Sphingosine 1-phosphate protects human umbilical vein endothelial cells from serum-deprived apoptosis by nitric oxide production
Sphingosine 1-phosphate protects human umbilical vein endothelial cells from serum-deprived apoptosis by nitric oxide production
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DOI：
10.1074/jbc.m011449200
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发表时间：
2001-04-06
影响因子：
4.8
通讯作者：
Kim, YM
中科院分区：
文献类型：
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作者：
Kwon, YG;Min, JK;Kim, YM
1-磷酸鞘氨醇(S1P)可防止内皮细胞凋亡。我们研究了S1P保护内皮细胞免受血清剥夺诱导的细胞凋亡的分子机制和信号通路。我们发现，人脐静脉内皮细胞(HUVECs)在血清剥夺24小时后发生了与DEVDase活性增加、caspase-3激活、细胞色素c释放和DNA片段化相关的凋亡。加入S1P、NO供体S-亚硝基-N-乙酰青霉胺(100um)或caspase-3抑制剂z-VAD-fmk可抑制这些凋亡标记物的表达。S1P的保护作用可被一氧化氮合酶抑制剂N-单甲基-L-精氨酸逆转，但不能被可溶性鸟苷酸环化酶抑制剂1H-(1，2，4)oxadiazolo[4，3-a]-quanoxaline-1-one，逆转，提示NO而不是cGMP对S1P具有保护作用。此外，S1P通过增强钙敏感的一氧化氮合酶活性而增加NO的产生，而不改变eNOS的蛋白水平。EDG-1反义寡核苷酸和EDG-3反义寡核苷酸可显著抑制S1P介导的细胞存活和NO的产生。加入G(I)蛋白抑制剂百日咳毒素、磷脂酶C(PLC)特异性抑制剂U73122和钙离子螯合剂BAPTA-AM可抑制S1P介导的NO生成。这些结果表明，S1P通过激活eNOS活性，主要通过EDG-1和-3/G(I)/PLC/Ca~(2+)信号通路来保护HUVEC免受凋亡的影响。
Sphingosine 1-phosphate (S1P) can prevent endothelial cell apoptosis. We investigated the molecular mechanisms and signaling pathways by which S1P protects endothelial cells from serum deprivation-induced apoptosis. We show here that human umbilical vein endothelial cells (HUVECs) undergo apoptosis associated with increased DEVDase activity, caspase-3 activation, cytochrome c release, and DNA fragmentation after 24 h of serum deprivation. These apoptotic markers were suppressed by the addition of S1P, the NO donor S-nitroso-N-acetylpenicillamine (100 muM), or caspase-3 inhibitor z-VAD-fmk. The protective effects of S1P were reversed by the nitric-oxide synthase (NOS) inhibitor N-monomethyl-L-arginine, but not by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)oxadiazolo[4,3-a]-quanoxaline-1-one, suggesting that NO, but not cGMP, is responsible for S1P protection from apoptosis. Furthermore, S1P increased NO production by enhancing Ca2+ sensitive NOS activity without changes in the eNOS protein level. S1P-mediated cell survival and NO production were suppressed significantly by pretreatment with antisense oligonucleotide of EDG-1 and partially by EDG-3 antisense. S1P-mediated NO production was suppressed by the addition of pertussis toxin, an inhibitor of G(i) proteins, the specific inhibitor of phospholipase C (PLC), U73122, and the Ca2+ chelator BAPTA-AM. These findings indicate that S1P protects HUVECs from apoptosis through the activation of eNOS activity mainly through an EDG-1 and -3/G(i)/PLC/Ca2+ signaling pathway.