Laboratory of Molecular Neurobiology, Academy of Biology and Biotechnology, Southern Federal University, 194/1 Stachky prospect, Rostov-on-Don, 344090, Russia.
Mol Neurobiol. 2017 Nov;54(9):6839-6856. doi: 10.1007/s12035-016-0191-x. Epub 2016 Oct 22.
In ischemic stroke, cell damage propagates from infarct core to surrounding tissue. To reveal proteins involved in neurodegeneration and neuroprotection, we explored the protein profile in penumbra surrounding the photothrombotic infarct core induced in rat cerebral cortex by local laser irradiation after Bengal Rose administration. Using antibody microarrays, we studied changes in expression of 224 signaling proteins 1, 4, or 24 h after photothrombotic infarct compared with untreated contralateral cortex. Changes in protein expression were greatest at 4 h after photothrombotic impact. These included over-expression of proteins initiating, regulating, or executing various apoptosis stages (caspases, SMAC/DIABLO, Bcl-10, phosphatidylserine receptor (PSR), prostate apoptosis response 4 (Par4), E2F1, p75, p38, JNK, p53, growth arrest and DNA damage inducible protein 153 (GADD153), glutamate decarboxylases (GAD65/67), NMDAR2a, c-myc) and antiapoptotic proteins (Bcl-x, p63, MDM2, p21WAF-1, ERK1/2, ERK5, MAP kinase-activated protein kinase-2 (MAKAPK2), PKCα, PKCβ, PKCμ, RAF1, protein phosphatases 1α and MAP kinase phosphatase-1 (MKP-1), neural precursor cell expressed, developmentally down-regulated 8 (NEDD8), estrogen and EGF receptors, calmodulin, CaMKIIα, CaMKIV, amyloid precursor protein (APP), nicastrin). Phospholipase Cγ1, S-100, and S-100β were down-regulated. Bidirectional changes in levels of adhesion and cytoskeleton proteins were related to destruction and/or remodeling of penumbra. Following proteins regulating actin cytoskeleton were over-expressed: cofilin, actopaxin, p120CTN, α-catenin, p35, myosin Va, and pFAK were up-regulated, whereas ezrin, tropomyosin, spectrin (α + β), β-tubulin and polyglutamated β-tubulin, and cytokeratins 7 and 19 were down-regulated. Down-regulation of syntaxin, AP2β/γ, and adaptin β1/2 indicated impairment of vesicular transport and synaptic processes. Down-regulation of cyclin-dependent kinase 6 (Cdk6), cell division cycle 7-related protein kinase (Cdc7 kinase), telomeric repeat-binding factor 1 (Trf1), and topoisomerase-1 showed proliferation suppression. Cytoprotection proteins AOP-1 and chaperons Hsp70 and Hsp90 were down-regulated. These data provide the integral view on penumbra response to photothrombotic infarct. Some of these proteins may be potential targets for antistroke therapy.
在缺血性中风中，细胞损伤从梗塞核心传播到周围组织。为了揭示与神经退行性变和神经保护有关的蛋白质，我们研究了在局部激光照射后，局部给予 Bengal Rose 诱导的大鼠大脑皮质光血栓性梗塞核心周围半影中的蛋白质图谱。使用抗体微阵列，我们研究了与未处理的对侧皮质相比，在光血栓性梗塞后 1、4 或 24 小时表达的 224 种信号蛋白的变化。光血栓性影响后 4 小时，蛋白质表达的变化最大。这些变化包括起始、调节或执行各种凋亡阶段的蛋白质的过表达（胱天蛋白酶、SMAC/DIABLO、Bcl-10、磷脂酰丝氨酸受体 (PSR)、前列腺凋亡反应蛋白 4 (Par4)、E2F1、p75、p38、JNK、p53、生长停滞和 DNA 损伤诱导蛋白 153 (GADD153)、谷氨酸脱羧酶 (GAD65/67)、NMDAR2a、c-myc) 和抗凋亡蛋白 (Bcl-x、p63、MDM2、p21WAF-1、ERK1/2、ERK5、MAP kinase-activated protein kinase-2 (MAKAPK2)、PKCα、PKCβ、PKCμ、RAF1、蛋白磷酸酶 1α 和 MAP kinase phosphatase-1 (MKP-1)、神经前体细胞表达、发育下调 8 (NEDD8)、雌激素和 EGF 受体、钙调蛋白、CaMKIIα、CaMKIV、淀粉样前体蛋白 (APP)、nicastrin)。磷酸酶 Cγ1、S-100 和 S-100β 下调。与半影破坏和/或重塑相关的黏附蛋白和细胞骨架蛋白的双向变化。以下是调节肌动蛋白细胞骨架的蛋白质过表达：丝切蛋白、actopaxin、p120CTN、α-连环蛋白、p35、肌球蛋白 Va 和 pFAK 上调，而 ezrin、原肌球蛋白、 spectrin (α + β)、β-微管蛋白和多聚谷氨酸化 β-微管蛋白以及细胞角蛋白 7 和 19 下调。突触小泡运输和突触过程受损的标志蛋白 syntaxin、AP2β/γ 和 adaptin β1/2 下调。细胞周期蛋白依赖性激酶 6 (Cdk6)、细胞分裂周期 7 相关蛋白激酶 (Cdc7 激酶)、端粒重复结合因子 1 (Trf1) 和拓扑异构酶-1 的下调表明增殖受到抑制。细胞保护蛋白 AOP-1 和伴侣分子 Hsp70 和 Hsp90 下调。这些数据提供了对半影对光血栓性梗塞反应的整体看法。其中一些蛋白质可能是抗中风治疗的潜在靶点。
