Naghibi Harat Z, Lakpour N, Sadeghipoor H R, Kamalinejad M, Eshraghian M R, Naghibi B, Akhondi M M, Binaafar S, Sadeghi M R
IVF Center, Arash Women Hospital, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Andrologia. 2015 Dec;47(10):1183-9. doi: 10.1111/and.12401. Epub 2015 Jan 28.
This study was designed to find out Ruta graveolens L. functional components, which have immobilisation effect on human spermatozoa for contraceptive use. A five-step fractionation method was used to derive different components from rue aqueous extract by using hexane, chloroform, ethanol, acetone and ultrapure water. Gas chromatography-mass spectrophotometery (GC-MS) of all fractions and the aqueous extract were performed to determine the chemical components. The immobilisation assay and membrane integrity test were also performed with four different coumarins, which were found in GC-MS in a concentration of 10 μm. Hexane, chloroform, acetone and ethanol fractions could significantly decrease motility of sperms within the first and the second hours. Hexane fraction had also significant immediate effect. The aqueous fraction had no effect on sperm motility. Meanwhile, GC-MS revealed that aqueous extract and effective fractions had similar coumarin compounds. We performed the immobilisation assay on four different coumarins, which were found in GC-MS in a concentration of 10 μm. Reduction of sperm motility was only significant for xanthotoxin. In the sperm viability and membrane integrity tests, hexane and ethanolic fractions could impair sperm vitality significantly, in contrast to coumarins. These results indicated that a part of immobilising effect of rue could be due to its coumarins. The possible mechanism could be blocking of spermatozoa potassium channels.
本研究旨在找出具有使人精子制动作用以供避孕使用的芸香功能成分。采用五步分馏法，用己烷、氯仿、乙醇、丙酮和超纯水从芸香水提取物中提取不同成分。对所有馏分和水提取物进行气相色谱 - 质谱联用（GC - MS）分析以确定化学成分。还用在GC - MS中浓度为10μm时发现的四种不同香豆素进行了制动试验和膜完整性测试。己烷、氯仿、丙酮和乙醇馏分在最初两小时内可显著降低精子活力。己烷馏分也有显著的即刻效应。水馏分对精子活力无影响。同时，GC - MS显示水提取物和有效馏分含有相似的香豆素化合物。我们对在GC - MS中浓度为10μm时发现的四种不同香豆素进行了制动试验。只有花椒毒素能显著降低精子活力。在精子活力和膜完整性测试中，与香豆素不同，己烷和乙醇馏分可显著损害精子活力。这些结果表明，芸香的部分制动作用可能归因于其香豆素。可能的机制可能是阻断精子钾通道。
