艳山姜挥发油通过调控巨噬细胞CD36和ABCA1表达抑制泡沫细胞的形成

目的:观察艳山姜挥发油对氧化低密度脂蛋白(ox-LDL)诱导巨噬细胞转化为泡沫细胞的抑制作用,并探索其机制。方法:使用佛波酯(PMA,100μg·L^-1)诱导人白血病单核细胞(THP-1)24 h后形成巨噬细胞,实验分为4组,分别为空白组(无血清RPMI 1640),模型组(80 mg·L^-1ox-LDL),艳山姜挥发油低剂量组(80 mg·L^-1ox-LDL+4μg·L^-1艳山姜挥发油),艳山姜挥发油高剂量组(80 g·L^-1ox-LDL+20μg·L^-1艳山姜挥发油)。噻唑蓝(MTT)比色法检测艳山姜挥发油对巨噬细胞的活性的影响,蛋白免疫印迹法(Western blot)检测巨噬细胞中白细胞分化抗原36(CD36)和三磷酸腺苷结合盒转运体A1(ABCA1)的表达,酶联免疫吸附测定(ELISA)检测巨噬细胞内胆固醇酯含量,油红O染色法检测巨噬细胞中脂质小滴的含量。结果:艳山姜挥发油对巨噬细胞无毒性。与空白组比较,模型组的巨噬细胞内脂滴和胆固醇酯的含量显著增加(P<0.01),CD36蛋白表达显著上升(P<0.01),ABCA1蛋白表达无显著变化;与模型组比较,艳山姜挥发油显著抑制巨噬细胞中脂滴和胆固醇酯的含量(P<0.01),下调CD36的蛋白表达(P<0.01),上调ABCA1蛋白的表达(P<0.01),艳山姜挥发油可抑制巨噬细胞向泡沫细胞的转化。结论:艳山姜挥发油对ox-LDL诱导的巨噬细胞向泡沫细胞的形成具有抑制作用,该药理作用与艳山姜挥发油下调巨噬细胞CD36和上调ABCA1蛋白的表达有关。

Essential Oil from Alpinia zerumbet Rhizome Inhibits Macrophage-derived Foam Cell Formation Via Modulating Expression of CD36 and ABCA1

Objective:To clarify the inhibitory effect of essential oil from Alpinia zerumbet rhizome(EOFAZ)on oxidized low-density lipoprotein(ox-LDL)-induced transformation of macrophage into foam cell and explore its possible mechanism.Method:THP-1 monocyte was incubated with 100μg·L^-1 phorbol myristate acetate(PMA)to grow into macrophage,experiment was divided into 4 groups as follows,control group,model group(80 mg·L^-1 ox-LDL),EOFAZ at low dose(80 mg·L^-1 ox-LDL+4μg·L^-1 EOFAZ)and EOFAZ at high dose(80 g·L^-1 ox-LDL+20μg·L^-1 EOFAZ).Mathye thiazolye telrazliurn(MTT)method was employed to examine the influence of EOFAZ on macrophage viability.Western blot was used to analyze the expression level of cluster of differentiation 36(CD36)and ATP-binding cassette transporter A1(ABCA1)protein in macrophage.Enzyme-linked immunosorbent assay(ELISA)was used to detect cholesteryl ester contents in macrophage.Oil red O staining was applied to determine the accumulation of lipids in macrophage.Result:EOFAZ showed non-toxic effect on macrophage.Compared to control group,macrophage in model group displayed higher level of cholesteryl ester and lipid droplet(P<0.01),as well as significant increasing of CD36 expression(P<0.01),but no effect on ABCA1 expression.EOFAZ notably reduced the contents of lipids and cholesteryl ester(P<0.01),down-regulated expression of CD36 and up-regulated expression of ABCA1 in macrophage in comparison with the model group(P<0.01),indicating that EOFAZ inhibited transformation of macrophage into foam cell.Conclusion:EOFAZ could inhibit ox-LDL-induced transformation of macrophage into foam cell,the underlying mechanism may involves its ability to increase CD36 expression and decrease ABCA1 expression in macrophage.