在小鼠脑出血性病变中对神经聚糖进行治疗性减少可促进少突胶质细胞生成和功能恢复。

Therapeutic reduction of neurocan in murine intracerebral hemorrhage lesions promotes oligodendrogenesis and functional recovery.

作者信息

Li Hongmin, Ghorbani Samira, Oladosu Olayinka, Zhang Ping, Visser Frank, Dunn Jeff, Zhang Yunyan, Ling Chang-Chun, Yong V Wee, Xue Mengzhou

机构信息

Department of Cerebrovascular Diseases, The Second Affiliated Hospital of Zhengzhou University, 2 Jingba Road, Zhengzhou, Henan, China.

Hotchkiss Brain Institute and Department of Clinical Neurosciences, University of Calgary, 3330 Hospital Drive, Calgary, AB, Canada.

出版信息

J Neuroinflammation. 2025 Jan 4;22(1):2. doi: 10.1186/s12974-024-03331-0.

DOI:10.1186/s12974-024-03331-0

PMID:39755654

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11699774/

Abstract

BACKGROUND

Intracerebral hemorrhage (ICH) causes prominent deposition of extracellular matrix molecules, particularly the chondroitin sulphate proteoglycan (CSPG) member neurocan. In tissue culture, neurocan impedes the properties of oligodendrocytes. Whether therapeutic reduction of neurocan promotes oligodendrogenesis and functional recovery in ICH is unknown.

METHODS

Mice were retro-orbitally injected with adeno-associated virus (AAV-CRISPR/Cas9) to reduce neurocan deposition after ICH induction by collagenase. Other groups of ICH mice were treated with vehicle or a drug that reduces CSPG synthesis, 4-4-difluoro-N-acetylglucosamine (difluorosamine). Rota-rod and grip strength behavioral tests were conducted over 7 or 14 days. Brain tissues were investigated for expression of neurocan by immunofluorescence microscopy and western blot analysis. Brain cryosections were also stained for microglia/macrophage phenotype, oligodendrocyte lineage cells and neuroblasts by immunofluorescence microscopy. Tissue structural changes were assessed using brain magnetic resonance imaging (MRI).

RESULTS

The adeno-associated virus (AAV)-reduction of neurocan increased oligodendrocyte numbers and functional recovery in ICH. The small molecule inhibitor of CSPG synthesis, difluorosamine, lowered neurocan levels in lesions and elevated numbers of oligodendrocyte precursor cells, mature oligodendrocytes, and SOX2 nestin neuroblasts in the perihematomal area. Difluorosamine shifted the degeneration-associated functional state of microglia/macrophages in ICH towards a regulatory phenotype. MRI analyses showed better fiber tract integrity in the penumbra of difluorosamine mice. These beneficial difluorosamine results were achieved with delayed (2 or 3 days) treatment after ICH.

CONCLUSION

Reducing neurocan deposition following ICH injury is a therapeutic approach to promote histological and behavioral recovery from the devastating stroke.

摘要

背景

脑出血（ICH）会导致细胞外基质分子显著沉积，尤其是硫酸软骨素蛋白聚糖（CSPG）成员神经聚糖。在组织培养中，神经聚糖会阻碍少突胶质细胞的特性。脑出血后神经聚糖的治疗性减少是否能促进少突胶质细胞生成和功能恢复尚不清楚。

方法

通过胶原酶诱导脑出血后，给小鼠经眶后注射腺相关病毒（AAV-CRISPR/Cas9）以减少神经聚糖沉积。其他脑出血小鼠组分别接受载体或一种减少CSPG合成的药物4-4-二氟-N-乙酰葡糖胺（二氟胺）治疗。在7天或14天内进行转棒和握力行为测试。通过免疫荧光显微镜和蛋白质印迹分析研究脑组织中神经聚糖的表达。脑冰冻切片也通过免疫荧光显微镜对小胶质细胞/巨噬细胞表型、少突胶质细胞谱系细胞和成神经细胞进行染色。使用脑磁共振成像（MRI）评估组织结构变化。

结果

腺相关病毒介导的神经聚糖减少增加了脑出血小鼠的少突胶质细胞数量并促进了功能恢复。CSPG合成的小分子抑制剂二氟胺降低了损伤部位的神经聚糖水平，并增加了血肿周围区域少突胶质细胞前体细胞、成熟少突胶质细胞和SOX2巢蛋白成神经细胞的数量。二氟胺使脑出血中与变性相关的小胶质细胞/巨噬细胞功能状态向调节表型转变。MRI分析显示二氟胺处理小鼠的半暗带纤维束完整性更好。这些二氟胺的有益结果是在脑出血后延迟（2或3天）治疗时实现的。