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Effects of Heliopsis longipes ethanolic extract on mouse spermatozoa in vitro

机译:长尾葵乙醇提取物对小鼠精子的体外作用

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Abstract Context: Heliopsis longipes (A. Gray) Blake (Asteraceae), a plant native to Mexico, is used in traditional medicine as analgesic and microbicide. The main component in the H. longipes ethanolic extract (HLEE) is affinin, as determined by HPLC/UV–visible and NMR measurement. To date, there is no documented evidence on the spermicidal activity of this extract. Objective: The objective of this study was to assess in vitro the effectiveness of HLEE as spermicide. Materials and methods: The spermicidal activity of HLEE was evaluated by the Sander–Cramer assay. Spermatozoa were incubated for 20 s with HLEE in concentrations ranging from 75 to 2000 μg/mL to determine the minimum effective concentration (MEC) value. The 50% effective concentration (EC50) of HLEE was estimated by assaying serial dilutions from the MEC. Additionally, sperms were incubated with 125, 250, or 500?μg/mL of HLEE to evaluate the viability and the integrity of sperm membrane. Lipid peroxidation was assessed by the thiobarbituric acid reactive substances assay. Results: HLEE caused an inhibition of 100% in spermatozoa motility at a MEC value of 2000?μg/mL; the EC50 value was 125?μg/mL. Additionally, exposure to HLEE at 125, 250, or 500?μg/mL for 30?min decreased sperm viability to 27%, 8%, and 2% of the control value, respectively, and significantly increased the percentage of sperms with structurally disorganized membrane. HLEE also increased significantly the level of lipid peroxidation in sperms with respect to controls. Discussion and conclusion: The results demonstrate the spermicidal activity of HLEE in vitro and suggest that this action is caused by oxidative damage and alterations in the spermatozoal membrane.
机译:摘要背景:墨西哥原住民植物长鳞葵(A. Gray)Blake(菊科)在传统医学中用作镇痛药和杀微生物剂。经HPLC / UV可见和NMR测量确定,长双螺旋菌乙醇提取物(HLEE)的主要成分是亲和素。迄今为止,尚无关于这种提取物杀精活性的文献记录。目的:本研究的目的是在体外评估HLEE作为杀精子剂的有效性。材料和方法:HLEE的杀精活性通过Sander–Cramer分析进行了评估。将精子与HLEE在75至2000μg/ mL范围内孵育20 s,以确定最小有效浓度(MEC)值。 HLEE的50%有效浓度(EC 50 )是通过分析来自MEC的系列稀释液来估算的。另外,将精子与125、250或500μg/ mL的HLEE孵育,以评估精子膜的活力和完整性。通过硫代巴比妥酸反应性物质测定评估脂质过氧化。结果:HLEE在2000?μg/ mL的MEC值下可抑制100%的精子活力。 EC 50 值为125?μg/ mL。此外,在HLEE中以125、250或500μg/ mL的浓度暴露30分钟,可使精子活力分别降低至对照值的27%,8%和2%,并显着增加结构紊乱的精子百分比膜。相对于对照,HLEE还显着增加了精子中脂质过氧化的水平。讨论与结论:结果证明了HLEE在体外具有杀精活性,并表明该作用是由氧化损伤和精子膜的改变引起的。

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