大鼠脑缺血再灌注损伤脑匀浆上清液对骨髓基质细胞的影响
发布时间：2018-11-05 14:22
【摘要】：目的 研究脑缺血再灌注损伤脑匀浆上清液对大鼠骨髓基质细胞形态、活性、神经元特异性烯醇化酶、电生理和神经营养因子表达情况的影响。 方法 将体外培养的大鼠骨髓基质细胞分别置于实验组(含脑缺血再灌注损伤脑匀浆上清液)、假手术组(含正常脑匀浆上清液)及空白对照组培养条件中,采用组织学、台盼蓝染色、免疫组化、膜片钳、SqRT-PCR等方法,观察细胞形态改变,测定活性,鉴定神经元特异性烯醇化酶表达,记录全细胞膜电流,检测NGF、GDNF、BDNF和bFGF在各时间点mRNA表达情况。 结果 实验组细胞形态发生胞体变圆变亮、突起增多伸长等“神经元样”改变,细胞活性良好;神经元特异性烯醇化酶阳性率及神经营养因子的表达水平高于其他组,差异有统计学意义;可引发出电压依赖性外向K~+电流,未见内向Na~+与Ca~(2+)电流,未能引发出动作电位。 结论 大鼠骨髓基质细胞在脑缺血再灌注损伤细胞微环境中生长良好,形态改变,类似“神经元样”,出现神经元特异性烯醇化酶,但无神经元的电生理特征,神经营养因子表达能力增强,在神经细胞损伤修复及损伤保护中发挥巨大的作用。
[Abstract]:Objective to study the effects of brain homogenate supernatant of cerebral ischemia-reperfusion injury on the morphology, activity, neuron-specific enolase, electrophysiology and neurotrophic factor expression of bone marrow stromal cells in rats. Methods Rat bone marrow stromal cells cultured in vitro were placed in the experimental group (containing brain homogenate supernatant of cerebral ischemia-reperfusion injury), sham operation group (including normal brain homogenate supernatant) and blank control group. By means of histology, trypan blue staining, immunohistochemistry, patch clamp and SqRT-PCR, the changes of cell morphology, activity, expression of neuron-specific enolase, recording of the whole cell membrane current and detection of NGF,GDNF, were observed. MRNA expression of BDNF and bFGF at different time points. Results in the experimental group, the morphogenetic cell body became round and bright, the process increased and elongated, and the "neuron-like" changes, and the cell activity was good. The positive rate of neuron-specific enolase and the expression of neurotrophic factor were significantly higher than those of other groups. It can induce voltage-dependent outward K ~ currents, but no inward Na~ and Ca~ (2) currents, but no action potential. Conclusion the rat bone marrow stromal cells grow well in the microenvironment of cerebral ischemia-reperfusion injury cells, and the morphology changes, similar to "neuron-like", with neuron-specific enolase, but no electrophysiological characteristics of neurons. The expression of neurotrophic factor plays an important role in the repair and protection of nerve cell injury.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329.28
本文编号：2312349
[Abstract]:Objective to study the effects of brain homogenate supernatant of cerebral ischemia-reperfusion injury on the morphology, activity, neuron-specific enolase, electrophysiology and neurotrophic factor expression of bone marrow stromal cells in rats. Methods Rat bone marrow stromal cells cultured in vitro were placed in the experimental group (containing brain homogenate supernatant of cerebral ischemia-reperfusion injury), sham operation group (including normal brain homogenate supernatant) and blank control group. By means of histology, trypan blue staining, immunohistochemistry, patch clamp and SqRT-PCR, the changes of cell morphology, activity, expression of neuron-specific enolase, recording of the whole cell membrane current and detection of NGF,GDNF, were observed. MRNA expression of BDNF and bFGF at different time points. Results in the experimental group, the morphogenetic cell body became round and bright, the process increased and elongated, and the "neuron-like" changes, and the cell activity was good. The positive rate of neuron-specific enolase and the expression of neurotrophic factor were significantly higher than those of other groups. It can induce voltage-dependent outward K ~ currents, but no inward Na~ and Ca~ (2) currents, but no action potential. Conclusion the rat bone marrow stromal cells grow well in the microenvironment of cerebral ischemia-reperfusion injury cells, and the morphology changes, similar to "neuron-like", with neuron-specific enolase, but no electrophysiological characteristics of neurons. The expression of neurotrophic factor plays an important role in the repair and protection of nerve cell injury.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329.28
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