目的 探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUcMSCs)移植对脑白质损伤(white matter damage,WMD)新生大鼠脑内少突胶质细胞及活化小胶质细胞的影响,以及新生大鼠远期神经行为学的改变. 方法 72只3日龄Sprague-Dawley大鼠行左侧颈总动脉结扎,并吸入6％O2和94％N2的混合气体4h,制成WMD模型.死亡12只,剩余60只随机分为实验组和对照组各30只.大鼠出生后0～24 h计为第1天,实验组大鼠于第3、4和5天每天腹腔注射HUcMSCs 1×106个(0.05 ml),对照组大鼠于同时间点腹腔注射磷酸盐缓冲液0.05 ml.2组均于第10和21天分别取8只大鼠,免疫组织化学方法检测左侧脑组织中髓鞘碱性蛋白(myelin basic protein,MBP)和单核巨噬细胞抗原(ectodermal dysplasia,ED)-1染色阳性细胞.2组均于第10和21天分别取3只大鼠,Western印迹技术检测MBP和ED-1蛋白表达.2组分别取8只大鼠测量体重,并于第28天行神经行为学评估.两样本均数比较采用t检验. 结果 实验组第10和21天MBP染色阳性细胞数[(11.8±4.1)和(23.8±8.1)个]均高于对照组[(6.7±3.1)和(11.5±5.8)个,t值分别为2.81和3.49,P值均＜0.05],ED-1染色阳性细胞数[(20.8±3.4)和(19.1±2.8)个]均低于对照组[(32.8±4.2)和(29.5±5.2)个,t值分别为6.23和4.94,P值均＜0.01].实验组第10和21天MBP的表达量(1.3±0.1和1.1±0.1)均高于对照组(0.8±0.0和0.6±0.1,t值分别为7.53和6.68,P值均＜0.01),ED-1的表达量(0.6±0.1和0.4±0.1)均低于对照组(1.0±0.1和0.8±0.1,t值分别为3.90和4.90,P值均＜0.01).实验组第7、10、14、21和28天体重分别为(15.0±1.2)、(16.6±0.9)、(27.0±1.6)、(44.2±2.3)和(68.1±4.2)g,均高于对照组[分别为(12.7±1.6)、(13.5±2.0)、(23.6±1.9)、(38.4±0.9)和(60.0±4.2)g,t值分别为-3.11、-3.97、-3.67、-6.52和-3.72,P值均＜0.05).第28天神经行为学测试:实验组旷场实验评分高于对照组[(36.5±2.9)与(24.3±3.6)分,t=7.36,P＜0.01],悬吊实验评分也高于对照组[(3.6±1.0)与(2.0±0.7)分,t=3.53,P＜0.01].实验组Cylinder实验左、右前足/双前足的比值[(49.8±13.3)％与(41.4±5.9)％]差异无统计学意义(t=0.86,P＞0.05);对照组左前足/双前足的比值高于右前足/双前足[(49.5±11.3)％与(31.2±3.2)％,t=4.38,P＜0.01].结论 HUcMSCs可提高新生大鼠WMD模型病变区域内少突胶质细胞的数量,减少小胶质细胞的活化,对体格发育有良好促进作用,并且对神经行为有改善作用.
