Geriatric Department, Hangzhou First Peoples Hospital, Hangzhou, Peoples Republic of China.
Cell Biol Int. 2012 May 1;36(5):497-502. doi: 10.1042/CBI20110466.
The role of connexins in atherogenesis has attracted increasing attention in recent years; however, whether oxidized LDL (low density lipoprotein) is one of the pro-atherosclerotic factors that can influence the expression of endothelial connexins remains unclear. To investigate the effect of oxidized LDL on the gene expression of the gap junction proteins connexin37, connexin40 and connexin43 in cultured HUVEC (human umbilical-vein endothelial cells) in vitro, HUVEC were cultured and divided into groups before being stimulated with various concentrations of oxidized LDL for different times. mRNA expression of connexin37, connexin40 and connexin43 in each group were determined semi-quantitatively by RT-PCR (reverse transcription-PCR), while protein expression of the three connexins was investigated by immunocytochemical staining. Various concentrations of oxidized LDL down-regulated mRNA and protein expression of connexin37, and up-regulated the expression of connexin40 and connexin43. Time-dependent dynamic alterations of the mRNA expression levels of connexin37, connexin40 and connexin43 were also found in HUVEC stimulated by oxidized LDL. We conclude that oxidized LDL can change the expression of connexin genes in cultured HUVEC within a short period, and thus may be involved in the pro-atherosclerotic effect of oxidized LDL.
近年来,缝隙连接蛋白在动脉粥样硬化形成中的作用受到越来越多的关注;然而,氧化型低密度脂蛋白(low density lipoprotein,LDL)是否是影响内皮细胞缝隙连接蛋白表达的促动脉粥样硬化因子之一尚不清楚。为了探讨氧化型 LDL 对体外培养的人脐静脉内皮细胞(human umbilical-vein endothelial cells,HUVEC)中缝隙连接蛋白 connexin37、connexin40 和 connexin43 基因表达的影响,将 HUVEC 进行培养并分组,然后用不同浓度的氧化型 LDL 刺激不同时间。用 RT-PCR(逆转录-PCR)半定量检测各组 connexin37、connexin40 和 connexin43 的 mRNA 表达,用免疫细胞化学染色法检测 3 种连接蛋白的蛋白表达。结果发现,不同浓度的氧化型 LDL 下调了 connexin37 的 mRNA 和蛋白表达,而上调了 connexin40 和 connexin43 的表达。用氧化型 LDL 刺激的 HUVEC 中还发现了 connexin37、connexin40 和 connexin43 的 mRNA 表达水平的时间依赖性动态变化。我们的结论是,氧化型 LDL 可以在短时间内改变培养的 HUVEC 中连接蛋白基因的表达,因此可能参与氧化型 LDL 的促动脉粥样硬化作用。