Recombinant Osteopontin Stabilizes Smooth Muscle Cell Phenotype via Integrin Receptor/Integrin-Linked Kinase/Rac-1 Pathway After Subarachnoid Hemorrhage in Rats.
Recombinant Osteopontin Stabilizes Smooth Muscle Cell Phenotype via Integrin Receptor/Integrin-Linked Kinase/Rac-1 Pathway After Subarachnoid Hemorrhage in Rats.
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DOI：
10.1161/strokeaha.115.011552
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发表时间：
2016-05
期刊：
影响因子：
8.3
通讯作者：
Zhang JH
中科院分区：
文献类型：
--
作者：
Wu J;Zhang Y;Yang P;Enkhjargal B;Manaenko A;Tang J;Pearce WJ;Hartman R;Obenaus A;Chen G;Zhang JH
据报道，重组骨桥蛋白（rOPN）在脑卒中动物模型中具有神经保护作用。本研究旨在探讨鼻腔给予rOPN在蛛网膜下腔出血（SAH）后早期脑损伤中对维持血管平滑肌表型的潜在作用及其机制。
使用192只成年雄性Sprague - Dawley大鼠。通过血管内穿刺法诱导SAH模型。在SAH前48小时向脑室注射整合素连接激酶（ILK）小干扰RNA。在诱导SAH前1小时给予整合素受体拮抗剂GRGDSP、黏着斑激酶（FAK）抑制剂Fib - 14和Rac - 1抑制剂NSC23766。SAH后通过脑室和鼻腔途径给予rOPN。采用SAH分级、神经功能评分、脑含水量、脑肿胀、苏木精 - 伊红染色、印度墨汁血管造影、蛋白质印迹法和免疫荧光等方法研究rOPN对血管平滑肌表型转化的机制。
SAH后24小时和72小时，脑动脉中血管平滑肌表型转化的标记蛋白α - 平滑肌肌动蛋白（α - SMA）显著减少，而SMemb显著增加。rOPN阻止了α - SMA和SMemb的变化，显著减轻了神经行为功能障碍，增加了脑动脉的横截面积和管腔直径，降低了脑含水量和脑肿胀，并改善了脑动脉的管壁厚度。GRGDSP、ILK小干扰RNA和NSC23766可消除rOPN的这些作用。SAH后3小时鼻腔应用rOPN也可减轻神经功能缺损。
rOPN可阻止血管平滑肌表型转化并改善神经功能预后，这可能是由整合素受体/ILK/Rac - 1通路介导的。
Recombinant Osteopontin (rOPN) has been reported to be neuroprotective in stroke animal models. The purpose of this study is to investigate a potential role and mechanism of nasal administration of rOPN on preserving the vascular smooth muscle phenotype in early brain injury after SAH. One hundred and ninety-two male adult Sprague-Dawley rats were used. The SAH model was induced by endovascular perforation. Integrin-linked kinase (ILK) siRNA was intracerebroventricularly injected 48 hours before SAH. The integrin receptor antagonist GRGDSP, FAK inhibitor Fib-14 and Rac-1 inhibitor NSC23766 were administered 1 hour before SAH induction. rOPN was administered via the intracerebroventricular and nasal route after SAH. SAH grade, neurological scores, brain water content, brain swelling, hematoxylin and eosin staining, India ink angiography, Western blots, and immunofluorescence were used to study the mechanisms of rOPN on the vascular smooth muscle phenotypic transformation. The marker protein of vascular smooth muscle phenotypic transformation alpha-SMA decreased and SMemb increased significantly at 24 and 72 hours in the cerebral arteries after SAH. rOPN prevented changes of alpha-SMA and SMemb, and significantly alleviated neurobehavioral dysfunction, increased the cross-section area and the lumen diameter of the cerebral arteries, reduced brain water content and brain swelling and improved the wall thickness of cerebral arteries. These effects of rOPN were abolished by GRGDSP, ILK siRNA and NSC23766. Intranasal application of rOPN at 3 hrs after SAH also reduced neurological deficits. rOPN prevented the vascular smooth muscle phenotypic transformation and improved the neurological outcome, which was possibly mediated by the integrin receptor/ILK/Rac-1 pathway.