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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。
Objective: To investigate the effects of macrophage colony stimulating factor (M-CSF) and protein kinase C inhibitor (STA) on the uptake of oxidized low density lipoprotein (OX-LDL) by murine peritoneal macrophages (MPM) METHODS: Membrane proteins were prepared by M-CSF and protein kinase C inhibitor STA pretreatment MPM. SDS-PAGE and Ligand blotting were performed to determine the value of [250 I] OX-LDL under different conditions. Results: The sialoprotein binding to [125I] OX-LDL was (2.45 ± 0.46) ΜG / G cell protein after pretreatment with sialidase, which was significantly lower than that of the control group .38 ± 1.78) ΜG / G cell protein. After MPM pretreatment with M-CSF and STA, the concentration-dependent saturation curve of [250 I] OX-LDL in the treatment group and the control group showed a linear curve similar to that of the M-CSF group Bmax (453.59 ± 15.39) ΜG / G protein was significantly higher than BMAX (322.77 ± 12.54) ΜG / G protein in the control group, but the KD value did not change much. The BMAX of STZ-treated group was (362.40 ± 15.31) MG / G protein, the KD value was (15.10 ± 2.67) MG / L and the control group BMAX was (264.76 ± 11.29) Protein, KD value was (17.43 ± 2.98) MG / L. CONCLUSIONS: Sialoprotein accepts up-regulation of M-CSF and STA, promotes uptake of OX-LDL by increasing its number on the MPM surface, and promotes foam cell formation.