摘要
目的:观察山莨菪碱对创伤性急性肺损伤家兔肺泡巨噬细胞中核因子κB活化和下游基因肿瘤坏死因子αmRNA表达的影响。方法:实验于2004-03/2005-02在解放军第四军医大学西京医院检验科临床分子生物学实验中心完成。将健康成年家兔72只抽签法随机分为3组(n=24):①山莨菪碱组:于撞击台上行胸壁撞击(冲击速度5.7m/s,力75.8N/cm2,冲撞能量约204J)后,立即静脉注射大肠杆菌脂多糖50μg/kg,建立创伤性急性肺损伤模型,造模后立即予以山莨菪碱2mg/kg静注,随后以1mg/(kg·h)静脉维持。②急性肺损伤组:造模同山莨菪碱组,造模后以等量生理盐水静注和维持。③正常对照组:不做胸壁撞击,静注等量生理盐水代替脂多糖,其余处理同急性肺损伤组。各组动物分别于模型建立后1,2,3,4h放血处死6只,留取支气管肺泡灌洗液,分离肺泡巨噬细胞,分别用凝胶电泳迁移率改变分析法和半定量反转录-聚合酶链反应法检测肺泡巨噬细胞中核因子κB的活性变化及肿瘤坏死因子αmRNA的表达水平,以相对吸光度表示。结果:67只兔进入结果分析。①核因子κB的活性:急性肺损伤组于模型建立后1～4h内均明显高于正常对照组(P<0.01),在2h达到高峰,其相对吸光度值,是同期正常对照组的8倍;山莨菪碱组高于正常对照组(P<0.01),显著低于急性肺损伤组(P<0.01),其中2h降幅最大,达30.19%。②肿瘤坏死因子αmRNA的表达水平:急性肺损伤组在模型建立后1h其表达即开始上升,两三小时达到高峰,3h最高,其相对吸光度值,是同期正常对照组的5倍,4h其表达较前有所下降,但仍明显高于正常对照组(P<0.01);山莨菪碱组各时间的表达显著低于急性肺损伤组(P<0.01,0.05),最大降幅出现在2h,达34.48%。结论:①机体遭受创伤及脂多糖刺激后,肺泡巨噬细胞被激活导致核因子κB的活化是肺泡巨噬细胞大量释放肿瘤坏死因子α等前炎细胞因子的关键�
AIM： To observe the effects of anisodamine on the activation of nuclear factor （NF）-κB and the expression of tumor necrosis factor （TNF）-α mRNA in pulmonary alveolar macrophage （PAM） in rabbits with traumatic acute lung injury （ALI）.
METHODS： The experiment was accomplished in the Experimental Center of Clinical Molecular Biology, Department of Clinical Medical Laboratory, Xijing Hospital of the Fourth Military Medical University of Chinese PLA from March 2004 to February 2005. Seventy-two healthy adult rabbits were randomly divided into three groups with 24 rats in each： （1）control group （C group）;（2）ALI group （I group）;（3）anisodamine treatment group （T group）. The rabbits of I and T groups were given colibacillus lipopolysaccharide （LPS） by intravenous injection at a dose of 50 μg/kg immediately after receiving an impact （velocity 5.7 m/s, force 75.8 N/cm^2, energy 204 J） on chest wall to make traumatic ALI. After the ALl model was made, the rabbits of T group were injected intravenously with 2 mg/kg anisodamine initially and then 1 mg/kg per hour for maintenance, while the rabbits of C group were treated with equal volume of saline by intravenous injection for maintenance. The rabbits of C group were injected intravenously with saline at the same dose instead, without any impact on chest wall. Every six rabbits of each group were killed at 1, 2, 3 and 4 hours respectively after the modeling to fetch bronchoalveolar lavage fluid （BALF） and isolate PAM. The activity of NF-κB and the expression of TNF-α mRNA in PAM were measured by electrophoretic mobility shift assay （EMSA） and semi-quantity reverse transcription polymerase chain reaction （RT-PCR）, and expressed as relative absorbance （RA）.
RESULTS： Totally 67 rabbits entered into the result analysis. （1）The activity of NF-κB： The activity was obviously increased in I group compared with that of C group during 1-4 hours after modeling （P 〈 0.01）, and reached the highest
出处
《中国临床康复》
CSCD
北大核心
2006年第40期94-96,共3页
Chinese Journal of Clinical Rehabilitation
