The Research Is The Effects Of Cystathionine-γ-lyase/Hydrogen Sulfide System Of Rat Skin Fibroblasts By Exogenous Hydrogen Sulfide

Objective The study is to explore the changes of in vitro culture of normal rat skinfibroblasts H2S concentration in the supernatant liquid and cystathionine-γ-lyasemRNA expression level which was added exogenous hydrogen sulfide and inhibitorof cystathionine-γ-lyase, to discuss the cystathionine-γ-lyase/hydrogen sulfide systemis or not exist in the normal rat skin fibroblasts.Methods Normal rat skin fibroblasts are the research object, cultivating andextending these fibroblasts with normal culture method, choosing the third generationof fibroblasts to divided into a blank group，a contrast group, PPG groups and NaHSgroups randomly. Setting up three deputy holes in cell culture plate of each group,each hole put in the same concentration and the same number of fibroblasts. Culturingcells for48hours, synchronize the cells and adding different concentration ofintervention, collecting cells and the supernatant after another24h.Each group ismeasured the H2S concentration in supernatant liquid by the sensitive sulphurelectrode assay, extracting the total RNA of fibroblasts with Trizol method andauthenticate total RNA, measured the expression level of cystathionine-γ-lyasemRNA by RT-PCR.Results The H2S concentration in supernatant liquid of NaHS groups is higher thanblank group and control group (P <0.05),the H2S concentration in supernatant liquidof PPG groups is lower than blank group and control group (P <0.05).There is nosignificant difference between the blank group, control group, NaHS group（N1,N2,N3）,PPG group（P1,P2,P3）of rats normal skin fibroblasts CSE mRNAexpression level.Conclusion H2S/cystathionine-γ-lyase system exist in the normal rat skin fibroblastswhich were cultivated extracorporeal. Exogenous hydrogen and PPG can not makechanges of CSEmRNA expression level in rat normal skin fibroblasts which wascultured outside body.