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目的研究Raw264.7细胞经低密度脂蛋白(LDL)诱导后脂质筏中脂类物质组分是否发生变化。方法蔗糖梯度超速离心得到对照组和实验组细胞中的脂质筏,气相色谱质谱(GC-MS)分析脂质筏中脂肪酸的变化;化学衍生脂质筏标志物-单唾液酸四己糖神经节苷脂(Ganglioside 1,GM1),高效液相色谱(HPLC)分析GM1的变化;胆固醇试剂盒检测脂质筏中胆固醇含量的变化。结果与对照组相比,经LDL诱导的Raw264.7细胞脂质筏中的单不饱和脂肪酸(MUFAs)组分中十六烯酸、油酸、二十四烯酸显著升高;多不饱和脂肪酸(PUFAs)组分中亚油酸、花生四烯酸和二十碳三烯酸显著降低;饱和脂肪酸组分中棕榈酸和硬脂酸显著升高,十四烷酸和二十烷酸含量无变化;GM1含量和胆固醇含量均显著增高,有统计学意义。说明LDL改变了细胞脂质筏的脂质微环境。结论 Raw264.7细胞脂质筏中的脂类物质可能与LDL氧化有关,有利于我们进一步研究LDL氧化过程中脂类物质的作用及其与蛋白质的相互作用。
Objective To investigate whether lipid components of lipid rafts in Raw264.7 cells are induced by low density lipoprotein (LDL). Methods Lipofluorescein and gas chromatography-mass spectrometry (GC-MS) were used to analyze the changes of fatty acids in lipid rafts by sucrose gradient ultracentrifugation. The chemical derivatives of lipid raft, monosialotetrahexose Ganglioside 1 (GM1) and high performance liquid chromatography (HPLC) were used to analyze the changes of GM1. Cholesterol kit was used to detect the changes of cholesterol in lipid rafts. Results Compared with the control group, the content of mono-unsaturated fatty acid (MUFAs) in Raw264.7 cell lipid rafts induced by LDL was significantly higher than that of the control group The content of linoleic acid, arachidonic acid and eicosatrienoic acid in PUFAs decreased significantly. The contents of palmitic acid and stearic acid in saturated fatty acids increased significantly. The content of tetradecanoic acid and eicosanoic acid No change; GM1 content and cholesterol content were significantly increased, with statistical significance. LDL has been shown to alter the lipid microenvironment of cellular lipid rafts. Conclusion The lipids in lipid rafts of Raw264.7 cells may be related to the oxidation of LDL, which is beneficial to the further study of lipid interactions and protein interactions in LDL oxidation.