摘要
目的探究全身亚低温联合人脐带血间充质干细胞(hCMNCs)移植改善新生大鼠缺氧缺血性脑损伤(HIBD)后认知障碍和能量衰竭的机制。方法7日龄新生SD乳鼠330只。随机分为11组,每组30只。空白对照组(A组)、HIBD常温组(B组)、HIBD亚低温持续72 h组(C组)、HIBD亚低温24 h干细胞移植组(D组)、HIBD亚低温48 h干细胞移植组(E组)、HIBD亚低温72 h移植组(F组)、HIBD亚低温24 h后持续3 d移植干细胞(G组)、HIBD 24 h干细胞移植组(H组)、HIBD 48 h干细胞移植组(I组)、HIBD 72 h干细胞移植组(J组)、HIBD 24 h、48 h、72 h干细胞移植组(K组)。乳鼠生长的第10天、第21天和第35天对其进行称重;通过Morris水迷宫实验检测乳鼠学习和记忆能力,超微电镜观察分析乳鼠脑组织神经元细胞及突触超微结构的变化,苏木精-伊红(HE)染色观察脑组织病理学变化,酶联免疫吸附试验检测乳鼠脑组织炎症因子和干细胞分化标记蛋白含量,免疫荧光染色检测脐带CD24和CD29阳性表达率,荧光显微镜测量线粒体膜电位。结果A、B、C、D、E、F、G、H、I、J、K组乳鼠生长第10天、第21天和第35天体重比较,采用重复测量设计的方差分析,结果:①不同时间点乳鼠体重有差异(F=13.285,P=0.000),各组乳鼠第21天与第35天体重比较,差异有统计学意义(P<0.05);②各组乳鼠体重有差异(F=20.097,P=0.000),B组乳鼠体重较A组降低(P<0.05),C~K组乳鼠体重较B组升高(P<0.05),G、I组乳鼠体重较B、C、D、E、F、H、J、K组均升高(P<0.05),且G组与I组乳鼠体重比较,差异无统计学意义(P>0.05);③各组乳鼠体重变化趋势有差异(F=28.712,P=0.000)。C~F组3种反射(悬崖调转反射、阴性趋地性、步态反射)时间较A组延长(P<0.05),但较B组缩短(P<0.05);C组3种反射时间较A、D~J组延长(P<0.05);G~K组3种反射时间与A组比较,差异无统计学意义(P>0.05);C~F组3种反射时间较G~K组延长(P<0.05)。HE染色结果显示,与A组比较,
Objective To explore the mechanism of mild whole-body hypothermia combined with human umbilical cord blood-derived mesenchymal stem cell transplantation in ameliorating cognitive impairment and energy failure after hypoxic-ischemic brain damage(HIBD)in neonatal rats.Methods A total of 3307-day-old neonatal SD rats were randomly divided into 11 groups,including blank control group(A),HIBD+normal temperature group(B),HIBD+72-hour mild hypothermia group(C),HIBD+24-hour mild hypothermia+stem cell transplantation group(D),HIBD+48-hour mild hypothermia+stem cell transplantation group(E),HIBD+72-hour mild hypothermia+stem cell transplantation group(F),HIBD+24-hour mild hypothermia+3-day stem cell transplantation group(G),HIBD+24-hour stem cell transplantation group(H),HIBD+48-hour stem cell transplantation group(I),HIBD+72-hour stem cell transplantation group(J),and HIBD+24-hour,48-hour,72-hour stem cell transplantation group(K),with 30 animals in each group.The rats were weighed and compared in the 10th,21st and 35th days of the growth.The learning and memory abilities of rats were detected by Morris water maze test.The ultrastructural changes of neurons and synapses in rat brain tissues were observed and analyzed by electron microscopy.The histopathological changes in the rat brain tissues were observed with H&E staining.Serum levels of inflammatory factors and stem cell differentiation markers were detected by enzyme-linked immunosorbent assay.The positive expression rates of CD24 and CD29 in the umbilical cord were detected by immunofluorescence.The mitochondrial membrane potential was detected by a fluorescence microscope.Results The rats in all the groups were weighed and compared on the 10th,21st and 35th days of growth.The repeated measures ANOVA showed that the body weight of rats at different time points was different(F=13.285,P=0.000),and that the body weight of rats at 21 d and 35 d in each group was significantly different(P<0.05).The body weight of rats was also different among the groups(F=20.097,P=0.000)
作者
祝巍
王立君
沈文生
王思思
李颖
李雨静
王静竹
高燕
方聪
赵寒阳
孙秀红
孙智勇
Wei Zhu;Li-jun Wang;Wen-sheng Shen;Si-si Wang;Ying Li;Yu-jing Li;Jing-zhu Wang;Yan Gao;Cong Fang;Han-yang Zhao;Xiu-hong Sun;Zhi-yong Sun(Deepartment of Neonatology,Jilin Women and Children Health Hospital,Changchun,Jilin 130012,China;Department of Pediatric Respiratory Medicine,The First Hospital of Jilin University,Changchun,Jilin 130061,China;College of Translational Medicine,The First Hospital of Jilin University,Changchun,Jilin 130061,China;Department of Pediatrics,Medical College of Yanbian University,Yanji,Jilin 133002,China)
出处
《中国现代医学杂志》
CAS
北大核心
2022年第18期13-24,共12页
China Journal of Modern Medicine
基金
吉林省科技发展计划项目(No:20190304110YY)
吉林省卫生与健康技术创新项目(No:2020J125)。
关键词
亚低温
干细胞移植
脑缺血损伤
能量衰竭
乳鼠
mild hypothermia
stem cell transplantation
cerebral ischemia injury
energy failure
neonatal rats
