反义p38α基因转染对缺氧复合烧伤血清处理心肌细胞炎性因子的表达  

Effects of antisense p38α gene transfection on expressions of inflammatory factors in cardiomyocytes following hypoxia and burn serum treatment

作　　者：郑军[1] 黄跃生[1] 黄晓元[2] 范鹏举[1] 贺伟峰[1] 张小容[1] 蒋建新[1] 粟永萍[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所 [2]中南大学湘雅医院烧伤整形外科,长沙430008

出　　处：《第三军医大学学报》2004年第23期2097-2101,共5页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划资助项目 ("973"项目 ) (G19990 54 2 0 2 );国家杰出青年科学基金资助项目 ( 30 12 50 4 0 )~~

摘　　要：目的　探讨反义p3 8α基因转染对缺氧复合烧伤血清处理心肌细胞炎性因子的表达。方法　建立缺氧复合烧伤血清培养心肌细胞模型 ,分对照组、缺氧复合烧伤血清组 (非转染组 )和反义p3 8α基因转染组 (转染组 )。烧伤血清采自 40 %TBSAⅢ度烧伤Wistar大鼠 ;采用含 1%O2 的混合气体造成缺氧模型 ;采用基因重组技术构建反义p3 8α基因重组体 ;用免疫印迹法和RT PCR法分别检测不同时相点p3 8α激酶蛋白及TNFα、IL 1βmRNA表达变化 ,并作统计学分析。 结果　成功构建反义p3 8α基因重组体 ;缺氧复合烧伤血清迅速、持续激活心肌细胞p3 8α激酶 ,心肌细胞TNFα、IL 1β表达增多 ;反义p3 8α基因显著抑制p3 8α激酶过度活化 (P <0 0 1) ,下调TNFα、IL 1β表达 (P <0 0 1)。 结论　p3 8α激酶途径介导了缺氧和烧伤血清所致心肌细胞损伤 ,抑制p3 8α激酶的持续活化能有效下调炎性因子TNFα、IL 1β表达 ,减轻该条件下心肌细胞损害。Objective To explore the effects of antisense p38α gene transfection on the expressions of inflammatory factors in cardiomyocytes following hypoxia and burn serum treatment. Methods Primary neonatal rat cardiomyocytes were cultured in vitro with hypoxia and burn serum treatment and divided into there groups: control group, hypoxia and burn serum treatment group (non-transfected group); antisense p38α gene transfection group (transfected group). Burn serum was collected from Wistar rats with 40% total body surface area (TBSA) Ⅲ degree burn. Cardiomyocytes inhaling mixed gas containing 1% O 2 were used as the hypoxia model. The antisense p38α gene recombinant was constructed by genetic recombination technique. p38α protein kinase expression level and mRNA level of TNFα and IL-1β were determined respectively at different time points by Western blot and RT-PCR. The results were processed statistically. Results The antisense p38α gene recombinant was constructed successfully. Hypoxia and burn serum induced rapid and long time activation of p38α kinase and up-regulated mRNA levels of TNFα and IL-1βin cardiomyocytes. Antisense p38α gene decreased p38α activity significantly (P<0.01) and down-regulated TNFα and IL-1β mRNA expressions (P<0.01). Conclusion These data suggest that p38α kinase pathway may play an important role in cardiomyocyte damage induced by hypoxia and burn serum. The inhibition of p38α kinase activation can down-regulate TNFα and IL-1β expressions and attenuate injury of cardiomyocytes exposed to burn and hypoxia.

关 键 词：反义p38α基因 心肌细胞 缺氧 烧伤血清 细胞因子 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学] R392.13[医药卫生—基础医学]