黑种草子不同组分抑制亚硝酸盐积累、血小板聚集和血管收缩的体外实验研究

The inhibitory effect of different fractions from Nigellae Semen in vitro on nitrite accumulation,platelet aggregation and vasoconstriction

目的研究黑种草子不同组分对巨噬细胞活化后含亚硝酸盐量、血小板诱导后聚集、离体血管经苯肾上腺素引起收缩的影响。方法黑种草子醇提物经萃取分别得到乙酸乙酯提取物、正丁醇提取物和水提取物,后经大孔树脂纯化分别得到组分Ⅰ(总黄酮类成分)、Ⅱ(总皂苷类成分)和Ⅲ(总多糖类成分);组分Ⅳ(总生物碱成分)为黑种草子经酸水提取后乙醇洗脱乙酸乙酯萃取。以脂多糖和干扰素γ诱导小鼠巨噬细胞活化引起细胞上清液中亚硝酸盐累积;以二磷酸腺苷、花生四烯酸、胶原诱导大鼠体外血小板聚集;以苯肾上腺素引起大鼠离体血管收缩,而后考查黑种草子不同组分对上述指标的影响。结果组分Ⅰ、Ⅱ和Ⅳ抑制亚硝酸盐形成,组分Ⅰ、Ⅱ和Ⅳ具有抗血小板聚集作用,组分Ⅳ具有部分内皮依赖性血管舒张活性。结论黑种草子Ⅰ、Ⅱ和Ⅳ组分可干预血管系统中炎症的多个环节,且各有偏重,这是其活血通经药效的物质基础。

AIM To study the effect of different fractions of Nigellae Semen on nitrite produced from macrophages,platelet aggregation and vasoconstriction.METHODS Three fractionsⅠ-Ⅲ（total flavones,total saponis,total polysaccharides） of Nigellae Semen were extracted by EtoAc,n-BuOH and water respectively,and then purified with microporous resin D101.Nigellae Semen was extracted by acid water and then isolated by ethanol to form the Fraction Ⅳ（total alkaloids）.Nitrite production of activated mouse macrophages by lipopolysaccharide plus IFN-γ was measured by Griess assay.Rat platelet aggregation was induced by adenosine diphosphate,arachidonic acid and collagen in vitro.The vasodilator effects of four fractions were investigated by isometric tension changes of aortic rings.RESULTS Fraction Ⅰ（total flavones）,Ⅱ（total saponis） and Ⅳ（total alkaloids） demonstrated the inhibition against macrophage nitric oxide（NO） production and platelet aggregation,and only Fraction Ⅳ（total alkaloids） reduced phenylephrine-induced vasoconstriction of pre-constricted aorta rings dependent on endothelium.CONCLUSION Fraction Ⅰ,Ⅱ and Ⅳ can work on the multi-nodes during the pathological progress of inflammation through their properties of inhibition of nitrite accumulation,anti-platelet aggregation and vasodilatation,which is the basic mechanism of its traditional effect of activating blood to promote menstruation partly.