Ghrelin对THP1源性泡沫细胞ATP结合盒转运子G1调控作用及途径.docVIP

Ghrelin对THP1源性泡沫细胞ATP结合盒转运子G1调控作用及途径 　 作者：王彦富 成蓓 万晶晶 梅春丽 刘玮 柯丽 何平 【摘要】 目的 探讨Ghrelin对人单核细胞系（THP1)源性泡沫细胞ATP结合盒转运子G1(ABCG1)表达的调控作用及其可能途径。方法 体外培养THP1，由佛波酯(PMA)作用使其分化为巨噬细胞，继而在氧化低密度脂蛋白(oxLDL)存在条件下进一步转变为泡沫细胞，油红O染色法鉴定泡沫细胞形成。运用RTPCR法和Western印迹法分别检测Ghrelin干预后及封闭过氧化物酶体增生物激活受体γ(PPARγ)后ABCG1 mRNA与蛋白表达。采用酶法，通过荧光分光光度计检测胆固醇含量及胆固醇流出率。结果 Ghrelin可明显减少细胞内脂滴的形成，增加胆固醇流出，并能显著增加单核／巨噬细胞泡沫化过程中ABCG1 mRNA水平和蛋白表达，此作用呈浓度依赖性。封闭PPARγ后，Ghrelin抑制泡沫细胞形成的效应被阻断，且PPARγ拮抗剂浓度越高，该阻断作用越明显。结论 Ghrelin可能通过上调ABCG1转录翻译水平延缓动脉粥样硬化的发生，且该效应可能通过PPARγ途径发挥作用。 【关键词】 Ghrelin；过氧化物酶体增生物激活受体γ；ATP结合盒转运子G1；泡沫细胞；动脉粥样硬化 【Abstract】 Objective To investigate the effects of Ghrelin on the expression of ATP binding cassette transporter G1 (ABCG1) in THP1 derived foam cells and its possible mechanism. Methods Human monocytic leukemia cell line (THP1) was chosen. The differentiation of THP1 cells into macrophages was induced by phorbol myristate acetate (PMA). Macrophages were then incubated with oxidized LDL (oxLDL) to generate foam cells. Ghrelin and antagonist of PPARγ were treated during foam cells formation. The ABCG1 protein and mRNA levels were detected by Western blotting and RTPCR. The effect of variance of cholesterol content was measured by zymochemistry viafluorospectrophotometer. Results Ghrelin increased the cholesterol efflux obviously. ABCG1 protein and mRNA levels were also decreased. The antagonist of peroxisome proliferatoractivated receptorγ (PPARγ) inhibited the effects of Ghrelin on ABCG1 expression in dosedependent manner. Conclusions Ghrelin might interfere atherosclerosis by upregulating the expression of ABCG1 via PPARγ pathway. 【Key words】 Ghrelin；Peroxisome proliferatoractivated receptorβ；ATP binding cassette transporter G1；Foam cell；Atherosclerosis 　　ATP结合盒转运子G1(ABCG1)是近年发现的膜蛋白。在动脉壁，ABCG1是单核巨噬细胞内胆固醇流出的重要途径，同时还能促进磷脂类及其他亲脂类化合物的流出，有助于清除脂质，防止泡沫细胞形成〔1,2〕。Ghrelin是一种促生长激素释放肽，与糖脂代谢及相关疾病有密切联系。流行病学研究发现血浆Ghrelin水平与代谢综合征的老年患者颈动脉粥样硬化（AS）呈负相关〔3〕，提示Ghrelin在AS的发展中扮演着有益的角色。本研究探讨了Ghrelin对单核／巨噬细胞泡沫化过程中脂质小