Anandamide induces apoptosis in human cells via vanilloid receptors -: Evidence for a protective role of cannabinoid receptors
Anandamide induces apoptosis in human cells via vanilloid receptors -: Evidence for a protective role of cannabinoid receptors
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DOI：
10.1074/jbc.m005722200
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发表时间：
2000-10-13
影响因子：
4.8
通讯作者：
Finazzi-Agrò, A
中科院分区：
文献类型：
--
作者：
Maccarrone, M;Lorenzon, T;Finazzi-Agrò, A
内源性大麻素anandamide（AEA）显示诱导凋亡小体形成和DNA片段化，程序性细胞死亡的标志，在人神经母细胞瘤CHP 100和淋巴瘤U937细胞。RNA和蛋白质合成抑制剂，如放线菌素D和放线菌酮AEA诱导的凋亡小体的数量减少到五分之一，而AEA转运蛋白抑制剂AM404或AEA水解酶抑制剂ATFMK显着增加死亡细胞的数量。此外，大麻素或香草素受体的特异性拮抗剂分别增强或抑制AEA诱导的细胞死亡。其他内源性大麻素，如2-花生四烯酰甘油，亚油酰乙醇酰胺，油酰乙醇酰胺，棕榈酰乙醇酰胺，在相同的实验条件下，没有促进细胞死亡。AEA诱导的凋亡小体的形成被细胞内钙离子的增加（3倍于对照组）、线粒体解偶联（6倍）和细胞色素c的释放（3倍）所抑制。细胞内钙离子螯合剂EGTA-AM可使凋亡小体的数量减少到对照组的40%，电转移的抗细胞色素c单克隆抗体可完全阻止AEA诱导的凋亡。此外，5-脂氧合酶抑制剂5，8，11，14-二十碳四炔酸和MK 886，环氧合酶抑制剂吲哚美辛，caspase-3和caspase-9抑制剂Z-DEVD-FMK和Z-LEHD-FMK，而一氧化氮合酶抑制剂N-硝基-L-精氨酸甲酯则不能显著降低AEA诱导的细胞死亡效应。所提供的数据表明大麻素受体通过香草素受体对AEA诱导的细胞凋亡具有保护作用。
The endocannabinoid anandamide (AEA) is shown to induce apoptotic bodies formation and DNA fragmentation, hallmarks of programmed cell death, in human neuroblastoma CHP100 and lymphoma U937 cells. RNA and protein synthesis inhibitors like actinomycin D and cycloheximide reduced to one-fifth the number of apoptotic bodies induced by AEA, whereas the AEA transporter inhibitor AM404 or the AEA hydrolase inhibitor ATFMK significantly increased the number of dying cells. Furthermore, specific antagonists of cannabinoid or vanilloid receptors potentiated or inhibited cell death induced by AEA, respectively. Other endocannabinoids such as 2-arachidonoylglycerol, linoleoylethanolamide, oleoylethanolamide, and palmitoylethanolamide did not promote cell death under the same experimental conditions. The formation of apoptotic bodies induced by AEA was paralleled by increases in intracellular calcium (3-fold over the controls), mitochondrial uncoupling (6-fold), and cytochrome c release (3-fold). The intracellular calcium chelator EGTA-AM reduced the number of apoptotic bodies to 40% of the controls, and electrotransferred anti-cytochrome c monoclonal antibodies fully prevented apoptosis induced by AEA, Moreover, 5-lipoxygenase inhibitors 5,8,11,14-eicosatetraynoic acid and MK886, cyclooxygenase inhibitor indomethacin, caspase-3 and caspase-9 inhibitors Z-DEVD-FMK and Z-LEHD-FMK, but not nitric oxide synthase inhibitor Nw-nitro-L-arginine methyl ester, significantly reduced the cell death-inducing effect of AEA. The data presented indicate a protective role of cannabinoid receptors against apoptosis induced by AEA via vanilloid receptors.