人胚胎骨髓间充质干细胞和单个核细胞静脉移植治疗大鼠脑梗死疗效的比较  

Comparsion between Intravenous Delivered Human Fetal Bone Marrow Mesenchymal Stromal Cells and Mononuclear Cells in the Treatment of Rat Cerebral Infarct

作　　者：黄爱华[1] 张萍萍[1] 张斌[1] 马步青[1] 关云谦[2] 周逸丹[1] HUANG Ai-hua ZHANG Ping-ping ZHANG Bin MA Bu-qing GUAN Yun-qian ZHOU Yi-dan(Department of Emergency, the Third People＇s Hospital of Hangzhou, Hangzhou 310000, China Department of Cell Biology, Xuanwu Hospital, Capital Medical University, Beijing 100053, China)

机构地区：[1]浙江省杭州市第三人民医院急诊科,杭州310000 [2]首都医科大学宣武医院细胞生物室,北京100053

出　　处：《中国医学科学院学报》2016年第5期497-506,共10页Acta Academiae Medicinae Sinicae

基　　金：杭州市卫生局重点项目(2011Z007);北京市科委健康培育项目(Z111107067311033)

摘　　要：目的比较人胚胎骨髓间充质干细胞(MSC)和单个核细胞(MNC)经静脉移植后对脑梗死的疗效。方法采用大脑中动脉远端凝断法制造大鼠脑梗死模型。将大鼠分为假手术组、缺血对照组(造模后经尾静脉给予生理盐水)、MSC组(造模后经尾静脉给予MSC)和MNC组(造模后经尾静脉给予MNC)。移植后2 d,取脑进行梗死体积测定;移植后1、3、5、7 d,采用肢体放置测验和平衡木实验进行行为学评价;移植后7 d,采用免疫荧光法测量梗死周边纹状体区激活的小胶质细胞Iba-1的染色丰度;移植后3、7 d,采用ELISA检测移植后梗死灶周边纹状体区胶质源性神经营养因子(GDNF)和脑源性神经营养因子(BDNF)的蛋白水平。结果缺血对照组、MSC组和MNC组的相对梗死体积分别为(37.85±4.40)%、(33.41±3.82)%和(30.23±3.63)%,其中,MSC组(t=2.100,P=0.034)和MNC组(t=2.109,P=0.0009)显著低于缺血对照组,MNC组显著低于MSC组(t=1.743,P=0.043)。移植后1 d,缺血对照组的肢体放置成功率为(4.32±0.71)%,明显低于假手术组的(9.73±0.36)%(t=2.178,P=8.61×10-11);MSC组和MNC组分别为(5.09±0.62)%(t=2.1009,P=0.024)和(5.90±0.68)%(t=2.1008,P=0.0001),明显高于缺血对照组;MNC组又明显高于MSC组(t=2.1009,P=0.0165)。MSC组和MNC组损伤对侧前肢错步数占总步数的百分比分别为(5.56±0.86)%(t=2.120,P=0.020)和(5.13±0.95)%(t=2.131,P=0.003),明显低于缺血对照组的(6.47±0.61)%。移植后3 d,MNC组的肢体放置成功率为(6.91±1.10)%,明显高于缺血对照组的(5.80±0.82)%(t=2.110,P=0.027);MSC组为(6.30±0.77)%,与缺血对照组差异无统计学意义(t=2.101,P=0.199)。MNC组损伤对侧前肢错步数占总步数的百分比为(4.34±0.58)%,明显低于缺血对照组的(5.31±0.65)%(t=2.100,P=0.006)和MSC组的(4.92±0.53)%(t=2.100,P=0.041),MSC组与缺血对照组差异无统计学意义(t=2.109,P=0.139)。假手术组、缺血对照组、MSC组和MNC组的Iba-1相对丰度分别为1.00+0.00、1.72±0.21、1.Objective To compare the effecacy of human mesenchymal stromal cell（ hM SC） with human mononuclear cell（ hM NC） in treating rat cerebral infarct.Methods The SD rat models of cerebral infarct were established by distal middle cerebral artery occlusion（ dM CAO）.Rats were divided into four groups：sham,ischemia vehicle,MSC,and MNC transplantation groups.For the transplantation group,1 × 106 hM SCs or hM NCs were intravascularly transplanted into the tail vein 1 hour after the ischemia onset.The ischemia vehicle group received dM CAO surgery and intravascular saline injection 1,3,5,and 7 days after the ischemia onset,and then behavioral tests were performed.At 48 h after the ischemia onset,the abundance of Iba-1,the symbol of activated microglia,was evaluated in the peri-ischemia striatum area; meanwhile,the neurotrophic factors such as glial cell line-derived neurotrophic factor（ GDNF） and brain-derived neurotrophic factor（ BDNF） in ipsilateral peri-ischemia striatum area were also measured.Results The relative infarct volume in ischemia vehicle group,hM SC group,and hM NC transplantation group were（ 37.85 ± 4.40） %,（ 33.41 ± 3.82） %,and（ 30.23 ± 3.63） %,respectively.The infarct volumes of MSC group（ t = 2.100,P = 0.034） and MNC group（ t = 2.109,P = 0.0009） were significantly smaller than that of ischemia vehicle group,and that of MNC group was significantly smaller than that of MSC group（ t = 1.743,P = 0.043）.One day after transplantation,the score of ischemia vehicle group in limb placing test was（ 4.32 ± 0.71） %,which was significantly lower than that in sham group（ 9.73 ± 0.36） %（ t =2.178,P = 8.61 × 10-11）.The scores of MSC and MNC group,which were（ 5.09 ±0.62） %（ t =2.1009,P =0.024） and（ 5.90 ± 0.68） %（ t = 2.1008,P = 0.0001）,respectively,were significantly higher than that of ischemia vehicle group; also,the score of MNC group was significantly higher than that of MSC group（ t = 2.1009,P = 0.0165）.The contralateral forelimb

关 键 词：脑梗死 间充质干细胞 单个核细胞 炎症 小胶质细胞 移植 

分 类 号：R392.28[医药卫生—免疫学]