补骨脂抗鱼类多子小瓜虫的活性成分研究
首发时间:2016-05-27
摘要:多子小瓜虫(Ichthyophthirius multifiliis,简称小瓜虫)是一种寄生鱼类体表的纤毛类原生动物,经常给水产养殖业造成重大经济损失。随着国家禁止孔雀石绿等特效药的使用,寻找安全有效的替代药物势在必行。本研究利用活性跟踪分离技术从中草药补骨脂(Psoralea corylifolia)中分离获得补骨脂定(psoralidin)和异补骨脂素(isopsoralen),并在离体和在体条件下评价其抗小瓜虫的活性。结果表明这两种化合物都具有良好的抗虫效果,并且补骨脂定的抗虫效果优于异补骨脂素。0.8 mg/L的补骨脂定可以在4小时内100%杀灭小瓜虫幼虫;0.9 mg/L和1.2 mg/L的补骨脂定可以导致小瓜虫包囊前体和包囊无法孵出小瓜虫幼虫;在体试验结果表明2.5 mg/L的补骨脂定浸泡患病鱼5小时可以显著降低小瓜虫孵化率。此外,利用细胞凋亡Annexin V-EGFP/PI双染法揭示2.5 mg/L的补骨脂定浸泡患病鱼导致小瓜虫孵化率显著降低可能是由于该浓度的补骨脂定引起了部分小瓜虫发生了细胞凋亡。由此可见,补骨脂抗小瓜虫的主要活性成分为补骨脂定,其独特的化学结构和优良的在体抗虫效果为研制新的抗虫药物提供了科学依据。
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In vitro and in vivo assessment of the effect of antiprotozoal compounds isolated from Psoralea corylifolia against Ichthyophthirius multifiliis in fish
Abstract: Ichthyophthirius multifiliis, an external fish parasite, often causes significant economic damage to the aquaculture industry. Since the use of malachite green was banned, the search of alternative substance to control I. multifiliis infections becomes stringent. In present study, in vitro and in vivo anti-ich efficacies of isopsoralen and psoralidin, two active compounds isolated from methanol extract of Psoralea corylifolia by bioassayguided fractionation based on the efficacy of anti-ich encysted tomonts,were evaluated. In vitro antiprotozoal efficacy of psoralidin is much better than that of isopsoralen. Psoralidin can kill all theronts at concentrations of 0.8 mg/L or more during 4 h exposure; and terminate reproduction of I. multifiliis post 6 h exposure of protomonts to 0.9 mg/L and encysted tomonts to 1.2 mg/L. In vivo trials showed that 5 h exposure of infected fish to 2.5 mg/L of psoralidin significantly reduced the number of theronts released from tomonts. Furthermore, we observed that a part of protomonts, collected from infected fish post treatment, presented characteristic morphological changes of apoptosis after staining with Annexin V-EGFP/propidium iodide, indicating the possible mechanism of psoralidin against I. multifiliis trophont in situ. On the basis of these results, psoralidin can be used as a potential lead compound for the developmentof commercial drug against I. Multifiliis.
Keywords: Ichthyophthirius multifiliis Goldfish Antiprotozoal activity Psoralidin Psoralea corylifolia
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No.4690871114989214****
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补骨脂抗鱼类多子小瓜虫的活性成分研究
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