缺氧-葡萄糖再灌注对星形胶质细胞间隙连接和葡萄糖摄取的影响

Xiang-yang Zhang, Cui Wang, W. Guo, Yan Chen
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摘要

目的探讨氧糖剥夺(OGD)和再灌注下星形胶质细胞间隙连接与葡萄糖摄取的关系。方法对新生SD大鼠1日龄的大脑皮层星形胶质细胞进行原代培养。用OGD法建立缺血细胞模型。实验分为对照组、OGD组和OGD+ CBX组。不同再灌注时间(0 h、12 h、24 h、48 h)后,流式细胞术检测2-NBDG对星形胶质细胞葡萄糖摄取的影响,免疫荧光法检测连接蛋白43(Cx43)间隙连接斑块。结果与对照组相比,OGD诱导的星形胶质细胞葡萄糖摄取随再灌注时间的增加而升高。再灌注24h时,OGD组葡萄糖摄食量为(2.32±0.43)nmol/μgDNA,对照组为(0.95±0.28)nmol/μgDNA。再灌注后24h,上调幅度高达2.63倍(t=13.99, P=0.0024)。与对照组相比,再灌注24h时Cx43缝隙连接数最多增加2.5倍(t=11.34, P=0.003),大小增加1.85倍(t=10.27, P=0.004)。CBX可使OGD后星形胶质细胞的葡萄糖摄取减少,再灌注后48 h减少42%。结论对干扰Cx43间隙连接的临床治疗值得考虑。关键词:星形胶质细胞;联接蛋白43;缝隙连接;Oxygen-glucose剥夺;葡萄糖的吸收
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Effects of oxygen glucose deprivation and reperfusion on gap junction and glucose uptake of astrocytes
Objective To explore the relationship between gap junction and glucose uptake of astrocytes under oxygen-glucose deprivation(OGD) and reperfusion. Methods Cerebral cortical astrocyte from 1 day newborn SD rats were undergone the primary culture. The ischemia cell model was established by OGD. This experiment were divided into control group, OGD group and OGD+ CBX group.After different reperfusion time (0 h, 12 h 24 h and 48 h), the glucose uptake of astrocyte was measured by 2-NBDG through flow cytometry analysis and connexin 43(Cx43) gap junction plaques was detected using immunofluorescene. Results Compared with the control group, the glucose uptake of astrocyte was up-regulated induced by OGD following different reperfusion time.The glucose uptake of OGD group was (2.32±0.43)nmol/μgDNA in 24 hours reperfusion time and was (0.95±0.28)nmol/μgDNA in control group. The up-regulation was up to 2.63-fold increase (t=13.99, P=0.0024) in 24 hours after reperfusion.Compared with the control group, the Cx43 gap junction number was up to 2.5- fold increase(t=11.34, P=0.003) and the size was 1.85-fold increase (t=10.27, P=0.004) in 24 h reperfusion. The glucose uptake of astrocyte after OGD was reduced by CBX and the decrease was 42% in 48 h after reperfusion. Conclusion Those results urges us consider the clinical treatment for interfering with Cx43 gap junction. Key words: Astrocyte; Connexin 43; Gap junction; Oxygen-glucose deprivation; Glucose uptake
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期刊介绍: "Chinese Journal of Behavioral Medicine and Brain Science" (CN 37-1468/R, ISSN 1674-6554) is a national academic journal under the supervision of the National Health Commission, sponsored by the Chinese Medical Association and Jining Medical College. The journal was founded in June 1992 and was formerly known as "Chinese Journal of Behavioral Medicine" (1992-1993) and "Chinese Behavioral Medical Science" (1994-2008). In 2009, it was renamed "Chinese Journal of Behavioral Medicine and Brain Science" with the approval of the State Administration of Press, Publication, Radio, Film and Television. The purpose of "Chinese Journal of Behavioral Medicine and Brain Science" is to implement the health and health policies of the Party and the State, implement the principle of combining theory with practice and popularization and improvement, and reflect the major progress in the theory and practical application of behavioral medicine and brain science in my country. It publishes academic papers and scientific research results in the field of behavioral medicine and brain science in my country, and has columns such as monographs/reviews, basic research, clinical research, health prevention, methods and techniques, psychological behavior and evaluation, and systematic evaluation.
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