Effect Of Egg Ylok Oil On The Proliferation Of HaCaT Cells Which Induced By LPS In Psoriasis Vulgaris Model And Bcl-2 Expression

Objective1.To explore the mechanism of egg yolk oil in treating the model of proliferation,which was regards LPS as inducer and HaCaT as model cell;2.To study the function of Bcl-2 in the model of proliferation,regarding LPS as inducer and HaCaT as model cell,which was treated by egg yolk oil.Methods1.The cytotoxicity of egg yolk oil was detected by MTT assay and cell morphology;2.The model of proliferation was established by LPS using in HaCaT,then detecting the proliferation via MTT assay,and the expression ofinflammatory gene IL-8、TNF-αwas detected by RT-PCR;3.To research the effect of apoptosis in the model of proliferatin,which was treated by egg ylok oil,by MTT assay、Hoechst and RT-PCR.The MTT assay was used to detect the inhibition of egg ylok oil in model cell,the Hoechst staining was used to detect the cell nucleus,RT-PCR was used to detect the expression of Bcl-2.Results1.The egg ylok oil and DMSO were acted on HaCaT cell for 24h,the absorbance was low compared with the blank group.P<0.05,the difference was no meaning in statistics.The form of cell which was acted by egg ylok oil and DMSO was no difference,compared with the blank group.The difference was statistically significant.The drug and DMSO was not toxic for cell within 24h;2.LPS stimulation HaCaT cell for 24h and 36h,the increment rate was 32.63%,and 49.61%respectively,Compared with the blank group,P<0.05.the difference was statistically significant.To consider the cost of time,chosing 24h for the follow-up expriment timing to stimulate the cell proliforation.The expression of IL-8、TNF-α was increased after the cell was stimulated by LPS,The proliferation raito was 583.7%,295.7%respectively.The former had statistically significant,P<0.01,compared with blank group.The expression of TNF-α in modling group was meaning in statistics,P<0.05,compared with blank group;3.The all of the egg yolk oil’s concentrations(250tg/mL、500μg/mL、1000μg/mL)had inhibitory effects on LPS-induced HaCaT cell.250μg/mL as the low concentration,500μg/mL as the middle concentration,1000μg/mL as the high concentration.The inhibiton raito was 8.3%,10.1%,16.7%.The low concentration was no meaning in statistic,P>0.05,Compared with the modle group.And the middle and high concentration was statistically significant compared with model group,P<0.05.The concentration was higher,The cell inhibition rate was higher.The expression of Bcl-2、IL-8 and TNF-α was affected by egg ylok oil with different concentration。And the inhibition rate increased with the concentration.The egg ylok oil from 250μg/mL to 1000μg/mL.The inhibition raito was 33.06%,29.43%,57.56%respectively,The inhibiton raito of low and middle concentration were no meaning in statistics,P<0.05,compared with modle group,only the high concentration had statistically significant.P<0.01.Conclusion1.The DMSO can be used as solvent of egg ylok oil,and the concentration of 200μg/ml to 1000μg/ml has no significant inhibitory effect on cell,which was the safe concentration range for drug to act on HaCaT cell;2.LPS can effectively proliferate cells and promote the expression of IL-8 and TNF-α,which can be used as a stimulus to the model of psoriasis in vitro.This model can be as a mature model to study psoriasis in vitro;3.The low、middle and high concentration of egg ylok oil has inhibitory effect acted on LPS-induced HaCaT cell proliferation,as well as reducing the expression of Bcl-2,egg ylok oil inhibition of cell proliferation,may be through reducing the expression of Bcl-2 and promote cell apoptosis.