不同濃度臭氧對大鼠離體腦片氧糖剝奪及再灌注損傷模型的保護作用

 師存偉 1，敬曉鵬 1，冶占福 1，宋濤 2
（ 1. 青海大學附屬醫院疼痛科 ，西寧 810001 ；2.  中國醫科大學附屬一院疼痛科 ，沈陽  110001）

摘要    目的   探討不同濃度臭氧對大鼠離體腦片氧糖剝奪及再灌注損傷模型的保護作用 。
方法    制備SD大鼠腦片，室溫下保存在人工腦脊液（ACSF）中，根據實驗分組情況選擇性經過下列程序：恢復期60 min、平衡期30 min、氧糖剝奪（OGD）期30 min  及再灌注期 90 min 。對照組（CTRL）不經過OGD期；模型組不施加臭氧干預（OGD/R）；實驗組于再灌注期分別給予 10，20，30， 40，50 μg/mL的臭氧，并且根據ACSF中是否含有人血白蛋白（HSA）分成2 個亞組 。再灌注期結束后分別測定各組ACSF中谷氨  酸（Glu）和乳酸脫氫酶（LDH）的含量 。結果   OGD 30 min后給予90 min的再灌注，模型組與對照組比較Glu 和LDH 釋放增加  （P < 0.01）。 不同濃度的臭氧在不同程度上拮抗了 Glu 和 LDH 的釋放，其中 40 μg/mL 的臭氧效應最明顯，低濃度（10~30  μg/mL）或高濃度（50 μg/mL）臭氧組與OGD/R組比較效應不明顯（P > 0.05）；在含有HAS組與不含HAS組比較，給予臭氧（40 μ  g/mL）干預能明顯減少Glu 和LDH 的釋放（P < 0.05）。

結論   40 μg/mL臭氧氣體對大鼠離體腦缺血再灌注損傷模型中的神經細  胞具有保護作用，并且HSA能增強臭氧的保護作用。
關鍵詞   臭氧；腦缺血；氧糖剝奪

Neuro?protective Effect of Different Concentration of Ozone on Cultured Rat Brain Slices Underwent Oxygen?glucose Deprivation and Reperfusion
SHICun?wei1，JING Xiao?peng1，YE Zhan?fu1，SONG Tao2
（1. Department of Pain Management，The Affiliated Hospital，Qinghai University，Xining 810001，China；2. Department of Pain Management，The First Hospital，Chi? na Medical University，Shenyang110001，China）
Abstract   Objective   To evaluate the neuro?protective effect of different concentration of ozone on cultured rat brain slices which were  subjected     to oxygen?glucose deprivation and reperfusion. Methods   In this study，brain slices of SD rats were cultured and subjected to oxygen?glucose depri?  vation and reperfusion. Slices were maintained in ACSF for 1 hour at room temperature（Recovery period）and thenequilibrated for an additional pe?      riod of 30 minutes（Equilibration）. Afterwards，the phase of oxygen/glucose deprivation（OGD）was carried out for 30 minutes. After the OGD peri?  od，ischemic solution was replaced by 2 mL of fresh，oxygenated ACSF for an additional 90?minute period（Reperfusion）. Slices were incubated in ACSF for 120 minutes（control conditions，CTRL）or subjected to oxygen/glucose deprivation for 30 minutes followed by 90?minute immersion in normally oxygenated ACSF（OGD/R）. Increasing concentrations of ozone（10?50 μg/mL）were administrated to ACSF during the reperfusion. Ozone treated group were divided into two subgroups according to the absence or in presence of Human Serum Albumin HSA（150 μg/mL）during     the reperfusion phase. Neuronal damage was assessed quantitatively by measuring the amount of both glutamate（Glu）and lactate dehydrogenase （LDH）released into the ACSF during 90?minutes?reperfusion period. Results   As expected，30 minutes of oxygen?glucose deprivation followed by 90 minutes of reperfusion（OGD/R）caused a highly significant release in LDH and Glu compared to the CTRL（P < 0.001）. Ozone（10?50  μg/mL）antagonized OGD/R?induced LDH and Glu release in different levels. In particular，40 μg/ml proved to be the most effective concentration compared with those of CTRL. While lower（10?30 μg/mL）or higher（50 μg/mL）ozone concentrations had not statistically significant effects.  The difference about the effects on LDH and Glu release was significance when comparing the groups with and without HAS（P < 0.01）. Conclusion Oxygen?ozone gaseous mixture in which the ozone ’sconcentration is 40 μg/mL appeared to be effective in reverting damage of rat brain slices which were subjected to oxygen?glucose deprivation；moreover，the presence of HSA in ACSF improved the ozone ’seffectiveness.
Key words    ozone；brain ischemia；oxygen and glucose deprivation


基金項目：國家自然科學基金（81271371）
作者簡介：師存偉（1972 -），男，副主任醫師，本科.
通信作者：宋濤，E-mail：songt2001@hotmail.com
收稿日期：2014-07-02 網絡出版時間：

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