目的 探討人臍帶間充質榦細胞(human umbilical cord mesenchymal stem cells,HUcMSCs)移植對腦白質損傷(white matter damage,WMD)新生大鼠腦內少突膠質細胞及活化小膠質細胞的影響,以及新生大鼠遠期神經行為學的改變. 方法 72隻3日齡Sprague-Dawley大鼠行左側頸總動脈結扎,併吸入6％O2和94％N2的混閤氣體4h,製成WMD模型.死亡12隻,剩餘60隻隨機分為實驗組和對照組各30隻.大鼠齣生後0～24 h計為第1天,實驗組大鼠于第3、4和5天每天腹腔註射HUcMSCs 1×106箇(0.05 ml),對照組大鼠于同時間點腹腔註射燐痠鹽緩遲液0.05 ml.2組均于第10和21天分彆取8隻大鼠,免疫組織化學方法檢測左側腦組織中髓鞘堿性蛋白(myelin basic protein,MBP)和單覈巨噬細胞抗原(ectodermal dysplasia,ED)-1染色暘性細胞.2組均于第10和21天分彆取3隻大鼠,Western印跡技術檢測MBP和ED-1蛋白錶達.2組分彆取8隻大鼠測量體重,併于第28天行神經行為學評估.兩樣本均數比較採用t檢驗. 結果 實驗組第10和21天MBP染色暘性細胞數[(11.8±4.1)和(23.8±8.1)箇]均高于對照組[(6.7±3.1)和(11.5±5.8)箇,t值分彆為2.81和3.49,P值均＜0.05],ED-1染色暘性細胞數[(20.8±3.4)和(19.1±2.8)箇]均低于對照組[(32.8±4.2)和(29.5±5.2)箇,t值分彆為6.23和4.94,P值均＜0.01].實驗組第10和21天MBP的錶達量(1.3±0.1和1.1±0.1)均高于對照組(0.8±0.0和0.6±0.1,t值分彆為7.53和6.68,P值均＜0.01),ED-1的錶達量(0.6±0.1和0.4±0.1)均低于對照組(1.0±0.1和0.8±0.1,t值分彆為3.90和4.90,P值均＜0.01).實驗組第7、10、14、21和28天體重分彆為(15.0±1.2)、(16.6±0.9)、(27.0±1.6)、(44.2±2.3)和(68.1±4.2)g,均高于對照組[分彆為(12.7±1.6)、(13.5±2.0)、(23.6±1.9)、(38.4±0.9)和(60.0±4.2)g,t值分彆為-3.11、-3.97、-3.67、-6.52和-3.72,P值均＜0.05).第28天神經行為學測試:實驗組曠場實驗評分高于對照組[(36.5±2.9)與(24.3±3.6)分,t=7.36,P＜0.01],懸弔實驗評分也高于對照組[(3.6±1.0)與(2.0±0.7)分,t=3.53,P＜0.01].實驗組Cylinder實驗左、右前足/雙前足的比值[(49.8±13.3)％與(41.4±5.9)％]差異無統計學意義(t=0.86,P＞0.05);對照組左前足/雙前足的比值高于右前足/雙前足[(49.5±11.3)％與(31.2±3.2)％,t=4.38,P＜0.01].結論 HUcMSCs可提高新生大鼠WMD模型病變區域內少突膠質細胞的數量,減少小膠質細胞的活化,對體格髮育有良好促進作用,併且對神經行為有改善作用.
목적 탐토인제대간충질간세포(human umbilical cord mesenchymal stem cells,HUcMSCs)이식대뇌백질손상(white matter damage,WMD)신생대서뇌내소돌효질세포급활화소효질세포적영향,이급신생대서원기신경행위학적개변. 방법 72지3일령Sprague-Dawley대서행좌측경총동맥결찰,병흡입6％O2화94％N2적혼합기체4h,제성WMD모형.사망12지,잉여60지수궤분위실험조화대조조각30지.대서출생후0～24 h계위제1천,실험조대서우제3、4화5천매천복강주사HUcMSCs 1×106개(0.05 ml),대조조대서우동시간점복강주사린산염완충액0.05 ml.2조균우제10화21천분별취8지대서,면역조직화학방법검측좌측뇌조직중수초감성단백(myelin basic protein,MBP)화단핵거서세포항원(ectodermal dysplasia,ED)-1염색양성세포.2조균우제10화21천분별취3지대서,Western인적기술검측MBP화ED-1단백표체.2조분별취8지대서측량체중,병우제28천행신경행위학평고.량양본균수비교채용t검험. 결과 실험조제10화21천MBP염색양성세포수[(11.8±4.1)화(23.8±8.1)개]균고우대조조[(6.7±3.1)화(11.5±5.8)개,t치분별위2.81화3.49,P치균＜0.05],ED-1염색양성세포수[(20.8±3.4)화(19.1±2.8)개]균저우대조조[(32.8±4.2)화(29.5±5.2)개,t치분별위6.23화4.94,P치균＜0.01].실험조제10화21천MBP적표체량(1.3±0.1화1.1±0.1)균고우대조조(0.8±0.0화0.6±0.1,t치분별위7.53화6.68,P치균＜0.01),ED-1적표체량(0.6±0.1화0.4±0.1)균저우대조조(1.0±0.1화0.8±0.1,t치분별위3.90화4.90,P치균＜0.01).실험조제7、10、14、21화28천체중분별위(15.0±1.2)、(16.6±0.9)、(27.0±1.6)、(44.2±2.3)화(68.1±4.2)g,균고우대조조[분별위(12.7±1.6)、(13.5±2.0)、(23.6±1.9)、(38.4±0.9)화(60.0±4.2)g,t치분별위-3.11、-3.97、-3.67、-6.52화-3.72,P치균＜0.05).제28천신경행위학측시:실험조광장실험평분고우대조조[(36.5±2.9)여(24.3±3.6)분,t=7.36,P＜0.01],현조실험평분야고우대조조[(3.6±1.0)여(2.0±0.7)분,t=3.53,P＜0.01].실험조Cylinder실험좌、우전족/쌍전족적비치[(49.8±13.3)％여(41.4±5.9)％]차이무통계학의의(t=0.86,P＞0.05);대조조좌전족/쌍전족적비치고우우전족/쌍전족[(49.5±11.3)％여(31.2±3.2)％,t=4.38,P＜0.01].결론 HUcMSCs가제고신생대서WMD모형병변구역내소돌효질세포적수량,감소소효질세포적활화,대체격발육유량호촉진작용,병차대신경행위유개선작용.
