缺氧/复氧条件下体外培养星形胶质细胞神经生长因子分泌的变化  

Production of Nerve Growth Factor in Primary Cultured Astrocytes under Hypoxia/Reoxygenation

作　　者：肖君[1] 张函[1] 谢敏杰[1] 王伟[1] 何丹[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030

出　　处：《神经损伤与功能重建》2015年第1期1-4,共4页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金(No.81301126;81030021);中国博士后科学基金(No.2013M531699)

摘　　要：目的:观察缺氧/复氧(H/R)条件下体外培养星形胶质细胞的活力变化及神经营养因子(NGF)的合成和分泌的变化。方法:采用原代培养的新生大鼠大脑皮质星形胶质细胞,分为正常(N)组及H/R组。在H/R不同时间点,应用倒置显微镜进行形态学观察,并结合台盼蓝染色,明确缺氧不同时间对星形胶质细胞形态和活力的影响。在H/R不同时间后,应用ELISA检测星形胶质细胞培养上清液中NGF蛋白的含量,应用q RT-PCR检测NGF m RNA的表达。结果:缺氧6 h、复氧48 h内,H/R组的星形胶质细胞的细胞活力未发生明显改变。在缺氧6 h、复氧12 h以及24 h后,H/R组的NGF m RNA表达量及蛋白的分泌较N组显著增加(P<0.01)。结论:H/R条件可促进体外培养星形胶质细胞NGF的分泌及合成量的增加。Objective： To investigate the secretion of nerve growth factor（NGF） in primary cultured astrocytes under hypoxia/reoxygenation（H/R）. Methods： Primary cultured rats cortical astrocytes were used, which were randomly assigned into groups of normal control（N group） and hypoxia/reoxygenation（H/R group）. Morphological observation of the astrocytes was detected by inverted phase contrast microscopy as an index of cellular damage under hypoxia. And tyrpan blue staining was applied to detect the cell injury rate at different time after reoxygenation. The levels of NGF protein were analyzed by enzyme-linked immune sorbent assay（ELISA）. And q RT-PCR was used to determine the levels of NGF m RNA. Results： The astrocytes viability of the H/R group has no significant changed after 6 h hypoxia and 48 h reoxygenation. Compared with the N group, the NGF m RNA level of the H/R group was remarkably increased after 24 h reoxygenation and 6 h hypoxia（P〈0.01）, and the level of NGF protein secretionof the H/R group increased significantly as well after 48 h reoxygenation and 6 h hypoxia（P〈0.01）.Conclusion： Hypoxia/reoxygenation effectively increased the expression of NGF in primary cultured astrocytes.

关 键 词：星形胶质细胞 神经生长因子 缺氧/复氧 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学]