【摘要】 目的:探讨巨噬细胞集落刺激因子(M-CSF)和蛋白激酶C抑制剂(STA)对小鼠腹腔巨噬细胞 (MPM)巨涎蛋白摄取氧化低密度脂蛋白(OX-LDL)的影响。方法:M-CSF与蛋白激酶C抑制剂STA预处理MPM后制备膜蛋白,开展SDS-PAGE电泳及配体印迹试验,测定不同条件下巨涎蛋白结合[125I]OX-LDL的值。结果:MPM 膜蛋白经唾液酸酶预处理后,巨涎蛋白与[125I]OX-LDL的结合值为(2．45±0．46)ΜG/G细胞蛋白,显著低于对照组的 (58．38±1．78)ΜG/G细胞蛋白。用M-CSF与STA预处理MPM后,处理组与对照组巨涎蛋白结合配体[125I]OX-LDL 呈现一趋向饱和的浓度曲线,经线性回归得两类似平行的直线,M-CSF组BMAX(453．59±15．39)ΜG/G蛋白,显著高于对照组的BMAX(322．77±12．54)ΜG/G蛋白,而KD值变化不大。STA处理组BMAX(362．40±15．31)ΜG/G蛋白,KD值为 (15．10±2．67)MG/L,对照组BMAX为(264．76±11．29)ΜG/G蛋白,KD值为(17．43±2．98)MG/L。结论:巨涎蛋白接受M- CSF和STA的上调作用,通过增加其在MPM表面数目而促进对OX-LDL的摄取,促进泡沫细胞的形成。更多还原

【Abstract】 ABM: To investigate the effects of human recombininant macrophage colonly - stimulating factor (M - CSF) and protein kinase C inhibitor, staurosporine (STA) , on the binding of macrosialin of the mouse peritoneal macrophages (MPM) to ox - LDL. METHODS: MPM was disrupted by using ultrasonic pulse method after preincubation of M - CSF and STA. The plasma membrane proteins were separated by SDS - PAGE under nonreducing condition. The separated proteins were transferred to a nitrocellulose membrane. The ligand blotting and immunoblotting were used. The effects of M - CSF and STA on expression of cell surface receptor were observed by means of autoradiography. RESULTS: Preincubation with medium containing neuraminidase dramatically decreased the binding of macrosialin to [125I] - ox - LDL [(2.45 ± 0.46) μg/g cell protein vs (58.38 ± 1.78) (μg/g cell protein]. When pretreated with M - CSF, macrosialin bound predominantly to ox - LDL. The Bmax of [125] - ox - LDL to macrosialin increased when macrophages were treated with M- CSF [(322.77± 12.54) vs (453.59± 15.39) μg/g] . Meanwhile, the dissociation constant of treated group showed little changes [(29.06±2.87) vs (28.26 ±4.10) mg/L]. The similar result was found when macrosialin was pretreated with STA, a protein kinase C inhibitor. The Bmax of [125I] - ox - LDL to macrosialin increased [(264.76± 11.29) vs (362.40± 15.31) μg/g]. The dissociation constant of treated group showed little changes either [(17.43±2.98) vs (15.10±2.67) mg/L]. CONCLUSION: The results indicate that M - CSF and STA enhance the binding of macrosialin to ox - LDL by increasing the cellular surface receptor number.更多还原