Effects Of Apolipoprotein A - I Mutant Recombinant High Density Lipoprotein On Transcription Factor ATF3

Objective: ATF3 as a TLR-induced repressor that plays an important role in inhibition of specific inflammatory signals. High density lipoprotein(HDL) or its major protein component Apo A-I has significant anti-inflammatory effect. Here we investigate ATF3 effect of Apolipoprotein A-I(Apo A-I) and its cysteine mutant recombinant high density lipoproteinmethod: A-I74, A-IM and A-IWT and A-I74 was expressed by the PET prokaryotic expression system. After purification by affinity chromatography, the recombinant high density lipoprotein was prepared by adding DPPC. Cμltured RAW264.7 cells, inflammatory cell model induced by LPS stimμlation. Then high density lipoprotein intervention. Detection of the expression level of inflammatory cytokines IL-6 and TNF-α by ELISA kit, observation of the cell morphological changes, Changes in the expression levels of m RNA ATF3 and protein were detected in the cells by RT-PCR and western blotting.Results: Cell morphological observation found that compared with the normal group, the LPS group and the rHDL228 group cell morphology tended to development macrophage and degradation, rHDL74,rHDLM and rHDLWT have different protective effects for LPS stimulated cells. ELISA detection showed that rHDL74, and rHDLWT rHDLM make TN-α and IL-6 expression levels returned to close to the blank control group, the level of TNF-α and IL-6 with rHDL 228 groups increased significantly(P < 0.01). Blotting Western results showed that compared with the control group, LPS group and rHDL 228 group ATF3 expression levels decreased, and the difference was significant(p<0.01). The expression levels of rHDL74, rHDLM and rHDLWT group were significantly higher and the difference was significant(p<0.05).Conclusion: Recombinant high density lipoprotein rHDL74, rHDLWT, rHDLM promotes the expression of ATF3 on m RNA and protein levels. And rHDL228 down its expression.