Soochow University Medical College, Soochow University, Suzhou, Jiangsu 215031, P.R. China.
Department of Neurology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou 563000, P.R. China.
Mol Med Rep. 2024 Jan;29(1). doi: 10.3892/mmr.2023.13128. Epub 2023 Nov 17.
Cerebrovascular diseases (CVDs) have become a global public health problem and ischemia‑reperfusion injury, the major cause of neurological impairment exacerbation, is closely related to excitotoxicity. The present study aimed to investigate the effects of changes in heat shock protein (HSP)90β expression and verify whether HSP90β regulates EAAT2 expression in a cerebral ischemia‑reperfusion injury model. Healthy adult Sprague‑Dawley (SD) male rats were used to establish a control group, sham‑operated group, middle cerebral artery occlusion (MCAO) group, empty virus group and lentivirus group. A model of cerebral ischemia‑reperfusion was established using the MCAO method. Lentivirus construction and injection were used to interfere with the expression of HSP90β. The modified neurological severity score was used to assess neurological deficits. Triphenyltetrazolium chloride staining was used to detect infarct areas. Immunofluorescence was used to detect HSP90β expression localization and the expression levels of HSP90β and EAAT2 were determined using western blotting and reverse transcription‑quantitative PCR. An MCAO model was successfully established and it was found that HSP90β, but not HSP90α, was upregulated after MCAO. HSP90β expression coincided with astrocyte markers in the ischemic penumbra area, while no expression was observed in microglia. Inhibition of HSP90β expression improved neurological deficits and alleviated brain injury by increasing EAAT2 expression. These results suggested that HSP90β is involved in the process of cerebral ischemia‑reperfusion injury in rats and that inhibition of HSP90β expression increases EAAT2 levels, conferring a neuroprotective effect in MCAO model rats.
脑血管疾病（CVDs）已成为全球公共卫生问题，而缺血再灌注损伤是导致神经功能恶化的主要原因，与兴奋性毒性密切相关。本研究旨在探讨热休克蛋白（HSP）90β表达变化的影响，并验证 HSP90β是否调节脑缺血再灌注损伤模型中的 EAAT2 表达。使用健康成年 Sprague-Dawley（SD）雄性大鼠建立对照组、假手术组、大脑中动脉闭塞（MCAO）组、空病毒组和慢病毒组。采用 MCAO 法建立脑缺血再灌注模型。慢病毒构建和注射用于干扰 HSP90β 的表达。采用改良神经功能缺损评分评估神经功能缺损。采用氯化三苯基四氮唑染色检测梗死面积。免疫荧光法检测 HSP90β表达定位及 HSP90β和 EAAT2 的表达水平采用 Western blot 和逆转录-定量 PCR 检测。成功建立 MCAO 模型，发现 MCAO 后 HSP90β（而非 HSP90α）上调。HSP90β表达与缺血半影区星形胶质细胞标志物一致，而在小胶质细胞中无表达。抑制 HSP90β表达通过增加 EAAT2 表达改善神经功能缺损并减轻脑损伤。这些结果表明，HSP90β参与大鼠脑缺血再灌注损伤过程，抑制 HSP90β表达可增加 EAAT2 水平，在 MCAO 模型大鼠中发挥神经保护作用。
