体外培养中IFN-γ、L-Arg及L-NNA对NO合成的影响及NO抗旋毛虫的作用

Influence of IFN-γ,L-Arg and L-NNA upon NO production and effects of NO resisting trichinella spiralis in vitro cultivation

目的 探讨体外培养中IFN γ、L Arg及L NNA对NO合成的影响及NO抗旋毛虫的作用。 方法 分离、纯化长爪沙鼠腹腔巨噬细胞 ,置RPMI16 4 0培养液中培养。设IFN γ组、L Arg组、L NNA组和对照组 ,每个实验组又分 5个不同的浓度组。分别向含有巨噬细胞的培养瓶中加入不同浓度的IFN γ、L Arg及L NNA进行体外培养。培养 2 4h后 ,用硝酸还原酶法分别测定培养液中的NO含量。将旋毛虫幼虫分别加入上述培养体系中进行体外培养 ,观察旋毛虫幼虫的活动及损伤。结果 ①体外培养中 ,激活的巨噬细胞能产生NO ,IFN γ和L Arg能促进NO的合成 ,L NNA则能抑制NO的合成 ,这种促进或抑制NO合成的作用均具有剂量依赖性 ,剂量越高作用越明显。②加入旋毛虫幼虫后 ,在IFN γ和L Arg培养体系中 ,随着NO浓度的升高及作用时间的延长 ,对虫体的抑制及杀伤作用越来越明显 ,导致其活动度减弱 ,虫体破裂 ,最终死亡 ;在L NNA培养体系中 ,L NNA浓度越高 ,对虫体的影响越小。结论 ①体外培养中 ,通过激活的巨噬细胞 ,IFN γ和L Arg能促进NO的合成 ,给予L NNA则能抑制NO的合成。

Objective To determine the Influence of IFN γ,L Arg and L NNA upon NO production and the effects of NO resisting Trichinella spiralis in vitro cultivation. Methods The macrophages were isolalated and purified from peritoneal cavity of jirds and cultured in medium of RPMI1640. IFN γ group, L Arg group, L NNA group and control group were established, and every experimental group was further divided into 5 groups of various concentrations respectively. Different doses of IFN γ, L Arg or L NNA were injected into culture bottles containing macrophages. After cultivation for 24 hours, the levels of NO content of culture medium by technique of Nitrate reductase were detected. The larvae of T. spiralis were added into above mentioned culture systems respectively, and the activity and damage were observed. Results ①In vitro culture activated macrophage can produce NO. The production of NO was increased by IFN γ and L Arg , but was decreased by L NNA, and the increasing or deccreasing of production of NO depended on the dosage. The begger the dosage, the more obvious the role. ②In culture systems of IFN γ and L Arg the inhibiting and killing and wounding to larvae became more and more obvious with the increasing of production of NO, the activity of larvae became weak, and tinally larvae became broken and dead. Conclusions ① IFN γ and L Arg can enhance NO production through activated macrophage, but L NNA significantly reduced NO. ②NO possesses the role of controlling and killing and wounding to larvae of T. spiralis.[