National Institute of Genetic Engineering and Biotechnology, P.O.Box: 149651/161, Tehran, Iran.
AAPS PharmSciTech. 2013 Mar;14(1):160-7. doi: 10.1208/s12249-012-9899-8. Epub 2012 Dec 19.
Anti-inflammatory effect of the alcoholic extracts of N. sativa seeds and its callus on mix glial cells of rat with regard to their thymoquinone (TQ) content was investigated. Callus induction was achieved for explants of young leaf, stem, petiole, and root of N. sativa on solid Murashige and Skoog (MS) medium containing 2,4-D (1 mg/l) and kinetin (2.15 mg/l). TQ content of the alcoholic extracts was measured by HPLC. Total phenols were determined using Folin-Ciocalteu method and antioxidant power was estimated using FRAP tests. The mix glial cells, inflamed by lipopolysaccharide, were subjected to anti-inflammatory studies in the presence of various amounts of TQ and the alcoholic extracts. Viability of the cells and nitric oxide production were measured by MTT and Griess reagent, respectively. The leaf callus obtained the highest growth rate (115.4 mg/day) on MS medium containing 2,4-D (0.22 mg/l) and kinetin (2.15 mg/l). Analyses confirmed that TQ content of the callus of leaf was 12 times higher than that measured in the seeds extract. However, it decreased as the calli aged. Decrease in the TQ content of the callus was accompanied with an increase in its phenolic content and antioxidant ability. Studies on the inflamed rat mix glial cells revealed significant reduction in the nitric oxide production in the presence of 0.2 to 1.6 mg/ml of callus extract and 1.25 to 20 μl/ml of the seed extracts. However, the extent of the effects is modified assumingly due to the presence of the other existing substances in the extracts.
研究了黑种草种子及其愈伤组织的醇提物的抗炎作用及其对大鼠混合神经胶质细胞的影响，因为它们含有一定量的胸腺醌（TQ）。通过固体 Murashige 和 Skoog（MS）培养基上的幼叶、茎、叶柄和根外植体诱导愈伤组织的形成，该培养基含有 2,4-D（1mg/L）和激动素（2.15mg/L）。通过 HPLC 测量醇提物中的 TQ 含量。使用 Folin-Ciocalteu 法测定总酚含量，使用 FRAP 试验测定抗氧化能力。在不同浓度的 TQ 和醇提物存在下，将脂多糖致炎的混合神经胶质细胞进行抗炎研究。通过 MTT 和 Griess 试剂分别测量细胞活力和一氧化氮的产生。含有 2,4-D（0.22mg/L）和激动素（2.15mg/L）的 MS 培养基上的叶愈伤组织获得了最高的生长速度（115.4mg/天）。分析证实，叶愈伤组织中的 TQ 含量是种子提取物中 TQ 含量的 12 倍。然而，随着愈伤组织的老化，其含量会下降。愈伤组织中 TQ 含量的减少伴随着其酚类含量和抗氧化能力的增加。对炎症大鼠混合神经胶质细胞的研究表明，在 0.2 至 1.6mg/ml 愈伤组织提取物和 1.25 至 20μl/ml 种子提取物存在的情况下，一氧化氮的产生显著减少。然而，由于提取物中存在其他现有物质，其效果的程度会发生变化。
