Jancinova Viera, Nosal Radomir, Lojek Antonin, Ciz Milan, Ambrozova Gabriela, Mihalova Danica, Bauerova Katarina, Harmatha Juraj, Perecko Tomas
Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia.
Neuro Endocrinol Lett. 2010;31 Suppl 2:79-83.
Formation of reactive oxygen species in neutrophils of rats with adjuvant arthritis and generation of nitric oxide in RAW 264.7 macrophages were analysed in the presence of pinosylvin.
The method of chemiluminescence was used for the detection of reactive oxygen species in blood of rats with adjuvant arthritis. Pinosylvin (50 mg/kg, daily, p.o.) and methotrexate (0.4 mg/kg, twice a week, p.o.) were applied separately or in a combination over a period of 28 days from the day of immunisation. Adjuvant arthritis was accompanied by a significantly increased number of neutrophils, by elevated concentration of oxidants in blood and by excessive responsiveness of neutrophils to stimulation with PMA. In rats treated with methotrexate, all these changes were significantly reduced and the inhibition became more pronounced when methotrexate was applied in the combination with pinosylvin; the monotherapy with pinosylvin did not induce any detectable changes in the parameters tested. Under in vitro conditions, pinosylvin inhibited formation of nitric oxide (NO) in macrophages, as demonstrated by the decreased concentration of nitrite - the end-product of NO metabolism (assessed by Griess' method), by the reduced expression of inducible NO synthase (detected by Western blot), and by the failure of pinosylvin to scavenge nitric oxide (measured amperometrically in cell-free system).
The observed ability of pinosylvin to decrease concentration of reactive oxygen and nitrogen species, along with its capacity to enhance the efficacy of methotrexate in arthritis treatment may shed more light into the pharmacological potential of this prospective natural substance.
在存在白皮杉醇的情况下，分析佐剂性关节炎大鼠中性粒细胞中活性氧的形成以及RAW 264.7巨噬细胞中一氧化氮的生成。
采用化学发光法检测佐剂性关节炎大鼠血液中的活性氧。从免疫当天起，连续28天分别或联合应用白皮杉醇（50毫克/千克，每日，口服）和甲氨蝶呤（0.4毫克/千克，每周两次，口服）。佐剂性关节炎伴有中性粒细胞数量显著增加、血液中氧化剂浓度升高以及中性粒细胞对佛波酯刺激的过度反应。在用甲氨蝶呤治疗的大鼠中，所有这些变化均显著减少，当甲氨蝶呤与白皮杉醇联合应用时抑制作用更为明显；单独使用白皮杉醇的单一疗法未引起所测参数的任何可检测到的变化。在体外条件下，白皮杉醇抑制巨噬细胞中一氧化氮（NO）的形成，这表现为亚硝酸盐（NO代谢终产物，通过格里斯法评估）浓度降低、诱导型一氧化氮合酶表达减少（通过蛋白质免疫印迹法检测）以及白皮杉醇未能清除一氧化氮（在无细胞系统中通过安培法测量）。
观察到的白皮杉醇降低活性氧和氮物种浓度的能力，以及其增强甲氨蝶呤治疗关节炎疗效的能力，可能会更深入地揭示这种潜在天然物质的药理潜力。
