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ALS转基因小鼠脊髓中miRNA-132表达减少BDNF表达增加

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ALS转基因小鼠脊髓中miRNA-132表达减少BDNF表达增加
Decrease in expression of miRNA-132 and increase in expression of BDNF in the spinal cord of ALS transgenic mice
上传时间:2019/4/18 15:10:37    作者:孙丹,王淑坤,丁康,刘美逸,蒋欣,陈燕春,周风华|下载

孙丹1,王淑坤1,丁康1,刘美逸2,蒋欣3,陈燕春3,周风华1*
(1潍坊医学院病理学教研室,2.潍坊医学院生物科学与技术学院,3.潍坊医学院组织学与胚胎学教研室,潍坊,261053)

Sun Dan1, Wang Shukun1, Ding Kang1, Liu Meiyi2, Jiang Xin3, ChenYanchun3, Zhou Fenghua1*
(1 Department of Pathology, Weifang Medical University, 2College of Biological Science and Technology, Weifang Medical University, 3Department of Histology and Embryology, Weifang Medical University, Weifang 261053, China)
摘要:目的  研究miRNA-132和脑源性神经营养因子(brain derived neurotrophic factor,BDNF)在肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)转基因小鼠脊髓中的表达变化,探讨miRNA-132、BDNF在ALS发病中的作用。方法 取SOD1-G93A ALS 转基因鼠发病早期(95d)、中期(108d)和晚期(122d)脊髓组织,应用qRT-PCR及原位杂交(hybridization in situ, ISH)技术检测miRNA-132的表达及定位,应用qRT-PCR 及Western blot技术检测BDNF在mRNA及蛋白水平变化,免疫荧光技术检测BDNF 在脊髓中的表达及分布,以同窝野生型鼠作为对照。结果 与野生型鼠比较,miRNA-132在ALS转基因鼠脊髓组织表达下降,miRNA-132阳性信号主要定位脊髓前角细胞胞体;在ALS转基因鼠脊髓组织BDNF mRNA及蛋白水平均增高,BDNF免疫阳性细胞主要表达于脊髓前角神经元,表达信号明显增强。结论 miRNA-132、BDNF可能在ALS发病过程中发挥了重要作用。
关键词:肌萎缩侧索硬化症;转基因鼠;miRNA-132;脑源性神经营养因子
〔中图分类号〕R741             〔文献标识码〕A             DOI:10.16705/ j. cnki. 1004-1850. 2019. 01. 003
Abstract: Objective To investigate the expression of miRNA-132 and brain derived neurotrophic factor (BDNF) in the spinal cord of amyotrophic lateral sclerosis (ALS) transgenic mice, and to explore the role of miRNA-132 and BDNF in the pathogenesis of ALS. Methods The spinal cord tissues of SOD1-G93A ALS transgenic mice were detected at early stage (95d), middle stage (108d) and late stage (122d). The expression and localization of miRNA-132 were detected by qRT-PCR and in situ hybridization (ISH). The changes of BDNF in mRNA and protein levels were detected by qRT-PCR and Western-blot. In addition, the expression of BDNF in the anterior horn of spinal cord and its co-localization with neurons were detected by immunofluorescence technique, compared with the wild-type mice from the same cages as controls. Results Compared with wild-type mice, miRNA-132 expression decreased in spinal cord tissue of ALS transgenic mice and its positive signal mainly localized the anterior horn cells of spinal cord. While the mRNA and protein levels of BDNF increased in spinal cord tissue. BDNF positive cells were mainly located in the anterior horn of spinal cord and its positive expression signal was significantly enhanced. The anterior horn is co-expressed with β-tubulin III labeled neurons. Conclusion miRNA-132 and BDNF may play an important role in the pathogenesis of ALS.
Keywords:Amyotrophic lateral sclerosis; transgenic mice; miRNA-132; BDNF

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