Ghazale Hussein, Ramadan Naify, Mantash Sara, Zibara Kazem, El-Sitt Sally, Darwish Hala, Chamaa Farah, Boustany Rose Mary, Mondello Stefania, Abou-Kheir Wassim, Soueid Jihane, Kobeissy Firas
Department of Biochemistry and Molecular Genetics, Faculty of Medicine, American University of Beirut, Beirut, Lebanon, Lebanon.
ER045, Laboratory of Stem Cells, DSST, Lebanese University, Beirut, Lebanon; Department of Biology, Faculty of Sciences-I, Lebanese University, Beirut, Lebanon.
Behav Brain Res. 2018 Mar 15;340:1-13. doi: 10.1016/j.bbr.2017.11.007. Epub 2017 Nov 7.
Traumatic Brain Injury (TBI) is a major cause of death and disability worldwide with 1.5 million people inflicted yearly. Several neurotherapeutic interventions have been proposed including drug administration as well as cellular therapy involving neural stem cells (NSCs). Among the proposed drugs is docosahexaenoic acid (DHA), a polyunsaturated fatty acid, exhibiting neuroprotective properties. In this study, we utilized an innovative intervention of neonatal NSCs transplantation in combination with DHA injections in order to ameliorate brain damage and promote functional recovery in an experimental model of TBI. Thus, NSCs derived from the subventricular zone of neonatal pups were cultured into neurospheres and transplanted in the cortex of an experimentally controlled cortical impact mouse model of TBI. The effect of NSC transplantation was assessed alone and/or in combination with DHA administration. Motor deficits were evaluated using pole climbing and rotarod tests. Using immunohistochemistry, the effect of transplanted NSCs and DHA treatment was used to assess astrocytic (Glial fibrillary acidic protein, GFAP) and microglial (ionized calcium binding adaptor molecule-1, IBA-1) activity. In addition, we quantified neuroblasts (doublecortin; DCX) and dopaminergic neurons (tyrosine hydroxylase; TH) expression levels. Combined NSC transplantation and DHA injections significantly attenuated TBI-induced motor function deficits (pole climbing test), promoted neurogenesis, coupled with an increase in glial reactivity at the cortical site of injury. In addition, the number of tyrosine hydroxylase positive neurons was found to increase markedly in the ventral tegmental area and substantia nigra in the combination therapy group. Immunoblotting analysis indicated that DHA+NSCs treated animals showed decreased levels of 38kDa GFAP-BDP (breakdown product) and 145kDa αII-spectrin SBDP indicative of attenuated calpain/caspase activation. These data demonstrate that prior treatment with DHA may be a desirable strategy to improve the therapeutic efficacy of NSC transplantation in TBI.
创伤性脑损伤（TBI）是全球范围内死亡和残疾的主要原因，每年有150万人受其影响。已经提出了几种神经治疗干预措施，包括药物给药以及涉及神经干细胞（NSC）的细胞疗法。所提出的药物中有一种是二十二碳六烯酸（DHA），一种多不饱和脂肪酸，具有神经保护特性。在本研究中，我们采用了一种创新的干预措施，即新生儿神经干细胞移植与DHA注射相结合，以改善脑损伤并促进TBI实验模型中的功能恢复。因此，将源自新生幼崽脑室下区的神经干细胞培养成神经球，并移植到实验性控制的皮质撞击TBI小鼠模型的皮质中。单独和/或与DHA给药联合评估神经干细胞移植的效果。使用爬杆和转棒试验评估运动功能障碍。使用免疫组织化学，移植的神经干细胞和DHA治疗的效果用于评估星形胶质细胞（胶质纤维酸性蛋白，GFAP）和小胶质细胞（离子钙结合衔接分子-1，IBA-1）的活性。此外，我们量化了神经母细胞（双皮质素；DCX）和多巴胺能神经元（酪氨酸羟化酶；TH）的表达水平。神经干细胞移植和DHA注射联合使用显著减轻了TBI诱导的运动功能障碍（爬杆试验），促进了神经发生，并伴随着损伤皮质部位胶质反应性的增加。此外，在联合治疗组中，发现腹侧被盖区和黑质中酪氨酸羟化酶阳性神经元的数量明显增加。免疫印迹分析表明，DHA +神经干细胞治疗的动物显示38kDa GFAP-BDP（降解产物）和145kDaαII-血影蛋白SBDP的水平降低，这表明钙蛋白酶/半胱天冬酶激活减弱。这些数据表明，预先用DHA治疗可能是提高神经干细胞移植在TBI中治疗效果的理想策略。
