Adipophilin Enhance Cellular Cholesteryl Ester By Inhibiting NCEH Activity Through PKCδ In Macrophages

[Objective]To obtain reliable evidence by adipophilin n CEH induce intracellular lipid accumulation by adipophilin, clarify the mechanism of n CEH induce intracellular lipid accumulation, find new targets for the prevention and treatment of atherosclerosis.[Methods] Adipophilin silencing and retroviral plasmid stable high expression vector was transfected into PA317 packaging cells, collect the virus infected RAW264.7 cells, screening, preparation, and high expression of adipophilin silencing cells. In the high and silencing of adipophilin expression in RAW264.7 cells were tested using m RNA and protein RT- PCR and Western blot to detect the expression of n CEH; Determination of intracellular cholesterol ester by( high performance liquid chromatography, HPLC). The high expression of adipophilin in RAW264.7 cells with PKCδ agonist PMA 0.1 mol/L 30 min after incubation with m RNA and RT- protein levels were PCR and Western blot to detect n CEH expression changes; to detect the contents of intracellular cholesterol ester by HPLC. The high expression of adipophilin in RAW264.7 cells with PKCδ inhibitor Rottlerin 0.1 mol/L 30 min after incubation with m RNA and RT- protein levels were PCR and Western blot to detect n CEH expression changes; to detect the contents of intracellular cholesterol ester by HPLC.[Results]in the high expression of adipophilin in RAW264.7 cells, the cholesterol content is increased significantly, in silence adipophilin expression in RAW264.7 cells, the cholesterol content is significantly reduced; at the same time, the expression of adipophilin can inhibit n CEH m RNA and protein; in addition, the high expression of adipophilin in RAW264.7 cells by PKCδ. Agonist PMA 0.1 mol/L 30 min after incubation with protein expression level of m RNA n CEH decreased significantly; in the high expression of adipophilin in RAW264.7 cells with PKCδ inhibitor Rottlerin 0.1 mol/L 30 min after incubation of m RNA and protein expression level of n CEH increased significantly.[ Conclusions ] The high expression of adipophilin promotes the accumulation of cholesteryl ester in macrophages, inhibiting the expression of n CEH m RNA and protein; Adipophilin promotes lipid accumulation in macrophages by PKCδ inhibitor n CEH.