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目的:探讨同型半胱氨酸(Hcy)通过影响脂代谢调控基因LXRα及其下游靶基因ABCA1和ABCG1的表达致人类急性单核细胞白血病细胞株(THP-1)源性巨噬细胞泡沫化在脂代谢紊乱中的作用。方法:培养THP-1单核细胞,佛波酯、氧化低密度脂蛋白(ox-LDL)共同诱导,采用中性红对巨噬细胞染色,并应用流式细胞技术检测巨噬细胞表面CD14的表达,测定巨噬细胞的含量。分为5组,分别加入0、10、50、100、200μmol/L的Hcy培养24h;采用RT-PCR法检测LXRα、ABCA1、ABCG1的mRNA表达;采用Western Blot法检测LXRα、ABCA1、ABCG1的蛋白表达。结果:实验组(Hcy 10、50、100、200μmol/L)泡沫细胞计数阳性百分率均较对照组升高(均P<0.05),其中以Hcy 100μmol/L组升高明显。在Hcy诱导浓度50μmol/L时巨噬细胞51.26%,CD14表达阳性率53.36%;诱导浓度100μmol/L时巨噬细胞98.5%,CD14表达阳性率73.37%。在不同浓度Hcy干预下,泡沫细胞内LXRα、ABCA1、ABCG1的mRNA及蛋白表达均低于对照组(均P<0.05)。结论:Hcy可降低THP-1源性泡沫细胞内LXRα、ABCA1、ABCG1mRNA及蛋白水平的表达,LXRα可能是Hcy影响脂代谢LXRα-ABCA1信号通路中的靶基因。
OBJECTIVE: To investigate the effect of homocysteine (Hcy) on the foaming of human acute myelomonocytic leukemia (THP-1) -derived macrophages by influencing the expression of lipid metabolism regulatory gene LXRα and its downstream target genes ABCA1 and ABCG1 The role of lipid metabolism disorders. Methods: THP-1 monocytes, phorbol esters and ox-LDL were induced to induce macrophages by using neutral red staining. Flow cytometry was used to detect the expression of CD14 on macrophages Expression, determination of macrophages content. The cells were divided into 5 groups and cultured in Hcy at 0, 10, 50, 100 and 200 μmol / L for 24 h. The mRNA expressions of LXRα, ABCA1 and ABCG1 were detected by RT-PCR. The protein expressions of LXRα, ABCA1 and ABCG1 expression. Results: The positive percentage of foam cells in experimental group (Hcy 10,50,100,200 μmol / L) was higher than that in control group (all P <0.05), especially in Hcy 100 μmol / L group. The macrophage 51.26% and CD14 positive rate were 53.36% when the Hcy concentration was 50μmol / L. The positive rate of CD14 was 98.5% and the positive rate of CD14 was 73.37% when the concentration was 100μmol / L. The mRNA and protein expressions of LXRα, ABCA1 and ABCG1 in foam cells were lower than those in control group (all P <0.05). CONCLUSION: Hcy can reduce the expression of LXRα, ABCA1 and ABCG1 mRNA and protein in THP-1 derived foam cells. LXRα may be the target of Hcy in LXRα-ABCA1 signaling pathway.