实时荧光定量PCR分析海水浸泡伤大鼠小肠组织NF-κB及IκB mRNA表达水平  

Analysis in the expression of NF-κB mRNA and IκB mRNA on small intestine tissues in opened abdominal injury with seawater immersion in rats by real-time PCR

作　　者：韩志海[1] 虞积耀[2] 王宇[1] 

机构地区：[1]海军总医院呼吸科,北京100048 [2]海军总医院病理科,北京100048

出　　处：《中国急救医学》2011年第1期40-43,共4页Chinese Journal of Critical Care Medicine

基　　金：解放军总后勤部医药卫生科研计划课题（No.06D004）

摘　　要：目的 建立检测大鼠小肠组织NF-κB及IκB-α mRNA表达水平的SYBR Green 实时定量（real time）PCR实验技术,观察海水浸泡伤大鼠小肠组织NF-κB及IκB-α mRNA表达水平的动态变化.方法 Wistar大鼠63只,随机分为三组：空白对照组（C组,n=7）,创伤组（W组,分为12、24、48、72 h四组,每组n=7）,创伤＋海水浸泡组（W＋S组,分为12、24、48、72 h四组,每组n=7）.W组采用腹部开放伤模型,W＋S组在腹部开放伤的基础上海水浸泡1 h.采用real-time PCR方法测定三组实验动物小肠组织NF-κB及IκB-α mRNA表达量并进行统计分析和组间比较.结果 与W组相比,W＋S组大鼠小肠组织NF-κB mRNA在72 h后出现显著表达水平上调（P〈0.01）,W组和W＋S组大鼠小肠组织IκB-α mRNA在48 h后都出现显著表达水平上调,但W＋S组在72 h IκB-α mRNA表达上调更为显著（P〈0.01）.结论 在海水浸泡伤的后期（＞72 h）,NF-κB mRNA表达明显上调,促进NF-κB蛋白水平的合成,从而进一步放大NF-κB信号转导途径,诱导炎症反应.同时, IκB-α mRNA表达水平的也明显上调,促使IκB-α蛋白表达增加,以尽量下调细胞核中NF-κB的活性.Objective To establish a SYBR Green quantitative real -time PCR method to monitor the dynamics for NF-κB mRNA and IκB mRNA of opened abdominal injury with seawater immersion in rats. Methods Wistar rats were randomly divided into three groups： the control group （n = 7）, the opened abdominal injury groups （n = 28, all rats were randomly divided into four groups separately , which groups were 12 h, 24 h, 48 h , and 72 h after wounded） and opened abdominal injury with seawater immersion groups （ n = 28, all rats were randomly divided into four groups separately , which groups were 12 h, 24 h, 48 h ,and 72 h after wounded）. Dynamic express of NF - κB mRNA and IκB mRNA of small intestine tissues was detected in every groups by real - time PCR method. Results NF- κB mRNA level was rise obviously after 72 h in the opened abdominal injury with seawater immersion groups than other groups in same time point（ P 〈 0.01 ）. The expression of IκB mRNA level was rise obviously after 48 h in the opened abdominal injury with seawater immersion groups compared with the opened abdominal injury groups after 48 h （ P 〈0.01 ）. Conclusion The NF - κB signal pathways was activated and magnified in latter periods of the opened abdominal injury with seawater immersion（ 〉 72 h） , Which led to the development of continued inflammatory course ultimately.

关 键 词：腹部开放伤 海水浸泡 实时定量PCR NF-ΚB IΚB mRNA 

分 类 号：R-332[医药卫生]