论文部分内容阅读
为了解脂蛋白对平滑肌细胞的单核细胞趋化蛋白-1mRNA和蛋白表达的影响，在牛主动脉平滑肌细胞培养基中分别加入低密度脂蛋白、氧化型低密度脂蛋白、极低密度脂蛋白和氧化型极低密度脂蛋白，培养24h，用异硫氰酸胍法提取细胞的总RNA，用γ-32P末端标记的单核细胞趋化蛋白-1的寡核苷酸探针进行狭缝杂交分析，检测平滑肌细胞的单核细胞趋化蛋白-1mRNA的表达。同时用夹心酶联免疫吸附试验检测条件培养基中单核细胞趋化蛋白-1的蛋白含量。结果发现。培养的牛主动脉平滑肌细胞能表达单核细胞趋化蛋白-1mRNA及蛋白，氧化型低密度脂蛋白和氧化型极低密度脂蛋白使其单核细胞趋化蛋白-1mRNA的表达明显增强，同时也使其条件培养基中单核细胞趋化蛋白-1的蛋白水平增加，而低密度脂蛋白和极低密度脂蛋白仅使平滑肌细胞中单核细胞趋化蛋白-1mRNA和蛋白的表达轻度增加。提示氧化型低密度脂蛋白和氧化型极低密度脂蛋白能诱导平滑肌细胞表达高水平的单核细胞趋化蛋白-1。
To understand the effect of lipoprotein on monocyte chemoattractant protein-1 mRNA and protein expression in smooth muscle cells, low density lipoprotein, oxidized low density lipoprotein, very low density lipoprotein And oxidized very-low-density lipoprotein. After culturing for 24 hours, the total RNA of the cells was extracted by guanidinium isothiocyanate method, and slits were made with oligonucleotide probes of γ-32P-labeled monocyte chemoattractant protein-1 Hybridization analysis was performed to detect the expression of monocyte chemoattractant protein-1 mRNA in smooth muscle cells. Simultaneous sandwich enzyme-linked immunosorbent assay was used to detect the protein content of monocyte chemoattractant protein-1 in conditioned media. turn out. Cultured bovine aortic smooth muscle cells can express monocyte chemoattractant protein-1 mRNA and protein, oxidized low-density lipoprotein and oxidized very-low density lipoprotein to enhance the expression of monocyte chemoattractant protein-1 mRNA Also increased the protein level of monocyte chemoattractant protein-1 in its conditioned medium, whereas low density lipoprotein and very low density lipoprotein caused only mild expression of monocyte chemoattractant protein-1 mRNA and protein in smooth muscle cells increase. It is suggested that oxidized low-density lipoprotein and oxidized very-low density lipoprotein can induce smooth muscle cells to express high level of monocyte chemoattractant protein-1.