摘要
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.
出处
《中国药物与临床》
CAS
2009年第B07期18-21,共4页
Chinese Remedies & Clinics