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背景:许旺细胞能分泌多种营养因子,有保护和营养神经元的作用,并对神经损伤后的修复起促进作用。目的:观察正常成年大鼠和不同胎龄鼠及幼鼠腰髓组织中许旺细胞源神经营养因子的表达,及其对脊髓不同发育时期的作用。设计:对照实验,动物实验。材料:实验于2003-09于南华大学动物实验室完成,采用Wister大鼠30只,雌雄不限,体质量为150～300g,由南华大学动物实验室提供。干预:①取材和切片:取体质量180～220g,成熟期为4～6周正常成年大鼠腰髓和背根神经节,继而行后固定。共8只雌鼠受孕,取不同孕龄母鼠中的胎鼠及幼鼠腰髓组织,分别行冰冻切片。②免疫组化染色:以抗许旺细胞源神经营养蛋白抗体作为一抗,行卵白素-生物素过氧化物酶复合物免疫组化染色,观察染色部位的灰度,通过电脑上扫描计算染色深度得出半定量值,分析许旺细胞源神经营养因子在腰髓的表达。主要观察指标:①正常腰髓免疫组化染色结果。②生长发育的腰髓免疫组化染色结果。结果:共8只雌鼠受孕,实验样本均取自8只受孕雌鼠的胎鼠及所产幼鼠。①正常腰髓免疫组化染色结果:许旺细胞源神经营养因子在成年大鼠腰髓的整个白质和灰质后角Ⅲ、Ⅳ、Ⅴ层有较低表达。②生长发育的腰髓免疫组化染色结果:许旺细胞源神经营养因子在胎龄14d和胎龄15d开始有表达,胚胎时期表达先增高后下降,其中胎龄21d最高。出生后许旺细胞源神经营养因子在腰髓中表达明显下降,其中第7和l2d最高。结论:许旺细胞源神经营养因子在成年大鼠腰髓及背根神经节有较低表达,在生长发育的腰髓中表达基本呈逐渐下降趋势。许旺细胞源神经营养因子对大鼠脊髓的发育可能起促进作用,此结果说明许旺细胞源神经营养因子在脊髓生长发育过程中,主要在胚胎发育时期起作用。
Background: Schwann cells can secrete many kinds of nutritional factors, protect and nourish neurons, and promote the repair after nerve injury. OBJECTIVE: To observe the expression of Schwann cell-derived neurotrophic factor (BDNF) in normal adult rats, different gestational age and juvenile rat’s lumbar spinal cord and its effect on different developmental stages of spinal cord. Design: control experiment, animal experiment MATERIALS: Experiments were performed at the Animal Laboratory of Nanhua University from September 2003 to September 2003. 30 Wister rats were used in this experiment. The average body weight was 150-300 g and provided by Animal Laboratory of Nanhua University. Intervention: ①Correction and section: The body weight of 180 ~ 220g, maturity of 4 to 6 weeks normal adult lumbar spinal cord and dorsal root ganglion, and then fixed after the line. A total of 8 female rats conceive, fetus and pups of different gestational age maternal lumbar spinal cord tissue, frozen section. ② Immunohistochemical staining: The anti-Schwann cell-derived neurotrophic protein antibody was used as the primary antibody to perform immunohistochemical staining of avidin-biotin peroxidase complex, and the grayscale of the stained part was observed. The staining was calculated by scanning on the computer Semi-quantitative value was obtained, the expression of Schwann cell-derived neurotrophic factor in the lumbar spinal cord was analyzed. MAIN OUTCOME MEASURES: ① normal lumbar spinal cord immunohistochemical staining results. ② growth and development of lumbar spinal cord immunohistochemical staining results. Results: A total of 8 female rats were pregnant. The experimental samples were obtained from the fetus and the young mice of 8 pregnant rats. ① Normal lumbar spinal cord immunohistochemical staining results: Schwann cell derived neurotrophic factor in the adult rat lumbar spinal cord white matter and gray matter posterior horn Ⅲ, Ⅳ, Ⅴ lower expression. ② The results of immunohistochemical staining of growth and development of the spinal cord: Schwann cell derived neurotrophic factor expression in gestational age 14d and gestational 15d, the expression of embryonic first increased and then decreased, of which the highest gestational age 21d. Schwann cells derived neurotrophic factor was significantly decreased in the lumbar spinal cord after birth, of which 7 and l2d highest. CONCLUSION: Schwann cell derived neurotrophic factor is less expressed in the lumbar spinal cord and dorsal root ganglion of adult rats, and gradually decreases in the developing lumbar spinal cord. Schwann cell derived neurotrophic factor may play an important role in the development of rat spinal cord. The results suggest that Schwann cell derived neurotrophic factor plays a role in embryonic development in the process of spinal cord growth and development.