重组腺病毒对烫伤大鼠肝组织核因子—kB活性的调控

目的 探索腺病毒介导IkBα基因对烫伤大鼠肝组织核转录因子NF－kB活性的调控作用。方法 应用重组缺陷型腺病毒载体（AdIkBα)预处理大鼠，烫伤后不同时相点取肝组织，提取组织核蛋白，与r^-32PATP标记的NF－kB特异性探针共同反应，利用凝胶电泳迁移率改变分析方法（EMSA）检测NF－kB/DNA结合性；提取肝组织总RNA，用RT－PCR法检测IL－1β,TNFα mRNA的表达。结果 与正常对照组相比，烫伤组致伤后0．5hNF-kB/DNA结合活性显增强，致伤后24h，仍保持较强结合活性。AdIkBα预处理组，大鼠NF－kB/DNA结合活性略高于正常，但显低于烫伤组。RT－PCR分析，大鼠烫伤后0．5h与对照组相比，IL－1β，TNFα表达显升高，持续致伤后24h;AdIkBα预处理组，IL－β和TNFα表达显低于烫伤组。结论 大鼠严重烫伤后，肝组织细胞内NF－kB迅速活化，IL－1β和TNFα的表达随之显增强，经AdIkBα预处理能显抑制大鼠严重烫伤后肝组织NF－kB的活化，并下调IL－1β、TNFα的表达。

The modulating effects of transfected IkappaBalpha gene on the NF-kappaB activity of hepatic tissue in scalded rats]

OBJECTIVE: To explore the modulating effects mediated by recombinant IkappaBalpha gene by adenovirus vector transfection on the NF-kappaB activity of hepatic tissue in scalded rats. METHODS: After being primed by recombinant defect adenovirus (AdIkappaBalpha) vector, the rats were scalded. The hepatic tissue was harvested at different time points and the nuclear protein was extracted and reacted with r(-23) P] ATP labelled NF-kB specific probe. NF-Kb/DNA combining activity was determined with electrophoretic mobility shifty assay (EMSA). The total RNA in hepatic tissue was extracted and the expressions of IL-1beta and TNFalpha mRNA were determined with RT-PCR. RESULTS: When compared to those in normal control, the NF-kappaB/DNA combining activity increased significantly in scalded rats at half an hour after scalding, and it lasted for 24 PBDs with rather strong activity. But in AdIkappaBalpha priming group, the rat NF-kappaB/DNA binding activity was slightly higher than that of normal control group, but was obviously lower than that of scalding group. CONCLUSION: The intracellular NF-kappaB in hepatic tissue was activated rapidly after that the rats were severely scalded, and the expression of IL-1beta and TNFalpha was enhanced significantly simultaneously. Priming of AdIkappaBalpha could evidently inhibit the activation of hepatic tissue NF-kB in rats injured by severe scalding, and down-regulated the expressions of IL-1beta and TNFalpha.