Effects of leptin on expression of acyl-coenzymeA: Cholesterol acyltransferases-1 in cultured human monocyte-macrophages

Abstract

To investigate the effects of leptin on expression of acyl-coenzymeA: cholesterol acyltransferases-1 (ACAT-1) in monocyte macrophage differentiation, human monocytic cells (THP-1) were cultured in RPMI 1640 and made to differentiate into macrophages under the incubation with phorbol myristate acetate (PMA) for 18 h. The cells were divided into 1 groups according to different intervention factors as follows: MCs cultured in RPMI1640 medium with 10% FBS for 48 h served as MC group (control group). MCs cultured in medium with serum-free RPMI1640 containing 5⁴r BSA. 100 nmol L PMA for 18 h as MP group. MCs cultured in RPMI1640 medium with 10%, FBS, 10 μmol/ml leptin for 18 h as leptin-MC group, and MCs cultured in medium with serum-free RPMI1640 containing 5% BSA. 100 nmol L PMA, and 10 μmol/ml leptin for 48 h as leptin MP group. Immunocytochemistry, reverse transcription polymerase chain reaction (RT-PCR) and Western blot were performed, respectively, to observe the effects of leptin on expression of ACAT-1 in the monocyte macrophage differentiation. Our results showed that expression of ACAT-1 protein and mRNA in MP-group is two times that in MC group (P<0.05), and the expression of ACAT 1 protein and mRNA increased by up to 1 folds in leptin-MP group as compared with that of MC group (P<0.01). Thus, our results support the idea that expression of ACAT-1 increases more in cultured human macrophages than in monocytes, and leptin can significantly promote ACAT 1 expression. It was concluded that high expression of ACAT-1 may accelerate the development of human atherogenesis, and leptin might participate in atherogenesis by increasing expression of ACAT 1.

Keywords

acyl CoA / cholesterol acyltransferases-1 / monocyte / macrophage

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