EPO对窒息诱导新生大鼠脑损伤和CoCl_(2)诱导人脑胶质瘤细胞系U251缺氧的改善作用  

Effects of EPO on brain injury after neonatal asphyxia in rats and hypoxia induced by CoCl_(2) in human glioma cell line U251

作　　者：呼格吉乐[1] 王婷[2] 苏慧敏[2] HU Gejile;WANG Ting;SU Huimin(Department of Laboratory Medicine,Affiliated Hospital of Inner Mengolia Medical University,Hohhot 010050,China;不详)

机构地区：[1]内蒙古医科大学附属医院检验科,呼和浩特010050 [2]内蒙古大学生命科学学院

出　　处：《山东医药》2023年第9期46-51,共6页Shandong Medical Journal

基　　金：内蒙古自治区自然科学基金项目(2022MS08004);内蒙古医科大学附属医院博士启动金(2022NYFYBS004)。

摘　　要：目的观察促红细胞生成素(EPO)对窒息诱导新生大鼠脑损伤和CoCl_(2)诱导人脑胶质瘤细胞系U251缺氧的改善作用。方法①7日龄SPF级新生Wistar大鼠随机分为对照组、窒息组、EPO组,每组22只。窒息组和EPO组新生大鼠采用常压窒息法构建脑损伤模型。EPO组脑损伤模型大鼠腹腔注射5000 IU/(kg•d)的EPO,连续注射14 d;窒息组脑损伤模型大鼠注射等体积的生理盐水,连续注射14 d;对照组不注射生理盐水和EPO。每组取10只大鼠,采用Morris水迷宫实验观察各组新生大鼠空间记忆能力,包括逃避潜伏期、上台前游泳路程、90 s内穿越原平台次数、平台内运动距离、平台滞留时间、穿越有效区域次数、有效区域内运动距离和有效区域内滞留时间。采用旷场实验观察各组新生大鼠自主运动能力,包括运动总路程、中央区域路程和平均速度。每组取6只大鼠,连续干预第3天和第7天时断头处死,取大鼠大脑称重并计算脑组织含水量,采用速率法检测血清脑损伤标志物乳酸脱氢酶(LDH)。②人脑胶质瘤细胞系U251分为空白对照组、CoCl_(2)组、EPO组,CoCl_(2)组加入CoCl_(2)制备缺氧模型,EPO组加入CoCl_(2)和EPO,空白对照组加入等量培养液,继续培养24 h。采用CCK-8法测算各组细胞存活率,采用流式细胞术测算各组细胞凋亡率,采用比色法测算各组细胞上清液LDH。结果①与对照组比较,窒息组新生大鼠的逃避潜伏期、上台前游泳路程、干预第3天和第7天脑组织含水量及血清LDH水平显著升高(P均<0.05);与窒息组相比,EPO组新生大鼠的逃避潜伏期、上台前游泳路程、干预第3天和第7天脑组织含水量及血清LDH水平显著降低(P均<0.05)。与对照组比较,窒息组新生大鼠90 s内穿越原平台次数、平台内运动距离、平台滞留时间、穿越有效区域次数、有效区域内运动距离、有效区域内滞留时间、运动总路程、中央区域路程、平�Objective To observe the effects of erythropoietin(EPO)on asphyxia-induced brain injury in neonatal rats and hypoxia induced by CoCl_(2) in human glioma cell line U251.Methods①7-day-old SPF newborn Wistar rats were randomly divided into the control group,asphyxia group,and EPO group,with 22 rats in each group.Neonatal rats in the asphyxia group and EPO group were subjected to normal pressure asphyxia to establish brain injury models.Rats in the EPO group were injected intraperitoneally with 5000 IU/(kg·d)EPO for 14 days,while rats in the asphyxia group were injected with the same volume of normal saline for 14 days.Rats in the control group were not injected with normal saline and EPO.Morris water maze test was used to observe the spatial memory ability of 10 rats in each group;it included escape latency,swimming distance before going on stage,the number of crossing the original platform in 90s,the movement distance in the platform,the residence time in the platform,the number of crossing the effective region,the movement distance in the effective region and the residence time in the effective region.The open field experiment was used to observe the autonomous movement ability of the newborn rats in each group,including the total distance,the distance in the central region and the average speed.Six rats in each group were decapitated and killed on the 3rd and 7th days after intervention.The rats'brains were weighed and the water content of brain tissue was calculated.The lactate dehydrogenase(LDH)of brain injury markers in serum were detected by velocity method.②Human glioma cell line U251 was divided into the blank control group,CoCl_(2) group and EPO group.Hypoxia models were made by adding CoCl_(2) in the CoCl_(2) group and EPO in the EPO group,and the same amount of culture medium was added in blank control group for 24 h.Cell survival rate was measured by CCK-8,apoptosis rate was measured by flow cytometry,and LDH in supernatant was measured by colorimetric method.Results①Compared with the control gro

关 键 词：促红细胞生成素 颅脑损伤 缺氧性脑损伤 新生儿脑损伤 星形胶质细胞 乳酸脱氢酶 

分 类 号：R563[医药卫生—呼吸系统]