摘要
目的以人的血管内皮细胞ECV304为靶点,研究Ox-LDL和8-Br-cAMP对ATP-结合盒转运子A1(ABCA1)、细胞间粘附因子-1(ICAM-1)及单核细胞趋化因子-1(MCP-1)基因mRNA和蛋白质表达的影响,阐明ABCA1基因在动脉粥样硬化(AS)形成中的可能机制。方法复苏培养血管内皮细胞,无血清培养基培养静止12 h后,分别加入Ox-LDL (30 ng/ml)、8-Br-cAMP (0.5 mmol/L)刺激3、6、12、24 h,设未加刺激同时孵育24 h的细胞为阴性对照组,以RT-PCR和Western蛋白印迹法检测ABCA1、ICAM-1及MCP-1 mRNA和蛋白质表达量。结果血管内皮细胞ECV304在给予Ox-LDL刺激后,ABCA1、ICAM-1、MCP-1的mRNA和蛋白质水平均增高,ICAM-1的高峰时间在12 h,其余高峰时间在6 h;在8-Br-cAMP的刺激下,ABCA1、ICAM-1、MCP-1的mRNA和蛋白质水平也增高,高峰时间均在6 h。结论公认的致AS因素Ox-LDL可通过引起血管内皮细胞ECV304中炎性细胞因子ICAM-1、MCP-1 mRNA及蛋白质水平增加,参与AS的形成,同时抗AS基因ABCA1在Ox-LDL刺激下代偿增加,具有AS保护作用。cAMP不仅可增加ABCA1的表达,而且可增加ICAM-1、MCP-1表达。
Objective To investigate the changes in the expressions of ATP-binding cassette transporter A1 (ABCA1), intercellular cell adhesion molecule type 1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) at mRNA and protein levels after the treatment of the vascular endothelial ECV304 cells with oxidized low-density lipoprotein (Ox-LDL) and 8-Br-cAMP, thereby to explore possible mechanisms by which ABCA1 affects the formation of atherosclerosis (AS). Methods Resuscitated and cultured ECV304 cells were incubated in serum-free medium for 12 h to induce the quiescence phase of growth, which were subsequently treated with Ox-LDL (30 ng/ml) and 8-Br-cAMP (0.5 mmol/L) respectively for 3, 6, 12, and 24 h. After the cells were harvested at the specified time points, the mRNA and protein levels of ABCA1, ICAM-1 and MCP-1 were detected by RT-PCR and Western blotting, respectively. Results The mRNA and protein expression levels of ABCA1, ICAM-1 and MCP-1 all increased after treatments with Ox-LDL and 8-Br-cAMP. The highest expression of ICAM-1 occurred in cells with a 12-hour treatment, and those of ABCA1 and MCP-1 occurred following 6-hour incubation with Ox-LDL. The expression peaks of ABCA1, ICAM-1 and MCP-1 all took place after 6-hour incubation with 8-Br-cAMP. Conclusions The mRNA and protein expressions of ICAM-1 and MCP-1 in vascular endothelial ECV304 cells increase in response to Ox-LDL treatment, in the event of which ABCA1 is also up-regulated to offer protective effects against AS. cAMP not only enhances the expression of ABCA1 but also those of ICAM-1 and MCP-1.
出处
《第一军医大学学报》
CSCD
北大核心
2004年第9期980-983,共4页
Journal of First Military Medical University
基金
国家自然科学基金(30171028)
广东省自然科学基金(010616)~~
