基质细胞衍生因子-1及CXC家族趋化因子-4在人脐带间充质干细胞干预新生大鼠脑白质损伤中的作用
기질세포연생인자-1급CXC가족추화인자-4재인제대간충질간세포간예신생대서뇌백질손상중적작용
Effects of stromal derived factor-1 and CXC chemokine receptor-4 in rats with white matter damage treated with human umbilical cord mesenchymal stem cells
  • 万方数据
    中华围产医学杂志 중화위산의학잡지
    CHINESE JOURNAL OF PERINATAL MEDICINE
    2014年 5期 329-336 ,共8页

    王诗雨%朱丽华%摆翔%蒋犁%李伟

    왕시우%주려화%파상%장리%리위

    脑白质病%趋化因子CXCL12%受体,CXCR4%间质干细胞%脐带%动物,新生 腦白質病%趨化因子CXCL12%受體,CXCR4%間質榦細胞%臍帶%動物,新生
    뇌백질병%추화인자CXCL12%수체,CXCR4%간질간세포%제대%동물,신생
    Leukoencephalopathies%Chemokine CXCL12%Receptors,CXCR4%Mesenchymal stem cells%Umbilical cord%Animals,newborn
    目的 探讨基质细胞衍生因子-l(stromal derived factor-1,SDF-1)及其受体——CXC家族趋化因子-4(CXC ehemokine receptor-4,CXCR-4)在人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)定向迁移治疗早产儿脑白质损伤的可能机制. 方法 108只3日龄清洁级Sprague-Dawley大鼠随机分为实验组、对照组和假手术组.实验组和对照组大鼠结扎左侧颈总动脉并吸人氧浓度为6％的氮氧混合气体4h,假手术组仅游离左侧颈总动脉.实验组大鼠缺氧处理后24 h腹腔注射0.5 mlhUC-MSCs(1×106个/ml).HE染色观察大鼠脑组织病理改变,分别采用免疫组织化学方法和实时荧光定量逆转录-聚合酶链反应技术检测各组大鼠5、7和14日龄时(各组各时间点6只大鼠)脑白质中SDF-1和CXCR-4蛋白及mRNA的表达情况.采用单因素方差分析及LSD检验进行统计学分析. 结果 HE染色显示,14日龄时实验组大鼠双侧脑室大小相似,未见明显细胞水肿、变性和坏死.假手术组大鼠双侧脑室形态大小均正常.对照组大鼠左侧脑室较对侧明显增大,脑室形状不规则,可见部分神经细胞变性、坏死.5、7和14日龄时,实验组大鼠脑组织SDF-1蛋白表达水平为0.15±0.06、0.24±0.01和0.12±0.01,CXCR-4蛋白表达水平为0.35±0 16、0.60±0.21和0.72±0.25;对照组大鼠脑组织SDF-1蛋白表达水平为0.13±0.01、0.16±0.01和0.08±0.01,CXCR-4蛋白表达水平为0.18±0.04、0.17±0.09和0.25±0.06;均高于假手术组(SDF-1蛋白表达水平为0.03±0.01、0.04±0.01和0.02±0.01,CXCR-4蛋白表达水平为0.04±0.02、0.05±0.03和0.05±0.03)(LSD检验,P值均＜0.05).实验组SDF-1蛋白在7日龄时表达达高峰,14日龄时下降,在7和14日龄时均高于对照组(LSD检验,P值均＜0.05).CXCR-4蛋白在5日龄即高于对照组,后逐渐升高,在7和14日龄表达高于5日龄,3个时间点表达水平均高于对照组(LSD检验,P值均＜0.05).5、7和14日龄时,实验组人鼠脑组织SDF-1 mRNA的表达水平分别为3.52±0.33、4.18±0.28和2.60±0.21,对照组分别为2.07±0.34、3.73±0.28和2.08±0.15,均高于假手术组(分别为0.99±0.17、1.00±0.16和1.31±0.32),实验组高于对照组,差异均有统计学意义(LSD检验,P值均＜0.05).实验组大鼠7日龄时SDF-1 mRNA表达达高峰,14日龄时下降至低于5日龄的水平(LSD检验,P值均＜0.05).对照组大鼠SDF-1 mRNA表达水平也在7日龄达高峰(LSD检验,P＜0.05),14日龄时下降至5日龄水平(LSD检验,P＞0.05).5、7和14日龄时,实验组大鼠脑组织CXCR 4 mRNA的表达水平分别为1.32±0.29、1.75±0.36和2.33±0.49,均高于假手术组(分别为1.00±0.16、0.94±0.16和0.81±0.14)和对照组(分别为0.97±0.14、0.97±0.15和1.07±0.25)(LSD检验,P值均＜0.05),实验组大鼠CXCR-4 mRNA表达水平在14日龄时达高峰,高于5和7日龄时(LSD检验,P值均＜0.05). 结论 SDF-1/CXCR-4表达改变可能在hUC-MSCs向早产儿脑白质损伤部位迁移过程中具有一定作用.
