The Influence Of Folium On Mitochondrial Cytochrome C Expression Of HaCaT Cells

Psoriasis is a common chronic inflammatory skin disease with keratinoCyte hyperplasia,inflammatory cell infiltration and neovascularization as its main pathological changes.It is a common chronic recurrent skin disease,and a typical skin loss of scaly erythema,which is difficult to be cured and easy to relapse.With the increasing of working pressure and change of environmental factors,the incidence of psoriasis showed an increasing trend.As the treatment with western medicine has side effect and the cost is very expensive,what's more,it will seriously affect the life quality of the patients,it is imminent to find better treatments.Objective:In this experiment,rhEGF is used to stimulate the excessive proliferation of HaCaT cells,the objective was to study the action mechanism and the role of Folium on inhibiting proliferation and promoting apoptosis of excessive proliferated HaCaT cells,observe the process of HaCat cell apoptosis induced by Folium,through endogenous pathway mediated by mitochondrial,and investigate the possible mechanism of Folium on the course of treatment of psoriasis.Methods:Culture cells in conventional way and induce excessive proliferation of keratinoCytes with lOng/ml rhEGF,using MTT to observe the effect of Folium extract on HaCat cell proliferation,using flow Cytometry to detect the mitochondrial transmembrane potential(??m)change;using Western blot to detect Cyt-c,Pro/Cleaved caspase-9?Bcl-xl protein expression.Results:(1)Folium extract inhibited HaCaT cell proliferation within the concentration range of 10?g/ml-80?g/ml(P<0.05),and the inhibiting effect is positively related with the concentration,i.e.increasing with the concentration.(2)According to the MTT results,taking Folium extract with the concentration of(0,20,40,60)ug/ml and acting on cells respectively.Folium promotes mitochondrial membrane potential with the increasing concentration.(3)the Western blot results showed the expression of Pro/Cleaved caspase-9?Bcl-xl decreases gradually when the concentration of Folium extract was 20?g/ml,40?g/ml,60?g/ml,while mitochondrial Cyt-c release increased.Conclusions:In a certain range,with the increasing concentration,Folium can prompt the mitochondrial membrane potential,decrease the proteins expression of Bcl-xl,release more Cytochrome c into the Cytosol,activate the Caspase family,and the Cleavedcaspase-9 expression increases.Our results showed that Folium can potently induce HaCat cell apoptosis in intrinsic pathway mediated by mitochondrial,it is a possible mechanism for psoriasis treatment.