The Role Of ATP-sensitive Potassium Channel In The Effect Of Sodium Hydrosulfide On The Intervention Of Burn Serum On Epidermal Cells

Objective:To investigate whether exogenous hydrogen sulfide（H₂S）plays a role in the process of injury to epidermal cells by burn serum by interfering with ATP-sensitive potassium channels(K_ATP channels).Methods:The epidermal cells extracted from the back of the newborn rats for 1 day were subcultured to the third generation and randomly divided into 6 groups:fetal bovine group,normal control group,burn control group,NaHS group,glibenclamide（Gli）group,Gli+NaHS group,inoculated in 96-well plate,and then cultured for 24hours,then synchronized.Fetal bovine serum,normal rat serum and burn serum medium were used in fetal bovine control group,normal control group and burn control group.The cultures were divided into 1,2,4,6 and 8 hours respectively.NaHS group and Gli group were treated with burn serum medium respectively,and then NaHS solution with concentration（50,100,150,200,250μmol/L）was used respectively.After intervention with 200μmol/L Gli solution,Gli+NaHS group was treated with 10%burned rat serum medium,and then treated with Gli and NaHS respectively.Finally,CCK-8 reagent was added and determined in the microplate reader at 450 nm.The OD（absorbance）value was obtained.After extracting the total protein of epidermal cells from each group,the expression level of KATP channel protein was detected by Western blot.Results:1.At the same time point,there was no statistically significant difference in OD value between the fetal calf group and the normal group（P>0.05）,and the difference between the normal group and the burn group was statistically significant after 4h（P<0.05）.2.Within the same concentration gradient,OD values of the burn group and the NaHS group were compared.When the intervention concentration of NaHS was C<150μmol/L,P>0.05,the difference was not statistically significant;when the intervention concentration of NaHS was C≥150μmol/L,P<0.05,the difference was statistically significant.3.At the same time,the OD value of burn group and Gli group was higher than that of Gli group（P<0.05）,and there was significant difference between burn group and Gli group.The OD value of NaHS group was higher than that of Gli+NaHS group（P<0.05）,and there was significant difference between NaHS group and Gli+NaHS group（P<0.05）.4.The expression level of K_ATP channel protein in the burn group was lower than that of the normal group and fetal bovine group,P<0.05,indicating a statistically significant difference.The expression level of K_ATP channel protein after the intervention of exogenous H₂S was higher than that of the burn group,and the expression level of K_ATP channel protein after the addition of Gli was lower than that of NaHS group,indicating a statistically significant difference.Conclusions:1.After the burn serum intervention time is more than 4h,it exerts obvious damage on the cells;2.Exogenous hydrogen sulfide NaHS intervention concentration can reach 150μmol/L,can play a significant cytoprotective effect;3.K_ATP channel blocker Gli can significantly inhibit the cytoprotective effect of NaHS and reduce the survival rate of cells.4.Exogenous H₂S antagonizes the injury of epidermal cells by inhibiting burn serum by regulating K_ATP channel protein level,and protects epidermal cells after burn.