人脐带间充质干细胞联合血肿抽吸术治疗大鼠脑出血的实验研究

Experimental study of human umbilical cord mesenchymal stem cells combined with minimal hematoma aspiration in the therapy of rats with intracerebral hemorrhage

目的观察脐带间充质干细胞(HUC-MSCs)移植联合血肿抽吸治疗大鼠脑出血后脑组织中p53基因的表达及大鼠神经功能恢复的情况。方法体外分离HUC-MSCs,移植前以BrdU标记。向尾状核注入2μLⅣ型胶原酶建立脑出血模型,将120只造模成功的SD大鼠随机分为脑出血组、血肿抽吸组、细胞移植组、细胞移植联合血肿抽吸组(联合治疗组),进行相应的处理后,于1、3、7、14和30 d 5个时间点分批处死大鼠,取脑组织,采用免疫组织化学法检测大鼠脑出血周围p53基因的表达及HUC-MSCs在脑组织的分布,并用改良的大鼠神经功能缺损评分(mNSS)评价大鼠的神经功能。结果与脑出血组比较,血肿抽吸组、细胞移植组和联合治疗组的mNSS评分、p53表达量均下降(P均<0.05);7 d时,联合治疗组的mNSS评分及p53表达量低于脑出血组和血肿抽吸组(P均<0.05),14 d、30 d时则低于其余3组(P均<0.05)。结论 HUC-MSCs移植联合血肿抽吸治疗可以显著减少大鼠脑出血后神经元的损伤,更有利于神经功能的恢复。

Objective To investigate the effect of human umbilical cord mesenchymal stem cells （HUC-MSCs） trans- plantation combined with hematoma aspiration in the therapy of rats with intracerebral hemorrhage by observing the changes of p53 gene expression and the recovery of neural functions. Methods HUC-MSCs were isolated in vitro and labeled by BrdU before transplantation. 2 μL IV collagenase was injected to the candate nucleus to establish intracerebral hemorrhage model. One hundred and twenty SD rats with successful modeling were randomly divided into 4 groups： intracerebral hemorrhage group, hematoma aspiration group, HUC-MSCs transplantation group and HUC-MSCs trans- plantation combined with hematoma aspiration group （combination group）. After the corresponding treatments, rats in each group were killed on 5 time points（days 1, 3, 7, 14 and 30）, then the brain tissues were isolated. The expres- sions of p53 in the cerebral tissues surrounding intracerebral hemorrhage on different time points and the distribution of HUC-MSCs were detected by immunohistochemical staining. Neural functional status of rats was assessed by modified neurological severity score（mNSS）. Results The mNSS scores and the expression of p53 in hematoma aspiration group, HUC-MSCs transplantation group and combination group were significantly lower than those in intracerebral hemorrhage group （ all P 〈 0.05 ）. The scores of mNSS and the expression of p53 in combination group were lower than those in intracerebral hemorrhage group and hematoma aspiration group on day 7 （ all P 〈 0.05）, and lower than those in the other three groups from day 14 to day 30. Conclusion The intervention of HUC-MSCs transplantation combined with hematoma aspiration can obviously reduce the damage of nerve cells in rats with intracerebral hemorrhage and it is beneficial to the recovery of neural function.