| Part OneEffects of ketamine on nitrogen monoxidum,oxygen Radicals and free intracellular calcium in Lipopolysaccharide-stimulated rat alveolar macrophage in vitroObjective:To investigate the effects of ketamine on production of Nitric oxide(NO),hydroxy radical(·OH),superoxide anion(O2·-)and free intracellular calcium([Ca2+]i)in the rat alveolar macrophage cell line (NR8383)activated by Lipopolysaccharide(LPS).Methods:Macrophage was distributed into five groups,group C, group L,group K1,group K2 and group K3.Group L,K1,K2 and K3 were stimulated with LPS.Before stimulation,Group L,K1,K2 and K3 were treated with phosphate buffer solution(PBS),10,100and 1000μmol/L ketamine.Group C was treated with PBS.The levels of NO,·OH and O2·- in the supernatant were assayed.The[Ca2+]i level was measured with fluorescent intensity by laser scanning confocal microscope.Results:The concentrations of NO,·OH,O2·-and[Ca2+]i in group L increased significantly(P＜0.01)when compared to Group C.The concentrations of NO,·OH and O2·- in Group K2,K3 were obviously lower than those of Group L(P＜0.05).There was no statistical difference between K2 and K3(P＞0.05).Conclusions:Ketamine(100 and 1000μmol/L)can inhibit LPS-induced production of NO,·OH,O2·- and[Ca2+]i. Part TwoEffects of Ketamine on OVA-induced the changes of Free Intracellular Calcium and Free-radical in Alveolar Macrophages in VitroObjective:To observe the effect of ketamine on OVA-induced the changes of free intracellular calcium([Ca2+]i)and free-radical in alveolar macrophages from sensitized rat in vitro.Methods:Macrophages were collected from Bronchoalveolar Lavage fluid of OVA-sensitized Sprague-Damley rats and treated with ketamine of concentration at 10μM,100μM and 1000μM before challenge with 100μg/mL OVA.The production of nitric oxide(NO),hydroxy radical (·OH)and superoxide anion(O2·-)in the supematant of culture was assayed by kits and the[Ca2+]i of macrophages was measured with mean fluorescent intensity(MFI)by laser scanning confocal microscope.Results:OVA challenge induced an elevation of[Ca2+]i and an increase of NO,·OH and O2·- in sensitized rat macrophages,which were inhibited by ketamine.Conclusions:Ketamine inhibited OVA-induced oxidative stress which involved in calcium. |
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