Effect Of Exogenous Hydrogen Sulfide On The H₂S/CSE System Of Epidermal Cells With The Intervention Of Burned Serum

Objective Takes epidermal cells of the rat cultured in vitro as experimental objects,use the rat serum after burn to simulat the growth environment of epidermal cells,to explore the content of hydrogen sulfide in epidermal cells and the expression level of CSEm RNA in the addition of exogenous hydrogen sulfide,explore the changes of H₂S/CSE system with exogenous hydrogen sulfide and burned rat serum intervened,and determine whether there is H₂S/CSE system in rat epidermal cells.Methods Use 10 normal rats and 42 Thirty percents of III degree burned rats after 18 hours of burn were taken blood from the abdominal aorta,the serum was taken after centrifugation.Takes the epidermal cells of rats cultured in vitro as experimental objects,using the primary epidermal cells of rats and cultured to the fourth generation,then randomly divided into three groups（normal control group,burn group,burn and NaHS group）,laying on the 6 hole plate and cultured for 48 hours,synchronized the epidermal cells with the basic medium.24 hours later,the normal group with intervention of normal rat serum,the burn group with the intervention of burned rat serum,burn and NaHS group with the intervention of burned rat serum and NaHS,then cultured in 1 hour,6 hours,12 hours and collect cells.The content of hydrogen sulfide in epidermal cells was tested by hydrogen sulfide testing kit.The total RNA of epidermal cells from each group was tested by centrifuge column method.After identification,the expression level of CSEmRNA in epidermal cells of each group was tested by RT-qPCRResults1.Take some damage skin from burned rats,and take a biopsy.The pathological report certificate the degree of damage reached the whole of the skin,be confirmed as third-degree burned.2.Comparison of the content of hydrogen sulfide in rat epidermal cells:In the same time,different interventions,the burn and NaHS group compared with the normal control group and the burn group,the content of hydrogen sulfide increased,P<0.05,the results have statistical significance;the content of hydrogen sulfide in the burn group is less than the normal control group,P<0.05,the results have statistical significance.Using the same intervention method,there was no significant change in the content of hydrogen sulfide,P>0.05,the result was not statistically significant.3.Comparison of the expression level of CSEmRNA in each group:According to the intervention time,in the normal control group,the expression level of CSEmRNA was no significant change,P>0.05,the results were not statistically significant.In the burns group and the burn and NaHS group,the expression level of CSEmRNA have differences,P<0.05,the results have statistical significance.According to the different intervention methods,the expression level of CSEmRNA in each group was different,P<0.05,the results were statistically significant.Conclusion1.H₂S/CSE system exist in epidermal cells of rats;2.With the stress of burn,the H₂S/CSE system in the epidermal cells of rats is reduced;3.Tiny amounts of the exogenous hydrogen sulfide can promote the effect of the H₂S/CSE system in the epidermal cells of rats.