锌指蛋白A20及其突变体质粒转染对小鼠内毒素血症的预防作用  

Preventive effect of Zinc finger protein mA20 and its mutant plasmid on endotoxemia

作　　者：左云龙[1] 吴丽娟[1] 匡凤梧[2] 蒋建新[3] 朱佩芳[3] 

机构地区：[1]第三军医大学附属大坪医院野战外科研究所检验科,重庆400042 [2]重庆医科大学附属儿童医院ICU [3]第三军医大学附属大坪医院第四研究室,重庆400042

出　　处：《中华创伤杂志》2007年第10期777-780,共4页Chinese Journal of Trauma

基　　金：军队“十一五”面上课题资助项目（06MA189）

摘　　要：目的 研究锌指蛋白A20（mA20）及其突变体mA20-ZnF4-7质粒转染对脂多糖（LPS）攻击所致小鼠内毒素血症的预防作用。方法用LPS攻击小白鼠制备内毒素血症动物模型。随机分为等渗盐水对照组（Ns组）、单纯LPS组（LPS组）、pCAGGS-flag组（质粒组）、pCAGGS-flag-mA20预防组（mA20组）和pCAGGS-flag-mA20-ZnF4-7预防组（mA20突变体组）。按10μg质粒：1．6ml等渗盐水比例溶于等渗盐水中，采用“基于流体动力学基因转染方法”对实验小鼠进行质粒预防性注射。以ELISA检测各组血浆和组织TNF-α、IL-1β表达并观察组织病理变化。结果 （1）mA20组和mA20-突变体组的TNF-α在各时相点表达分别为：6h：（211．942±9．133）pg／ml、（163．536±0．502）pg／ml，12h：（195．130±26．318）pg／ml、（206．446±34．797）pg／ml，24h：（182．963±14．527）pg／ml、（158．029±19．176）pg／ml，均显著低于LPS组和质粒组（P〈0．05）。（2）mA20组和mA20突变体组的IL-1β在各时间点表达分别为：6h：（137．908±4．487）Pg／ml、（138．598±8．733）pg／ml，12h：（137．678±3．539）pg／ml、（137．448±7．953）pg／ml，24h：（129．862±11．602）pg／ml、（126．873±6．942）pg／ml，均显著低于LPS组和质粒组（P〈0．05）。（3）mA20组和mA20突变体组内各时间点之间TNF-α及IL-1β表达比较，差异无统计学意义（P〉0．05）。（4）病理观察结果显示，mA20组和mA20突变体组的肝、肺损伤均较LPS组和质粒组轻。结论 基于流体动调控炎症反应力学的mA20及其突变体mA20-ZnF4-7质粒转染对实验性内毒素血症有相似的预防作用，mA20有功能丰余性，为临床应用及mA20突变体研究提供了一定的实验基础。Objective To study the preventive effect of zinc finger protein mA20 and its mutant mA20-ZnF4-7 on endotoxemia of lipopolysaccharide （LPS） -assaulting mice. Methods We prepared the endotoxemia animal model by assaulting mice with LPS at 2.5 mg/kg. These animals were divided randomly into NS group, LPS-treatment group, pCAGGS-flag group, pCAGGS-flag-mA20 prevention group and pCAGGS-flag-mA20-ZnF4-7 prevention group. By using the method of hydrodynamics-based gene transfection, we intravenously injected mice with 10 μg of plasmid DNA in 1.6 ml saline via the tail vein. The contents of TNF-α, IL-1β in blood plasma were detected by ELISA, when inflammatory reaction was also observed pathologically. Results （ 1 ） The TNF-α expressions of pCAGGS-flag-mA20 and pCAGGS-flag-mA20-ZnF4-7 prevention groups were （201. 797 ± 4. 789） pg/ml and （235. 710 ± 6. 108） pg/ml at the 6th hour, （236. 580 ± 7. 497） pg/ml and （ 255. 130 ± 7. 827 ） pg/ml at the 12th hour, （ 154.551± 16.460） pg/ml and （ 223. 246± 11.316） pg/ml at the 24th hour, which were significantly lower than those of LPS-treatment and pCAGGS-flag groups. （2）The IL-1 [5 expressions of pCAGGS-flag-mA20 and pCAGGS-flag-mA20-ZnF4-7 prevention groups were （87. 103 ± 3.84） pg/ml and （0. 101. 586 ±2. 486） pg/ml at the 6th hour, （ 83. 655± 4.973 ） pg/ml and （94.460 ± 3.110 ） pg/ml at the 12th hour, （77.678±6.555） pg/ml and （87. 103±4. 826） pg/ml at the 24th hour, which were also significandy lower than those of LPS-treatment and pCAGGS-flag groups. （ 3 ） Microscope observation of pathological section showed that the inflammatory damage of pCAGGS-flag-mA20 and pCAGGS-flag-mA20- ZnF4-7 prevention groups was lighter than those of LPS-treatment and pCAGGS-flag groups. No signifi- cant difference was found between pCAGGS-flag-mA20, pCAGGS-flag-mA20-ZnF4-7 prevention groups and LPS and pCAGGS-flag groups. Conclusions The method of hydrodynamics-based gene transfection can efficiently delivery plasmid DNA

关 键 词：内毒素类 转染 小鼠 锌指蛋白A20 

分 类 号：R686[医药卫生—骨科学]