常压高浓度氧治疗对脑缺血-再灌注大鼠星形胶质细胞活性的影响

doi:10.3969/j.issn.1006-7795.2015.05.009

首都医科大学学报 ›› 2015, Vol. 36 ›› Issue (5): 709-713.doi: 10.3969/j.issn.1006-7795.2015.05.009

• 脑血管病、脑老化的基础及临床研究 • 上一篇下一篇

常压高浓度氧治疗对脑缺血-再灌注大鼠星形胶质细胞活性的影响

师文娟, 梁佳, 董雯, 戚智锋, 刘克建

首都医科大学宣武医院北京市老年病医疗研究中心脑血管病研究室, 北京 100053

收稿日期:2015-07-14 出版日期:2015-10-21 发布日期:2015-10-20
基金资助:
国家自然科学基金(81171242,81200928)资助。

Effects of normobaric hyperoxia on activation of astrocyte following cerebral ischemia-reperfusion in rats

Shi Wenjuan, Liang Jia, Dong Wen, Qi Zhifeng, Liu Kejian

Cerebrovascular Diseases Research Institute, Xuanwu Hospital, Capital Medical University, Beijing Geriatric Medical Research Center, Beijing 100053, China

Received:2015-07-14 Online:2015-10-21 Published:2015-10-20
Contact: 刘克建 E-mail:kliu@salud.unm.edu
Supported by:
This study was supported by National Natural Science Foundation of China (81171242, 81200928).

摘要/Abstract

摘要： 目的观察常压高浓度氧治疗(normobaric hyperoxia, NBO)对脑缺血-再灌注大鼠星形胶质细胞活化的标志物——胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)以及缝隙连接蛋白43(connexin 43, Cx43)表达的影响,初步探讨其作用机制。方法将健康成年雄性SD大鼠15只(280~320 g)采用数字表法随机分为假手术组(Sham)、正常氧浓度组(Normoxia)和NBO组,应用线栓法制备大鼠大脑中动脉阻塞模型,缺血1.5 h 再灌注24 h。Sham组和Normoxia组术后呼吸普通空气,NBO 组于缺血后5 min至再灌注前呼吸100%常压氧气。采用免疫荧光染色和Western blotting检测方法,研究NBO治疗对脑缺血皮质和基底节区星形胶质细胞GFAP和Cx43表达的影响。结果免疫荧光结果显示:与Normoxia组相比,NBO组缺血侧皮质GFAP阳性细胞数目明显减少,细胞突起减少、荧光强度减弱。Western blotting法分析结果显示:与Normoxia组相比,NBO组皮质和基底节区GFAP水平均有所降低,其中皮质著降低(P<0.05),表达变化与免疫荧光结果一致;与Sham组相比,Normoxia组缺血侧皮质和基底节区Cx43水平均有所降低,其中缺血侧皮质Cx43表达显著减少(P<0.01);与Normoxia组相比,NBO组缺血皮质Cx43蛋白显著升高(P<0.01),NBO组缺血基底节区Cx43蛋白水平差异无统计学意义。结论 NBO治疗可能通过调节缺血侧皮质缝隙连接蛋白Cx43的表达水平来抑制脑缺血星形胶质细胞过度活化,从而实现脑神经保护作用。

关键词: 脑缺血, 常压高浓度氧治疗, 星形胶质细胞, 皮质, 基底节区

Abstract: Objective To observe the effect of normobaric hyperoxia (NBO) on GFAP (the activation marker of astrocyte) and connexin43 expression induced by cerebral ischemia-reperfusion injury in rats, and preliminarily explore the effect of NBO treatment on activation of astrocytes. Methods A total of 15 healthy adult male Sprague-Dawley (SD) rats (280-320 g) were randomly divided into Sham group (n=3), normoxia group (n=6) or NBO group (n=6). A model of middle cerebral artery occlusion for 1.5 h and reperfusion for 24 h was induced by using the intraluminal suture method. The sham group and the normoxia group rats breathed normal air, and instead NBO group rats were exposed to 100% oxygen after ischemia from 5 minutes until reperfusion. Immunofluorescence staining and Western blot were used to observe the expression of glial fibrillary acidic protein and connexin 43 in basal ganglia and cortex after cerebral ischemia.Results Immunofluorescence staining results showed that compared with the normoxia group, the number of GFAP positive cells in the ischemic cortex of NBO group was significantly reduced with the processes and fluorescence intensity decreased. Western blot results showed that compared with the normoxia group, the expression of GFAP protein was decreased in NBO group, and GFAP expression in the ischemic cortex of NBO group was statistically significantly reduced (P<0.05), and it was consistent with the immunofluorescence staining results. Compared with the sham group, the Cx43 protein expression was decreased in the normoxia group and that in the ischemic cortex of the normoxia group was statistically significantly reduced (P<0.01). Compared with the normoxia group, the Cx43 protein expression was increased in NBO group and that in the ischemic cortex of NBO group was statistically significantly increased (P<0.01). The Cx43 protein expression in basal ganglia did not change significantly. Conclusion NBO treatment decreased the excessive activation of astrocyte probably through increasing Cx43, therefore, NBO may play a protective role in the ischemia-reperfusion injured brain.

