【摘要】：目的：建立缺氧缺糖神經(jīng)細胞損傷模型，探討EPO發(fā)揮神經(jīng)保護作用的可能機制，希望為EPO在神經(jīng)系統(tǒng)疾病的臨床應(yīng)用提供理論依據(jù)。 方法：將出生后24h內(nèi)C57BL/6小鼠的大腦皮質(zhì)神經(jīng)元原代培養(yǎng)7-10d后，神經(jīng)元特異性烯醇化酶（Neuron specific enolase, NSE）免疫組化染色方法鑒定神經(jīng)元的純度；用含連二亞硫酸鈉的無糖Earle’s液模擬大腦缺血缺氧性損傷，建立缺氧缺糖神經(jīng)細胞損傷模型。分組：1、正常組；2、模型組（缺氧缺糖神經(jīng)細胞損傷模型）；3、EPO組(EPO組干預(yù)的缺氧缺糖神經(jīng)細胞損傷模型）。MTT檢測細胞存活率、FITC-Annexin V/PI熒光染色方法檢測細胞凋亡率、測定LDH漏出率，觀察EPO對缺氧缺糖損傷神經(jīng)元的保護作用。 結(jié)果：1、成功原代培養(yǎng)神經(jīng)細胞，經(jīng)NSE免疫組化方法鑒定神經(jīng)元純度大于90%；2、建立缺氧缺糖神經(jīng)細胞損傷模型，模型組中神經(jīng)細胞凋亡率為（15.2±0.37），LDH漏出率為（20.4±0.33），細胞存活率為（54.1±0.51），與正常組相比差異有統(tǒng)計學(xué)意義（P0.01）；3、與模型組相比，EPO組的細胞凋亡率、LDH漏出率低于模型組，細胞存活率高于模型組（P0.01）。 結(jié)論：原代培養(yǎng)小鼠神經(jīng)細胞，成功構(gòu)建缺氧缺糖神經(jīng)細胞損傷模型。EPO能夠減輕缺氧缺糖對神經(jīng)元的損傷，，該保護作用可能是通過抑制神經(jīng)細胞凋亡途徑介導(dǎo)完成。
[Abstract]:Objective: to establish a model of hypoxia-glucose deficiency nerve cell injury and to explore the possible mechanism of neuroprotective effect of EPO in order to provide a theoretical basis for the clinical application of EPO in nervous system diseases. Methods: after primary culture of cerebral cortical neurons of C57BL/6 mice within 24 hours after birth for 7-10 days, the purity of neurons was identified by immunohistochemical staining with neuron-specific enolase (Neuron specific enolase, NSE). A model of hypoglycemic neuronal injury was established by simulating cerebral ischemia and hypoxia injury with Earle's solution containing sodium bisulfite. Groups: 1, normal group, 2, model group (hypoxia and glucose deficiency nerve cell injury model); (3). MTT, FITC-Annexin V/PI fluorescence staining and LDH leakage were used to detect cell survival rate, apoptosis rate and LDH leakage rate in EPO group (hypoxia / glucose deficient nerve cell injury model). To observe the protective effect of EPO on neurons injured by hypoxia and glucose deficiency. Results: 1. The primary cultured neurons were successfully cultured, and the purity of neurons was higher than 90 by NSE immunohistochemical method. 2. In the model group, the apoptosis rate was (15.2 鹵0.37), LDH leakage rate was (20.4 鹵0.33), and the cell survival rate was (54.1 鹵0.51). Compared with the normal group, the difference was statistically significant (P0.01). 3Compared with the model group, the apoptosis rate and LDH leakage rate in EPO group were lower than those in the model group, and the cell survival rate was higher than that in the model group (P0.01). Conclusion: EPO can attenuate the neuronal damage induced by hypoxia and glucose deficiency, which may be mediated by inhibiting neuronal apoptosis.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R741

【參考文獻】

相關(guān)期刊論文 前9條

1 張廣云;胡曉;袁平;王建怡;;谷氨酸誘導(dǎo)新生小鼠皮層神經(jīng)細胞損傷體外模型的建立[J];神經(jīng)解剖學(xué)雜志;2012年04期

2 金寶;張育才;崔云;;促紅細胞生成素對發(fā)育期小鼠腦損傷后腦源性神經(jīng)營養(yǎng)因子和酪氨酸受體激酶B表達的影響[J];上海交通大學(xué)學(xué)報(醫(yī)學(xué)版);2012年01期

3 王丹丹;王建;彭穎;王茜;馬驍;唐丹霞;夏厚林;曾南;;四味芳香開竅藥的揮發(fā)性成分對缺血缺氧PC12細胞及細胞內(nèi)Ca~(2+)的影響[J];西安交通大學(xué)學(xué)報(醫(yī)學(xué)版);2012年03期

4 吉訓(xùn)明;段麗芬;朱榆紅;李淑婷;姜玲玲;羅玉敏;;大鼠頸動脈重度狹窄合并腦梗死后腦促紅細胞生成素的變化[J];中國藥物與臨床;2009年01期

5 侯麗淳;關(guān)雪蓮;韓鳳;楊慧;王昆祥;;促紅細胞生成素對大鼠腦出血周邊組織谷氨酸及腦水含量的影響[J];中風(fēng)與神經(jīng)疾病雜志;2012年02期

6 鄭敏;郭蓮軍;呂青;何治;徐旭林;喻欣;宗賢剛;;大鼠皮質(zhì)及海馬神經(jīng)元體外缺氧缺糖模型建立[J];中國老年學(xué)雜志;2006年03期

7 孫曉佳;孫宇;孫霓;王勝軍;李秀江;;EPO對心肺復(fù)蘇后大鼠海馬神經(jīng)元的細胞凋亡、Bcl-2及Bax表達的影響[J];中國實驗診斷學(xué);2012年06期

8 石瑞麗;胡金鳳;孔令雷;牛非;吳苗苗;何鑫;李培鋒;陳乃宏;;瓜子金皂苷己對連二亞硫酸鈉致氧糖剝奪/復(fù)供誘導(dǎo)PC12細胞凋亡的抑制作用[J];中國藥理學(xué)通報;2013年03期

9 胡海燕;崔志慧;陳翔;王文花;張曉艷;;清心開竅方含藥腦脊液對連二亞硫酸鈉所致PC12細胞損傷的保護機制研究[J];中國中藥雜志;2013年09期

本文編號：2345861