Microparticles induce cell cycle arrest through redox-sensitive processes in endothelial cells: implications in vascular senescence.
Microparticles induce cell cycle arrest through redox-sensitive processes in endothelial cells: implications in vascular senescence.
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DOI：
10.1161/jaha.112.001842
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发表时间：
2012-06
影响因子：
5.4
通讯作者：
Touyz RM
中科院分区：
文献类型：
--
作者：
Burger D;Kwart DG;Montezano AC;Read NC;Kennedy CR;Thompson CS;Touyz RM
慢性疾病加速衰老过程中的内皮功能障碍，这是一个与细胞衰老相关的过程。然而，这一过程背后的机制尚不清楚。我们研究了内皮细胞（EC）衍生的微粒（MPs）是否促进了EC衰老，并质疑活性氧在这一过程中的作用。小鼠原代ECs顺序传代诱导衰老。从传代4到传代21，细胞保持了ECs的表型特征。21代ECs表现出衰老特征，衰老相关β-半乳糖苷酶（SA-βgal）染色增加，细胞周期G1/G0期细胞比例增加，p66Shc磷酸化增加（P<0.05）。与传代4相比，第21代ECs的微粒形成增加（与传代4相比增加约2.2倍，P<0.05）， Rho激酶抑制剂法舒地尔阻断了这种增加。细胞周期分析和衰老相关β-半乳糖苷酶染色表明，将传代4 ECs暴露于MPs后，细胞从增殖表型转变为非增殖表型。MPs增加了O2•−（~ 2.7倍）和H2O2（~ 2.6倍）的EC生成，这种作用被夹克花碱（烟酰胺腺嘌呤二核苷酸磷酸氧化酶抑制剂）和鱼藤酮（线粒体氧化酶抑制剂）阻断，但不被别嘌呤醇（黄嘌呤氧化酶抑制剂）阻断。MPs增加了细胞周期蛋白p21 cip1和p16ink4a的表达，并刺激了ECs中p66Shc的磷酸化（与未治疗的ECs相比p <0.05）。用活性氧清除剂4,5-二羟基苯-1,3-二磺酸钠（铁）预处理可消除MPs的衰老作用。MPs通过烟酰胺腺嘌呤二核苷酸磷酸氧化酶和线粒体衍生的活性氧促进EC衰老。这种氧化还原敏感过程可能在衰老血管功能障碍中起重要作用。[J] .中华心脏杂志，2012;1:e001842 . doi: 10.1161/JAHA.112.001842.]
Chronic disease accelerates endothelial dysfunction in aging, a process associated with cell senescence. However, the mechanisms underlying this process are unclear. We examined whether endothelial cell (EC)-derived microparticles (MPs) facilitate EC senescence and questioned the role of reactive oxygen species in this process. Senescence was induced by sequential passaging of primary mouse ECs. Cells retained phenotypic characteristics of ECs from passage 4 through passage 21. Passage 21 ECs exhibited features of senescence, including increased staining of senescence-associated β-galactosidase (SA-βgal), a greater percentage of cells in G1/G0 phase of the cell cycle, and increased phosphorylation of p66Shc (P<0.05). Microparticle formation from passage 21 ECs was increased versus passage 4 ECs (∼2.2-fold increase versus passage 4, P<0.05), and the Rho kinase inhibitor fasudil blocked this increase. Exposure of passage 4 ECs to MPs shifted cells from a proliferating to a nonproliferating phenotype, as indicated by cell cycle analysis and increased senescence-associated β-galactosidase staining. MPs increased EC generation of O2•− (∼2.7-fold) and H2O2 (∼2.6-fold), effects blocked by apocynin (nicotinamide adenine dinucleotide phosphate oxidase inhibitor) and rotenone (mitochondrial oxidase inhibitor) but not by allopurinol (xanthine oxidase inhibitor). MPs increased expression of cell cycle proteins p 21 cip1 and p16ink4a and stimulated phosphorylation of p66Shc in ECs (P<0.05 versus untreated ECs). Pretreatment with the reactive oxygen species scavenger sodium 4,5-dihydroxybenzene-1,3-disulfonate (tiron) abrogated the prosenescent effects of MPs. MPs promote EC senescence through nicotinamide adenine dinucleotide phosphate oxidase- and mitochondrial-derived reactive oxygen species. Such redox-sensitive processes may be important in vascular dysfunction in aging. (J Am Heart Assoc. 2012;1:e001842 doi: 10.1161/JAHA.112.001842.)