Yu G, Deng Z, Qu Z
Department of Pathology, Tongji Medical University, Wuhan 430030.
Zhonghua Bing Li Xue Za Zhi. 1998 Jun;27(3):174-6.
To clarify whether lipoproteins, particularly oxidatively modified low density lipoprotein (OX-LDL) and very low density lipoprotein (OX-VLDL) play a role in the express of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein in endothelial cells (ECs).
After a 24-hour exposure to LDL, OX-LDL, VLDL and OX-VLDL respectively the total RNA in call aorta ECs was extracted by means of guanidinium isothiocyanate method. MCP-1 mRNA expression in ECs was examined by dot blot analysis using a gamma-32P-end-labelled 35 mer oligonucleotide probe of MCP-1. Meanwhile, MCP-1 protein in EC conditioned media (EC-CM) of each group was determined by sandwich ELISA, and MCP-1 protein in ECs was examined immunocytochemically as well.
Cultured calf aorta ECs expressed MCP-1 mRNA and protein, and OX-LDL and OX-VLDL induced further a strong expression of MCP-1 mRNA, and an increased MCP-1 protein level in EC-CM. The expression of MCP-1 mRNA and protein was only slightly increased when exposed to LDL and VLDL, and immunohistological staining with polyclonal MCP-1 antibody give a similar result.
OX-LDL and OX-VLDL are able to induce a strong expression of MCP-1 in ECs.
阐明脂蛋白，尤其是氧化修饰的低密度脂蛋白（OX-LDL）和极低密度脂蛋白（OX-VLDL）是否在内皮细胞（ECs）中单核细胞趋化蛋白-1（MCP-1）mRNA和蛋白的表达中发挥作用。
分别将主动脉内皮细胞暴露于低密度脂蛋白、氧化修饰的低密度脂蛋白、极低密度脂蛋白和氧化修饰的极低密度脂蛋白24小时后，采用异硫氰酸胍法提取细胞总RNA。使用γ-32P末端标记的35聚体MCP-1寡核苷酸探针通过斑点印迹分析检测内皮细胞中MCP-1 mRNA的表达。同时，采用夹心酶联免疫吸附测定法测定每组内皮细胞条件培养基（EC-CM）中的MCP-1蛋白，并通过免疫细胞化学方法检测内皮细胞中的MCP-1蛋白。
培养的小牛主动脉内皮细胞表达MCP-1 mRNA和蛋白，氧化修饰的低密度脂蛋白和氧化修饰的极低密度脂蛋白进一步诱导MCP-1 mRNA的强烈表达，并使内皮细胞条件培养基中MCP-1蛋白水平升高。当暴露于低密度脂蛋白和极低密度脂蛋白时，MCP-1 mRNA和蛋白的表达仅略有增加，用多克隆MCP-1抗体进行免疫组织化学染色得到类似结果。
氧化修饰的低密度脂蛋白和氧化修饰的极低密度脂蛋白能够诱导内皮细胞中MCP-1的强烈表达。
