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目的:研究早期干预对缺氧缺血性脑损伤(Hypoxic-ischemic brain injury)新生大鼠内源性神经干细胞分化能力的影响。方法:共75只7日龄SD(Sprague Dawley)大鼠,随机分成模型组(n=50)和假手术组(n=25),模型组用Rice法制作50只缺氧缺血性脑病模型,分成HIBD组(n=25)和干预组(n=25)。对干预组进行早期干预,HIBD组和假手术组常规饲养,每组选取1天,7天,14天,21天,28天5个时间点处死5只,每只鼠处死前都进行腹腔BrdU注射标记新生细胞,应用荧光免疫双标技术检测各时间点海马齿状(DG)GFAP/BrdU和NeuN/BrdU阳性细胞数。结果:(1)3组大鼠DG区均有BrdU/NeuN和BrdU/GFAP阳性双标记染色;(2)与假手术组比较,干预组及HIBD组大鼠DG区BrdU/NeuN和BrdU/GFAP阳性双标记染色在术后14,21,28天均有显著增高(P<0.01)。(3)早期干预组大鼠术后14,21,28天时的BrdU/NeuN和BrdU/GFAP阳性双标记染色均显著高于HIBD组(P<0.01)。结论:缺氧缺血致大鼠脑损伤可诱导内源性神经干细胞分化;早期干预可提高脑损伤鼠内源性神经干细胞分化能力。
Objective: To investigate the effect of early intervention on the differentiation of endogenous neural stem cells in neonatal rats with hypoxic-ischemic brain injury (Hypoxic-ischemic brain injury). Methods: A total of 75 7-day-old Sprague Dawley rats were randomly divided into model group (n = 50) and sham operation group (n = 25) , Divided into HIBD group (n = 25) and intervention group (n = 25). Intervention groups were intervened early, and HIBD group and sham operation group were routinely housed. Five rats in each group were sacrificed at 1, 7, 14, 21 and 28 days. Neonatal cells were injected with fluorescent labeled double immunofluorescence technique to detect GFAP / BrdU and NeuN / BrdU positive cells in hippocampus dentate (DG) at each time point. Results: (1) BrdU / NeuN and BrdU / GFAP positive double staining were all detected in the DG area of the 3 groups. (2) Compared with the sham operation group, BrdU / NeuN and BrdU / GFAP Positive double-labeled staining in 14,21,28 days after surgery were significantly higher (P <0.01). (3) The positive BrdU / NeuN and BrdU / GFAP double staining in the early intervention group was significantly higher than that in the HIBD group at 14, 21 and 28 days after operation (P <0.01). Conclusion: Hypoxic-ischemic brain damage can induce the differentiation of endogenous neural stem cells in rats. Early intervention can improve the differentiation of endogenous neural stem cells in rats after brain injury.