Department of Anesthesiology, University of Texas Medical Branch, Galveston, TX, USA.
Department of Pharmacology, University of Texas Medical Branch, Galveston, TX, USA.
Br J Pharmacol. 2018 Jan;175(2):284-300. doi: 10.1111/bph.13642. Epub 2016 Nov 15.
We hypothesized that an in vitro, stretch-based model of neural injury may be useful to identify compounds that decrease the cellular damage in neurotrauma.
We screened three neural cell lines (B35, RN33B and SH-SY5Y) subjected to two differentiation methods and selected all-trans-retinoic acid-differentiated B35 rat neuroblastoma cells subjected to rapid stretch injury, coupled with a subthreshold concentration of H O , for the screen. The model induced marked alterations in gene expression and proteomic signature of the cells and culminated in delayed cell death (LDH release) and mitochondrial dysfunction [reduced 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) conversion]. Follow-up studies utilized human stem cell-derived neurons subjected to rapid stretch injury.
From screening of a composite library of 3500 drugs, five drugs (when applied in a post-treatment regimen relative to stretch injury) improved both LDH and MTT responses. The effects of rifampicin were investigated in further detail. Rifampicin reduced cell necrosis and apoptosis and improved cellular bioenergetics. In a second model (stretch injury in human stem cell-derived neurons), rifampicin pretreatment attenuated LDH release, protected against the loss of neurite length and maintained neuron-specific class III β-tubulin immunoreactivity.
We conclude that the current model is suitable for medium-throughput screening to identify compounds with neuroprotective potential. Rifampicin, when applied either in pre- or post-treatment, improves the viability of neurons subjected to stretch injury and protects against neurite loss. Rifampicin may be a candidate for repurposing for the therapy of traumatic brain injury.
This article is part of a themed section on Inventing New Therapies Without Reinventing the Wheel: The Power of Drug Repurposing. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.2/issuetoc.
我们假设一种体外拉伸模型的神经损伤可能有助于发现减少神经创伤细胞损伤的化合物。
我们筛选了三种神经细胞系（B35、RN33B 和 SH-SY5Y），并采用了两种分化方法，选择了全反式视黄酸分化的 B35 大鼠神经母细胞瘤细胞，进行快速拉伸损伤，并结合亚阈值浓度的 H2O2 进行筛选。该模型诱导细胞的基因表达和蛋白质组学特征发生明显改变，并最终导致延迟细胞死亡（LDH 释放）和线粒体功能障碍[减少 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴盐（MTT）转化]。后续研究利用快速拉伸损伤的人干细胞源性神经元进行。
从 3500 种药物的复合文库中筛选出 5 种药物（与拉伸损伤后的治疗方案相比）可改善 LDH 和 MTT 反应。进一步研究了利福平的作用。利福平可减少细胞坏死和凋亡，并改善细胞的生物能量学。在第二个模型（人干细胞源性神经元的拉伸损伤）中，利福平预处理可减轻 LDH 释放，防止轴突长度丢失，并维持神经元特异性 III 类β-微管蛋白免疫反应性。
我们得出结论，目前的模型适合于高通量筛选，以确定具有神经保护潜力的化合物。利福平无论是在预处理还是后处理中应用，都可以提高受到拉伸损伤的神经元的活力，并防止轴突丢失。利福平可能是治疗创伤性脑损伤的候选药物。
本文是关于“创新治疗方法，无需重新发明轮子：药物再利用的力量”这一主题部分的文章。要查看本部分中的其他文章，请访问 http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.2/issuetoc.
