Preliminarily Investigate The Effect Of DDAVP And IL-4 On The Expression Of LPCAT And PAF-AH In The Macrophage

Objectives To explore the effect of DDAVP combined with IL-4 on the expression of LPCAT and PAF-AH m RNA in the RAW264.7 cell line.Methods RAW 264.7 cells were treated in four different ways.After cultured,mRNA of the cell was extract.In the following step,the m RNA extracted was transformed to c DNA.At last,Real-time PCR was used to assess the expression of LPCAT,PAF-AH m RNA.The expression of LPCAT,PAF-AH m RNA was calculated by the fomula expression level=(Etarget)(CTcontrol-CTsample)/(Eref)(CTcontrol-CTsample).CTcontrol is the expansion cycles of actin,CTsample is the expansion cycles of PAF-AH ? LPCAT m RNA,Eref is the of expansion efficiency of actin m RNA, Etarget is the expansion efficiency of PAF-AH ?LPCAT m RNA.The expression level was analysised statistically by SPSS 17.0. Levene method was for homogeneity of variance and LSD t test and Dunnett T3 t test were for difference between the two groups,(P<0.05).The cell line of IL-4 and DDAVP group was cultured in the medium with IL-4(10 ng /ml) for 12 h,then was cultured in the medium with DDAVP(100ng /ml) for 30 min and m RNA of the cell line was extracted.DDAVP group was cultured in the medium without IL-4 and DDAVP for 12 h,then was cultured in the medium with DDAVP(100ng /ml) for 30 min and m RNA of the cell line was extracted.IL-4group was cultured in the medium with IL-4(10 ng /ml) for 12.5h,then m RNA of the cell line was extracted.Control group was cultured in the medium without IL-4 and DDAVP for12.5h,then m RNA was extracted.Results In the IL-4 and DDAVP group,the expression level of LPCAT m RNA was 1.23±0.41.DDAVP group was 0.77±0.35.IL-4 group was 0.95±0.42.Control group was 0.70±0.23.Among four groups,F=2.58,P>0.05,so There is no significant difference in the expression level of LPCAT in these four groups.The expression level of PAF-AH in the IL-4+DDAVP group was 0.54±0.34,DDAVP group was 0.30±0.15,IL-4 group was 0.30±0.11,control group was 0.44±0.34.Among four groups,F=1.237,P>0.05,so there is no significant difference in the expression level of PAF-AH in these four groups.Conclusions DDAVP conbined with IL-4 may up-regulate the expression of LPCAT mRNA.