Hribljan Valentina, Salamon Iva, Đemaili Arijana, Alić Ivan, Mitrečić Dinko
Dinko Mitrečić, Laboratory for Stem Cells, Croatian Institute for Brain Research, University of Zagreb School of Medicine, Šalata 12, Zagreb, Croatia,
Croat Med J. 2018 Oct 31;59(5):203-212. doi: 10.3325/cmj.2018.59.203.
To analyze how neural stem cells (NSC) transplantation in the stroke-affected mouse brain influences the expression of genes involved in apoptosis-inducing factor (AIF)-mediated cell death – apoptosis inducing factor mitochondria associated 1 (), ring finger protein 146 (), and cyclophilin A (); necroptosis –receptor interaction protein kinase 1 (), , and mixed-lineage kinase domain-like protein (); and apoptosis – Caspase 3 () and .
Four groups of animals were used to obtain mRNA for quantitative reverse transcription polymerase chain reaction analysis: healthy animals (n = 3), animals with stroke (n = 4), animals with stroke treated by stem cell transplantation (n = 7), and animals with stroke treated by proliferation-supporting medium (n = 5). Ischemic brain injury was induced by transient left middle cerebral artery occlusion. Statistical analysis was performed using one-way analysis of variance with Tukey test.
NSC transplantation in the stroke-affected mouse brain significantly increased the expression of ( < 0.05), a gene-encoding protein with well-known protective effects on hypoxic damage, and significantly down-regulated the expression of damage-supportive genes, .01) and ( < 0.001). We were able to distinguish between the effect produced by stem cell transplantation (, , , ) and the effect produced by supporting the tissue with proliferation-supporting medium (, ).
Beside revealing some clearly positive effects of stem cells transplantation on the stroke-affected brain, our results suggest that the tissue response triggered by stem cells points toward the desired, regeneration-supporting levels of expression of a certain gene at a certain time point.
分析神经干细胞（NSC）移植到中风小鼠脑内如何影响参与凋亡诱导因子（AIF）介导的细胞死亡相关基因——凋亡诱导因子线粒体相关蛋白1（AIFM1）、环指蛋白146（RNF146）和亲环素A（CypA）的表达；坏死性凋亡相关基因——受体相互作用蛋白激酶1（RIPK1）、RIPK3和混合谱系激酶结构域样蛋白（MLKL）的表达；以及凋亡相关基因——半胱天冬酶3（Caspase 3）和Bax的表达。
使用四组动物获取mRNA用于定量逆转录聚合酶链反应分析：健康动物（n = 3）、中风动物（n = 4）、接受干细胞移植治疗的中风动物（n = 7）和接受增殖支持培养基治疗的中风动物（n = 5）。通过短暂性左大脑中动脉闭塞诱导缺血性脑损伤。采用单因素方差分析和Tukey检验进行统计学分析。
将NSC移植到中风小鼠脑内显著增加了AIFM1（P < 0.05）的表达，AIFM1是一种对缺氧损伤具有众所周知保护作用的编码蛋白基因，并且显著下调了损伤支持基因RNF146（P < 0.01）和Caspase 3（P < 0.001）的表达。我们能够区分干细胞移植产生的效应（AIFM1、RNF146、Caspase 3、Bax）和用增殖支持培养基支持组织产生的效应（RIPK1、RIPK3、MLKL）。
除了揭示干细胞移植对中风脑的一些明显积极作用外，我们的结果表明，干细胞引发的组织反应指向了特定时间点特定基因表达的理想的、支持再生的水平。
