Biochemistry Department, Science College, King Saud University, Riyadh, Saudi Arabia.
Appl Biochem Biotechnol. 2013 Mar;169(6):1858-69. doi: 10.1007/s12010-013-0113-6. Epub 2013 Jan 24.
The best known physiologic function of secreted phospholipase A2 (sPLA2) group IIA (sPLA2-IIA) is defense against bacterial infection through hydrolytic degradation of bacterial membrane phospholipids. In fact, sPLA2-IIA effectively kills Gram-positive bacteria and to a lesser extent Gram-negative bacteria and is considered a major component of the eye's innate immune defense system. The antibacterial properties of sPLA2 have been demonstrated in rabbit and human tears. In this report, we have analyzed the bactericidal activity of dromedary tears and the subsequently purified sPLA2 on several Gram-positive bacteria. Our results showed that the sPLA2 displays a potent bactericidal activity against all the tested bacteria particularly against the Staphylococcus strains when tested in the ionic environment of tears. There is a synergic action of the sPLA2 with lysozyme when added to the bacteria culture prior to sPLA2. Interestingly, lysozyme purified from dromedary tears showed a significant bactericidal activity against Listeria monocytogene and Staphylococcus epidermidis, whereas the one purified from human tears displayed no activity against these two strains. We have also demonstrated that Ca(2+) is crucial for the activity of dromedary tear sPLA2 and to a less extent Mg(2+) ions. Given the presence of sPLA2 in tears and intestinal secretions, this enzyme may play a substantial role in innate mucosal and systemic bactericidal defenses against Gram-positive bacteria.
分泌型磷脂酶 A2(sPLA2)IIA 亚型(sPLA2-IIA)最为人所知的生理功能是通过水解细菌膜磷脂来抵抗细菌感染。事实上,sPLA2-IIA 能有效杀死革兰氏阳性菌,在一定程度上也能杀死革兰氏阴性菌,被认为是眼睛先天免疫防御系统的主要组成部分。sPLA2 的抗菌特性已在兔和人泪液中得到证实。在本报告中,我们分析了单峰驼泪液的杀菌活性以及随后从泪液中分离纯化的 sPLA2 对几种革兰氏阳性菌的作用。结果表明,sPLA2 在离子环境中对所有测试的细菌均显示出强大的杀菌活性,特别是对金黄色葡萄球菌菌株。当 sPLA2 与溶菌酶一起添加到细菌培养物中时,会产生协同作用。有趣的是,从单峰驼泪液中纯化的溶菌酶对李斯特菌和表皮葡萄球菌表现出显著的杀菌活性,而从人泪液中纯化的溶菌酶对这两种菌株均无活性。我们还证明了 Ca2+ 对单峰驼泪液 sPLA2 的活性至关重要,而 Mg2+ 离子的作用则较小。鉴于 sPLA2 存在于泪液和肠道分泌物中,这种酶可能在针对革兰氏阳性菌的先天黏膜和全身杀菌防御中发挥重要作用。