Key words: cerebral ischemia, normobaric hyperoxia, astrocyte, cortex, basal ganglia

中图分类号:

R743.31

师文娟, 梁佳, 董雯, 戚智锋, 刘克建. 常压高浓度氧治疗对脑缺血-再灌注大鼠星形胶质细胞活性的影响[J]. 首都医科大学学报, 2015, 36(5): 709-713.

Shi Wenjuan, Liang Jia, Dong Wen, Qi Zhifeng, Liu Kejian. Effects of normobaric hyperoxia on activation of astrocyte following cerebral ischemia-reperfusion in rats[J]. Journal of Capital Medical University, 2015, 36(5): 709-713.

参考文献

[1] Liu W, Hendren J, Qin X J, et al. Normobaric hyperoxia attenuates early blood-brain barrier disruption by inhibiting MMP-9-mediated occludin degradation in focal cerebral ischemia[J]. J Neurochem, 2009, 108(3):811-820.
[2] Yuan Z, Liu W, Liu B, et al. Normobaric hyperoxia delays and attenuates early nitric oxide production in focal cerebral ischemic rats[J]. Brain Res, 2010, 1352:248-254.
[3] Liang J, Qi Z, Liu W, et al. Normobaric hyperoxia slows blood-brain barrier damage and expands the therapeutic time window for tissue-type plasminogen activator treatment in cerebral ischemia[J]. Stroke, 2015, 46(5):1344-1351.
[4] Pekny M, Nilsson M. Astrocyte activation and reactive gliosis[J]. Glia, 2005, 50(4):427-434.
[5] Sofroniew M V, Vinters H V. Astrocytes: biology and pathology[J]. Acta Neuropathol, 2010, 119(1):7-35.
[6] Nakase T, Fushiki S, Naus C C. Astrocytic gap junctions composed of connexin 43 reduceapoptotic neuronal damage in cerebral ischemia[J]. Stroke, 2003, 34:1987-1993.
[7] Tajiri N, Dailey T, Metcalf C, et al. In vivo animal stroke models: a rationale for rodent and non-human primate models[J]. Transl Stroke Res,2013, 4(3):308-321.
[8] Alvarez J I, Katayama T, Prat A. Glial influence on the blood brain barrier[J]. Glia, 2013, 61(12):1939-1958.
[9] Fitch M T, Silver J. CNS injury, glial scars, and inflammation: inhibitory extracellular matrices and regeneration failure[J]. Exp Neurol, 2008, 209(2):294-301.
[10] 高志,赵海苹,罗玉敏,等. 缺血后适应抑制大鼠脑缺血-再灌注后星形胶质细胞的增生 [J]. 首都医科大学学报,2014, 35(3):310-314.
[11] 王振华, 刘云会,马腾,等. 大鼠局灶性脑缺血再灌注早期血脑屏障通透性变化的研究[J]. 中国医科大学学报,2012,41(12) :1073-1076.
[12] 夏傲, 刘向荣,罗玉敏,等. 低温对脑缺血后血-脑屏障完整性保护机制的研究进展[J]. 中国脑血管病杂志,2012,9(8) :445-448.
[13] 闫荣, 罗晓光,张尧,等. 星形胶质细胞条件培养液诱导骨髓基质细胞向神经元样细胞分化的神经营养机制探讨[J]. 中国医科大学学报,2013,42(8) :989-995.
[14] Tong X H, Gu Y C, Jiao H, et al. Inhibition of gap junctional intercellular communication protects astrocytes from hypoxia/reoxygenation injury[J]. Zhejiang Da Xue Xue Bao Yi Xue Ban, 2015, 44(1):67-73.
[15] Ben Achour S, Pont-Lezica L, Bechade C, et al. Isastrocyte calcium signaling relevant for synapticplasticity? [J]. Neuron Glia Biol, 2010, 2:1-9.
[16] Márquez-Rosado L, Solan J L, Dunn C A, et al.Connexin43 phosphorylation in brain, cardiac, endothelial and epithelial tissues[J]. Biochim Biophys Acta, 2012, 1818(8):1985-1992.

常压高浓度氧治疗对脑缺血-再灌注大鼠星形胶质细胞活性的影响

Effects of normobaric hyperoxia on activation of astrocyte following cerebral ischemia-reperfusion in rats

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