蓖麻蛋白对人外周血B淋巴细胞的作用及相关基因表达的研究  

Study on the Effect of Ricin on Human Peripheral Blood B Lymphocyte and the Expression of Related Genes

作　　者：甄二东 黄素娟[1] 化朝举 陶聪[1] 杨述林[1] 王彦芳[1] 周荣[1] 敖红[1] 李奎[1] ZHEN Er-dong HUANG Su-juan HUA Chao-ju TAO Cong YANG Shu-lin WANG Yan-fang ZHOU Rong AO Hong~ LI Kui(Key Laboratory of Farm Animal Genetic Resources and Gerrnplam Innovation, Ministry of Agriculture, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,农业部畜禽遗传资源与种质创新重点实验室,北京100193

出　　处：《中国畜牧兽医》2016年第11期2803-2810,共8页China Animal Husbandry & Veterinary Medicine

基　　金：转基因新品种培育重大专项(2014ZX08011-006);中国农业科学院科技创新工程(ASTIP-IAS05)

摘　　要：试验旨在研究蓖麻蛋白对人外周血B淋巴细胞(IM-9)毒性作用及对相关基因表达的影响。将提取的蓖麻蛋白按不同浓度添加到培养基培养6、12h,研究剂量-时间效应,确定半数抑制浓度,然后以半数抑制浓度培养细胞2、6、10、12h,在每个时间段检测免疫相关基因CD40、IL-1β、TNF-α和凋亡相关基因Bcl-2、Bax、Caspase-3的表达情况。结果表明,蓖麻蛋白对IM-9细胞的毒性作用随浓度和时间的增加而增强。取接近半数抑制浓度20ng/mL作用IM-9细胞2、6、10、12h,免疫相关基因中,TNF-α和CD40基因在6h试验组表达量与对照组相比显著升高(P<0.05),10、12h达到极显著水平(P<0.01);IL-1β基因表达量试验组、对照组间差异不显著(P>0.05)。凋亡基因中,与对照比相比,试验组Bax基因表达量10h极显著降低(P<0.01),12h显著降低(P<0.05);Bcl-2基因表达量4个时间段均达到显著水平(P<0.05),2、12h达到极显著水平(P<0.01);Caspase-3基因除6h表达量降低外,其他时间段表达量均显著升高(P<0.05),2h为极显著水平(P<0.01)。表明蓖麻蛋白能显著影响IM-9细胞的基因表达,TNF-α和CD40基因是两个潜在用IM-9细胞评价蛋白毒性的检测基因。This study was amied to learn the toxic effects of ricin on human peripheral blood B lymphocytes（IM-9）and the expression of related genes.The extracted ricin of different concentrations were added to culture cells for 6and 12 hto make sure the median inhibitory concentration,then culture cells at the median inhibitory concentration for 2,6,10 and 12h,at each time point detect the expression of immune related genes CD40,IL-1β,TNF-αand apoptosis related genes Bcl-2,Bax,Caspase-3.The results showed that the inhibitory effect of ricin on IM-9cells was increased with the increase of concentration and time.The expression of TNF-αand CD40 genes increased significantly in experimental group than control group at 6h（P〈0.05）.10and12 hreached extremely significant level（P〈0.01）;There was no significance of IL-1βgene expression between experimental and control groups（P〉0.05）.About apoptosis gene,the Bax gene expression decreased extremely significantly in experimental group at 10h（P〈0.05）and decreased significantly at 12h（P〈0.05）;Bcl-2gene had reached significant levels of four timeperiods（P〈 0.05）,and 2,12 h were extremely significant（P〈 0.01）;The expression of Caspase-3reduced at 6h,the other time points increased significantly（P〈0.05）,at 2hreached extremely significant level（P〈0.01）.It showed that the gene expression of IM-9cells could be significantly affected by ricin,TNF-αand CD40 genes were two potential genes to evaluate the toxicity by IM-9cells.

关 键 词：蓖麻蛋白 外周血B淋巴细胞 基因表达 

分 类 号：Q78[生物学—分子生物学]