PDF(Pubmed)
Abstract：
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
摘要：
■脑出血（ICH），第二种最常见的中风类型，会导致患病患者长期残疾。这项研究是为了检查ICH后内源性神经干细胞（eNSC）和再生微环境的变化模式，观察细胞在微环境中的迁移与免疫细胞极化状态变化的关系,为临床神经修复研究提供研究依据。
■使用胶原酶注射方法进行建模。通过向大鼠脑组织中注射VII型胶原酶（2U），在成年雌性Sprague-Dawley（SD）大鼠中诱导ICH模型。所有实验大鼠体重为280-300g。为了模拟不同时间点的ICU，包括急性期（1周内），亚急性期（1-3周），和慢性期（超过3周），在注射后3天收获脑组织（3DPI），10DPI，20DPI，和30DPI进行建模效果评价。用DCX抗体对脑组织切片进行免疫荧光染色,观察不同时间点eNSC在脑组织中迁移的变化规律。用CD206抗体和CD86抗体进行脑组织切片的免疫荧光染色,以分别观察ICM后脑组织再生微环境中促炎(M1型)和抗炎(M2型)免疫细胞的变化模式。
■在SD大鼠脑组织中注射VII型胶原酶成功诱发自发性ICH。形成的血肿的体积在3DPI时开始逐渐增加,并在10DPI时达到其最大值。之后,血肿逐渐吸收，30DPI完全吸收。对脑组织中eNSCs变化的模式分析显示，在3个DPI时，有少量的eNSCs被激活，但很快他们的数量开始减少。到10DPI，eNSC逐渐开始增加。在20DPI时,大量的eNSC迁移到出血部位。然后在30DPI时，eNSCs的数量显着减少（P<0.01）。对脑组织免疫微环境的分析表明，促炎（M1型）免疫细胞在10和20DPI时明显增加（P<0.01），在30DPI时降低。3DPI时抗炎（M2型）免疫细胞开始逐渐增多，在20DPI时显著降低(P<0.05)，然后在30DPI时显示增加。
■大鼠ICH后，向ICH位点迁移的eNSC先增加后减少。免疫微环境表现出一种变化模式，其中炎症首先被抑制，然后晋升，最后再次镇压。炎症可能对神经干细胞的迁移有刺激作用，但过度的炎症激活对神经干细胞的分化和进一步激活有抑制作用。ICH之后,修复和保护的早期阶段（10d）和亚急性期（20d）可能为干预提供最佳机会。
■使用胶原酶注射方法进行建模。通过向大鼠脑组织中注射VII型胶原酶（2U），在成年雌性Sprague-Dawley（SD）大鼠中诱导ICH模型。所有实验大鼠体重为280-300g。为了模拟不同时间点的ICU，包括急性期（1周内），亚急性期（1-3周），和慢性期（超过3周），在注射后3天收获脑组织（3DPI），10DPI，20DPI，和30DPI进行建模效果评价。用DCX抗体对脑组织切片进行免疫荧光染色,观察不同时间点eNSC在脑组织中迁移的变化规律。用CD206抗体和CD86抗体进行脑组织切片的免疫荧光染色,以分别观察ICM后脑组织再生微环境中促炎(M1型)和抗炎(M2型)免疫细胞的变化模式。
■在SD大鼠脑组织中注射VII型胶原酶成功诱发自发性ICH。形成的血肿的体积在3DPI时开始逐渐增加,并在10DPI时达到其最大值。之后,血肿逐渐吸收，30DPI完全吸收。对脑组织中eNSCs变化的模式分析显示，在3个DPI时，有少量的eNSCs被激活，但很快他们的数量开始减少。到10DPI，eNSC逐渐开始增加。在20DPI时,大量的eNSC迁移到出血部位。然后在30DPI时，eNSCs的数量显着减少（P<0.01）。对脑组织免疫微环境的分析表明，促炎（M1型）免疫细胞在10和20DPI时明显增加（P<0.01），在30DPI时降低。3DPI时抗炎（M2型）免疫细胞开始逐渐增多，在20DPI时显著降低(P<0.05)，然后在30DPI时显示增加。
■大鼠ICH后，向ICH位点迁移的eNSC先增加后减少。免疫微环境表现出一种变化模式，其中炎症首先被抑制，然后晋升，最后再次镇压。炎症可能对神经干细胞的迁移有刺激作用，但过度的炎症激活对神经干细胞的分化和进一步激活有抑制作用。ICH之后,修复和保护的早期阶段（10d）和亚急性期（20d）可能为干预提供最佳机会。
