神经系统疾病已成为全球第二大死因,其中脑卒中位居所有神经系统疾病之首[1].在我国部分脑血管疾病高发的城市,脑出血占脑卒中患者的26.7%~51.5%,是我国致死亡和残疾的主要疾病之一[2].然而目前对脑出血的治疗仅限于脱水降颅压等对症治疗,无法修复、填充脑坏死区域死亡的神经细胞,导致脑出血预后不良,往往造成难以治愈的瘫痪甚至危及生命[3].因此,很多学者将注意力转移至脑出血后的神经保护与修复治疗,干细胞因其自我更新和多向分化潜能得到了广泛关注.人脐血间充质干细胞(human umbilical cord blood mesenchymal stem cells, hUCB-MSCs)不仅来源广泛、采集方便,而且细胞含量更丰富、更原始,移植抗宿主反应轻[4-5],相对无菌,被视为细胞治疗的最理想来源.目前hUCB-MSCs移植的实验性治疗已经应用于许多神经系统疾病,如阿尔茨海默病、帕金森病[6-7]、自身免疫性脑脊髓膜炎、肌萎缩侧索硬化[8-9]和脊髓损伤[10].一些研究发现,hUCB-MSCs对大鼠脑缺血损伤有治疗作用[11],而对于脑出血的研究却鲜有报道.我们旨在观察局部hUCB-MSCs移植治疗脑出血大鼠的疗效,为hUCB-MSCs移植治疗脑出血提供更多的理论依据.
Objective To observe the expression of nestin nestin mRNA in umbilical cord mesenchymal stem cells (MSCs) from the perspective of gene .Methods The fresh umbilical cord tissues underwent MSCc separation and then were cultured and purified .Cells from the umbilical cord were counted by enzymatic digestion , and the activity was detec-ted by flow cytometry .The primary, first-generation and second-generation cellular mRNA were extracted to detect the ex-pression of nestin mRNA by real-time PCR.Results The cells isolated from umbilical cord grew in good condition , in which a large part of the cells grew in fusiform , cells after passage had the similar morphology with primary cells and grew faster.There were individual cells which were inneural-like growth.Nestin mRNA expression was found in umbilical cord MSCs, and the expression level after the passage was significantly decreased , meanwhile, the nestin protein changes were consistent with that of differentiation from the neural precursor cells into neural cells .Conclusion The nestin mRNA ex-pression was found in the MSCs tissue genes .Even the expression level of nestin mRNA decreases after passage , it still can be used as an alternative seed cells for transplantation .
背景:研究报道在特定的体内外环境下,脐血间充质干细胞能够诱导分化成为包括神经干细胞在内的多种组织细胞.目的:评价人脐血单个核细胞经腰穿途径移植后治疗急性一氧化碳中毒后迟发性脑病的疗效及安全性.方法:一氧化碳中毒后迟发性脑病患者60例随机分为2组.对照组给予高压氧及药物治疗;治疗组采用鞘内注射法将经密度梯度离心法分离出的人脐血单个核细胞移植到一氧化碳中毒性脑病患者的蛛网膜下腔,余治疗方法同对照组.分别于人脐血单个核细胞移植前、移植后3,9,12个月对患者进行简易精神状态检查法、改良Asworth肌肉痉挛程度分级及日常生活量表评分检查;比较两组患者MRI变化;同时对随诊患者行胸片、心电图及血生化检查,客观评价人脐血单个核细胞移植的安全性.结果与结论:人脐血单个核细胞移植3,9,12个月,治疗组Asworth肌肉痉挛程度分级评分均显著低于对照组(P=0.032);移植后9,12个月简易智力状况检查法及日常生活量表评分均显著高于对照组(P＜0.05);两组患者神经功能在各时间点的变化趋势相似.人脐血单个核细胞移植后12个月MRI检查结果显示,治疗组患者MRI改善程度较对照组明显.提示鞘内注射移植人脐血单个核细胞治疗一氧化碳中毒后迟发脑病疗效优于高压氧治疗.
