Intervention Of Hydrogen Sulfide On Apoptosis In Renal Tubular Epithelial Cell Induced By Cisplatin

Objective:To study the intervation of hydrogen sulfide on apoptosis in renal tubular epithelial cell by cisplatin.Reserch machine processed on HK-2 cell after affecting by DDP and Na HS.Probe new foundation about H2 S protect DDP injured HK-2 cells in the clinic.Methods:The cell were grouping into three groups according to randomized controlled principle:the normal control group,cisplatin exposure group(In MTT arrey will be 0mg/L,1 mg/L,2mg/L,5 mg/L,10mg/L,20 mg/L,40 mg/L,in the following it will be 20mg/L) and various concentration hydrogen sulfide treatment group(In MTT arrey will be0mmol/L,0.2 mmol/L,0.4 mmol/L,0.5 mmol/L,0.8 mmol/L,1.0 mmol/L?2.0mmol/L Na HS and 20mg/L DDP,in the following it will be 0.5mmol/L or 1.0mmol/L Na HS and 20mg/L DDP).Propagate HK-2 cell of renal tubular epithelial in vitro.The cisplatin-mediated cytotoxic effects were determined by MTT analysis. Using MTT analysis test the apoptosis after different concentrations of hydrogen sulfide and cisplatin combination on HK-2 cells.Morphological changes of the cell can be assessed by DAPI?Annexin V-FITC/PI staining.Annexin V-FITC/PI fiow cytometryanalyzed HK-2 cell's apoptosis.Test the expression product of Daxx m RNA and Fas m RNA,in which study relationship of the drug and apoptosis of HK-2 cells.Results:1.According to MTT,various concentrations of DDP(1?2?5?10?20?40mg/L) accelerate apoptosis of HK-2 cell,the OD value is0.229±0.022;0.198±0.024;0.189±0.069;0.188±0.030;0.165±0.012;0.105±0.0358.And have statistical significance between 20mg/L DDP and 0mg/L DDP(OD value 0.270±0.064)(p<0.05). Add various concentrations Na HS(0.2?0.4?0.5?0.8?1.0mmol/L) into 20mg/L DDP,the OD value is 0.173±0.051;0.186±0.023;0.259±0.050;0.263±0.033;0.268±0.098;0.196±0.025?And have statistical significance between20mg/L DDP and 20mg/L DDP+0.5mmol LNa HS(OD value0.153±0.017)(p<0.05).But when Na HS up 1mmol/L,cell protection down,the OD value will be 0.196±0.025.2.Through the DAPI?Annexin V-FITC/PI staining,under the effect of 20mg/L DDP,The average visual field apoptosis cell number of HK-2cell is 35.02±6.079,while normal negative control group is 4.23±1.015,it has statistical significance between them(p<0.01). Add various concentrations of Na HS(0.5 ? 1.0mmol/L) can reduce apoptosis,the average visual field apoptosis cell number will be 22.55±2.912 ?9.78±2.063.Both of the have statistical significance between DDP group(p<0.05).3.Flow cytometry's result demonstrated that cisplatin induced under the effect of 20mg/LDDP,The apoptosis rate of HK-2 cell is( 31.598±1.014) %,while normal negative control group is(5.167±0.612)%,it has statistical significance between them(p<0.01).Add various concentrations of Na HS(0.5 ? 1.0mmol/L) can reduce apoptosis,the apoptosis rate will be( 18.375±2.239) % ?(11.636±1.233)%.Both of the have statistical significance between DDP group(p<0.05).4.The Real Time PCR sugguest:after HK-2 cells were treated with cisplatin, the DAXX m RNA?Fas m RNA expression product increased compared with those in the control group(p< 0. 05),after combine with hydrogen sulfide of various concentrations,the apoptosis rate significantly declined.(p<0.05)Conclutions:The apoptosis of human renal tubular epithelial cells induced by cisplatin can be protected by hydrogen sulfide,it's of the concentration dependence?...