| Objective To observe the effect of ketamine (10μM /100μM /1000μ M ) on activation of nuclear factor-kappa B(NF- κB )and the expression of TNF- α in LPS-induced lung injury cell model and to investigate the effect of Ketamine on ALI/ARDS.Methods LPS-induced PAM model of mice was got from culture media of PAM added to LPS(10μg/ml) . It was divided into four groups:(l) LPS groups (induced by LPS 10μg/ml);(2) Ketamine(10μM )+LPS groups (induced by LPS 10μg/ml and KET 10μM ) (3) Ketamine (100μM )+LPS groups (induced by LPS 10μg/ml and KET 100μM ) (4) Ketamine (1000μM )+LPS groups (induced by LPS 10μg/ml and KET 1000μM). The activity of NF-kappaB and the concentration of TNF-α in the supernatant were measured by immunohistochemistry, image analysis and ELISA after induced in 0.5,l,4,6hour.Result ( 1) In LPS added ketamine(10μM,100μM ,1000μM ) groups, the activation of NF-kappaBp50 is obviously lower at the time point of lh and 4h than in LPS group(P<0.05), at the same time dependenting on volume. The more density of ketamine, the less activation of NF-kappaBp50.There are different between all groups(P<0.05).(2) The activation test of NF-kappaBp50 is the same as NF-kappaBp65.(3) after adding the different density of ketamine in LPS, the content of TNF-α from 0.5h to 6h is the less than LPS(P<0.05).The content of LPS+ketamine 1000μM have a apparently drop than LPS at 0.5h and 1h(p<0.01).The density of ketamine is getting more, the content of TNF-α is getting less at 0.5h,lh and 4h, there are very different between all groups(P<0.05).Conclusion Based on these results, we conclude that LPS might active NF-kappaB in the PAMs and lead to the increasement of transcription and expression of TNF-α gene, which may be the key point of releasing of various inflammatory medium in ALI; Ketamine could lower the release of TNF-α by inhibiting the activation of NF-kappaB to improve the LPS-induced ALL it may be helpful in lung protection. |
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