Toyama Daniela O, Ferreira Marcelo J P, Romoff Paulete, Fávero Oriana A, Gaeta Henrique H, Toyama Marcos H
UNESP, Campus Experimental do Litoral Paulista, PraÇa Infante Dom Henrique s/no, Bairro Parque Bitaru, 11330-900 São Vicente, SP, Brazil.
Departamento de Botânica, Instituto de Biociências, Universidade de São Paulo, RuadoMatão 277, 05508-090 São Paulo, SP, Brazil.
Biomed Res Int. 2014;2014:726585. doi: 10.1155/2014/726585. Epub 2014 Sep 2.
The aim of this paper was to investigate the effect of chlorogenic acid (5-caffeoylquinic acid, 5CQA), isolated from Baccharis oxyodonta, on the structure and pharmacological effect of secretory phospholipase A2 (sPLA2) from Crotalus durissus terrificus. All in vitro and in vivo experiments were conducted using a purified sPLA2 compared under the same experimental conditions with sPLA2 : 5CQA. 5CQA induced several discrete modifications in the secondary structure and the hydrophobic characteristics of native sPLA2 that induced slight changes in the α-helical content, increase in the random coil structure, and decrease of fluorescence of native sPLA2. Moreover, 5CQA significantly decreased the enzymatic activity and the oedema and myonecrosis induced by native sPLA2. As the catalytic activity of sPLA2 plays an important role in several of its biological and pharmacological properties, antibacterial activity was used to confirm the decrease in its enzymatic activity by 5CQA, which induced massive bacterial cell destruction. We found that 5CQA specifically abolished the enzymatic activity of sPLA2 and induced discrete protein unfolding that mainly involved the pharmacological site of sPLA2. These results showed the potential application of 5CQA in the snake poisoning treatment and modulation of the pathological effect of inflammation induced by secretory PLA2.
本文旨在研究从尖齿酒神菊中分离出的绿原酸(5-咖啡酰奎尼酸,5CQA)对杜氏响尾蛇分泌型磷脂酶A2(sPLA2)的结构和药理作用的影响。所有体外和体内实验均使用纯化的sPLA2,并在相同实验条件下与sPLA2:5CQA进行比较。5CQA诱导了天然sPLA2二级结构和疏水特性的几种离散变化,导致α-螺旋含量略有变化、无规卷曲结构增加以及天然sPLA2荧光降低。此外,5CQA显著降低了天然sPLA2诱导的酶活性、水肿和肌坏死。由于sPLA2的催化活性在其多种生物学和药理特性中起重要作用,因此利用抗菌活性来证实5CQA导致其酶活性降低,这会引起大量细菌细胞破坏。我们发现5CQA特异性地消除了sPLA2的酶活性,并诱导了主要涉及sPLA2药理位点的离散蛋白质解折叠。这些结果表明5CQA在蛇毒中毒治疗以及调节分泌型磷脂酶A2诱导的炎症病理效应方面具有潜在应用价值。