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<p>Promoting nerve regeneration is a crucial approach in treating ischemic stroke, with neural stem cells (NSCs) playing a vital role in this process. Qixian Tongluo Formula (QXTLF) has shown effectiveness in treating ischemic stroke, yet its impact on regulating the directional differentiation of NSCs during ischemic stroke remains unclear. A middle cerebral artery occlusion (MCAO) model was induced in Sprague-Dawley rats, and primary NSCs were isolated from these rats. By gavage QXTLF (7.83&#xa0;g/kg/d or 31.32&#xa0;g/kg/d) was administered intragastrically to the MCAO rats. In vitro experiments confirmed the successful isolation of NSCs from rats, capable of differentiating into immature neurons, mature neurons and astrocytes. In vitro, NSCs were exposed to fibrinogen (Fg) and induced by low glucose-oxygen (LGO) conditions to simulate the NSC niche in ischemic stroke. Various assessments were conducted, including neurological score evaluation in MCAO rats, analysis of infarct size using 2,3,5-triphenyltetrazolium chloride (TTC) staining, and examination of the expression of markers like bromodeoxyuridine (BrdU), doublecortin (DCX), microtubule-associated protein 2 (MAP-2), glial fibrillary acidic protein (GFAP), and bone morphogenetic protein type I receptor (BMPRI)/inhibitor of DNA binding 3 (ID3) by immunofluorescence and western blot analysis. The Fg content was quantified by enzyme-linked immunosorbent assay. Results demonstrated that QXTLF reduced cerebral infarction and decreased the neurological score of MCAO rat in a dose-dependent manner. Additionally, QXTLF promoted the directional differentiation of NSCs into neurons within ipsilateral hippocampus following cerebral ischemia. Furthermore, the study revealed that QXTLF facilitated the directional differentiation of LGO-induced NSCs into neurons and implicated Fg in mediating this effect. QXTLF was found to regulate the directional differentiation of LGO-induced NSCs through the Fg-mediated BMPRI/ID3 signal axis, highlighting its role in promoting the differentiation of NSCs into neurons post-ischemic stroke.</p>

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Qixian Tongluo Formula promotes the directional differentiation of neural stem cells into neurons via fibrinogen-mediated BMPRI/ID3 signal axis after ischemic stroke

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年10月30日

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  • Shengqiang Zhou,
  • Bo Li,
  • Guo Mao,
  • Wen Zeng,
  • Yanjun Chen,
  • Jia Huang,
  • Lingjuan Tan,
  • Dahua Wu,
  • Fang Liu

摘要

Promoting nerve regeneration is a crucial approach in treating ischemic stroke, with neural stem cells (NSCs) playing a vital role in this process. Qixian Tongluo Formula (QXTLF) has shown effectiveness in treating ischemic stroke, yet its impact on regulating the directional differentiation of NSCs during ischemic stroke remains unclear. A middle cerebral artery occlusion (MCAO) model was induced in Sprague-Dawley rats, and primary NSCs were isolated from these rats. By gavage QXTLF (7.83 g/kg/d or 31.32 g/kg/d) was administered intragastrically to the MCAO rats. In vitro experiments confirmed the successful isolation of NSCs from rats, capable of differentiating into immature neurons, mature neurons and astrocytes. In vitro, NSCs were exposed to fibrinogen (Fg) and induced by low glucose-oxygen (LGO) conditions to simulate the NSC niche in ischemic stroke. Various assessments were conducted, including neurological score evaluation in MCAO rats, analysis of infarct size using 2,3,5-triphenyltetrazolium chloride (TTC) staining, and examination of the expression of markers like bromodeoxyuridine (BrdU), doublecortin (DCX), microtubule-associated protein 2 (MAP-2), glial fibrillary acidic protein (GFAP), and bone morphogenetic protein type I receptor (BMPRI)/inhibitor of DNA binding 3 (ID3) by immunofluorescence and western blot analysis. The Fg content was quantified by enzyme-linked immunosorbent assay. Results demonstrated that QXTLF reduced cerebral infarction and decreased the neurological score of MCAO rat in a dose-dependent manner. Additionally, QXTLF promoted the directional differentiation of NSCs into neurons within ipsilateral hippocampus following cerebral ischemia. Furthermore, the study revealed that QXTLF facilitated the directional differentiation of LGO-induced NSCs into neurons and implicated Fg in mediating this effect. QXTLF was found to regulate the directional differentiation of LGO-induced NSCs through the Fg-mediated BMPRI/ID3 signal axis, highlighting its role in promoting the differentiation of NSCs into neurons post-ischemic stroke.