维生素C对染镍肺泡巨噬细胞iNOS的影响
Effects of vitamin C on expression of nitric oxide synthase of alveolar macrophage treated with Ni2O3
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摘要:目的 探讨镍在大鼠肺泡巨噬细胞诱导型一氧化氮合成酶(iNOS)基因诱导表达中的作用, 以及维生素C对巨噬细胞iNOS诱导表达的影响及其机制。方法 采用体外细胞培养法测定在维生素C(25, 50, 100μmol/L)作用下, 体外染镍肺泡巨噬细胞的存活率及活力。同时观察细胞内一氧化氮(nitric oxide, NO)和活性氧的产生, 以及一氧化氮合酶(nitric oxide synthase, NOS)活力的变化。以逆转录-聚合酶链反应(RT-PCR)法测定iNOSmRNA的表达。结果 在体外染镍肺泡巨噬细胞中加入不同浓度的维生素C后可提高该细胞的存活率和活力, 可减少NO和活性氧的产生, 降低NOS和活性, 还可抑制镍诱导的iNOSmRNA的表达水平。结论 镍可通过诱导大鼠肺泡巨噬细胞iNOS基因表达, 产生大量NO, 导致细胞脂质过氧化, 而维生素C可通过抑制iNOSmRNA的表达, 拮抗镍对肺泡巨噬细胞的细胞毒作用。Abstract:Objective To study the effects of vitamin C on the expression of NOS of alveolar macrophages exposure to Ni2O3 in vitro.Methods The alveolar macrophages were cultured by exposure to Ni2O3 in vitro.Meanwhile, vitamin C(25, 50 and 100 μmol/ L)were added in the medium, respectively.The detection of active oxygen, nitric oxide(NO)and activity of nitric oxide synthase (NOS)in alveolar macrophages were carr ied out.Reverse transcription polymerase chain reaction was used to check the mRNA level of induced nitric oxide synthase.Results Vitamin C could decrease mortality and increase survival activity of alveolar macrophages.Active oxygen, NO and NOS activity decreased in the same time.The expression of iNOS was dramatically suppressed in the same time.Conclusion T he expression of iNOS was induced by Ni2O3 in alveolar macrophages, which gave rise to producing a great amount of NO and resulted in the lipid peroxidation of cells.Vitamin C could antagonize the lipid peroxidation of alveolar macrophage induced by Ni2O3, which probably due to its antioxidant property.
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