标题：鼠神经生长因子对新生大鼠缺氧缺血性脑损伤后巢蛋白表达的影响

      作者：许锋，姚文静，曹春霞，薛媛媛，李玉运，马立吉，付海波    淄博市中心医院儿科，山东 淄博 255036

      卷次： 2020年31卷21期

      【摘要】 目的 观察鼠神经生长因子(mNGF)对新生大鼠缺氧缺血性脑损伤(HIBD)后巢蛋白(nestin)表达水平的影响，研究内源性神经干细胞增殖情况，探讨mNGF可能的神经保护机制。方法 新生7 d龄Wistar大鼠108只按完全随机法分为对照组、HIBD组和治疗组，每组36只。结扎大鼠左侧颈总动脉和8%低氧暴露2 h，制作新生大鼠HIBD模型。对照组仅暴露游离左侧颈总动脉，不结扎左侧颈总动脉和低氧暴露。治疗组大鼠于左侧股二头肌部位肌肉注射mNGF (10 μg/kg)，1次/d，连用3 d。HIBD组大鼠于相同部位注射等量生理盐水3 d。每组随机取12只大鼠分别于术后4 d、7 d、14 d处死后，制备大鼠海马部位脑组织HE及免疫组化切片。利用计算机图像分析技术检测各组nestin阳性细胞计数。结果 对照组海马部位椎体细胞排列整齐，HIBD组海马部位椎体细胞排列紊乱，可见较多凋亡神经细胞，治疗组可见海马部位椎体细胞排列较整齐，可见少量凋亡神经细胞；对照组、HIBD组及治疗组大鼠术后4 d nestin阳性细胞个数分别为(15.25±3.02)个、(24.67±3.37)个、(40.33±2.68)个，差异有统计学意义(P<0.05)；对照组、HIBD组及治疗组大鼠术后 7 d nestin阳性细胞个数分别为(17.33±2.71)个、(30.58±4.03)个、(48.75±2.99)个，差异有统计学意义(P<0.05)；对照组、HIBD组及治疗组大鼠术后 14 d nestin阳性细胞个数分别为(14.58±1.83)个、(22.75±2.05)个、(32.17±2.52)个，差异有统计学意义(P<0.05)。结论 新生HIBD大鼠经mNGF干预后位于海马齿状回nestin表达增加，提示可能促进内源性神经干细胞的增殖、再生，在HIBD后神经再生、修复中可能发挥一定保护作用。
      【关键词】 Wistar大鼠；鼠神经生长因子；缺氧缺血；脑；巢蛋白；干细胞
      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2020）21—2721—04

Effects of mouse nerve growth factor on nestin expression in newborn rats with hypoxia-ischemic brain damage.XU Feng, YAO Wen-jing, CAO Chun-xia, XUE Yuan-yuan, LI Yu-yun, MA Li-ji, FU Hai-bo.
Department of Pediatrics,the Central Hospital of Zibo City, Zibo 255036, Shandong, CHINA
【Abstract】 Objective To observe the effects of mouse nerve growth factor (mNGF) on nestin expression inneonatal rats after hypoxia-ischemic brain damage (HIBD), to study the proliferation of endogenous neural stem cellsand explore the possible neuroprotective mechanism. Methods One hundred and eight 7-day-old newborn Wistar ratswere randomly assigned into control group, HIBD group, and treatment group (n=36 each). HIBD rat models were estab-lished by ligation of the left carotid artery along with 2-hour exposure to 8% oxygen in the latter two groups. The con-trol group only exposed the left carotid artery, without ligation or hypoxia-exposure. The treatment group received an in-tramuscular injection of mNGF (10 μg/kg) once a day for three days after insult, while the other two groups received nor-mal saline at the same time. In each group the rats were randomly executed immediately, on the 4th, 7th and 14th day afterthe operation (n=12). The changes of nestin expression in hippocampus were examined by Immunohistochemistry andHE staining, and nestin positive cells were detected by computer image analysis. Results In the control group, the ver-tebral cells in the hippocampus were arranged orderly. In the HIBD group, the vertebral cells in the hippocampus were ar-ranged disorderly, with more apoptotic neurons. In the treatment group, the vertebral cells in the hippocampus were ar-ranged orderly, and a small number of apoptotic neurons were visible. The number of nestin positive cells in controlgroup, HIBD group and treatment group on the 4th day after insult was 15.25±3.02, 24.67±3.37, 40.33±2.68, respective-ly, and the difference was statistically significant (P<0.05). The number of nestin positive cells in control group, HIBDgroup, and treatment group on the 7th day after insult were 17.33±2.71, 30.58±4.03, 48.75±2.99 respectively, and the dif-ference was statistically significant (P<0.05). The number of nestin positive cells in control group, HIBD group, andtreatment group on the 14th day after insult were 14.58±1.83, 22.75±2.05, 32.17±2.52, respectively, and the differencewas statistically significant (P<0.05). Conclusion mNGF could enhance the number of nestin positive cells in hippo-campal dentate gyrus region of neonatal rats with HIBD, and possibly promote the proliferation of neural stem cells. Itmay play an important role in regeneration of neurocytes damaged by hypoxia-ischemia.
      【Key words】 Wistar rat; Mouse nerve growth factor; Hypoxia-ischemia; Brain; Nestin; Stem cells