天冬氨酸-谷氨酸载体1调节脑室周围白质软化早产鼠的髓鞘形成  

Aspartate-glutamate carrier 1 regulates myelination in preterm rats with periventricular leukomalacia

作　　者：王丽珍 羊才进 陈蓉[2] 李玲 Wang Lizhen;Yang Caijin;Chen Rong;Li Ling(Department of Pediatrics,Hainan Western Central Hospital,Danzhou 571700,Hainan Province,China;Department of Neurology,The First Affiliated Hospital of Hainan Medical University,Haikou 570102,Hainan Province,China)

机构地区：[1]海南西部中心医院儿科,海南省儋州市571700 [2]海南医学院第一附属医院神经内科,海南省海口市570102

出　　处：《中国组织工程研究》2023年第33期5263-5269,共7页Chinese Journal of Tissue Engineering Research

基　　金：2017年海南自然科学基金项目(817325),项目负责人:王丽珍。

摘　　要：背景:脑室周围白质软化是早产儿最严重的医疗并发症之一,目前尚无有效治疗措施。目的:探讨天冬氨酸-谷氨酸载体1(aspartate-glutamate carrier 1,AGC1)对脑室周围白质软化早产鼠髓鞘形成的调节作用。方法:60只出生后第2天雄性SD幼鼠随机分为假手术组和脑室周围白质软化组(n=30)。体外分离5只2日龄SD大鼠室周白质组织,制备白质祖细胞,建立实验性缺氧葡萄糖剥夺模型。分别在建立脑室周围白质软化、缺氧葡萄糖剥夺模型前,采用AGC1质粒(pcDNA3-AGC1)处理大鼠或细胞。在造模后不同时间点通过实时荧光定量PCR检测室周白质组织和细胞中AGC1表达。在脑室周围白质软化后14 d,采用免疫荧光法检测髓鞘碱性蛋白、别藻蓝蛋白表达,电子显微图像观察放射冠和胼胝体的超微结构。结果与结论:(1)在脑室周围白质软化后14 d,与假手术组相比,脑室周围白质软化组大鼠白质中髓鞘碱性蛋白表达、别藻蓝蛋白阳性少突胶质细胞数量以及有髓轴突数量显著减少(P<0.01);(2)与0 h相比,体内脑室周围白质软化开始后12-24 h时AGC1 mRNA表达显著升高(P<0.05),而在72 h-14 d时显著降低(P<0.05);(3)在体外对照条件下,与对照组相比,在缺氧葡萄糖剥夺后24-48 h时AGC1 mRNA表达显著升高(P<0.05),而在缺氧葡萄糖剥夺后7-14 d时AGC1 mRNA表达以及髓鞘碱性蛋白^(+)/AGC1^(+)少突胶质细胞形成显著降低(P<0.05);(4)pcDNA3-AGC1处理组在脑室周围白质软化后14 d处髓鞘碱性蛋白染色、别藻蓝蛋白阳性少突胶质细胞数量和有髓轴突数量比pcDNA3-NC对照组显著增多(P<0.05);(5)在体外实验中,增强AGC1表达进一步促进了缺氧葡萄糖剥夺后72h、7 d和14 d时少突胶质细胞前体细胞分化为髓鞘碱性蛋白^(+)/AGC1^(+)少突胶质细胞(P<0.05);提示脑室周围白质软化早产鼠模型和体外缺氧葡萄糖剥夺细胞模型中AGC1表达下调,通过上调AGC1可促进少突胶质细胞分BACKGROUND:Pe riventricular leukomalacia is one of the most serious medical complications in preterm infants and there is yet no effective treatment for periventricular leukomalacia.OBJECTIVE:To investigate the effects of aspartate-glutamate carrier 1(AGC1)on myelination in preterm rats with periventricular leukomalacia.METHODS:Sixty male Sprague-Dawley rat pups on postnatal day 2 were randomly divided into sham group(n=30)and periventricular leukomalacia group(n=30).For in vitro studies,periventricular white matter tissues were isolated from five rat pups on postnatal day 2 to prepare white matter progenitor cells and establish an expe rimental oxygen-glucose deprivation model.Befo re the establishment of periventricular leukomalacia and oxygen-glucose deprivation models,pcDNA3-AGC1 plasmids were used to treat rats or cells.RT-qPCR was used to detect the expression of AGC1 in periventricular white matter tissue and cells at different time points after modeling.The expression of myelin basic protein and allophycocyanin was detected by immunofluorescence,and the ultrastructure of the corona radiate and corpus callosum was observed by electron microscopy at 14 days after periventricular leukomala cia.RESULTS AND CONCLUSION:(1)Compared with the sham group,the expression of myelin basic protein and the number of allophycocyanin-positive oligodendrocytes and myelinated axons were significantly decreased in the periventricular leukomalacia group at 14 days after periventricular leukomalacia(P<0.01).(2)AGC1 mRNA expression began to significantly increase in the white matter tissue at 12-24 hours(P<0.05)and decreased significantly at 72 hours-14days(P<0.05)after periventricular leukomalacia in vivo,compared with 0 hour.(3)Under control conditions in vitro,AGC1 mRNA expression was increased significantly at 24-48 hours after oxygen-glucose deprivation(P<0.05),and the AGC1 mRNA expression and the formation of myelin basic protein^(+)/AGC1^(+)oligodendrocytes decreased significantly at 7-14 days after oxygen-glucose depriva

关 键 词：天冬氨酸-谷氨酸载体1 脑室周围白质软化 早产 少突胶质细胞 髓鞘 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R722