Human leukocytes differentially express endocannabinoid‐glycerol lipases and hydrolyze 2‐arachidonoyl‐glycerol and its metabolites from the 15‐lipoxygenase and cyclooxygenase pathways
Human leukocytes differentially express endocannabinoid‐glycerol lipases and hydrolyze 2‐arachidonoyl‐glycerol and its metabolites from the 15‐lipoxygenase and cyclooxygenase pathways
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人类白细胞差异表达内源性大麻素-甘油脂肪酶，并水解 2-花生四烯酰-甘油及其来自 15-脂氧合酶和环加氧酶途径的代谢物
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发表时间：
2019
影响因子：
5.5
通讯作者：
N. Flamand
中科院分区：
文献类型：
--
作者：
Caroline Turcotte;Élizabeth Dumais;Anne;C. Martin;Marie;É. Bissonnette;L. Boulet;M. Laviolette;V. Di Marzo;N. Flamand
2-花生四烯酸-甘油（2-AG）是一种具有抗炎特性的内源性大麻素。阻断2-AG水解以增强CB 2信号传导已被证明在小鼠炎症模型中有效。然而，2-AG脂肪酶的表达从未在人白细胞中进行过彻底研究。在此，我们研究了人血白细胞和肺泡巨噬细胞（AM）中7种2-AG水解酶的表达，发现了以下蛋白质表达模式：（MAG脂肪酶;嗜酸性粒细胞，AM，单核细胞），羧酸酯酶（CES 1;单核细胞，AM），棕榈酰蛋白硫酯酶（PPT 1; AM）、α/β-水解酶结构域（ABHD 6;主要是AM）、ABHD 12（全部）、ABHD 16 A（全部）和LYPLA 2（溶血磷脂酶2;单核细胞、淋巴细胞、AM）。我们接下来发现，所有白细胞都可以水解2-AG及其衍生自环氧合酶-2（前列腺素E2-甘油[PGE 2-G]）和15-脂氧合酶（15-羟基-二十碳四烯酰-甘油[15-HETE-G]）的代谢产物。中性粒细胞和嗜酸性粒细胞在水解2-AG及其代谢产物方面始终优于单核细胞和淋巴细胞。此外，白细胞水解2-AG及其代谢产物的效力为每个白细胞的2-AG ≥ 15-HETE-G >> PGE 2-G。使用抑制剂甲基花生四烯酸-氟膦酸酯（MAFP），4-硝基苯基-4-（二苯并[d][1，3]间二氧杂环戊烯-5-基（羟基）甲基）哌啶-1-羧酸酯（JZL 184），帕莫司他丁B，4′-氨基甲酰基联苯-4-基甲基（3-（吡啶-4-基）苄基）氨基甲酸酯，N-甲基-N-[[3-（4-甲基吡啶-4-基）苄基]氨基甲酸酯（4-吡啶基）苯基]甲基]-4 ′-（氨基羰基）[1，1 '-联苯]-4-基酯氨基甲酸（WWL 70），4'-[甲基[[3-（4-吡啶基）苯基]甲基]氨基]羰基]氧基]-[1，1 ′-联苯基]-4-羧酸乙酯（WWL 113）、四氢lipstatin和ML 349，我们不能确定负责水解2-AG、PGE 2-G、和15-HETE-G此外，选择性MAG脂肪酶抑制剂JZL 184可阻断中性粒细胞对2-AG、PGE 2-G和15-HETE-G的水解，以及淋巴细胞对PGE 2-G和15-HETE-G的水解，这两种细胞类型具有有限的/无MAG脂肪酶。使用基于活性的蛋白质谱（ABPP）探针标记白细胞中的水解酶，我们发现它们表达许多MAFP敏感的水解酶和一种未知的JZL 184敏感的水解酶，分子量为1.52 kDa。总之，我们的结果表明，人白细胞擅长通过多种脂肪酶水解2-AG及其代谢产物，可能还通过一种尚未表征的52 kDa水解酶。阻断人体中的2-AG水解可能会消除人白细胞降解2-AG及其代谢产物的能力，并增加其体内抗炎作用。
2‐Arachidonoyl‐glycerol (2‐AG) is an endocannabinoid with anti‐inflammatory properties. Blocking 2‐AG hydrolysis to enhance CB2 signaling has proven effective in mouse models of inflammation. However, the expression of 2‐AG lipases has never been thoroughly investigated in human leukocytes. Herein, we investigated the expression of seven 2‐AG hydrolases by human blood leukocytes and alveolar macrophages (AMs) and found the following protein expression pattern: monoacylglycerol (MAG lipase; eosinophils, AMs, monocytes), carboxylesterase (CES1; monocytes, AMs), palmitoyl‐protein thioesterase (PPT1; AMs), α/β‐hydrolase domain (ABHD6; mainly AMs), ABHD12 (all), ABHD16A (all), and LYPLA2 (lysophospholipase 2; monocytes, lymphocytes, AMs). We next found that all leukocytes could hydrolyze 2‐AG and its metabolites