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儿茶素多酚以及植物寄生线虫大豆异皮线虫提取物对三种线虫蛋白酶活性和行为的影响

Effects of catechin polyphenols and preparations from the plant-parasitic nematode Heterodera glycines on protease activity and behaviour in three nematode species.

作者信息

Masler E P

机构信息

Nematology Laboratory United States,Department of Agriculture,Agricultural Research Service, 10300 Baltimore Avenue, BARC-West,BeltsvilleMD20705-2350,USA.

出版信息

J Helminthol. 2014 Sep;88(3):349-56. doi: 10.1017/S0022149X13000254. Epub 2013 May 2.

Abstract

Protease activities in preparations from the plant-parasitic nematodes Heterodera glycines and Meloidogyne incognita and the free-living nematode Panagrellus redivivus were inhibited by exposure to a series of eight catechin polyphenol analogues, (+)-catechin, (-)-epicatechin (EC), (-)-gallocatechin (GC), (-)-epigallocatechin (EGC), (-)-catechin gallate (CG), (-)-gallocatechin gallate (GCG), (-)-epicatechin gallate (ECG) and (-)-epigallocatechin gallate (EGCG) (1 mm each), and by a preparation from H. glycines cysts. General protease activity detected with the FRET-peptide substrate QXL520-KSAYMRF-K(5-FAM)a and proteasome chymotrypsin-like (CTL) activity detected with succinyl-LLVY-AMC were each inhibited significantly more (P < 0.05) by the gallated form of the polyphenol than by the corresponding non-gallated form. Species differences in response to inhibition across all analogues were revealed with the CTL substrate, but CG was a consistently potent inhibitor across all three species and with each substrate. A heat-stable component (CE) from H. glycines cysts inhibited M. incognita CTL activity by 92.07 ± 0.68%, significantly less (P < 0.05) in H. glycines (52.86 ± 2.77%), and by only 17.24 ± 0.55% (P < 0.05) in P. redivivus preparations. CTL activity was, however, inhibited more than 60% in all preparations by the proteasome-specific inhibitor MG-132. Hatching of M. incognita infective juveniles exposed to 1 mm CG, ECG, GCG or EGCG was reduced by 83.88 ± 4.26%, 69.98 ± 9.14%, 94.93 ± 1.71% and 87.93 ± 2.89%, respectively, while hatching of H. glycines was reduced less than 25% by each analogue. CE had no effect on nematode hatch, but did cause a 60% reduction in mobility of H. glycines infective juveniles exposed overnight to CE in vitro, which was more (P < 0.05) than the reduction of M. incognita infective juvenile mobility (20%).

摘要

植物寄生线虫大豆胞囊线虫(Heterodera glycines)、南方根结线虫(Meloidogyne incognita)以及自由生活线虫美丽新杆线虫(Panagrellus redivivus)提取物中的蛋白酶活性,在暴露于一系列八种儿茶素多酚类似物,即(+)-儿茶素、(-)-表儿茶素(EC)、(-)-没食子儿茶素(GC)、(-)-表没食子儿茶素(EGC)、(-)-儿茶素没食子酸酯(CG)、(-)-没食子儿茶素没食子酸酯(GCG)、(-)-表儿茶素没食子酸酯(ECG)和(-)-表没食子儿茶素没食子酸酯(EGCG)(各1 mM)以及大豆胞囊线虫孢囊提取物后受到抑制。用FRET肽底物QXL520-KSAYMRF-K(5-FAM)a检测到的一般蛋白酶活性以及用琥珀酰-LLVY-AMC检测到的蛋白酶体胰凝乳蛋白酶样(CTL)活性,与相应的非没食子酸化形式相比,多酚的没食子酸化形式对其抑制作用均显著更强(P < 0.05)。使用CTL底物揭示了所有类似物在抑制反应方面的物种差异,但CG对所有三个物种以及每种底物而言都是一种始终有效的抑制剂。大豆胞囊线虫孢囊中的一种热稳定成分(CE)抑制南方根结线虫CTL活性达92.07±0.68%,在大豆胞囊线虫中抑制作用显著较弱(P < 0.05,为52.86±2.77%),而在美丽新杆线虫提取物中仅抑制17.24±0.55%(P < 0.05)。然而,蛋白酶体特异性抑制剂MG-132在所有提取物中对CTL活性的抑制均超过60%。暴露于1 mM CG、ECG、GCG或EGCG的南方根结线虫感染性幼虫的孵化率分别降低了83.88±4.26%、69.98±9.14%、94.93±1.71%和87.93±2.89%,而每种类似物对大豆胞囊线虫孵化率的降低均小于25%。CE对线虫孵化没有影响,但确实导致体外过夜暴露于CE的大豆胞囊线虫感染性幼虫的运动能力降低了60%,这一降低幅度比南方根结线虫感染性幼虫运动能力的降低幅度(20%)更大(P < 0.05)。

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