苦瓜蛋白对THP-1巨噬细胞源性泡沫细胞的形成及三磷酸腺苷结合盒转运体A1的影响

为研究苦瓜蛋白对泡沫细胞形成的作用及其机制 ,用离心、超滤和色谱技术从苦瓜 (MomordicacharantiaL)果肉中提取苦瓜蛋白 (Momordicin) ,THP 1细胞经 16 0nmol/LPMA孵育 2 4h ,诱导其分化成巨噬细胞 ,然后用 2 5mg/L乙酰化低密度脂蛋白孵育 4 8h ,使其泡沫化 ,以观察苦瓜蛋白对泡沫细胞形成的影响。用高效液相色谱法分析细胞内胆固醇和胆固醇酯的变化 ,用逆转录—聚合酶链反应技术检测苦瓜蛋白对三磷酸腺苷结合盒转运体A1的影响。结果发现 ,经苦瓜蛋白作用后 ,细胞内总胆固醇及胆固醇酯均显著减少 (对照组总胆固醇为 6 34± 2 1,苦瓜蛋白处理组为 2 5 7± 2 6 ,P <0 .0 1;对照组胆固醇酯为 339± 2 1,苦瓜蛋白处理组为 93± 9,P <0 .0 1)。胆固醇酯与总胆固醇比值从对照组的 6 2 .9%下降到 36 .2 %。同时 ,细胞内三磷酸腺苷结合盒转运体A1表达显著上调 (P <0 .0 5 )。提示苦瓜蛋白抑制THP 1单核巨噬细胞源性泡沫细胞的形成可能与上调三磷酸腺苷结合盒转运体A1有关。

Effects of Momordicin on THP-1 Marcrophage-Derived Foam Cells and ATP Binding Cassette Transporter A1

Aim To study the inhibitory effect and mechanisms of Momordicin on foam cell. Methods Momordicin (28 kDa) was purified from the juice of Momordica charantial by eccentricity, super-filtration and chromatography technology. After incubated with 160 nmol/L PMA for 24 h, THP-1 was induced to macrophage, this macrophage was induced to foam cell by incubated with 25 mg/L ac-LDL for 48 h, and the effect of Momordicin on foam cell was measured, cholesteryl and cholesteryl ester within THP-1 were measured by high performance liquid chromatography (HPLC), and regulated effect of Momordicin on ATP binding cassette transporter A1 (ABCA1) was measured by RT-PCR. Results A positive inhibitory effect of Momordicin on foam cell was found. Total cholesterol (TC) and cholesteryl ester (CE) were reduced remarkably by Momordicin(P<0.01), CE/TC was reduced from 62.9% to 36.2% by Momordicin, ABCA1 was up-regulated by Momordicin (P=0.035) remarkably. Conclusions The inhibitory effect of Momordicin on foam cell may related to it's activity of up-regulating ABCA1.