脑梗死模型大鼠缺血脑组织中胰岛素样生长因子结合蛋白3表达水平及其与血管新生的关系

doi:10.13481/j.1671-587x.20200415

摘要/Abstract

摘要： 目的：探讨脑梗死大鼠缺血脑组织中胰岛素样生长因子结合蛋白3（IGFBP-3）表达水平及其与血管新生的关系，阐明IGFBP-3在脑梗死血管新生中的作用。方法：将SD大鼠随机分为对照组和模型组，模型组大鼠又分为模型1d组、模型3d组和模型7d组3个亚组。采用线栓法建立大脑中动脉栓塞缺血大鼠模型。采用神经行为学评分评估大鼠神经行为，2，3，5-氯化三苯基四氮唑（TTC）染色观察脑梗死情况，HE染色观察脑组织病理形态表现，免疫组织化学染色检测大鼠脑组织中IGFBP-3和血管内皮生长因子（VEGF）蛋白表达水平，逆转录-聚合酶链反应（RT-PCR）检测大鼠脑组织中IGFBP-3和VEGF mRNA表达水平，免疫荧光染色检测大鼠脑组织中缺血半暗区微血管密度。结果：对照组大鼠无神经功能缺损和脑梗死灶；模型组大鼠24h神经行为学评分为（1.86±0.73）分，脑梗死体积为（28.34±3.57）%。HE染色，对照组大鼠脑组织正常，模型1d组大鼠脑组织出现水肿，模型3d组大鼠脑组织水肿和坏死明显，模型7d组大鼠脑组织水肿和坏死减轻。与对照组比较，各模型组大鼠脑组织中IGFBP-3和VEGF蛋白及mRNA表达水平和缺血半暗区微血管密度均明显升高（P<0.01），模型3d组大鼠脑组织上述各指标最高，模型7d组大鼠脑组织中IGFBP-3和VEGF蛋白及mRNA表达水平和缺血半暗区微血管密度降低。大鼠脑组织中IGFBP-3 mRNA表达水平与VEGF mRNA表达水平和微血管密度均呈正相关关系（r=0.563，P<0.05；r=0.612，P<0.05）。结论：脑梗死大鼠缺血脑组织中IGFBP-3表达水平升高可能促进脑梗死缺血后血管新生。

关键词: 脑梗死, 胰岛素样生长因子结合蛋白3, 血管内皮生长因子, 血管新生

Abstract: Objective: To investigate the expression level of insulin-like growth factor binding protein-3(IGFBP-3) in ischemic brain tissue of the rats with cerebral infarction and its relationship with angiogenesis,and to elucidate the role of IGFBP-3 in angiogenesis of cerebral infarction. Methods: The SD rats were randomly divided into control group and model group,and the rats in model group were divided into three subgroups of model 1 d group, model 3 d group and model 7 d group, with 30 rats in each group.The rat model of middle cerebral artery occlusion ischemia was established by suture method.Neurobehavioral score method was used to assess the neural behavior of the rats.Cerebral infarctions were observed by 2,3,5-triphenyltetrazolium chloride (TTC) staining.The pathomorphology of brain tissue of the rats was observed by HE staining.Immunohistochemical staining was used to measure the expression levels of IGFBP-3 and vascular endothelial growth factor (VEGF) proteins in brain tissue of the rats.Reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression levels of IGFBP-3 and VEGF mRNA in brain tissue of the rats.Immunofluorescence staining was used to measure the microvessel density in the ischemic penumbra of the brain tissue of the rats. Results: There were no neurological deficits and cerebral infarction of the rats in control group.In model group,the neurobehavioral score of the rats was (1.86±0.73) scores and the volume of cerebral infarction was (28.34±3.57)%.The HE staining results showed that the brain tissue of the rats in control group was normal;in model 1 d group,edema occurred in the brain tissue of the rats; in model 3 d group,edema and necrosis of brain tissue of the rats were obvious; in model 7 d group, edema and necrosis of brain tissue were alleviated.Compared with control group, the expression levels of IGFBP-3 and VEGF proteins and mRNA in brain tissue of the rats in various model groups and the microvessel densities in ischemic penumbra were increased (P<0.01);the above indexes in brain tissue of the rats in model 3 d group were the highest;the expression levels of IGFBP-3 and VEGF proteins and mRNA in brain tissue of the rats in model 7 d group and the microvessel density in ischemic penumbra were decreased.There were positive correlation between the expression levels of IGFBP-3 mRNA and the VEGF mRNA and microvessel density in brain tissue of the rats (r=0.563,P<0.05;r=0.612,P<0.05). Conclusion: The expression level of IGFBP-3 in ischemic brain tissue of the rats with cerebral infarction is increased, which may promote angiogenesis after cerebral infarction.

Key words: cerebral infarction, insulin-like growth factor binding protein-3, vascular endothelial growth factor, angiogenesis

中图分类号:

R743.3

卜一, 张硕, 钱旭东, 王红梅, 窦志杰. 脑梗死模型大鼠缺血脑组织中胰岛素样生长因子结合蛋白3表达水平及其与血管新生的关系[J]. 吉林大学学报(医学版), 2020, 46(04): 759-764.

BU Yi, ZHANG Shuo, QIAN Xudong, WANG Hongmei, DOU Zhijie. Expression of insulin-like growth factor binding protein-3 in ischemic brain tissue of cerebral infarction model rats and its relationship with angiogenesis[J]. Journal of Jilin University(Medicine Edition), 2020, 46(04): 759-764.

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