摘要
目的了解氧化修饰的低密度脂蛋白（ＯＸＬＤＬ）和极低密度脂蛋白（ＯＸＶＬＤＬ）对内皮细胞（ＥＣ）和单核细胞趋化蛋白１（ＭＣＰ１）ｍＲＮＡ和蛋白表达的影响。方法在牛主动脉ＥＣ培养基中分别加入ＬＤＬ、ＯＸＬＤＬ、ＶＬＤＬ和ＯＸＶＬＤＬ，培养２４小时，用异硫氰酸胍法提取细胞的总ＲＮＡ，用γ３２Ｐ末端标记的ＭＣＰ１寡核苷酸探针进行ｄｏｔｂｌｏｔ分析，检测ＥＣ的ＭＣＰ１ｍＲＮＡ的表达。同时用夹心ＥＬＩＳＡ检测各组ＥＣ条件培养基（ＣＭ）中的ＭＣＰ１蛋白含量。用免疫细胞化学法检测ＥＣ的ＭＣＰ１蛋白的表达。结果培养的牛ＥＣ能表达ＭＣＰ１ｍＲＮＡ及蛋白，ＯＸＬＤＬ和ＯＸＶＬＤＬ使其ＭＣＰ１ｍＲＮＡ的表达明显增强，同时使其ＣＭ中ＭＣＰ１蛋白水平增加，而ＬＤＬ和ＶＬＤＬ仅使ＥＣ的ＭＣＰ１ｍＲＮＡ和蛋白的表达轻度增加。免疫细胞化学显示，对照组、ＬＤＬ组和ＶＬＤＬ组ＥＣ胞浆对ＭＣＰ１抗体均呈弱阳性反应，而ＯＸＬＤＬ组和ＯＸＶＬＤＬ组则呈强阳性反应。
Objective 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 chemoattactant protein 1 (MCP 1) mRNA and protein in endothelial cells (ECs). Methods After a 24 hour exposure to LDL, OX LDL, VLDL and OX VLDL respectively the total RNA in calf aorta ECs was extracted by means of guanidinium isothiocyanate method. MCP 1 mRNA expression in ECs was examined by dot blot analysis using a γ 32 P 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. Results 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 exposured to LDL and VLDL, and immunohistological staining with polyclonal MCP 1 antibody give a similar result. Conclusion OX LDL and OX VLDL are able to induce a strong expression of MCP 1 in ECs.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
1998年第3期174-176,共3页
Chinese Journal of Pathology
基金
国家自然科学基金
关键词
脂蛋白
血管内皮
趋化因子
巨噬细胞
冠心病
Lipoproteins Endothelium, vascular Chemotactic factors, macrophage
