核因子-кB靶向性寡核苷酸“圈套”策略对创伤性炎症大鼠肝脏TNF-α的作用  

Effects of decoy strategy targeted to NF-κB on TNF-α in rat liver after traumatic inflammation

作　　者：杨文军[1] 梁华平[1] 徐祥[1] 时德[2] 郝天智[1] 国华[1] 李德卫[2] 

机构地区：[1]第三军医大学大坪医院野战外科研究所 [2]重庆医科大学第一附属医院血管外科,重庆市400016

出　　处：《中华肝胆外科杂志》2004年第3期177-180,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家重点基础研究发展规划项目 (G19990 5 42 0 3 );国家自然基金资助项目 ( 3 0 170 3 67);全军十五课题项目 ( 2 0 0 2 2 0 0 4)

摘　　要：目的　探讨NF кB靶向性寡核苷酸 (ODN)“圈套”策略对创伤性炎症大鼠肝脏TNF α的作用。方法　Wistar大鼠 96只 ,随机分成对照组、创伤性炎症组、“圈套”ODN组和变异“圈套”ODN组。运用凝胶阻滞实验检测创伤性炎症术后肝脏组织NF кB的活性及合成“圈套”ODN的体外竞争抑制试验 ,用RT PCR检测大鼠肝脏组织TNF αmRNA水平 ,ELISA检测大鼠肝脏组织TNF α的蛋白水平。结果　大鼠创伤性炎症术后 3h肝脏NF кB的活性开始升高 ,术后 12h达高峰。肝脏组织TNF αmRNA水平和蛋白水平也明显上升 ,与NF кB的活性改变一致 ,“圈套”ODN在体外能有效地抑制NF кB活性。“圈套”ODN治疗后大鼠肝脏组织TNF α的表达明显下降 ,肝脏功能明显好转 ,而变异“圈套”ODN没有效果。结论　NF кB靶向性“圈套”ODN通过特异性抑制NF кB活性 ,可以有效地抑制创伤性炎症大鼠肝脏炎症介质TNF α的释放 ,从而改善肝脏功能。Objective To study the effects of decoy strategy targeted to NF-κB on TNF-α in rat liver after traumatic inflammation. Methods Ninety-six rats were randomized into 4 groups: control group, traumatic inflammation group, traumatic inflammation plus decoy ODN group and traumatic inflammation plus mutant decoy ODN group. The rats were sacrificed in 3, 6, 12, 24, 48 and 72 h after traumatic inflammation, respectively, and the contents of plasma ALT were determined. After hepatocytes were isolated and nuclear protein extracted, DNA binding activity of NF-κB was measured by EMSA. Decoy ODN's competition-inhibiting effects were assayed by EMSA. TNF-α expression in liver tissue was assayed by RT-PCR and TNF-α level determined by ELISA. Results After traumatic inflammation, DNA binding activity of NF-κB increased. TNF-α expression and level also significantly increased and was positively correlated to the activity of NF-κB. Decoy ODN could effectively inhibit the activity of NF-κB in vitro. After using decoy ODN, the expression of TNF-α and plasma ALT significantly decreased. Conclusions Decoy strategy targeted to NF-κB could effectively inhibit the release of TNF-α through specifically inhibiting the activity of NF-κB.

关 键 词：核因子—κB 靶向性寡核苷酸 “圈套”策略 创伤性炎症 大鼠 肝脏 TNF—α 

分 类 号：R641[医药卫生—外科学]