Protease activated receptor-1 mediates cytotoxicity during ischemia using in vivo and in vitro models.
Protease activated receptor-1 mediates cytotoxicity during ischemia using in vivo and in vitro models.
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DOI：
10.1016/j.neuroscience.2014.09.038
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发表时间：
2014-12-05
期刊：
影响因子：
3.3
通讯作者：
Bai, J.
中科院分区：
文献类型：
--
作者：
Rajput, P. S.;Lyden, P. D.;Chen, B.;Lamb, J. A.;Pereira, B.;Lamb, A.;Zhao, L.;Lei, I. -F.;Bai, J.
关键词：
蛋白酶激活受体（PARs）存在于大脑的神经元和星形胶质细胞中。丝氨酸蛋白酶凝血酶在中风后的最初几个小时内激活PAR - 1，它似乎与细胞毒性有关。凝血酶拮抗剂和PAR - 1抑制剂与中风后细胞死亡减少和行为保护相关，但尚无数据支持PAR - 1作用与益处之间存在机制联系。我们试图确定PAR - 1在介导缺血性损伤中的关键作用。我们利用在慢病毒载体中与绿色荧光蛋白包装在一起的短发夹RNA，在大鼠大脑中动脉闭塞（MCAo）之前敲低尾状核内侧的PAR - 1，并在氧 - 葡萄糖剥夺之前敲低大鼠神经元中的PAR - 1。我们还在标准的MCAo模型中比较了老年PAR - 1基因敲除小鼠与老年PAR - 3、PAR - 4小鼠以及年轻的野生型小鼠。在MCAo过程中，沉默PAR - 1显著减轻了神经功能缺损，减少了内皮屏障渗漏，并降低了体内神经元变性。在缺血的尾状核内侧敲低PAR - 1使细胞能够在缺血损伤中存活；被感染的细胞对TUNEL和c - Fos损伤标志物呈阴性。感染了PAR - 1 shRNA的原代培养神经元在缺氧/无糖条件下，无论是否添加凝血酶，都显示出更强的神经保护作用。与老年对照组或年轻野生型相比，老年PAR - 1基因敲除小鼠的梗死和血管破坏减少。我们证明了PAR - 1在缺血过程中起着关键作用。沉默或去除PAR - 1可显著保护神经元和星形胶质细胞。进一步开发作用于PAR - 1或其下游通路的药物可能会产生强大的中风治疗方法。
Protease activated receptors (PARs) populate neurons and astrocytes in the brain. The serine protease thrombin, which activates PAR-1 during the first hours after stroke, appears to be associated with the cytotoxicity. Thrombin antagonists and PAR-1 inhibitors have been correlated with reduced cell death and behavioral protection after stroke, but no data yet supports a mechanistic link between PAR-1 action and benefit. We sought to establish the essential role of PAR-1 in mediating ischemic damage. Using a short hairpin mRNA packaged with green fluorescent protein in a lentivirus vector, we knocked downPAR-1 in the medial caudate nucleus prior to rat middle cerebral artery occlusion (MCAo) and in rat neurons prior to oxygen-glucose deprivation. We also compared aged PAR-1 knockout mice with aged PAR-3, PAR-4 mice and young wild-type mice in a standard MCAo model. Silencing PAR-1 significantly reduced neurological deficits, reduced endothelial barrier leakage, and decreased neuronal degeneration in vivo during MCAo. PAR-1 knock-down in the ischemic medial caudate allowed cells to survive the ischemic injury; infected cells were negative for TUNEL and c-Fos injury markers. Primary cultured neurons infected with PAR-1 shRNA showed increased neuroprotection during hypoxic/aglycemic conditions with or without added thrombin. The aged PAR-1 knockout mice showed decreased infarction and vascular disruption compared to aged controls or young wild types. We demonstrated an essential role for PAR-1 during ischemia. Silencing or removing PAR-1 significantly protected neurons and astrocytes. Further development of agents that act at PAR-1or its downstream pathways could yield powerful stroke therapy.
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影响因子：
8.3
作者：
Chen B;Friedman B;Cheng Q;Tsai P;Schim E;Kleinfeld D;Lyden PD
通讯作者：
Lyden PD
影响因子：
8.3
作者：
Adams, Harold P., Jr.;Effron, Mark B.;Hacke, Werner
通讯作者：
Hacke, Werner
影响因子：
8.3
作者：
BEDERSON, JB;PITTS, LH;BARTKOWSKI, H
通讯作者：
BARTKOWSKI, H
影响因子：
8.3
作者：
Fisher M;Feuerstein G;Howells DW;Hurn PD;Kent TA;Savitz SI;Lo EH;STAIR Group
通讯作者：
STAIR Group
DOI：
10.1073/pnas.85.10.3440
发表时间：
1988-05-01
影响因子：
11.1
作者：
GURWITZ, D;CUNNINGHAM, DD
通讯作者：
CUNNINGHAM, DD