Ruan Q, Deng Z, Song J
Department of Pathology, Tongji Medical University, Wuhan.
Chin Med J (Engl). 1996 Mar;109(3):206-9.
To understand whether very low density lipoprotein (VLDL) and oxidatively modified very low density lipoprotein (OX-VLDL) can stimulate vascular smooth muscle cells (SMCs) to express high level of monocyte chemotactic protein (MCP-1) mRNA.
LDL and VLDL were isolated from normal blood donors by density gradient ultracentrifugation and oxidatively modified by adding CuCl2 and incubation at 37 degrees C for 24 hours. Rabbit aortic SMCs were cultured by a substrate-attached explant method, and their total RNA was extracted by guanidinium isothiocyanate method. The expression of MCP-1 mRNA in SMCs was examined by dot blotting analysis using a probe of gamma-22P-end-labeled 35-mer oligonucleotide.
Dot blotting analysis showed that cultured SMCs can express MCP-1 mRNA and the level of MCP-1 mRNA was increased significantly in SMCs after 24-hour coculture with VLDL (6-fold) and OX-VLDL (20-fold).
VLDL and OX-VLDL both can induce MCP-1 mRNA expression in cultured rabbit aortic SMCs and the latter is more effective than the former.
了解极低密度脂蛋白（VLDL）和氧化修饰的极低密度脂蛋白（OX-VLDL）是否能刺激血管平滑肌细胞（SMC）高水平表达单核细胞趋化蛋白（MCP-1）mRNA。
通过密度梯度超速离心从正常献血者血液中分离低密度脂蛋白（LDL）和极低密度脂蛋白（VLDL），并通过添加CuCl2和在37℃孵育24小时进行氧化修饰。采用贴壁外植体法培养兔主动脉平滑肌细胞，并用异硫氰酸胍法提取其总RNA。使用γ-22P末端标记的35聚体寡核苷酸探针通过斑点印迹分析检测平滑肌细胞中MCP-1 mRNA的表达。
斑点印迹分析显示，培养的平滑肌细胞可表达MCP-1 mRNA，与VLDL（6倍）和OX-VLDL（20倍）共培养24小时后，平滑肌细胞中MCP-1 mRNA水平显著升高。
VLDL和OX-VLDL均可诱导培养的兔主动脉平滑肌细胞中MCP-1 mRNA表达，且后者比前者更有效。