    목적 탐토기질세포연생인자-l(stromal derived factor-1,SDF-1)급기수체——CXC가족추화인자-4(CXC ehemokine receptor-4,CXCR-4)재인제대간충질간세포(human umbilical cord mesenchymal stem cells,hUC-MSCs)정향천이치료조산인뇌백질손상적가능궤제. 방법 108지3일령청길급Sprague-Dawley대서수궤분위실험조、대조조화가수술조.실험조화대조조대서결찰좌측경총동맥병흡인양농도위6％적담양혼합기체4h,가수술조부유리좌측경총동맥.실험조대서결양처리후24 h복강주사0.5 mlhUC-MSCs(1×106개/ml).HE염색관찰대서뇌조직병리개변,분별채용면역조직화학방법화실시형광정량역전록-취합매련반응기술검측각조대서5、7화14일령시(각조각시간점6지대서)뇌백질중SDF-1화CXCR-4단백급mRNA적표체정황.채용단인소방차분석급LSD검험진행통계학분석. 결과 HE염색현시,14일령시실험조대서쌍측뇌실대소상사,미견명현세포수종、변성화배사.가수술조대서쌍측뇌실형태대소균정상.대조조대서좌측뇌실교대측명현증대,뇌실형상불규칙,가견부분신경세포변성、배사.5、7화14일령시,실험조대서뇌조직SDF-1단백표체수평위0.15±0.06、0.24±0.01화0.12±0.01,CXCR-4단백표체수평위0.35±0 16、0.60±0.21화0.72±0.25;대조조대서뇌조직SDF-1단백표체수평위0.13±0.01、0.16±0.01화0.08±0.01,CXCR-4단백표체수평위0.18±0.04、0.17±0.09화0.25±0.06;균고우가수술조(SDF-1단백표체수평위0.03±0.01、0.04±0.01화0.02±0.01,CXCR-4단백표체수평위0.04±0.02、0.05±0.03화0.05±0.03)(LSD검험,P치균＜0.05).실험조SDF-1단백재7일령시표체체고봉,14일령시하강,재7화14일령시균고우대조조(LSD검험,P치균＜0.05).CXCR-4단백재5일령즉고우대조조,후축점승고,재7화14일령표체고우5일령,3개시간점표체수평균고우대조조(LSD검험,P치균＜0.05).5、7화14일령시,실험조인서뇌조직SDF-1 mRNA적표체수평분별위3.52±0.33、4.18±0.28화2.60±0.21,대조조분별위2.07±0.34、3.73±0.28화2.08±0.15,균고우가수술조(분별위0.99±0.17、1.00±0.16화1.31±0.32),실험조고우대조조,차이균유통계학의의(LSD검험,P치균＜0.05).실험조대서7일령시SDF-1 mRNA표체체고봉,14일령시하강지저우5일령적수평(LSD검험,P치균＜0.05).대조조대서SDF-1 mRNA표체수평야재7일령체고봉(LSD검험,P＜0.05),14일령시하강지5일령수평(LSD검험,P＞0.05).5、7화14일령시,실험조대서뇌조직CXCR 4 mRNA적표체수평분별위1.32±0.29、1.75±0.36화2.33±0.49,균고우가수술조(분별위1.00±0.16、0.94±0.16화0.81±0.14)화대조조(분별위0.97±0.14、0.97±0.15화1.07±0.25)(LSD검험,P치균＜0.05),실험조대서CXCR-4 mRNA표체수평재14일령시체고봉,고우5화7일령시(LSD검험,P치균＜0.05). 결론 SDF-1/CXCR-4표체개변가능재hUC-MSCs향조산인뇌백질손상부위천이과정중구유일정작용.
