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作 者:钱俊超 周臣 QIAN Junchao;ZHOU Chen(School of Medicine,Anhui University of Science and Technology,Huainan Anhui 232001,China;Institute of Health and Medical Technology,Chinese Academy of Sciences,Hefei Anhui 230031,China;Hefei Cancer Hospital,Chinese Academy of Sciences,Hefei Anhui 230031,China)
机构地区:[1]安徽理工大学医学院,安徽淮南232001 [2]中国科学院合肥物质科学研究院,健康与医学技术研究所,安徽合肥230031 [3]中国科学院合肥肿瘤医院,安徽合肥230031
出 处:《安徽理工大学学报(自然科学版)》2024年第1期96-102,共7页Journal of Anhui University of Science and Technology:Natural Science
基 金:国家自然科学基金资助项目(U1932158,81871085,82271519);山东省自然科学基金肿瘤防治联合基金重点项目(ZR2019LZL018);安徽省自然科学基金杰出青年项目(2208085J10);中国博士后科学基金面上项目(2019M652403);山东省博士后创新项目(202002048)。
摘 要:目的检测SD大鼠脊髓损伤后急性期不同脑区内谷氨酰胺合成酶(GS)的活性和蛋白表达变化,并探讨可能的机制。方法取雄性6~8周龄SD大鼠32只,随机分为对照组(n=8)和实验组(n=24)。对大鼠施行右侧脊髓半横断术建立脊髓损伤模型。随后分别在脊髓损伤前和损伤后1、3、7d处死两组大鼠,取大脑并显微分割后做组织匀浆。使用GS活性检测试剂盒和蛋白印迹测定相关脑区的GS活性和表达以及胶质纤维酸性蛋白(GFAP)的表达水平。结果脊髓一侧损伤后,两侧丘脑的GS活性在1d后明显降低,随后持续增高至损伤后7d。GS蛋白表达变化与活性变化趋势类似,但在损伤后1d无显著改变;对侧下丘脑的GS活性和蛋白表达在损伤后1d略有下降,随后逐渐上升并在损伤后7d较1d显著升高。相较于未损伤对照组,脊髓损伤后GFAP表达仅在对侧下丘脑有显著变化。在小脑、初级运动皮层和同侧下丘脑均未检测到明显的GS和GFAP改变。结论脊髓损伤会导致大脑谷氨酰胺合成酶的活性和表达的区域特异性改变,这可能与星形胶质细胞有关。Objective To investigate the longitudinal changes in glutamine synthetase(GS)activity and protein expression in different brain regions of a rat spinal cord hemisection injury(SCI)model during acute phases and interpret the underlying mechanisms.Methods Thirty-two male Sprague-Dawley rats(6~8 weeks of age)were randomly divided into the control group(n=8)and experimental group(n=24).A SCI model was established by right-side spinal cord hemisecthon.Subsequently,the rats were euthanized at different times(day1,day3 and day7 post-injury),and the brains were collected and dissected.GS activity and protein levels,as well as glial fibrillary acidic protein(GFAP)level,in relevant brain regions were determined by using the GS Activity Assay Kit and western blot analysis.Results With the spinal cord hemisection injury,GS activity in the thalamus on both sides decreased significantly after 1day and then increased continually until day7 post-injury.Changes trend both in GS protein expression and activity was similar to each other,but the protein expression did not change significantly at day1 post-injury.In the contralateral hypothalamus,GS activity and protein expression slightly decreased at day1 post-injury,and then restored gradually and increased significantly at day7 post-injury than that at day1.GFAP protein expression was significantly changed only in the contralateral hypothalamus following SCI.Finally,no significant GS and GFAP changes were detected in the cerebellum,primary motor cortex and ipsilateral hypothalamus.Conclusion Overall,these findings elucidate the region-specific changes in GS activity and protein expression in the brain after SCI,which may be associated with activated astrocytes.
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