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目的建立新生鼠低血糖脑损伤模型,并观察损伤后不同脑区神经元变性的动态变化。方法 160只新生Wistar大鼠随机分为胰岛素诱导短时间低血糖组(INS-S组,n=40)、胰岛素诱导长时间低血糖组(INS-P组,n=40)、饥饿组(FAS组,n=40)和对照组(CON组,n=40)。在大鼠出生后第2、4、6天分别采用皮下注射胰岛素(15U/kg)和饥饿的手段,诱导新生鼠低血糖;在生后第6天诱导低血糖后2h、6h、1d、3d、7d和14d采用Fluoro-JadeB(FJB)染色法观察7个脑区变性神经元的动态变化,并应用特异性神经核抗原(NeuN)荧光免疫双标对FJB阳性细胞进行细胞类型鉴定。结果 INS-S组、FAS组和CON组新生大鼠各脑区各时间点均未见FJB阳性细胞,而INS-P组在旁矢状面、梨状皮层、海马齿状回、丘脑和下丘脑可见大量的FJB阳性细胞,并同时显示NeuN免疫源性。结论胰岛素(15U/kg)皮下注射可建立稳定可靠的新生鼠低血糖脑损伤模型。新生鼠反复严重低血糖可引起广泛的神经元变性,皮层、丘脑、下丘脑和海马齿状回对低血糖损害更为敏感。
Objective To establish neonatal hypoglycemic brain injury model and observe the dynamic changes of neuron degeneration in different brain regions after injury. Methods 160 newborn Wistar rats were randomly divided into three groups: insulin-induced short-term hypoglycemia group (INS-S group, n = 40), insulin induced long-term hypoglycemia group (INS-P group, n = 40), starvation group Group, n = 40) and control group (CON group, n = 40). Hypoglycemia was induced in neonatal rats by hypodermic injection of insulin (15U / kg) and starvation on days 2, 4 and 6 after birth. The hypoglycemia was induced at 2, 6, 1, 3d The dynamic changes of degenerative neurons in 7 brain regions were observed by Fluoro-JadeB (FJB) staining on day 7, day 7 and day 14. The cell types of FJB positive cells were identified by fluorescent double immunofluorescence staining with specific NeuN antigen. Results There were no FJB positive cells in INS-S group, FAS group and CON group at each time point in brain regions of rats, but there was no significant difference in INS-P group between the sagittal plane, piriform cortex, hippocampal dentate gyrus, Thalamus shows a large number of FJB positive cells, and at the same time shows NeuN immunogenicity. Conclusion Insulin (15U / kg) subcutaneously injected can establish a stable and reliable neonatal hypoglycemic brain injury model. Repeated severe neonatal hypoglycemia can cause a wide range of neuronal degeneration, cortical, thalamic, hypothalamic and hippocampal dentate gyrus is more sensitive to hypoglycemia.