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【摘要】：目的 研究腦缺血再灌注損傷腦勻漿上清液對大鼠骨髓基質細胞形態(tài)、活性、神經元特異性烯醇化酶、電生理和神經營養(yǎng)因子表達情況的影響。 方法 將體外培養(yǎng)的大鼠骨髓基質細胞分別置于實驗組(含腦缺血再灌注損傷腦勻漿上清液)、假手術組(含正常腦勻漿上清液)及空白對照組培養(yǎng)條件中,采用組織學、臺盼藍染色、免疫組化、膜片鉗、SqRT-PCR等方法,觀察細胞形態(tài)改變,測定活性,鑒定神經元特異性烯醇化酶表達,記錄全細胞膜電流,檢測NGF、GDNF、BDNF和bFGF在各時間點mRNA表達情況。 結果 實驗組細胞形態(tài)發(fā)生胞體變圓變亮、突起增多伸長等“神經元樣”改變,細胞活性良好;神經元特異性烯醇化酶陽性率及神經營養(yǎng)因子的表達水平高于其他組,差異有統(tǒng)計學意義;可引發(fā)出電壓依賴性外向K~+電流,未見內向Na~+與Ca~(2+)電流,未能引發(fā)出動作電位。 結論 大鼠骨髓基質細胞在腦缺血再灌注損傷細胞微環(huán)境中生長良好,形態(tài)改變,類似“神經元樣”,出現(xiàn)神經元特異性烯醇化酶,但無神經元的電生理特征,神經營養(yǎng)因子表達能力增強,在神經細胞損傷修復及損傷保護中發(fā)揮巨大的作用。
[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.
【學位授予單位】：遼寧醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R329.28

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