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目的观察并检测脂肪型脂肪酸结合蛋白(FABP4)在THP-1单核细胞经巨噬细胞向泡沫细胞分化过程中的表达改变及其DNA启动子区甲基化变化,探讨FABP4在单核源性泡沫细胞形成中的作用。方法体外培养THP-1单核细胞,分别经佛波酯(PMA)和氧化低密度脂蛋白(ox-LDL)诱导分化为巨噬细胞和泡沫细胞,采用实时荧光定量PCR(qRT-PCR)和Western blot检测三种细胞中FABP4的mRNA和蛋白表达;巢式降落式甲基化特异性PCR(nt-MSP)法检测FABP4 DNA甲基化水平。结果 THP-1单核细胞经巨噬细胞向泡沫细胞分化过程中,与单核细胞相比FABP4 mRNA表达量分别增加了3.87倍和2.08倍(P<0.01);蛋白表达量分别增加了1.21倍和0.69倍(P<0.01);各组之间FABP4甲基化水平无明显差异(P>0.05)。结论FABP4可能参与了THP-1单核细胞向巨噬细胞和泡沫细胞的分化过程,其机制可能与FABP4 DNA甲基化无关。
Objective To observe and detect the expression of FABP4 in the process of THP-1 monocyte-derived macrophages differentiating into foam cells and the methylation of DNA promoter region, and to explore the role of FABP4 in monocyte-derived The role of foam cells in the formation. Methods THP-1 monocytes were cultured in vitro and differentiated into macrophages and foam cells by phorbol ester (PMA) and ox-LDL respectively. Real-time quantitative PCR (qRT-PCR) The expression of FABP4 mRNA and protein in the three cell lines was detected by Western blot. The DNA methylation level of FABP4 was detected by nested-drop-specific methylation-specific PCR (nt-MSP). Results Compared with monocytes, FABP4 mRNA expression in THP-1 monocytes increased by 3.87-fold and 2.08-fold (P <0.01) compared with monocytes, respectively. The protein expression of THP-1 monocytes increased by 1.21-fold And 0.69 times (P <0.01) respectively. There was no significant difference in the methylation level of FABP4 between the two groups (P> 0.05). Conclusion FABP4 may be involved in the differentiation of THP-1 monocytes into macrophages and foam cells. The mechanism may be related to FABP4 DNA methylation.