诱导性神经干细胞移植通过 Crry 和 Akt 在闭合性颅脑损伤小鼠模型中的相互作用发挥神经保护作用。
Induced neural stem cell grafts exert neuroprotection through an interaction between Crry and Akt in a mouse model of closed head injury.
机构信息
Department of Neurosurgery, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, 610072, China.
Department of Neurosurgery, The PLA General Hospital, Beijing, 100853, China.
出版信息
Stem Cell Res Ther. 2021 Feb 12;12(1):128. doi: 10.1186/s13287-021-02186-z.
BACKGROUND
Recently, growing evidence has indicated an important role of the complement system, a crucial component of immunity, in mediating neuroinflammation and promoting neuronal apoptosis following closed head injury (CHI). We previously reported that transplanted induced neural stem cells (iNSCs) pre-treated with CHI mouse serum could enhance complement receptor type 1-related protein y (Crry) expression and ameliorate complement-mediated damage in mouse CHI models. However, the mechanism underlying the elevated levels of Crry expression remains elusive.
METHODS
CHI models were established using a standardized weight-drop device. We collected CHI mouse serum at 12 h post-trauma. RT-QPCR assay, western blot analysis, complement deposition assay, Akt inhibition assay, flow cytometry, cell transplantation, and functional assay were utilized to clarify the mechanism of Crry expression in iNSCs receiving CHI mouse serum treatment.
RESULTS
We observed dramatic increases in the levels of Crry expression and Akt activation in iNSCs receiving CHI mouse serum treatment. Remarkably, Akt inhibition led to the reduction of Crry expression in iNSCs. Intriguingly, the treatment of iNSC-derived neurons with recombinant complement receptor 2-conjugated Crry (CR2-Crry), which inhibits all complement pathways, substantially enhanced Crry expression and Akt activation in neurons after CHI mouse serum treatment. In subsequent in vitro experiments of pre-treatment of iNSCs with CR2-Crry, we observed significant increases in the levels of Crry expression and Akt activation in iNSCs and iNSC-derived astrocytes and neurons post-treatment with CHI mouse serum. Additionally, an in vivo study showed that intracerebral-transplanted iNSCs pre-treated with CR2-Crry markedly enhanced Crry expression in neurons and protected neurons from complement-dependent damage in the brains of CHI mice.
CONCLUSION
INSCs receiving CR2-Crry pre-treatment increased the levels of Crry expression in iNSCs and iNSC-derived astrocytes and neurons and attenuated complement-mediated injury following CHI.
背景
最近的研究证据表明，补体系统在介导神经炎症和促进闭合性颅脑损伤（CHI）后的神经元凋亡方面起着重要作用，补体系统是免疫系统的一个关键组成部分。我们之前的研究报道，预处理 CHI 小鼠血清的诱导多能神经干细胞（iNSCs）可增强补体受体 1 相关蛋白 y（Crry）的表达，并改善小鼠 CHI 模型中的补体介导损伤。然而，Crry 表达水平升高的机制仍不清楚。
方法
使用标准化重物坠落装置建立 CHI 模型。在创伤后 12 小时收集 CHI 小鼠血清。利用 RT-QPCR 检测、Western blot 分析、补体沉积检测、Akt 抑制检测、流式细胞术、细胞移植和功能检测来阐明 iNSCs 接受 CHI 小鼠血清处理后 Crry 表达的机制。
结果
我们观察到 iNSCs 接受 CHI 小鼠血清处理后 Crry 表达和 Akt 激活水平显著增加。值得注意的是，Akt 抑制导致 iNSCs 中 Crry 表达减少。有趣的是，用重组补体受体 2 结合的 Crry（CR2-Crry）处理 iNSC 衍生神经元，该物质可抑制所有补体途径，在 CHI 小鼠血清处理后显著增强神经元中 Crry 的表达和 Akt 的激活。在随后的 iNSC 用 CR2-Crry 预处理的体外实验中，我们观察到 iNSCs 和 iNSC 衍生的星形胶质细胞和神经元在接受 CHI 小鼠血清处理后 Crry 表达和 Akt 激活水平显著增加。此外，体内研究表明，CR2-Crry 预处理的脑内移植 iNSCs 可显著增加 CHI 小鼠大脑中神经元的 Crry 表达，并保护神经元免受补体依赖性损伤。
结论
接受 CR2-Crry 预处理的 iNSCs 增加了 iNSCs 和 iNSC 衍生的星形胶质细胞和神经元中的 Crry 表达，并减轻了 CHI 后补体介导的损伤。
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