氧化苦参碱对HaCaT细胞核转录因子κB表达及一氧化氮分泌的影响  

Effects of oxymatrine on expression of nuclear factor-kappa B and nitric oxide secretion in HaCat cells

作　　者：张彩晴[1] 郭宏林[2] 冯捷[1] 闫小宁[1] 李文彬[1] 胡刚[1] 

机构地区：[1]西安交通大学第二附属医院皮肤科,陕西省西安市710004 [2]解放军第三二三医院医务处,陕西省西安市710054

出　　处：《中国组织工程研究与临床康复》2008年第20期3849-3852,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:P物质与细胞因子之间存在互相调节的作用。实验拟验证氧化苦参碱、P物质对HaCaT细胞核转录因子κB蛋白和基因表达及一氧化氮分泌量的影响。方法:实验于2007-05/10在西安交通大学中心实验室完成。HaCat细胞株为ATCC公司产品;氧化苦参碱粉剂由中国药品检验所提供;P物质为Alexis公司产品。选择生长良好的HaCat细胞用于实验,分为空白对照组,小、中、大剂量氧化苦参碱组(分别加入10,50,100mg/L氧化苦参碱)和P物质组。氧化苦参碱各剂量药物预处理24h后加入10-5mol/L的P物质作用存量30min。采用western-blot和反转录-聚合酶链反应法分别检测HaCaT细胞核转录因子κB蛋白和基因的表达情况;并检测一氧化氮的含量。结果:与空白对照组相比,P物质组细胞P65蛋白相对表达量、细胞核转录因子κB的mRNA相对表达量明显增高(P<0.05),与P物质组相比,氧化苦参碱组P65蛋白相对表达量、细胞核转录因子κB的mRNA相对表达量明显降低(P<0.01)。P物质组一氧化氮分泌量增加,氧化苦参碱组一氧化氮分泌量减少,并呈剂量依赖关系。结论:氧化苦参碱可下调人角质细胞株HaCaT细胞核转录因子κB的表达,并使一氧化氮的分泌量下降,且该效应呈剂量依赖关系。AIM： Substance P has a mutual regulative effect with the cytokines, and this study was designed to verify the effect of oxymatrine and substance P on the protein and gene expression of nuclear factor-kappa B （NF- κB） as well as the nitric oxide （NO）＇s secretion of the HaCaT cells. METHODS： The experiment was carried out in the Central Laboratory of Xi＇an Jiaotong University from May to October in 2007. HaCaT cell line was produced by ATCC Company; oxymatrine powder was offered by the Chinese Institute for Drug Control; substance P was the produced of Alexis Company. The well-cultured HaCaT cells were divided into blank control group, three oxymatrine groups pretreated with different concentrations （10, 50 and 100 mg/L） of oxymatrine, and substance P group. After cultured for 24 hours, 10^ 5 mol/L substance P was added to the cells. Thirty minutes later, NF- κB P65 protein expressions were evaluated by western-blot, and mRNA expressions were evaluated by reverse transcription polymerase chain reaction; NO level in the supernatant of cell culture medium was also detected. RESULTS： Compared with the blank control group, the expressions of P65 protein and mRNA of NF- κ B enhanced, the secretions of nitric oxide were obviously increased in the substance .P group （P 〈 0.05）. But the oxymatrine inhibited these phenomena in a dose-dependent manner （P 〈 0.0 l）. CONCLUSION： Oxymatrine can down-regulate the expression of NF- κB of human keratinocyte HaCaT cell line and reduce the release of NO, these effects show a dose dependence.

关 键 词：氧化苦参碱 HACAT细胞 P物质 NF-κ B 一氧化氮 组织构建 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]