间充质干细胞不同移植途径对修复大鼠缺氧缺血性脑损伤作用的研究

目的]比较研究大鼠骨髓间充质干细胞(rMSCs)经三条不同途径移植对其修复大鼠缺氧缺血性脑损伤(HIBD)作用的影响。方法]113只Wistar大鼠随机分成脑局部定位移植rMSCs组、尾静脉移植rMSCs组、鞘内移植rMSCs组、假手术组、模型组。造模后3d各移植组分别脑局部定位注射rMSCs、尾静脉注射rMSCs、鞘内注射rMSCs。造模后24d行Morris水迷宫实验检测大鼠学习、记忆能力,移植后1d、4d、7d、14d、21d、28d分别取脑组织作病理学检查,并应用双标免疫组化检测rMSCs的分布及神经元特异性烯醇化酶(NSE)和胶质纤维酸性蛋白(GFAP)表达情况。结果 ]各移植组的逃避潜伏期较模型组显著缩短(P<0.05),穿越平台次数显著增加(P<0.05),但移植组之间无显著性差异(P>0.05)。免疫组化显示各移植组脑组织内都出现rMSCs,脑局部移植的细胞随时间推移以注射点为中心向周围移行,尾静脉移植的细胞分布相对广泛,鞘内移植的细胞随时间推移自脑实质外周向深处移行,移植后1d、4d、7d、14d脑rMSCs组与鞘rMSCs组均显著多于尾rMSCs组(P<0.05)。NSE阳性率在三组之间无显著性差异(P>0.05),GFAP阳性率以移植后21d尾rMSCs组最高(P<0.05)。结论]rMSCs经三条途径移植均能改善HIBD大鼠学习、记忆能力,但无明显差异;脑局部定位移植与鞘内移植均可使较多rMSCs早期进入脑实质,经尾静脉移植的rMSCs向星形胶质细胞分化较多。

The Optimal Transplanting Way for Rat Mesenchymal Stem Cells Repairing Hypoxic-ischemic Brain Damage in Rats

Objective] Comparative study on the effect of rat mesenchymal stem cells（rMSCs） transplanted by three different ways on repairing hypoxic-ischemic brain damage（HIBD） in rats.Methods] 113 Wistar rats were randomly divided into 5 groups,named respectively by letters from A～E.Excepting 14 rats in Group A were sham-operated,the other 99 were made into HIBD model by Rice method.The rats in Groups A and B were treated with nothing,the others were injected with rMSCs respectively in the local brain tissue with stereotaxis instrument,the caudal vein and the spinal subarachnoid space 3 days after hypoxic-ischemic（H-I） were inujred.Learning and memory abilities were assessed by Morris water maze 24 days after H-I insult;Distribution and migration of rMSCs,expression of neuron-specific enolase（NSE） or glial fibers acid protein（GFAP） on rMSCs were shown by double-labeled immunohistochemistry（IHC）.Results] There are no significance differences of average escape latency and ring crossing times among Groups C,D and E.IHC shows rMSCs presenting in the brains of all transplanted groups which migrated from the injected point to surrounding tissues in Groups C,distributed relatively wide in Group D,and migrated from the peripheral region to the center of the brain in Group E.RMSCs in Groups C and E are significantly more than that in Group D at the 1st,4th,7th,14th day.GFAP expressing rate is the highest in Group D at the 21st day.Conclusions] The learning and memory abilities of HIBD in rats are improved with rMSCs transplantation by three different transplanting ways,but the differences are not obvious.There are more rMSCs migrating into brain tissue early by lacol injection and intrathecal injection,and there are more rMSCs differentiating into astrocytes by caudal vein injection.