高压氧治疗促进HIBD新生大鼠内源性神经干细胞的迁移与分化

Hyperbaric oxygen promotes the migration and differentiation of endogenous neural stem cells in neonatal rats with hypoxic-ischemic brain damage

目的该研究探讨了高压氧(HBO)对缺氧缺血性脑损伤(HIBD)新生大鼠内源性神经干细胞(NSCs)的迁移与分化的影响。方法7日龄Sprague-Dawley新生大鼠随机分为对照组,模型组及HBO组。采用Rice-Vannucci方法制成HIBD模型。造模后3 h内行HBO治疗。分别于HBO治疗后7 d、14 d、28 d,采用BrdU/DCX,BrdU/β-tubulin,BrdU/GFAP和BrdU/O4免疫荧光双标法,用共聚焦显微镜动态检测侧脑室室管膜下区(sub-ventricular zone,SVZ)与大脑皮层内源性NSCs的迁移与分化。结果治疗后7 d,HBO组损伤侧SVZ区BrdU+DCX+细胞数(84±21个/mm2)增加,多于对照组(39±14个/mm2)与模型组(68±17个/mm2)(P<0.05);治疗后14 d,SVZ区BrdU+DCX+细胞数减少,而皮层区BrdU+DCX+细胞数增加,HBO组明显多于对照组(P<0.01);治疗后28 d,各组大脑皮层BrdU+DCX+细胞数减少,大脑皮层出现BrdU+β-tubulin+,BrdU+GFAP+,BrdU+O4+细胞。HBO组BrdU+β-tubulin+与BrdU+O4+细胞数显著高于对照组与模型组(P<0.05)。结论高压氧可促进HIBD新生大鼠内源性NSCs迁移到大脑皮层并分化为成熟的神经细胞。

Objective To explore the effects of hyperbaric oxygen （HBO） treatment on the migration and differentiation of endogenous neural stem cells （NSCs） in neonatal rats with hypoxie-ischemie brain damage （HIBD）. Methods Seven-day-old Sprague-Dawley rats were randomly divided into the normal control （CON）, the HIBD model and the HBO groups （ HBO treatment was administered at 2 ATA, once daily for 7 days within 3 hrs after HIBD）. HIBD model was prepared according to the classic Rice-Vannucci method. BrdU/DCX, BrdU/β-tubulin, BrdU/GFAP and BrdU/O4 immunofluoreseence were examined by eonfocal microscopy in the subventricular zone （SVZ） and the cortex 7, 14 and 28 days after HBO treatment. Results The BrdU^＋ DCX^＋ cells in the SVZ （84 ± 21 eells/mm^2 ） in the HBO group were significantly higher than those in the CON group （39 ± 14 cells/mm^2） （P 〈0.05） and the HIBD model group （68 ± 17 cells/mm^2） （P 〈 0.05 ） 7 days after HBO treatment. Fourteen days after HBO treatment, the BrdU^＋ DCX^＋ cells decreased in the SVZ and more cells were observed in the cortex in the HBO group as compared with the CON group （ P 〈 0.01 ）. The BrdU^＋ β-tubulin^＋ , BrdU^＋ GFAP^＋ and BrdU^＋ O4^＋ cells were observed in the cortex, and more BrdU^＋ β-tubulin^＋ and BrdU^＋O4^＋ cells were observed in the HBO group as compared with the CON and the HIBD model groups （P 〈 0.05） 28 days after HBO treatment. Condnsions HBO treatment may promote endogenous NSCs to migrate to the cortex and differentiate into mature neuroeytes in neonatal rats with HIBD.