    Objective To investigate the effects and the migration mechanisms of stromal derived factor-1 (SDF-1) and CXC chemokine receptor-4 (CXCR-4) in rats with white matter damage treated with human umbilical cord mesenchymal stem cells (hUC-MSCs).Methods A total of 108 three-day old Sprague-Dawley rats were randomly divided into the experimental group,control group and sham group.The left common carotid artery was ligated and then exposed to hypoxia of 6％ O2 and 94％ N2 in rats in the experimental and control groups.Rats in sham group were neither ligation nor hypoxia.After 24 hours,rats in the experimental group were administered 0.5 ml hUC-MSCs (1 × 106/ml) intraperitoneally,and rats in control and sham groups were administered 0.5 ml saline by the same way.Immunohistochemistry and reverse transcription-polymerase chain reaction were used to determine the expression of SDF-1 and CXCR-4 protein and mRNA in 5-,7-and 14-day-old rats.Analysis of variance and the LSD test were used for statistical analysis of the data.Results HE staining showed that,in 14 day-old rats of the experimental group,bilateral cerebral ventricles were similar with no cellular edema or necrocytosis.In the sham group,bilateral cerebral ventricles were also normal.However,in the control group,ventriculomegaly,cellular degeneration and necrocytosis were observed on the left side.On the 5th,7th and 14th day,SDF-1 protein levels were 0.15±0.06,0.24±0.01 and 0.12±0.01,and CXCR-4 protein levels were 0.35±0.16,0.60±0.21 and 0.72±0.25,respectively,in the experimental group; SDF-1 protein levels were 0.13 ± 0.01,0.16± 0.01 and 0.08± 0.01,and CXCR-4 protein levels were 0.18 ± 0.04,0.17 ± 0.09 and 0.25 ± 0.06,respectively,in the control group,and all were higher than those in the sham group (SDF-1 protein levels were 0.03 ± 0.01,0.04± 0.01 and 0.02±0.01; and CXCR-4 protein levels were 0.04±0.02,0.05±0.03 and 0.05±0.03,respectively) (LSD test,all P＜0.05).SDF-1 protein increased to a peak on the 7th day and decreased on the 14th day in the experimental group,however,these values were both higher than those in the control group (LSD test,both P＜0.05).CXCR-4 protein increased on the 5th day and continued to increase up to the 14th day in the experimental group,and these values were higher than those in the control group at the three time points (LSD test,all P＜0.05).In 5-,7-and 14-day-old rats,SDF-1 mRNA levels were 3.52 ± 0.33,4.18± 0.28 and 2.60± 0.21,respectively,in the experimental group,which were higher than those in the control group (2.07± 0.34,3.73 ± 0.28 and 2.08± 0.15,respectively),and were even higher than those in the sham group (0.99±0.17,1.00±0.16 and 1.31 ±0.32,respectively) (LSD test,all P＜0.05).In the experimental group,SDF-1 mRNA levels reached a peak on the 7th day,and on the 14th day,it decreased to the level lower than that on the 5th day (LSD test,all P＜0.05).In the control group,SDF-1 mRNA levels also reached a peak in 7-day-old rats,but not in 14-day-old rats,which was similar to 5-day old rats (LSD test,9＞0.05).In 5-,7 and 14-day-old rats of the experimental group,CXCR-4 mRNA levels were 1.32±0.29,1.75±0.36 and 2.33±0.49,respectively,higher than those in the sham group (1.00±0.16,0.94±0.16 and 0.81±0.14,respectively) and the control group (0.97±0.14,0.97±0.15 and 1.07±0.25,respectively) (LSD test,all P＜0.05).In the experimental group,CXCR-4 mRNA levels were higher in 14-day-old rats than that in 5-and 7-day-old rats (LSD test,both P＜0.05).Conclusions SDF-1/CXCR-4 may play a vital role in the migration of hUC-MSCs homing to damaged brain.
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