摘要
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。
Aim To understand whether low density lipopro-teins (LDL).very low density lipoprotein(VLDL),especially oxidized low density lipoprotein(ox-LDL)and oxidized very low density lipoprotein(ox-VLDL)have effects on the expression of monocyte chemoat-tractant protein-1(MCP-1)mRNA and protein in cul-tured calf aortic smooth muscle cells(SMCs).Methods After a 24-hour exprosure to LDL,ox-LDL,VLDL and ox-VLDL respectively.the total RNA in calf aortic SMCs was extracted by using the guanidinium isothiocyanate method.The MCP-1mRNA expression in SMCs was examined by slot blot analysis using a γ-32 p-end-labeled 35 mer oligonu-cleotides probe of MCP-1.Meanwhile,MCP-1 pro-tein content in the SMC conditioned media(SMC-CM)of each group was determined by sandwich ELISA.Results Cultured calf aortic SMCs were able to ex-press MCP-1 mRNA and protein,the lever of MCP-1mRNA in SMCs and the concentration of MCP-1 pro-tein in SMC-CM were increased significantly after a 24hour-exposure to ox-LDL and ox-VLDL,respective-ly,whereas the expression of MCP-1 mRNA and pro-tein was only slightly increased when exposed to LDL and VLDL.Conclusion ox-LDL and ox-VLDL can induce a strong expression of MCP-1 in cultured SMCs.
出处
《中国动脉硬化杂志》
CAS
CSCD
1998年第4期301-304,共4页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金!39470289
关键词
脂蛋白
动脉粥样硬化
血管平滑肌
Lipoprotein
Muscle,smooth,vascular
Monocyte chemoattractant protein
Atherosclerosis
