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林容旭, 樊朝凤, 崔文耀, 等. 大鼠脑出血后内源性神经干细胞迁移和微环境中免疫细胞表型变化规律的关系[J]. koko体育app 学报（医学版）, 2024, 55(3): 635-640. DOI:
引用本文: 林容旭, 樊朝凤, 崔文耀, 等. 大鼠脑出血后内源性神经干细胞迁移和微环境中免疫细胞表型变化规律的关系[J]. koko体育app 学报（医学版）, 2024, 55(3): 635-640. DOI:
LIN Rongxu, FAN Chaofeng, CUI Wenyao, et al. Relationship Between the Migration of Endogenous Neural Stem Cells and the Pattern of Change in Immune Cell Phenotypes in the Microenvironment After Intracerebral Hemorrhage in Rats[J]. Journal of Sichuan University (Medical Sciences), 2024, 55(3): 635-640. DOI:
Citation: LIN Rongxu, FAN Chaofeng, CUI Wenyao, et al. Relationship Between the Migration of Endogenous Neural Stem Cells and the Pattern of Change in Immune Cell Phenotypes in 🌄the Microenvironment After Intracerebral Hemorrhage in Rats[J]. Journal of Sichuan University (Medical Sciences), 2024, 55(3): 635-640. DOI:

大鼠脑出血后内源性神经干细胞迁移和微环境中免疫细胞表型变化规律的关系

Relationship Between the Migration of Endogenous Neural Stem Cells and the Pattern of Change in Immune Cell Phenotypes in the Microenvironment After Intracerebral Hemorrhage in Rats

  • 摘要:
    目的 研究脑出血（intracerebral hemorrhage, ICH）后内源性神经干细胞（endogenous neural stem cells, eNSCs)及再生微环境的变化规律，观察eNSCs迁移与微环境免疫细胞极化状态变化规律之间的关系，为临床神经修复研究提供研究基础。
    方法 采用胶原酶注射法造模，在体质量为280～300 g的成年雌性Sprague-Dawley大鼠脑组织注射Ⅶ型胶原酶（2 U）诱导脑出血，为模拟急性期（1周内）、亚急性期（1～3周）及慢性期（>3周）脑出血的时间点，在注射后第3天（3-day post injection, 3 DPI）、10 DPI、20 DPI、30 DPI取脑组织评价造模效果。采用DCX抗体对脑组织切片进行免疫荧光染色，观察不同时间脑组织中eNSCs迁移变化规律；采用CD206抗体、CD86抗体对脑组织切片进行免疫荧光染色，分别观察脑组织促炎型（M1型）免疫细胞和抑炎型（M2型）免疫细胞在脑出血后再生微环境变化规律。
    结果 在SD大鼠脑组织内注射Ⅶ型胶原酶可成功诱导大鼠自发性脑出血，血肿体积自3 DPI开始逐渐增加，到10 DPI时血肿体积达到最大。此后血肿逐渐吸收，在30 DPI时全部吸收。对脑组织eNSCs变化规律分析显示，在3 DPI时有少量eNSCs被激活但很快便减少，在10 DPI 时eNSCs逐渐开始增多，到20 DPI时有大量eNSCs迁移至脑出血部位，而在30 DPI 时eNSCs又明显减少（P<0.01）。对脑组织免疫微环境分析显示，促炎型（M1型）免疫细胞在10、20 DPI时明显增加（P<0.01），在30 DPI 时减少。抑炎型（M2型）免疫细胞在3 DPI 时开始逐渐增加，到20 DPI 时明显减少（P<0.05），而在30 DPI 时又出现增加。
    结论 大鼠脑出血后，向脑出血部位迁移的eNSCs先增多后减少，免疫微环境表现出先抑炎、再促炎、最后抑炎的变化规律。炎症对eNSCs迁移可能有刺激作用，但是过强的炎症激活对eNSCs的分化及进一步激活有抑制作用。脑出血后修复及脑保护早期（10 d内）及亚急性期（20 d内）可能是最佳的干预时机。
     
    Abstract:
    Objective Intracerebral hemorrhage (ICH), the second most common type of stroke, can cause long-lasting disability in the afflicted patients. The study was conducted to examine the patterns of change in endogenous neural stem cells (eNSCs) and in the regenerative microenvironment after ICH, to observe the relationship between the migration of eNSCs and the pattern of change in the polarization state of immune cells in the microenvironment, and provide a research basis for research on clinical nerve repair.
    Methods The collagenase injection method was used for modeling. The ICH model was induced in adult female Sprague-Dawley (SD) rats by injecting type VII collagenase (2 U) into the brain tissue of rats. All the experimental rats weighed 280-300 g. In order to simulate the ICU at different time points, including the acute phase (within 1 week), subacute phase (1-3 weeks), and the chronic phase (over 3 weeks), brain tissues were harvested at 3 day post injection (3 DPI), 10 DPI, 20 DPI, and 30 DPI to evaluate the modeling effect. Immunofluorescence staining of the brain tissue sections was performed with DCX antibody to observe the pattern of change in the migration of eNSCs in the brain tissue at different time points. Immunofluorescence staining of brain tissue sections was performed with CD206 antibody and CD86 antibody for respective observation of the pattern of change in pro-inflammatory (M1-type) and anti-inflammatory (M2-type) immune cells in the regenerative microenvironment of the brain tissue after ICM.
    Results Spontaneous ICH was successfully induced by injecting type Ⅶ collagenase into the brain tissue of SD rats. The volume of the hematoma formed started to gradually increase at 3 DPI and reached its maximum at 10 DPI. After that, the hematoma was gradually absorbed and was completely absorbed by 30 DPI. Analysis of the pattern of changes in eNSCs in the brain tissue showed that a small number of eNSCs were activated at 3 DPI, but very soon their number started to decrease. By 10 DPI, eNSCs gradually began to increase. A large number of eNSCs migrated to the hemorrhage site at 20 DPI. Then the number of eNSCs decreased significantly at 30 DPI (P<0.01). Analysis of the immune microenvironment of the brain tissue showed that pro-inflammatory (M1 type) immune cells increased significantly at 10 and 20 DPI (P<0.01) and decreased at 30 DPI. Anti-inflammatory (M2 type) immune cells began to increase gradually at 3 DPI, decreased significantly at 20 DPI (P<0.05), and then showed an increase at 30 DPI.
    Conclusion After ICH in rats, eNSCs migrating toward the site of ICH first increase and then decrease. The immune microenvironment demonstrates a pattern of change in which inflammation is suppressed at first, then promoted, and finally suppressed again. Inflammation may have a stimulatory effect on the migration of eNSCs, but excessive inflammatory activation has an inhibitory effect on the differentiation and further activation of eNSCs. After ICH, the early stage of repair and protection (10 d) and the subacute phase (20 d) may provide the best opportunities for intervention.
     
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