研究移植人羊膜间充质干细胞（hAMSCs）是否能促进脑出血大鼠神经功能恢复，探索其可能作用机制。

138只雄性Wistar大鼠按照随机数字表法分为人皮肤成纤维细胞（Fibroblasts）治疗组（46只）、磷酸盐缓冲液（PBS）治疗组（46只）和hAMSCs治疗组（46只）。脑出血模型采用颅内注射Ⅶ型胶原酶制作，24 h后颅内移植hAMSCs、Fibroblasts和PBS。应用流式细胞仪检测hAMSCs表面抗原。免疫荧光检测细胞分化、血管再生和神经再生。酶联免疫吸附剂测定试剂盒检测脑源性神经营养因子（BDNF）和血管内皮生长因子（VEGF），改良神经功能评分（mNSS）检测行为学。

分离培养的hAMSCs呈现成纤维细胞样细胞形态，表达CD29、CD44、CD90、CD105和CD166，不表达CD19、CD34和CD45。mNSS评分显示，颅内出血（ICH）7 d后，hAMSCs组评分明显高于Fibroblasts治疗组和PBS治疗组，差异具有统计学意义（P<0.05）。ICH后14 d，hAMSCs组BrdU⁺细胞数目（55.33±6.59）高于Fibroblasts组（26.66±3.58）和PBS组（25.50±2.90），差异具有统计学意义（P<0.05）。hAMSCs组Brdu⁺/DCX⁺细胞数目（30.66±3.65）高于Fibroblasts组（11.83±2.70）和PBS组（10.00±2.54），差异具有统计学意义（P<0.05）。hAMSCs组血管性血友病因子⁺血管数目[（67.50±6.76）/mm²]高于Fibroblasts组[（31.66±6.33）/mm²]和PBS组[（28.83±5.58）/mm²]，差异具有统计学意义（P<0.05）。ICH后7 d和14 d，hAMSCs组BDNF表达水平[（63.16±2.31）pg/mg，（38.57±3.69）pg/mg]高于Fibroblasts组[（46.18±2.45）pg/mg，（20.52±2.81）pg/mg]和PBS组[（43.18±2.30）pg/mg，（18.28±2.39）pg/mg]，差异具有统计学意义（P<0.05）。ICH后7、14、21 d，hAMSCs组VEGF表达水平[（43.32±3.65）pg/mg，（30.90±2.62）pg/mg，（24.89±2.85）pg/mg]高于Fibroblasts组[（22.88±2.62）pg/mg，（15.41±2.40）pg/mg，（10.57±1.54）pg/mg]和PBS组[（20.81±3.06）pg/mg，（14.44±2.32）pg/mg，（11.42±1.48）pg/mg]，差异具有统计学意义（P<0.05）。移植的hAMSCs在颅内存活最少27 d，不表达神经元标志物β-tubulin Ⅲ和星型胶质细胞标志物胶质纤维酸性蛋白。

移植hAMSCs能促进脑出血大鼠神经功能恢复，其作用机制可能是通过增加神经营养因子表达，促进血管再生和神经再生。因此hAMSCs是治疗脑出血的理想干细胞。

We assessed whether transplantation of human amniotic mesenchymal stem cells (hAMSCs) promotes neurological recovery in an intracerebral hemorrhage rat model. In addition, the potential mechanisms underlying the possible benefits of this therapy were investigated.

A total of 138 male Wistar rats were divided into fibroblasts group, phosphate-buffered saline(PBS) group and hAMSCs group (n=46). The intracerebral hemorrhage (ICH) models were induced by intrastriatal injection of VII collagenase and then intracerebrally administered hAMSCs, Fibroblasts, or PBS at 24 h after ICH. hAMSCs were characterized using flow cytometry. Immunohistochemistry were performed to assay differentiation, angiogenesis and neurogenesis. The expressions of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) were analyzed by ELISA. Neurological function was assessed with the modified neurological severity score (mNSS).

hAMSCs displayed a fibroblast-like morphology, were positive for CD29, CD44, CD90, CD105, and CD166, and negative for CD19, CD34, and CD45. From 7 d after ICH, mNSS scores of the hAMSCs group were significantly higher than that of PBS group and Fibroblasts group (P<0.05). Fourteen days after ICH, the number of Brdu⁺ cells in the hAMSCs group (55.33±6.59) was significantly higher than that in PBS group (25.50±2.90) and Fibroblasts group (26.66±3.58) (P<0.05). The number of Brdu⁺/DCX⁺ cells (30.66±3.65) in the hAMSCs group was significantly higher than that in PBS group (10.00±2.54) and fibroblasts group (11.83±2.70) (P<0.05). The number of von Willebrand factor (vWF⁺) blood vessels [(67.50±6.76)/mm²] in the hAMSCs group was significantly higher than that in PBS group [(28.83±5.58)/mm²] and Fibroblasts group [(31.66±6.33)/mm²] (P<0.05). Seven and fourteen days after ICH, the levels of BDNF[(63.16±2.31) pg/mg, (38.57±3.69) pg/mg] of the hAMSCs group were significantly higher than that of PBS group [(43.18±2.30) pg/mg, (18.28±2.39) pg/mg] and Fibroblasts group[(46.18±2.45) pg/mg, (20.52±2.81) pg/mg](P<0.05). Seven, fourteen and twenty-one days after ICH, the levels of VEGF [(43.32±3.65) pg/mg, (30.90±2.62) pg/mg, (24.89±2.85) pg/mg] of the hAMSCs group were significantly higher than that of PBS group [(20.81±3.06) pg/mg, (14.44±2.32) pg/mg, (11.42±1.48) pg/mg] and Fibroblasts group [(22.88±2.62) pg/mg, (15.41±2.40) pg/mg, (10.57±1.54) pg/mg]. The transplanted hAMSCs survived for at least 27 d and were negative for fi-tubulin Ⅲ and glial fibrillary acidic protein.

hAMSCs treatment significantly promotes neurological recovery in rats after ICH. The mechanism of action could be mediated by increasing neurotrophic factor expression, and promotion of neurogenesis and angiogenesis. Thus, hAMSCs are candidate stem cells for the treatment of ICH.