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作 者:章顺荣[1] 高越[1] 洪鸣鸣[1] 王晓楠[1] 封菲[1] 郭士遵[2]
机构地区:[1]杭州市第一人民医院干部科,310006 [2]杭州市第一人民医院心内科,310006
出 处:《浙江医学》2010年第11期1607-1609,共3页Zhejiang Medical Journal
基 金:浙江省医药卫生科技计划项目(2008B145)
摘 要:目的研究氧化低密度脂蛋白(Ox—LDL)对体外培养的人脐静脉内皮细胞缝隙连接蛋白Connexin37基因表达的影响。方法常规人脐静脉内皮细胞体外培养并传代。根据Ox—LDL的不同浓度分为对照组(不加Ox—LDL)及50、100、200mg/L干预组,干预24h后通过RT-PCR方法检测并比较各组内皮细胞Connexin37mRNA的表达。根据Ox—LDL干预的不同时间分为对照组(不加Ox—LDL)以及50mg/LOx—LDL干预6、12、24、48、72h组,检测比较各组Connexin37mRNA的表达情况;并检测比较对照组和100mg/L Ox-LDL干预组间Connexin37蛋白的表达水平。结果50、100、200mg/L的Ox—LDL干预24h后均能引起内皮细胞Connexin37mRNA表达的显著减少(P〈0.01),以50mg/L的Ox—LDL干预效果最明显(P〈0.01).50mg/L的Ox-LDL干预12h后Connexin37mRNA的表达开始显著下降(P〈0.01),24h下降最明显(P〈0.01),48h仍下降(P〈0.05),至72h时恢复到刺激前水平。100mg/L的Ox—LDL干预24h后内皮细胞Connexin37蛋白表达水平显著降低(P〈0.01)。结论Ox-LDL可以抑制人脐静脉内皮细胞Connexin37mRNA和Connexin37蛋白的表达。Objective To investigate the effect of oxidized LDL (Ox-LDL) on the expression of gap junction protein connexin37 gene in cultured human umbilical vein endothelium cell s (HUVEC). Methods Human umbilical vein endothelial cell cultured in normal condition were treated with 0mg/L, 50mg/L,100mg/L and 200mg/L Ox-LDL respectively for 24 h; then the expression of connexin37 gene mRNA in HUVEC was detected by RT-PCR method. The cultured HUVECs were treated with 50mg/L Ox-LDL for 0h,6h, 12h, 24h, 48h and 72h respectively; then the expression of connexin37 mRNA was detected. The expression of connexin 37protein level was also detected by immunocytochemistry in 0mg/L and 100mg/L Ox-LDL groups. Results Ox-LDL at concentration of 50mg/L, 100mg/L and 200mg/L down-regulated mRNA expression level of Connexin37 gene in cultured HUVEC after 24 h intervention (P 〈0.01),while the 50mg/L Ox-LDL was most predominant. After 12 h intervention with 50mg/L Ox-LDL the mRNA expression level began to decrease (P〈0.01), reached the lowest level at 24 h (P〈0.01) and maintained for 48 h (P 〈0.05), then the mRNA expression recovered after 72 h intervention (P 〉0.05). After 24h treatment with 100 mg/L Ox-LDL the connexin37 .protein expression in HUVECs was decreased compared to the control group (P 〈0.01). Conclusion Ox-LDL can down-regulate the protein and mRNA expression of Connexin37 gene in cultured human vein endothelium cells,which indicates that connexin37 may be involved in pro-atherosclerotic effect of Ox-LDL.
关 键 词:缝隙连接蛋白 Connexin37 氧化低密度脂蛋白 人脐静脉内皮细胞 基因表达
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