Doligalska Maria, Jóźwicka Kinga, Donskow-Łysoniewska Katarzyna, Kalinowska Małgorzata
Department of Parasitology, Faculty of Biology University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland.
Department of Plant Biochemistry, Faculty of Biology University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland.
Vet Parasitol. 2017 Jul 15;241:5-13. doi: 10.1016/j.vetpar.2017.05.003. Epub 2017 May 11.
Avena sativa L., 1753 (Poaceae) is used as feed for livestock and as a crop rotation agent. The purpose of the study was to examine the molecular mechanisms behind the antihelminth activity of the oat saponins avenacoside B (AveB) and 26-desglucoavenacoside B (26DGAveB) by evaluating their effect on Heligmosomoides bakeri, a parasitic nematode of mice. The avenacosides AveB and 26DGAveB were separated and purified from A. sativa green leaves, and their mycotoxic activity was confirmed against the fungus Trichoderma harzianum. The anti-nematode activity of the avenacosides was measured by egg hatching assay. In the surviving L3 larvae exposed to avenacosides, the expression of CED-9, a protein of the apoptosis pathway, was identified by Western blotting. The protein profile of L3 larvae was monitored by High Performance Liquid Chromatography (HPLC). The action of saponins on glycoprotein pump (Pgp) activity in L3 larvae was compared to that of the pump blocker Verapamil (VPL). A mouse model was used to measure the infectivity of L3 larvae exposed to AveB and 26DGAveB, and the outcome of the immune response. Both compounds induced morphological changes in larvae and blocked Pgp activity; however, only 26DGAveB provoked expression of CED-9. The infected mice displayed changes in the molecular pattern of larval proteins and enhanced IL-4 production, indicating that avenacosides reduced the infectivity of H. bakeri larvae. In avenacosides, the residue without glucose at the C26 position demonstrated greater anti-nematode activity. Our findings indicate that A. sativa compounds are natural products with anti-parasitic activity.
燕麦（Avena sativa L.，1753，禾本科）被用作牲畜饲料和轮作作物。本研究的目的是通过评估燕麦皂苷燕麦皂苷B（AveB）和26 - 去葡萄糖燕麦皂苷B（26DGAveB）对小鼠寄生线虫巴氏类圆线虫（Heligmosomoides bakeri）的作用，来研究其抗蠕虫活性背后的分子机制。从燕麦绿叶中分离并纯化出燕麦皂苷AveB和26DGAveB，并证实它们对哈茨木霉（Trichoderma harzianum）具有抗真菌毒素活性。通过虫卵孵化试验测定燕麦皂苷的抗线虫活性。在暴露于燕麦皂苷的存活L3幼虫中，通过蛋白质免疫印迹法鉴定凋亡途径蛋白CED - 9的表达。通过高效液相色谱法（HPLC）监测L3幼虫的蛋白质谱。将皂苷对L3幼虫糖蛋白泵（Pgp）活性的作用与泵阻滞剂维拉帕米（VPL）的作用进行比较。使用小鼠模型来测量暴露于AveB和26DGAveB的L3幼虫的感染性以及免疫反应的结果。两种化合物均诱导幼虫形态变化并阻断Pgp活性；然而，只有26DGAveB能引发CED - 9的表达。受感染的小鼠幼虫蛋白质分子模式发生变化，IL - 4产生增加，表明燕麦皂苷降低了巴氏类圆线虫幼虫的感染性。在燕麦皂苷中，C26位无葡萄糖的残基表现出更强的抗线虫活性。我们的研究结果表明，燕麦化合物是具有抗寄生虫活性的天然产物。
