Summary: 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 an...Summary: 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 48 h. The cells were divided into 4 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 5 % BSA, 100 nmol/L PMA for 48 h as MP group, MCs cultured in RPMI1640 medium with 10 % FBS, 10 μmol/ml leptin for 48 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 4 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.展开更多
文摘Summary: 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 48 h. The cells were divided into 4 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 5 % BSA, 100 nmol/L PMA for 48 h as MP group, MCs cultured in RPMI1640 medium with 10 % FBS, 10 μmol/ml leptin for 48 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 4 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.