derived from cyclooxygenase‐2 (prostaglandin E2‐glycerol [PGE2‐G]) and the 15‐lipoxygenase (15‐hydroxy‐eicosatetraenoyl‐glycerol [15‐HETE‐G]). Neutrophils and eosinophils were consistently better at hydrolyzing 2‐AG and its metabolites than monocytes and lymphocytes. Moreover, the efficacy of leukocytes to hydrolyze 2‐AG and its metabolites was 2‐AG ≥ 15‐HETE‐G >> PGE2‐G for each leukocyte. Using the inhibitors methylarachidonoyl‐fluorophosphonate (MAFP), 4‐nitrophenyl‐4‐(dibenzo[d][1,3]dioxol‐5‐yl(hydroxy)methyl)piperidine‐1‐carboxylate (JZL184), Palmostatin B, 4′‐carbamoylbiphenyl‐4‐yl methyl(3‐(pyridin‐4‐yl)benzyl)carbamate, N‐methyl‐N‐[[3‐(4‐pyridinyl)phenyl]methyl]‐4′‐(aminocarbonyl)[1,1′‐biphenyl]‐4‐yl ester carbamic acid (WWL70), 4′‐[[[methyl[[3‐(4‐pyridinyl)phenyl]methyl]amino]carbonyl]oxy]‐[1,1′‐biphenyl]‐4‐carboxylic acid, ethyl ester (WWL113), tetrahydrolipstatin, and ML349, we could not pinpoint a specific hydrolase responsible for the hydrolysis of 2‐AG, PGE2‐G, and 15‐HETE‐G by these leukocytes. Furthermore, JZL184, a selective MAG lipase inhibitor, blocked the hydrolysis of 2‐AG, PGE2‐G, and 15‐HETE‐G by neutrophils and the hydrolysis of PGE2‐G and 15‐HETE‐G by lymphocytes, two cell types with limited/no MAG lipase. Using an activity‐based protein profiling (ABPP) probe to label hydrolases in leukocytes, we found that they express many MAFP‐sensitive hydrolases and an unknown JZL184‐sensitive hydrolase of ∼52 kDa. Altogether, our results indicate that human leukocytes are experts at hydrolyzing 2‐AG and its metabolites via multiple lipases and probably via a yet‐to‐be characterized 52 kDa hydrolase. Blocking 2‐AG hydrolysis in humans will likely abrogate the ability of human leukocytes to degrade 2‐AG and its metabolites and increase their anti‐inflammatory effects in vivo.
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影响因子：
2.9
作者：
Wang, Ran;Borazjani, Abdolsamad;Matthews, Anberitha T.;Mangum, Lee C.;Edelmann, Mariola J.;Ross, Matthew K.
通讯作者：
Ross, Matthew K.
RAW 264.7 细胞和 H1819 细胞中前列腺素 E2 和前列腺素 F2α 甘油酯钙动员的结构决定因素。
影响因子：
2.9
作者：
Richie-Jannetta,Robyn;Nirodi,ChaitanyaS;Crews,BrendaC;Woodward,DavidF;Wang,JennyW;Duff,PeterT;Marnett,LawrenceJ
通讯作者：
Marnett,LawrenceJ
影响因子：
5.8
作者：
Crow, J. Allen;Bittles, Victoria;Borazjani, Abdolsamad;Potter, Philip M.;Ross, Matthew K.
通讯作者：
Ross, Matthew K.
活化的有机磷杀虫剂对人 THP1 单核细胞/巨噬细胞中脂质甘油酯代谢的灭活：羧酸酯酶 1 和 2 的作用。
影响因子：
4.1
作者：
Xie S;Borazjani A;Hatfield MJ;Edwards CC;Potter PM;Ross MK
通讯作者：
Ross MK
影响因子：
29.4
作者：
Cao Z;Mulvihill MM;Mukhopadhyay P;Xu H;Erdélyi K;Hao E;Holovac E;Haskó G;Cravatt BF;Nomura DK;Pacher P
通讯作者：
Pacher P