目的 观察人脐带间充质干细胞(human umbilical cord mesenchymal stem cells, hUCMSCs)移植及联用神经节苷酯GM1治疗新生大鼠缺氧缺血性脑损伤(hypoxic-ischemic brain damage, HIBD)的效果。方法 RICE法制备七日龄新生大鼠HIBD模型, 随机分为hUCMSCs移植组(n=15)、hUCMSCs+GM1组(n=15), 72 h后经颈静脉移植等量的hUCMSCs, hUCMSCs+GM1组再腹腔注射GM1, 两组均在移植后第31、32、33、34、35天用Morris水迷宫对两组大鼠行为学测试, 比较两组大鼠脑功能恢复情况。免疫荧光染色观察DiI标记的hUCMSCs移植后在两组大鼠脑中的分布情况;神经元前体细胞标记DCX特异性染色观察hUCMSCs移植后分化成神经元前体细胞的情况。结果 Morris水迷宫测试:hUCMSCs+GM1组逃避潜伏期明显短于hUCMSCs移植组, 差异具有统计学意义(P＜0.05)。移植后hUCMSCs能迁徙到脑组织中, 并分布于缺血区的小血管旁, 能分化成神经元前体细胞。hUCMSCs+GM1组大鼠脑片干细胞分布明显多于hUCMSCs移植组(P＜0.05)。结论 hUCMSCs移植对HIBD新生大鼠脑功能有修复作用, 联用GM1, 有协同作用。
Abstract
Objective To observe the effect of human umbilical cord mesenchymal stem cells (hUCMSCs) combined with ganglioside (GM1) on the treatment of hypoxic-ischemic brain damage (HIBD) in neonate rats. Methods First, the models of 7-day-old neonate rats with HIBD brain were established.Then, after 72 hours, the same amount hUCMSCs were transplanted into the hUCMSCs transplantation group and the hUCMSCs+GM1 group, at the same time, GM1 was injected into the hUCMSCs+GM1 group through abdominal cavity.Spatial learning was assessed using a Morris water maze test from 31 to 35 days after the transplantation.Finally, after the behavioral tests, brain tissues of rats were removed after 4% paraformaldehyde was perfused in cardiac to fix brain and successive sections were made using a cryomicrotome.Immunofluorescence staining was performed to examine the distribution of DiI-labeled hUCMSCs and expression of DCX. Results The hUCMSCs+GM1 group showed more significant neurological functions improvement than the hUCMSCs transplantation group as revealed by Morris water maze test.The transplantation descendants of hUCMSCs could migrate to the brain tissue, and could differentiate into neuronal precursor cells, and the hUCMSCs+GM1 group showed more DiI-labeled hUCMSCs in brain tissue. Conclusion Transplanted hUCMSCs could improve the recovery of neurological functions after HIBD and promote rats' behavioral function improvement.And GM1 exerts a synergistic effect in this treatment.
关键词
人脐带间充质干细胞 /
神经节苷酯 /
缺氧缺血性脑损伤 /
移植
Key words
human umbilical cord mesenchymal stem cells /
hypoxic-ischemic brain damage /
transplantation /
ganglioside
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