【目的】以猪中性粒细胞为靶细胞,研究白芍制剂对其相关功能(呼吸爆发和弹性蛋白酶释放)及磷酸二酯酶活性的影响。【方法】中性粒细胞分离纯化后,以细胞色素C还原法检测中性粒细胞呼吸爆发,以弹性蛋白酶底物的减少反映弹性蛋白酶的释放量,以HPLC检测底物cAMP的变化反映磷酸二酯酶的活性等。【结果】白芍制剂呈剂量依赖性抑制中性粒细胞的呼吸爆发(P〈0.001),3个不同浓度的抑制率分别为33.15%、45.96%和62.22%;对弹性蛋白酶的释放在低浓度时表现抑制作用(P〈0.05),抑制率为23.22%,在高浓度时表现促进作用;对中性粒细胞磷酸二酯酶活性具有抑制作用,低、高浓度抑制率分别为13.24%和25.87%(P〈0.01);HPLC指纹图谱分析表明白芍制剂主要存在5种成分。【结论】白芍制剂对猪中性粒细胞呼吸爆发具有明显的抑制作用,对弹性蛋白酶释放在低浓度有显著抑制作用,其作用机制可能与白芍制剂对磷酸二酯酶活性的抑制有关。
[Objective] The objective of this experiment is to study the influence ofBaishao (Paeonia lactiflora) preparation on respiratory burst and elastase release of neutrophils with porcine neutrophils as the target cells. [ Method] after the separation and purification of neutrophils, respiratory burst of neutrophils was detected with cytochrome C reduction method, elastase release was indicated with the substrate reduction of elastase, and phosphodiesterase (PDE) activity was calculated by cAMP change examined with HPLC before and after the reaction, [Result] Baishao preparation could significantly inhibit neutrophils respiratory burst dose dependently (P〈0.001), the inhibition ratios of three different concentrations were 33.15%, 45.96% and 62.22%, respectively; the adjustment of Baishao preparation on elastase release was duplicate, having inhibition in low concentration, inhibition ratio was 23.22%, while having promotion in high concentration. Baishao preparation could obviously inhibit PDE activity in neutrophils, and the inhibition ratios of low and high concentration were 13.24% and 25.87% (P〈0.01), respectively. Analysis of HPLC finger print showed that there were five ingredients in Baishao preparation. [ Conclusion ] Baishao preparation has significant inhibition on respiratory burst of neutrophils, and has obvious inhibition on elastase release at low concentration, the mechanism of which possibly has a close relation with the inhibition on PDE activity.