骨髓单个核细胞移植对脑出血大鼠的影响

Bone marrow mononuclear cell transplantation in the treatment of intracerebral hemorrhage in rats

目的:探讨骨髓单个核细胞(BMMNCs)移植对脑出血(ICH)大鼠的影响。方法:通过密度梯度离心法提取大鼠BMMNCs并经流式细胞仪鉴定;采用立体定向技术向尾状核内注射Ⅶ型胶原酶构建大鼠ICH模型;24只大鼠分为PBS组和BMMNCs组,每组12只,于移植后第1、3、7和14天进行神经功能缺损评分(mNSS)。90只大鼠分为假手术组、PBS组和BMMNCS组,每组30只,分别于移植后第0、1、3、7和14天处死6只大鼠检测脑组织含水量;移植后第7天行甲酚紫染色观察大脑皮层神经元数量;移植后第28天在荧光显微镜下观察BMMNCs在脑内的存活、分布和分化情况。结果:BMMNCs移植后不同时间点,与PBS组相比,大鼠神经功能恢复较好(F时间=15.243,F组间=22.826,F交互=4.932,P均<0.05),大鼠脑组织含水量明显降低(F=27.518、106.965、45.006,P均<0.001),大脑皮层神经元数量增多。BMMNCs能够迁移分布至大鼠脑损伤区,并分化为神经元和星形胶质细胞。结论:静脉移植BMMNCs可减轻ICH大鼠脑水肿,BMMNCs能够在出血周边存活并分化为神经元和星形胶质细胞,改善ICH大鼠神经功能。

Aim： To investigate the effect of transplanted bone marrow mononuclear cells （BMMNCs） in a rat model of intracerebral hemorrhage（ICH）. Methods： BMMNCs were isolated by Ficoll density gradient centrifugation. Cell surface antigens of CD34, CD45 and CD90 were identified by flow cytometry. Intracerebral hemorrhage models were established by stereotactic injection of collagenase VII into caudate putamen. Rats were randomly divided into sham group, PBS group and BMMNCs group. Functional deficits were assessed with the modified neurological severity score on the 1 st, 3rd, 7th, and 14th day after transplantation. The number of neurons around hematoma was observed by Niss＇s staining on day 7 after transplantation. The survival, migration, distribution and differentiation of BMMNCs were assessed by immunofluorescence double staining on day 28 after transplantation. Results： At different times after transplantation,BMMNC transplantation significantly improved neurological function（ Ftime = 15. 243, Fgroup = 22. 826, Finteraction= 4. 932, P 〈 0.05 ） , reduced brain edema（F = 27. 518,106. 965,45. 006, P 〈 0. 001 ）, and increased the number of neurons compared with other groups. BMMNCs were able to migrate into perihematoma region and differentiated into neurons and astrocytes. Conclusion ： BMMNCs transplantation can significantly reduce brain edema and promote neurological recovery by differentiating into neurons and astrocytes in a rat model of ICH.