尾加压素Ⅱ诱导巨噬细胞表达单核细胞趋化蛋白1上调及机制研究  

UrotensinⅡinduces upregulation of monocyte chemoattractant protein-1 expression in macrophages and its mechanism

作　　者：邸北冰[1] 丁文惠[1] 赵静[1] 宋娜娜[1] 董晓[1] 褚松筠[1] 唐朝枢[2] 

机构地区：[1]北京大学第一医院心内科,100034 [2]北京大学第一医院分子心血管学教育部重点实验室,100034

出　　处：《中华老年心脑血管病杂志》2011年第10期925-928,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(30871066)

摘　　要：目的观察尾加压素Ⅱ(UⅡ)对小鼠巨噬细胞(RAW264.7细胞)表达和分泌单核细胞趋化蛋白1(MCP-1)的影响。方法 RAW264.7细胞用UⅡ10^(-10)、10^(-9)、10^(-8)和10^(-7)mol/L刺激后,检测MCP-1 mRNA和蛋白的水平;还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶抑制剂二苯基碘(DPI)或抗氧化剂N-乙酰半胱氨酸(NAC)预处理细胞后,再加入UⅡ10^(-7)mol/L刺激,检测MCP-1 mRNA表达和蛋白的分泌;用UⅡ刺激细胞不同时间段后,检测NADPH氧化酶亚基的表达水平;用UⅡ刺激细胞后,流式细胞仪检测细胞活性氧的生成。结果与不加UⅡ时比较,UⅡ10^(-7)mol/L刺激12 h后,巨噬细胞MCP-1 mRNA表达和蛋白分泌分别提高了100.5%和57.0%(P<0.01)。UⅡ刺激12 h后,p47phox mRNA表达为不加UⅡ时的1.96倍(P<0.01)。UⅡ刺激6 h后,巨噬细胞活性氧为不加UⅡ时的1.8倍(P<0.01)。与单用UⅡ刺激比较,用DPI或NAC预处理后,MCP-1mRNA表达分别下调21.8%和36.2%(P<0.01),蛋白分泌分别下降22.3%和19.5%(P<0.05)。结论 UⅡ可能通过NADPH氧化酶依赖的活性氧介导了小鼠巨噬细胞MCP-1的表达和分泌上调。Objective To investigate the role of reactive oxygen species （ROS） in urotensin Ⅱ（UⅡ）-induced monocyte chemoattractant protein-1 （MCP-1） expression in RAW264. 7 cells. Methods RAW264.7 cells were stimulated by UⅡ（10 10 to 10^-7 mol/L） for 12 h（MCP-1 mRNA expression） and 18 h（MCP-1 protein secretion） respectively. The cells were pretreated with reduced nicotinamide adenine dinucleotide phosphate（NADPH） oxidase inhibitor diphenyle- neiodonium（DPI 10 μmol/L） or antioxidant N-acetylcysteine（NAC 3 mmol/L） for 0.5 h, then were stimulated by 10^-7 mol/L U Ⅱ for 12 h（MCP-I mRNA expression） and 18 h（MCP-1 protein secretion） respectively. The expression of p47phox, a subunit of NADPH oxidase, was detected by RT-PCR after treated with U Ⅱ for different periods. ROS levels were assessed by flow cytometer after exposure to UⅡ for different hours. Results MCP-1 protein secretion and mRNA expres- sion were increased by 57.0% and 100.5% after stimulated by UⅡ 10.7 mol/L（P〈0.01） ,respectively. The p47phox mRNA expression was increased by U Ⅱ , and U Ⅱ -induced ROS production by macrophage reached the maximal effect at 6 h. Both DPI and NAC were able to inhibit U Ⅱ- induced MCP-1 secretion and mRNA expression. Conclusion U Ⅱ-induced MCP-1 mRNA expression and protein secretion may be mediated by NADPH oxidase-derived ROS.

关 键 词：尿紧张素类 巨噬细胞 趋化因子CCL2 NAD 活性氧 逆转录聚合酶链反应 

分 类 号：R543.5[医药卫生—心血管疾病]