Objective To investigate the alterations in oligodendrocyte and microglia and changes in neurobehavior after human umbilical cord mesenchymal stem cells (HUcMSCs) intervention on the newborn rat model of white matter damage (WMD) induced by hypoxia-ischemia.Methods After the operation of left common carotid artery ligation and 4 h hypoxia (6％ O2 and 94％ N2),twelve three-day-old Sprague-Dawley rats died and the remaining sixty rats were randomly divided into the experimental group and the control group.The first day was 0 to 24 h after birth.Rats of the experimental group were intraperitoneally injected the fourth generation HUcMSCs 1 × 106 (0.05 ml) on the third,fourth and fifth day respectively.At the same time,rats of the control group were intraperitoneally injected phosphate buffer (0.05 ml).Eight rats of each group were executed on the tenth and twenty first day respectively to detect the number of cells positive for myelin basic protein (MBP) and ectodermal dysplasia-1 (ED-1) staining by immunohistochemistry.Three rats of each group were executed on the tenth and twenty first day respectively to detect MBP and ED-1 expression by western blot.Eight rats of each group were weighed and underwent the neurobehavioral evaluation on the twenty eighth day.Data were analyzed using t test.Results On the tenth and twenty first day,the numbers of MBP-positive cells in the experimental group (11.8 ± 4.1 and 23.8± 8.1) were significantly higher than those in the control group (6.7±3.1 and 11.5 ± 5.8,t=-2.81 and 3.49,both P＜0.05) ; and the numbers of ED-1 positive cells in the experimental group (20.8 ± 3.4 and 19.1 ± 2.8) were significantly lower than those in the control group (32.8±4.2 and 29.5±5.2,t=6.23 and 4.93,both P＜0.01).On the tenth and twenty first day,MBP expressions in the experimental group (1.3 ± 0.1 and 1.1 ± 0.1) were higher than those in the control group (0.8±0.0 and 0.6±0.1,t=-7.53 and 6.68,both P＜0.01) ; and the ED-1 expressions in the experimental group (0.6±0.1 and 0.4±0.1) were lower than those in the control group (1.0±0.1 and 0.8±0.1,t=3.09 and 4.90,both P＜0.01).Weight on the seventh,tenth,fourteenth,twenty first and twenty eighth day in the experimental group [(15.0± 1.2),(16.6±0.9),(27.0± 1.6),(44.2±2.3) and (68.1 ±4.2) g] was significantly higher than that in the control group [(12.7 ± 1.6),(13.5 ± 2.0),(23.6 ± 1.9),(38.4± 0.9) and (60.0± 4.2) g,t=-3.11,-3.97,3.67,-6.52 and-3.72,all P＜0.05].On the twenty eighth day,the score of open field test in the experimental group was significantly higher than that in the control group (36.5 ± 2.9 vs 24.3 ± 3.6,t=7.36,P＜0.01).So was the hanging test (3.6± 1.0 vs 2.0±0.7,t=3.53,P＜0.01).In Cylinder test,the ratio of left/both and right/both forefeet in the experimental group were similar [(49.8± 13.3) ％ vs (41.4±5.9) ％,t=0.86,P＞0.05],but the ratio of left/both forefeet in the control group was higher than right/both [(49.5 ± 11.3) ％ vs (31.2±3.2) ％,t=4.38,P＜0.01].Conclusions HUcMSCs are able to enhance the number of oligodendrocytes while weaken the activity of microglias in the WMD newborn rat model,and to promote the physical development and improve the rat neurobehavior.
