七叶皂苷精氨酸盐抗炎作用的量效、时效关系及对NO合成的影响

目的研究七叶皂苷精氨酸盐(Aescine-Arginine,ACA)抗炎作用的量效关系及时效关系,并探讨ACA对脂多糖(LPS)诱导的巨噬细胞一氧化氮(NO)合成的影响。方法70只♂小鼠随机分成7组,即生理盐水组、双氯芬酸钠(10mg·kg-1)组、ACA(0.5、1、2、4和8mg·kg-1)组,采用小鼠腹腔毛细血管通透性模型,以小鼠腹腔渗出液中伊文思兰的渗出量为指标,观察ACA抗炎作用的量效关系;90只♂小鼠随机分成9组,即生理盐水组、双氯芬酸钠(10mg·kg-1)组及ACA(2mg·kg-1)给药,1、2、4、8、12、16和24h组,采用小鼠腹腔毛细血管通透性模型,以小鼠腹腔渗出液中伊文思兰的渗出量为指标,观察ACA抗炎作用的时效关系;用脂多糖(LPS)作为巨噬细胞(RAW264.7)的NO合成诱导剂,加入不同浓度的ACA,培养后取上清液,用Griess试剂测定NO产生量。结果ACA的2,4和8mg·kg-13个剂量皆能明显降低腹腔渗出液中伊文思兰的渗出量,且具有明显的剂量依赖关系;ACA在给药后4,8,12,16和24h能够明显降低腹腔渗出液中伊文思兰的渗出量;LPS在1mg·L-1浓度作用于RAW 264.7细胞,可诱导大量NO合成。ACA对LPS诱导的NO合成有明显的抑制作用,其抑制作用具有明显的量效关系。结论ACA的抗炎作用在4h起效,药效可持续24h,且具有明显的剂量依赖性;ACA降低LPS诱导的炎性因子NO的产生,可能是ACA减轻炎症反应机制之一。

Dose-and time-effect relationship on anti-inflammation of Aescine-Arginine and its effect on nitric oxide synthesis

Aim To study the dose-and time-effect relationship on anti-inflammation of ACA and the its effect on LPS-induced nitric oxide synthesis in macrophages.Methods For dose-effect relationship study,seventy male Kunming white mice were randomly divided into seven groups:control,diclofenac sodium(10 mg· kg-1) and ACA(0.5,1,2,4 and 8 mg·kg-1) group.The capillary permeability was measured by detecting Evans Blue absorbance after glacial acetic acid was injected intraperitoneally.For time-effect relationship study,ninety male Kunming white mice were randomly divided into nine groups: control,diclofenac sodium(10 mg· kg-1),and ACA(2 mg·kg-1;1,2,4,8,12,16 and 24 h,respectively) group.Also the capillary permeability was measured by detecting Evans Blue absorbance after glacial acetic acid was injected intraperitoneally.Nitric oxide(NO) production in RAW264.7 cells was induced by LPS in the presence or absence of ACA.The content of NO in the supernatant was detected with Griess reagent.Results The capillary hyperpermeability in mice was significantly decreased in a dose-dependent manner at different doses of ACA(2,4 and 8 mg·kg-1).When the mice were given ACA at dose of 2 mg·kg-1,the capillary hyperpermeability in mice was obviously decreased after 4,8,12,16 and 24 h of administration.LPS at the concentration of 1 mg·L-1 could induce a large ampunt of NO synthesis in RAW264.7 cells.The ACA showed a significant inhibitory effect on LPS-induced NO production in a dose dependent manner.Conclusions The anti-inflammatory action of ACA was observed after 4 h of administration,and the effect could persist at least 24 h in a dose dependent manner.ACA could reduce LPS-induced production of inflammtory factors resulting in the anti-inflammatory effect.