磷脂酶A2酶对大鼠体外心脏PUFA释放和氧基化蛋白形成的不同影响

Anne Manson , Tanja Winter , Harold M. Aukema
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引用次数: 1

摘要

磷脂酶A2(PLA2)酶切割细胞膜磷脂并释放多不饱和脂肪酸(PUFA),这些脂肪酸可以转化为氧基化物。然而,人们对PLA2对PUFA的偏好知之甚少,对这如何进一步影响oxylpin的形成也知之甚少。因此,我们研究了不同PLA2基团在大鼠心脏PUFA释放和oxylipin形成中的作用。Sprague-Dawley大鼠心脏匀浆在没有或与varespladib(VAR)、甲基花生四烯酸氟磷酸盐(MAFP)或EDTA一起孵育。通过HPLC-MS/MS测定游离PUFA和oxylipins,并通过RT-qPCR测定异构体表达。VAR对sPLA2 IIA和/或V的抑制降低了ARA和DHA的释放,但只有DHA氧基化酶受到抑制。MAFP减少了ARA、DHA、ALA和EPA的释放,以及ARA、LA、DGLA、DHA、ALA和EPA oxylipins的形成。有趣的是,环氧合酶和12-脂氧合酶氧合酶没有被抑制。sPLA2和iPLA2亚型的mRNA表达水平最高,而cPLA2的水平较低,与活性一致。总之,sPLA2酶导致DHA氧基化酶的形成,而iPLA2可能负责健康大鼠心脏中大多数其他氧基化蛋白的形成。PUFA的释放不能暗示氧化脂质的形成,因此,两者都应在PLA2活性研究中进行评估。
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Phospholipase A2 enzymes differently impact PUFA release and oxylipin formation ex vivo in rat hearts

Phospholipase A2 (PLA2) enzymes cleave cell membrane phospholipids and release polyunsaturated fatty acids (PUFA), which can be converted into oxylipins. However, little is known about PLA2 preference for PUFA, and even less is known about how this further impacts oxylipin formation. Therefore, we investigated the role of different PLA2 groups in PUFA release and oxylipin formation in rat hearts. Sprague-Dawley rat heart homogenates were incubated without or with varespladib (VAR), methyl arachidonyl fluorophosphonate (MAFP) or EDTA. Free PUFA and oxylipins were determined by HPLC-MS/MS, and isoform expressions by RT-qPCR. Inhibition of sPLA2 IIA and/or V by VAR reduced the release of ARA and DHA, but only DHA oxylipins were inhibited. MAFP reduced the release of ARA, DHA, ALA, and EPA, and the formation of ARA, LA, DGLA, DHA, ALA, and EPA oxylipins. Interestingly, cyclooxygenase and 12-lipoxygenase oxylipins were not inhibited. mRNA expression levels of sPLA2 and iPLA2 isoforms were highest whereas levels of cPLA2 were low, consistent with activity. In conclusion, sPLA2 enzymes lead to the formation of DHA oxylipins, while iPLA2 is likely responsible for the formation of most other oxylipins in healthy rat hearts. Oxylipin formation cannot be implied from PUFA release, thus, both should be evaluated in PLA2 activity studies.

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来源期刊
Prostaglandins, leukotrienes, and essential fatty acids
Prostaglandins, leukotrienes, and essential fatty acids Clinical Biochemistry, Endocrinology, Diabetes and Metabolism
CiteScore
5.30
自引率
0.00%
发文量
0
审稿时长
64 days
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