NF-κB反义寡核苷酸对博莱霉素致小鼠急性肺损伤的保护作用

Protective Effect of Antisense Oligonucleotides to Nuclear Factor-κB on Bleomycin-induced Acute Lung Injury in Mouse

目的研究转录因子NF-κB在博莱霉素(BLM)致小鼠急性肺损伤病理过程中的重要作用,及其反义寡核苷酸对肺损伤的防治作用。方法小鼠随机分为5组,于0d经小鼠尾静脉分别注射150mg/kg或300mg/kgBLM,在注射前6h和注射后第5d,150组和300组分别注射生理盐水(NS);anti150组和anti300组分别注射p65的硫甙反义寡核苷酸(900μg/只);同时设立正常对照组,分别在3个时间点(注射前6h、0d、第5d),经尾静脉注射NS,观察各组小鼠体质量变化、死亡情况和肺组织病理改变。结果(1)150组和anti150组死亡率分别为53%和0%(P<0.05);而300组和anti300组死亡率分别为100%和60%(P<0.05)。p65反义寡核苷酸抑制了BLM所致小鼠死亡;(2)150组和anti150组小鼠的最低体质量分别减轻至原来的(65.5±7.8)%和(82.1±3.4)%(P<0.01)。p65反义寡核苷酸改善了BLM所致小鼠体质量减轻;(3)肺组织HE染色显示,BLM致死的150组小鼠,可见肺组织血管结构丧失,出血性水肿,肺泡腔内大量炎性渗出;非BLM致死小鼠注射BLM后14d,可见肺组织大片实变,肺泡结构丧失。而anti150组14d明显减轻了BLM导致的肺组织损伤。结论NF-κB可能在BLM致小鼠急性肺损伤过程中具有重要作用。NF-κB反义寡核苷酸对BLM所致小鼠急性肺损伤具有保护作用。

Objective To investigate the role of nuclear factor-κB（NF-κB） as well as the preventive and therapeutic effect of its antisense oligenucleofides on bleomycin-induced acute lung injury in mouse. Methods The animals were injected 150 mg/kg or 300 mg/kg bleomycin dissolved in 200 μl of 0.9% NaCl solution intravenously via the tail vein. Antisense phosphorotioate oligenucleotides to the p65 subunit of NF-κB were dissolved in 200 μl of 0.9% NaCl solution and injected via the tail vein at 6.h before and 5 d after the bleomycin injection,and the animals of 150 group and 300 group were injected 200 μl of 0.9% NaCl solution at 6 h before,0 d and 5 d after the bleomycin injection,respectively. At the same time,the 200 μl of 0.9% NaCl solution was administrated at 6 h before,0 d and 5 d in the normal control group,respectively. The effects of the antisense oligonucleotides on the mortality,body weight and pulmonary histopathology were observed. Resuits （1） The mortality of 150 group and anti 150 group was 53% and 0% （P 〈 0.01 ）,respectively. However,the mortality of the 300 group and anti 300 group was 100% and 60% （P 〈 0.05）,respectively,which indicated that the antisense oligonucleotides significantly lowered the mortality of the mouse induced by bleomycin. （2） The animals of 150 group and anti 150 group lost 34.5% and 17.9% body weight （P 〈 0.01 ） ,respectively ,suggesting that the antisense oligonucleotides significantly reversed the loss of body weight.（3 ）The toss of vascular integrity with hemorrhagic edema was shown by pulmonary histopathology in the dead animals of the 150 group;and the diffuse consolidation of lung parenchyma with loss of alveolar arch/tecture and increased cellularity was seen in the animals killed 14 days after 150 mg/kg bleomycin injection. However,a significant inhibition on this lesions was shown in the animals of the anti150 group killed at 14 day,indicating that p65 antisense oligonucleotides significantly lessened the pathological changes induced by bleomycin in the lung. Conclusion NF-κB plays a very important role in bleomycin-induced acute lung injury in mouse,which is protected by its antisense oligonucleotides.