氯胺酮对内毒素小鼠肺泡巨噬细胞核因子-κB活化的影响

Effect of ketamine on activation of nuclear factor-kappa B in LPS-induced pulmonary alveolar macrophage

目的观察不同浓度的氯胺酮对脂多糖(LPS)诱发的小鼠肺泡巨噬细胞核因子(NF)-κB的活化,进一步探讨氯胺酮对急性肺损伤(ALI)/急性呼吸窘迫综合征(ARDS)的应用前景。方法在小鼠肺泡巨噬细胞(PAM)培养液中加入脂多糖(LPS)10μg/ml得到内毒素肺损伤细胞模型。实验分为LPS刺激组(加LPS10μg/ml)、氯胺酮干预组1(同时加入LPS10μg/ml和氯胺酮10μmol/L)、氯胺酮干预组2(同时加入LPS10μg/ml和氯胺酮100μmol/L)、氯胺酮干预组3(同时加入LPS10μg/ml和氯胺酮1000μmol/L)4组。分别于刺激后0.5,1,4,6h留取细胞,用免疫组织化学染色图像分析法检测细胞核中NF-κB活性。结果①LPS+不同浓度的氯胺酮(10,100,1000μmol/L)后,NF-κBp50活性与LPS组相比在1h和4h处有明显的下降(P<0.05),且有剂量依赖性,氯胺酮浓度越大,NF-κBp50活性下降越明显,各组之间差异有统计学意义(P<0.05)。②NF-κBp65活性的检测与NF-κBp50的检测结果相似。结论氯胺酮通过对PAM内NF-κB活化的抑制,可阻止和延缓LPS诱发的ALI的发生,可能有一定的肺保护作用。

Objective To observe the effect of ketamine （10,100,1000μmol/L） on activation of nuclear factor-kappa B （NF-κB） in LPS-induced lung injury cell model and to investigate the effect of Ketamine on ALI/ARDS. Me,otis LPS-induced PAM model of mice was got from culture media of PAM added to LPS （10μg/ml）. It was divided into four groups：① LPS groups （induced by LPS 10 μg/ml）;②Ketamine（10μmol/L）＋LPS groups （induced by LPS 10μg/ml and KET 10μmol/L）;③Ketamine （100μmol/L）＋LPS groups （induced by LPS 10μg/ml and KET 100μmol/L）;④ Ketamine （1000μmol/L）＋LPS groups （induced by LPS 10μg/ml and KET 1000μmol/L）. The activity of NF-κB and were measured by immunohistochemistry, image analysis after induced in 0.5,1,4,6 hour. Result （1） In LPS added ketamine（10, 100, 1000μmol/L）groups, the activation of NF-κB p50 is obviously lower at the time point of 1 h and d h than in LPS group（P〈0.05）, at the same time dependenting on volume. The more density of ketamine, the less activation of NF-κB p50. There are different between all groups（P〈0.05）.（2） The activation test of NF-κB p50 is the same as NF-κB p65. Conclusion LPS might active NF-κB in the PAMs and lead to the increasement of transcription. Ketamine could inhibit the activation of NF-κB to improve the LPS-induced ALI, it may be helpful in lung protection.