Interactions Of The Adipophilin With ACAT1/nCEH In Lipid-loaded Macrophages Induced By Oxidized Low Density Lipoprotein

Objective: To explore whether adipophilin interact with ACAT1and nCEH in lipid-loadedRAW264.7cell induced by oxidized low density lipoprotein, try to clarificate themechanism of the change of lipid content in macrophages which overexpressedadipophilin.Method: Bioinformatic predict the relationship between adipophilin and ACAT1/nCEH；RAW264.7cells were incubated with50μg/ml Ox-LDL for for different time. The lipidsaccumulation of cells were stained by Oil red O staining, the expression of mRNA andproteins of adipophilin, ACAT1and nCEH were detected by semi-quantitative reversetranscription-polymerase chain reaction(RT-PCR) and western blot respectively.Interactions between adipophilin and ACAT1/nCEH were detected byCo-immunoprecipitation.Result: Adipophilin were connected with ACAT1and nCEH by Bioinformatic prediction;Oil Red O Dye showed that there were lot of lipid droplets accumulated in RAW264.7mouse macrophages with ox-LDL treatment atfer6hours. As the incubation time ofOx-LDL was extended in RAW264.7cells, the expression of mRNA and proteins ofadipophilin,ACAT1and nCEH were significantly increased, compared with0h,(P＜0.05，n=3).Co-immunoprecipitation showed that there were interactions between adipophilinand ACAT1in RAW264.7mouse macrophages with ox-LDL treatment before3h；therewere not interactions between adipophilin and ACAT1in6hours. There were notinteractions between adipophilin and nCEH in RAW264.7mouse macrophages with ox-LDL treatment before1h, detected by Co-immunoprecipitation； there were interactionsbetween adipophilin and ACAT1in3and6hours.Conclusion: There were interactions between adipophilin and ACAT1/nCEH in RAW264.7mouse macrophages with ox-LDL treatment. It suggests that adipophilin may havesynergistic effects of ACAT1/nCEH in lipid-loaded RAW264.7cell induced by oxidizedlow density lipoprotein.