, underwent the above experiments.Experimental datas were statistical analysised by SPSS19.0statistical software. Results:1. Effects of ketamine on the ratio of cell apoptosisThe SV-HUC-1cells were cultured under varying concentrations of ketamine (Ommol/L,0.5mmol/L,1mmol/L,2mmol/L) for24hours, compared with the control group, flow cytometry showed that the ratio of cell apoptosis in each group were (3.33±0.24)%,(5.16±0.57)%,(9.39±0.52)%,(11.33±0.40)%, P values<0.05. Correlation analysis showed that, the ratio of cell apoptosis and the ketamine concentration was positively correlated (r=0.829, P values<0.05).Culture SV-HUC-1cells at different duration of times (Oh,12h,24h,48h),with the ketamine concentration of2mmol/L, compared with the control group, flow cytometry showed that the ratio of cell apoptosis in each group were (3.72±0.22)%,(6.45±0.32)%,(11.18±0.45)%,(12.24±0.40)%, P values<0.05. Correlation analysis showed that, the ratio of cell apoptosis and the duration of stimulate time were positively correlated (r=0.762, P values<0.05).2. Effects of ketamine on the relative protein levels of Bax, Bcl-2, pro-caspase-3, cleaved Caspase-3in SV-HUC-1cellsThe SV-HUC-1cells were cultured under varying concentrations of ketamine (Ommol/L,0.5mmol/L,1mmol/L,2mmol/L) for24hours, the expression of Bax and Cleaved Caspase-3(17KD>19KD) protein was significantly increased compared with the control group, Bax/Bcl-2ratio was increased (P values<0.05), and with the increase of the concentration of ketamine increased protein expression (rBax=0.986, r,9KD=0.888, r17KD=0.769, rBax/Bcl-2=0.982, P values<0.05); Bcl-2and pro-caspase-3protein expression decreased compared with the control group(P values<0.05), and with the increase of the concentration of ketamine decreased expression (rBc1-2=-0.963, rpro_caspase-3=-0.894, P <0.05).Culture SV-HUC-1cells at different duration of times (Oh,12h,24h,48h),with the ketamine concentration of2mmol/L, the expression of Bax and Cleaved Caspase-3(17KD、19KD) protein was significantly increased compared with the control group, Bax/Bcl-2ratio was increased (P values<0.05), and with the extension of time increased protein expression (rBax=0.951, r19KD=0.787, r17KD=0.942, rBax/Bcl-2=0.979, P values<0.05); Bcl-2and pro-caspase-3protein expression decreased compared with the control group(P values<0.05), and with the extension of time decreased expression (rBcl-2=-0.949, rpro-caspase-3=-0.914, P <0.05).Conclusions:Ketamine can induce SV-HUC-1cells apoptosis significantly, occurred in a dose-dependent and time-dependent manner. Illustrates that ketamine may induce apoptosis of bladder epithelial cells, leading to the epithelial cell layer become thinner, so further to damage the subcutaneous muscle tissue, which may be an important mechanism leading to the early ketamine-associated urinary tract injury.