人白介素-10基因转染对大鼠脑缺血再灌注损伤半影区炎性因子基因和蛋白表达的影响  

Effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats

作　　者：傅军[1] 张一[2] 张成生[1] 杨常春[2] 王士雷[4] 林其谁[5] 宣海星[5] 周岱 

机构地区：[1]四川省内江市第一人民医院神经外科,641000 [2]江苏省苏州大学附属第三医院神经外科 [3]附属第一医院神经外科 [4]山东省青岛大学附属医院麻醉科 [5]中国科学院上海生命科学研究院生物化学与细胞生物学研究所

出　　处：《中华老年医学杂志》2008年第10期775-779,共5页Chinese Journal of Geriatrics

摘　　要：目的观察人白介素（IL）-10基因转染对大鼠局灶脑缺血再灌注损伤半影区肿瘤坏死因子-α（TNF-α）和白介素-1β（IL-1β）基因和蛋白表达的影响。方法建立大脑中动脉栓塞（MCAO）模型，采用立体定向脑室内注射的方式进行转染，逆转录聚合酶链式反应（RT—PCR）和酶联免疫吸附实验（ELISA）检测其转染效果，氯化三苯四氮唑（TTC）染色测定脑梗死体积，荧光实时定量PCR检测半影区TNF-α和IL-1β基因表达情况，ELISA法检测半影区TNF-α和IL-1β蛋白的含量。结果正常对照组、缺血对照组、空质粒组和IL-10基因转染组半影区TNF-α蛋白含量分别为（0．66±0．04）、（1．16±0．26）、（1．15±0．26）ng／g和（0．84±0．05）ng／g，IL-1β蛋白含量分别为（0．37±0．05）、（1．25±0．39）、（1．21±0．58）ng／g和（0．62±0．05）ng／g。与正常对照组比较，其余各组TNF-α和IL-1β蛋白含量明显升高（P〈0．01），而IL-10基因转染组TNF-α和IL-1β含量则较缺血对照组和空质粒组显著降低（P〈0．01）；正常对照组、缺血对照组、空质粒组和IL-10基因转染组半影区TNF-α mRNA的表达量分别为1．00±0．53、9．42±1．83、9．69±1．96和3．53±1．09；IL-10 mRNA的表达量分别为1．00±0．51、27．81±4．84、23．96±4．90和13．55±4．45。与正常对照组比较，其余各组TNF-α和IL-1β mRNA表达最明显升高（P〈0．01），而IL-10基因转染组TNF-α和IL-1β mRNA表达量则较缺血对照组和空质粒组显著降低（P〈0．01）。结论人IL-10基因转染后能抑制大鼠局灶脑缺血再灌注损伤半影区TNF-α和IL-1β基因和蛋白的表达，可能是其发挥缺血脑保护作用的机制之一。Objective To observe the effects of human IL-10 gene transfeetion on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemiareperfusion injury in rats and to investigate its neuroprotective mechanism. Methods Rats were divided into four groups： normal control group, isehemic control group, empty plasmid group and human IL-10 gene transfeeted group. The mRNA and protein expressions of IL-1β and TNF-α in the penumbra area were detected by fluorescence real-time quantitative PCR and ELISA respectively. Results In normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group, the levels of protein expression of TNF-α in penumbra area were（0.66 ± 0.04） ,（1.16±0.26） ,（1. 15±0.26）ng/g and（0.84±0.05）ng/g, and the levels of protein expression of IL-1β in penumbra area were（0.37±0.05）, （1.25±0.39）, （1.21 ±0.57）ng/g and（0.62±0.05） ng/g, respectively. Compared with normal control group, the levels of protein expression of TNF-α and IL-1β were significantly higher in other three groups（all P〈0.01）, and lower in human IL-10 gene transfeeted group than in ischemic control group and empty plasmict group（all P〈0.01）. In normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group, the levels of mRNA expression of TNF-α in penumbra area were 1.00±0.53,9.42±1.83,9. 69±1.96 and 3.53±1, 09, and the levels of mRNA expression of IL-1β in penumbra area were 1.00 ±0.51,27.81 ± 4.84,23.96 ± 4.90 and 13.55 ± 4.45, respectively. Compared with normal control group, the levels of mRNA expression of TNF-α and IL-1β were significantly higher in other three groups（all P〈0.01 ）, and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group（all P〈0. 01）. Conclusions The human IL-10 gene transfection may play an protective effect on cerebral ischemia through inhibiting mRNA and protein ex

关 键 词：白细胞介素类 基因表达 肿瘤坏死因子Α 脑缺血 再灌注损伤 

分 类 号：R686[医药卫生—骨科学]