Guan Jingxia, Zhang Shaofeng, Zhou Qin, Yuan Zhenhua, Lu Zuneng
a Department of Neurology , Renmin Hospital of Wuhan University , Wuhan , China.
Neurol Res. 2016 Sep;38(9):809-16. doi: 10.1080/01616412.2016.1210356. Epub 2016 Jul 23.
To investigate the effect of thrombin preconditioning (TPC) on the intracerebral hemorrhage (ICH)-induced proliferation, migration, and function of subventriclular zone (SVZ) cells and to find new strategies that enhance endogenous neurogenesis after ICH.
Male Sprague-Dawley rats were randomly divided into 3 groups (ICH, TPC, and control group). Rats of each group were randomly divided into 5 subgroups (3-d, 7-d, 14-d, 21-d, and 28-d subgroup). ICH was caused by intrastrial stereotactic administration of collagenase type IV. Brdu was used to label newborn SVZ cells. Organotypic brain slices were cultured to dynamically observe the migration of SVZ cells at living brain tissue. Migration of Dil-labeled SVZ cells in living brain slices was traced by time-lapse microscopy. To assess whether SVZ cells migrating to injured striatum had the ability to form synapses with other cells, brain slices from each group were double immunolabeled with Brdu and synapsin I.
The number of Brdu-positive cells markedly increased in the ipsilateral SVZ and striatum 3 days after TPC, peaked at 14 days (P < 0.01), continued to 21 days, and then gradually decreased at 28 days with significant difference compared to the ICH group at each time point (P < 0.01). Migration of Dil-labeled SVZ cells in brain slices in each group was observed and imaged during a 12-h period. Dil-labeled SVZ cells in the TPC group were observed to migrate laterally toward striatum with time with a faster velocity compared to the ICH group (P < 0.01). Our study also demonstrated that TPC induced strong colocalization of Brdu and synapsin I in the ipsilateral striatum between 3 and 28 days after injury.TPC made colocalization of Brdu and synapsin I appear earlier and continue for a longer time compared to the ICH group.
Our results demonstrated that TPC could promote proliferation, migration, and function of SVZ cells after ICH, which may provide a new idea for enhancing endogenous neurogenesis and developing new therapeutic strategies against ICH-induced brain injury.
探讨凝血酶预处理（TPC）对脑出血（ICH）诱导的脑室下区（SVZ）细胞增殖、迁移及功能的影响，并寻找增强ICH后内源性神经发生的新策略。
将雄性Sprague-Dawley大鼠随机分为3组（ICH组、TPC组和对照组）。每组大鼠再随机分为5个亚组（3天亚组、7天亚组、14天亚组、21天亚组和28天亚组）。通过脑内立体定向注射IV型胶原酶诱导脑出血。用溴脱氧尿苷（Brdu）标记新生的SVZ细胞。培养脑片以动态观察活脑组织中SVZ细胞的迁移。通过延时显微镜追踪活脑片中Dil标记的SVZ细胞的迁移。为评估迁移至损伤纹状体的SVZ细胞是否具有与其他细胞形成突触的能力，对每组脑片进行Brdu和突触素I双重免疫标记。
TPC后3天，同侧SVZ和纹状体中Brdu阳性细胞数量显著增加，14天时达到峰值（P<0.01），持续至21天，然后在28天时逐渐下降，与ICH组在各时间点相比差异有统计学意义（P<0.01）。在12小时内观察并记录了每组脑片中Dil标记的SVZ细胞的迁移情况。与ICH组相比，TPC组中Dil标记的SVZ细胞随时间向纹状体横向迁移的速度更快（P<0.01）。我们的研究还表明，TPC在损伤后3至28天诱导同侧纹状体中Brdu和突触素I强烈共定位。与ICH组相比，TPC使Brdu和突触素I的共定位出现更早且持续时间更长。
我们的结果表明，TPC可促进ICH后SVZ细胞的增殖、迁移及功能，这可能为增强内源性神经发生和开发针对ICH所致脑损伤的新治疗策略提供新思路。
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