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
{"title":"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","authors":"Anne Manson ,&nbsp;Tanja Winter ,&nbsp;Harold M. Aukema","doi":"10.1016/j.plefa.2023.102555","DOIUrl":null,"url":null,"abstract":"<div><p>Phospholipase A<sub>2</sub> (PLA<sub>2</sub>) enzymes cleave cell membrane phospholipids and release polyunsaturated fatty acids (PUFA), which can be converted into oxylipins. However, little is known about PLA<sub>2</sub> preference for PUFA, and even less is known about how this further impacts oxylipin formation. Therefore, we investigated the role of different PLA<sub>2</sub> 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 sPLA<sub>2</sub> 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 sPLA<sub>2</sub> and iPLA<sub>2</sub> isoforms were highest whereas levels of cPLA<sub>2</sub> were low, consistent with activity. In conclusion, sPLA<sub>2</sub> enzymes lead to the formation of DHA oxylipins, while iPLA<sub>2</sub> 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 PLA<sub>2</sub> activity studies.</p></div>","PeriodicalId":94179,"journal":{"name":"Prostaglandins, leukotrienes, and essential fatty acids","volume":null,"pages":null},"PeriodicalIF":3.0000,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Phospholipase A2 enzymes differently impact PUFA release and oxylipin formation ex vivo in rat hearts\",\"authors\":\"Anne Manson ,&nbsp;Tanja Winter ,&nbsp;Harold M. Aukema\",\"doi\":\"10.1016/j.plefa.2023.102555\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Phospholipase A<sub>2</sub> (PLA<sub>2</sub>) enzymes cleave cell membrane phospholipids and release polyunsaturated fatty acids (PUFA), which can be converted into oxylipins. However, little is known about PLA<sub>2</sub> preference for PUFA, and even less is known about how this further impacts oxylipin formation. Therefore, we investigated the role of different PLA<sub>2</sub> 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 sPLA<sub>2</sub> 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 sPLA<sub>2</sub> and iPLA<sub>2</sub> isoforms were highest whereas levels of cPLA<sub>2</sub> were low, consistent with activity. In conclusion, sPLA<sub>2</sub> enzymes lead to the formation of DHA oxylipins, while iPLA<sub>2</sub> 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 PLA<sub>2</sub> activity studies.</p></div>\",\"PeriodicalId\":94179,\"journal\":{\"name\":\"Prostaglandins, leukotrienes, and essential fatty acids\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.0000,\"publicationDate\":\"2023-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Prostaglandins, leukotrienes, and essential fatty acids\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0952327823000248\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Prostaglandins, leukotrienes, and essential fatty acids","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0952327823000248","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
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.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
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
期刊最新文献
Gallein but not fluorescein enhances the PGD2-stimulated synthesis of osteoprotegerin and interleukin-6 in osteoblasts Omega-3 fatty acids mitigate skin damage caused by ultraviolet-B radiation The disparate effects of omega-3 PUFAs on intestinal microbial homeostasis in experimental rodents under physiological condition Resolvin D4 mitigates lipopolysaccharide-induced lung injury in mice Blood EPA and DHA status among people living in the United States from 2000 to 2023
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1