中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (2): 277-284.doi: 10.4103/1673-5374.265562

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

选择性IIA类组蛋白脱乙酰基酶抑制剂-TMP269对脑卒中的神经保护机制

  

  • 出版日期:2020-02-15 发布日期:2020-05-25
  • 基金资助:
    国家自然科学基金（81501134）

Neuroprotective mechanism of TMP269, a selective class IIA histone deacetylase inhibitor, after cerebral ischemia/reperfusion injury

Lu Su1, Dan Liang2, Shen-Yi Kuang1, Qiang Dong1, Xiang Han1, Zheng Wang1   

  1. 1 Department of Neurology, Huashan Hospital, Fudan University, Shanghai, China
    2 Department of Rehabilitation Medicine, Huashan Hospital, Fudan University, Shanghai, China
  • Online:2020-02-15 Published:2020-05-25
  • Contact: Xiang Han, PhD,hansletter@163.com; Zheng Wang, PhD,zheen_wang@163.com.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81501134 (to ZW).

PDF

125

摘要：

研究表明，多种组蛋白脱乙酰基酶抑制剂可以保护机体免受脑缺血损伤，其中TMP269是一种选择性IIA类组蛋白脱乙酰基酶抑制剂，其中枢神经保护作用在以前的基础实验中已得到证实，但具体机制尚不清楚。为此，实验拟揭示TMP269保护脑缺血再灌注损伤的最佳浓度，及其神经保护机制。将雄性SD大鼠随机分成6组，假手术组(sham)、单纯脑缺血再灌注组(I/R)、脑缺血再灌注+TMP269 1，4，10，16 mg/kg组。脑缺血再灌注损伤通过大脑中动脉闭塞(MCAO)模型诱导，此外不同剂量TMP269注射组，均在诱导缺血前0.5 h通过腹腔注射给药。(1)用Western blot和免疫组织化学染色检测TMP269对组蛋白2乙酰化的作用发现，注射TMP269 24 h后，大鼠运动皮质中组蛋白2的乙酰化水平增加；(2)用TTC染色检测TMP269对脑缺血再灌注损伤面积的影响发现，不同剂量的TMP269能减少大鼠脑梗死面积；(3)用Western blot、免疫组织化学染色和EB染色检测TMP269对血脑屏障的作用发现，TMP269抵消了脑缺血再灌注导致的异常内皮细胞通透性改变；(4)用Western blot和免疫组织化学染色检测TMP269对组织型缓激肽释放酶的调控作用发现，TMP269可上调内源性组织型缓激肽释放酶(TK)的表达；(5)通过Western blot和免疫组织化学染色进一步确定TMP269最佳药物浓度，发现与TMP269 1，10，16 mg/kg相比，大鼠腹腔注射TMP269发挥脑缺血后神经保护作用的最佳浓度是4 mg/kg；(6)作者认为，TMP269可上调组蛋白2的乙酰化水平，减轻脑缺血再灌注引起的内皮细胞损伤，同时上调内源性组织型缓激肽释放酶的表达，发挥神经保护作用。实验方案经复旦大学实验动物科学系批准（批准号：20140143C001）。

orcid: 0000-0003-4608-2083 (Xiang Han) 

         0000-0002-3595-3069 (Zheng Wang)

关键词: 缺血性脑卒中, 组蛋白脱乙酰基酶抑制剂, TMP269, 药物治疗, 组织激肽释放酶, TK, 血脑屏障, BBB, 内皮细胞通透性, 神经保护作用, 神经再生

Abstract: TMP269 is a selective class IIA histone deacetylase inhibitor that has a protective effect on the central nervous system, whose specific mechanism of action is unclear. We aimed to reveal the optimal concentration of TMP269 for protecting against cerebral ischemia/reperfusion injury and its neuroprotective mechanism. Male Sprague-Dawley rats were randomly divided into sham, ischemia/reperfusion, and 1, 4, 10 and 16 mg/kg TMP269 groups. Cerebral ischemia/reperfusion injury was induced by middle cerebral artery occlusion. TMP269 was intraperitoneally administered at different doses 0.5 hours before ischemia induction. Western blot assay and immunohistochemistry were used to detect effects of TMP269 on histone 2 acetylation. The results showed that the level of histone 2 acetylation was increased 24 hours after TMP269 injection. 2,3,5-Triphenyltetrazolium chloride staining was utilized to examine effect of TMP269 on infarct volume. The results found that different doses of TMP269 could reduce the infarct volume. Western blot assay, immunohistochemistry and Evans blue staining were employed to measure the effect of TMP269 on blood-brain barrier. The results showed that TMP269 counteracted the abnormal endothelial cell permeability changes caused by cerebral ischemia/reperfusion. Western blot assay and immunohistochemistry were used to determine the effect of TMP269 on tissue kallikrein. The results found that TMP269 up-regulated the expression of tissue kallikrein. Western blot assay further determined the optimal concentration to be 4 mg/kg. In conclusion, TMP269 plays a neuroprotective role by up-regulating the level of histone 2 acetylation, alleviating endothelial cell injury after cerebral ischemia/reperfusion, and up-regulating the expression of tissue kallikrein. The experimental protocol was approved in 2014 by the Department of Laboratory Animal Science, Fudan University, China (approval No. 20140143C001).

Key words: blood-brain barrier, drug treatment, endothelial cell permeability, histone deacetylase inhibitor, neuroprotection, stroke, tissue kallikrein, TMP269