含IκB基因的腺病毒转染对大鼠油酸和内毒素诱发多器官功能障碍综合征的影响

Effects of adenovirus transfected with IκB on multiple organ dysfunction syndrome induced with oleic acid and lipopolysaccharide in rats

目的评价含IκB基因的腺病毒转染对大鼠油酸和内毒素（LPS）诱发多器官功能障碍综合征（MODS）的影响。方法成年SD大鼠50只，雌雄不拘，体重100～120g，随机分为3组：对照组（C组，n=10）、MODS组（M组，n=20）和IκB基因转染组（T组，n=20）。C组尾静脉注射生理盐水0．25ml／kg，M组和T组尾静脉分别注射油酸0．25ml／kg；4h后，C组颈内静脉注射生理盐水1ml，M组和T组分别注射LPS3．5mg／kg（1ml）；同时T组注射含boB基因1ml的腺病毒（滴度1×10^9pfu）进行转染。C组于给予生理盐水后7d、M组和T组于注射LPS或基因转染后1、7d时，腹主动脉采血样，测定血浆肌酸磷酸肌酶、谷丙转氨酶、总胆红素和肌酐的水平，并进行血气分析；然后处死大鼠，取肝、肺组织，根据阳性细胞百分比和阳性细胞染色程度进行免疫组化染色评分，并测定NF—κBp65蛋白表达水平，以反映NF—κB活性。结果与C组比较，M组血浆肌酐、谷丙转氨酶、总胆红素和肌酸磷酸肌酶的水平升高，pH值降低，M组和T组PaO2降低，肺、肝组织免疫组化染色评分和NF-κB活性升高（P〈0．05）；与M组注射LPS后1d时比较，M组注射LPS后7d时pH值降低，T组基因转染后1d时血浆谷丙转氨酶、总胆红素和肌酸磷酸肌酶的水平降低，肝组织NF-κB活性降低（P〈0．05）；与M组注射LTS后7d时比较，T组基因转染后7d时血浆肌酐、谷丙转氨酶、总胆红素和肌酸磷酸肌酶的水平降低，PaO2升高，肺、肝组织免疫组化染色评分降低，NF-κB活性降低（P〈0．05）。结论含IκB基因的腺病毒转染可抑制大鼠NF—κB活化，阻断其调控炎性因子的合成，从而减轻油酸和内毒素诱发的MODS。

Objective To investigate the effects of adenovirus transfected with IκB （Ad-IκB） on multiple organ dysfunction syndrome （MODS） induced with oleic acid and lipopolysaccharide （LPS） in rats. Methods Fifty pathogen-free SD rats of both sexes weighing 100-120 g were randomly divided into 3 groups ： group I control （C,n= 10）; groupⅡ MODS （M, n=20） and group Ⅲ Ad-IκB ＋ MODS （T,n =20）. The animals were anesthetized with intraperitoueal （IP） chloral hydrate 0.3-0.4 ml/100 g. MODS was induced with iv oleic acid 0.25 ml/kg and iv LPS 3.5 mg/kg at an interval of 4 h in group M and T. In group T （ Ⅲ ） Ad-bcB was given iv simultaneously with LPS. Blood samples were taken from abdominal aorta at 1 and 7 days after LPS or IκB transfected administration for determination of plasma creatine phosphokinase, glutamic pyruvic transaminase （GPT）, total bilirubin and blood creatinine and blood gas analysis. The animals were then sacrificed and the liver and lungs were removed at 1 and 7 days after blood samples were obtained for determination of expression of NF-κB p65 in lung and liver tissue using immuno-histochemical staining and Western blotting. Immunohistochemical staining was scored （0 = no positive cells, 4 = 81%-100% positive cells）. Results Creatinine,GPT, total bilirubin and creatine phosphokinase were significantly higher in group M than in group C and boB transfected significantly attenuated the MODS-induced increase. PaO2 at day 7 was significantly lower in group M and T than in group C and was significantly higher in group T than in group M. The expression of NF-κB p65 protein was significantly higher in group M than in group C and was significantly decreased by IκB transfected in group T. The immuno-histochemical staining scores were highest in group M and lowest in group C. Conclusion Ad-κB treatment can inhibit the activation of NF-κB and synthesis of inflammatory factors, resulting in amelioration of MODS induced with oleic acid and LPS.