骨髓间充质干细胞与上清液治疗大鼠脑梗死的疗效

目的探讨骨髓间充质干细胞与上清液治疗大鼠脑梗死的疗效。方法将40只脑梗死大鼠模型用随机数字表法分为氯化钠注射液组(9 g.L-1氯化钠注射液2 mL)、骨髓间充质干细胞(BMSCs)移植组(2.0×109L-1BM-SCs悬液2 mL)、上清液移植组(BMSCs上清液2 mL)及混悬液移植组(上清液与BMSCs混悬液2 mL),每组10只。在模型再灌注24 h后,用10 g.L-15-溴脱氧尿核苷(BrdU)溶液标记,连续标记28 d,并于最后1次注射后2 h处死大鼠。观察各组病灶增殖期神经前体细胞的数目变化及神经损伤严重程度(NSS)评分。结果治疗后28 d BrdU、Br-dU+抗人神经元特异性烯醇化酶、BrdU+胶质纤维酸性蛋白标记的阳性细胞数BMSCs移植组、上清液移植组和混悬液移植组较氯化钠注射液组明显增多,差异有统计学意义(P<0.05);上清液移植组和混悬液移植组较BMSCs移植组增多,差异有统计学意义(P<0.05);混悬液移植组较上清液移植组增多,差异有统计学意义(P<0.05)。与移植前比较,各组大鼠动物模型移植后第7、14、28天的神经功能情况均有不同程度的恢复,NSS评分逐渐降低,差异均有统计学意义(P<0.05或P<0.01);从第7天起,与氯化钠注射液组比较,BMSCs移植组、上清液移植组和混悬液移植组NSS评分均明显降低,差异有统计学意义(P<0.05或P<0.01);混悬液移植组NSS评分明显低于BMSCs移植组和上清液移植组,差异有统计学意义(P<0.01)。结论上清液与BMSCs混悬液能够诱导病灶区内源性神经前体细胞的增殖、分化,这可能与上清液内的神经营养因子及细胞分化诱导因子诱导BMSCs分化为神经细胞有关。

Effects of bone marrow mesenchymal stem cells and its supernate on rats with cerebral infarction

Objective To explore the effect of bone marrow mesenchymal stem cells(BMSCs) and its supernate on rats with cerebral infarction.Methods Forty rats were randomly divided into sodium chloride injection group(9 g·L-1 sodium chloride injection 2 mL),BMSCs transplantation group(2.0×109 L-1 BMSCs suspension 2 mL),BMSCs supernate transplantation group(BMSCs supernate 2 mL) and suspension transplantation group(supernatant and BMSCs suspension 2 mL);ten rats in each group.Twenty-four hours after reperfusion,the endogenous neural precursor cells were labeled by 10 g·L-1 bromodeoxyuridine(BrdU) solution for twenty-eight days,the rats were executed after the last injection two hours.The number of neural precursor cells of multiplication period and neurological severity scores(NSS) were observed in each group.Results The positive cells labeled by BrdU,BrdU+neurone specific enolase,and BrdU+glial fibrillary acidic protein(GFAP) in BMSCs transplantation group,BMSCs supernate transplantation group and suspension transplantation group were more than those in sodium chloride injection group(P<0.05);and the positive cells in BMSCs supernate transplantation group and suspension transplantation group were more than those in BMSCs transplantation group(P<0.05);and the positive cells in suspension transplantation group were more than those in BMSCs supernate transplantation group(P<0.05).Compared with pretransplant,at the seventh,fourteenth and twenty-eighth day after transplantation,the nerve function of rats recovered in different degrees,and the NSS scores were decreased gradually,the differences had statistical significance(P<0.01).Starting from the seventh day,the NSS score in BMSCs transplantation group,BMSCs supernate transplantation group and suspension transplantation group was significantly lower than that in sodium chloride injection group(P<0.05 or P<0.01);the NSS score in suspension transplantation group was significantly lower than that in BMSCs transplantation group and BMSCs supernate transplantation group(P<0.05).Conclusion The supernatant and suspension of BMSCs can induce the proliferation and differentiation of endogenous neural precursor cells in focal zone,which may be related to the neurotrophic factors and cellular differentiation inducing factor in supernatant induced BMSCs differentiated into neural cell.