移植人脐带间充质干细胞治疗缺氧缺血性脑损伤的两种方法比较

目的比较人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs)经颈静脉及经腹腔两种途径移植后在缺氧缺血性脑损伤(hypoxic-ischemic brain damage,HIBD)新生大鼠脑内的分布、分化情况,寻找简单易行且有效的移植途径。方法 RICE法制备7日龄新生大鼠HIBD模型,随机分组:HIBD组(n=10)、腹腔移植组(n=15)、颈静脉移植组(n=15),另随机取未造模的10只为正常对照组。造模后72h后分别对颈静脉移植组和腹腔移植组移植等量的hUCMSCs(1×106个/只),而HIBD组及正常对照组不移植。于移植后36d免疫荧光染色比较Dil标记的hUCMSCs移植后在各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。结果移植后hUCMSCs能迁徙到脑组织中,能分化成神经元前体细胞。经颈静脉移植途径优于经腹腔移植(P0.01)。结论 hUCMSCs移植后可以在HIBD新生大鼠脑内存活,并向神经元前体细胞分化,对HIBD新生大鼠脑功能有修复作用,尤以经颈静脉移植明显。

Study on two approaches of transplantation of human umbilical cord mesenchymal stem cells

Objectiv To compare the distribution and differentiation of human umbilical cord mesenchymal stem cells(hUCMSCs) after the transplantation via the jugular vein and via the intraperitioneal injection,and investigate the effect of hUCMSCs transplantation on the recovery of neurological functions after hypoxic-ischemic brain damage (HIBD). Methods The models of 7-day-old neonate rats with HIBD brain were established,and randomly assigned to:HIBD group (n=10),the intravenous injection group (n=15),the intraperitioneal injection group (n=15) and control group (n=10),the HIBD group and control group were not transplanted.The same amount hUCMSCs(1×10⁶,0.2 mL)were transplanted into the intravenous injection group and the intraperitioneal injection group after 72 hours.Finally,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 Dil-labeled hUCMSCs and expression of DCX. Results The transplantation descendants of hUCMSCs could migrate to the brain tissue,and could differentiate into neuronal precursor cells,and the intravenous injection group showed more Dil-labeled hUCMSCs in brain tissue. Conclusions Transplanted hUCMSCs can survive and differentiate in the HIBD rats' brain.The HUCMSCs can effectively accelerate the recovery of neurological functions after HIBD.And the experiment proved intravenous injection is better than intraperitioneal injection in repairing rats' brain function.