人脐血干细胞移植治疗大鼠脑出血实验探究.docVIP

人脐血干细胞移植治疗大鼠脑出血实验探究 　 作者：潘凤华，丁新生Δ，李晓波，丁海霞，张炜民，邓小萱，姚 娟 目的 研究人脐血干细胞（human umbilical cord blood cells，HUCBC）移植治疗脑出血大鼠的效果及HUCBC在出血大鼠脑内的状况。方法 采集足月新生儿脐带血60～100ml，分离出其中的单个核细胞，体外培养并予5-溴脱氧嘧啶尿苷（Brdu）标记48h。用VII型胶原酶注入到大鼠右侧苍白球诱导脑出血模型，24h后经尾静脉注射3×106HUCBC入大鼠体内，同时设立生理盐水组及对照组，对移植后大鼠的神经损害严重程度评分（neurological severity scores，NSS），移植后35天杀死大鼠，用免疫组织化学技术研究移植后的HUCBC的存活、迁徙、分化状况。结果 HUCBC在体外具有增殖能力；HUCBC移植组大鼠自移植后21天起其NSS显著低于生理盐水组及对照组（Plt;0.05）；移植后35天脑组织切片中可见Brdu染色阳性细胞，出血侧明显多于对侧，神经上皮干细胞蛋白（Nestin）以及神经元标记神经元烯醇化酶（NSE）染色呈弱阳性，胶质纤维酸性蛋白（GFAP）染色呈强阳性；生理盐水组及对照组未见各染色阳性细胞。结论 HUCBC植入大鼠出血脑内能存活并向损伤区域迁徙，分化为星形胶质细胞和神经元，从而改善其神经功能。HUCBC移植可作为脑出血的有效治疗手段。 【关键词】 人脐血干细胞；移植；脑出血 【Abstract】 Objective To explore the effect of the human umbilical cord stem blood cells(HUCBC) transplantation to treat intracerebral hemorrhagic rats and the status of transplanted HUCBC in the hemorrhagic brain tissue of these rats.Methods HUCBC got from the placenta of full-term babies were cultured and marked with Brdu(5μmol/l) for two days.Intracerebral hemorrhage(ICH)models was induced in rats by injection of VⅡtype collagenase into the right globus pallidus and the HUCBC were transplanted into the tail vein 24 hours later. Neurological Severity Scores(NSS) tests were undertaken and immunohistochemistry method was used to check the migration and differentiation of HUCBC in rats.Results The HUCBC had the capacity of proliferation in vitro . Twenty-one days later，the neurological function of rats that received transplantation recovered much better than the rats without transplantation，as evidenced by NSS(Plt;0.05).Within the brain tissues ， 5-bromodeoxyuridine(Brdu ) reactive HUCBC were distributed throughout the lesions in recipient rats 35 days after transplantation.The vast majority of cells localized in the hemorrhagic hemisphere and few cells were observed in the contralateral hemisphere. A few HUCBC were reactive for the neural stem cells markers neuroepthelial stem cell protein(Nestin) and the neuronal markers neuron-specific endas