摘要
目的研究骨髓单个核细胞(BMMNCs)对脑梗死小鼠神经元变性死亡的影响。方法将雄性昆明小鼠随机分为PBS治疗组和BMMCs治疗组。线栓法制作小鼠大脑中动脉栓塞(MCAO)模型;梯度离心法提取BMMNCs;酶联免疫吸附法(ELISA)评价常氧及低氧培养条件下BMMNCs分泌IL-10、VEGF和IGF-1的能力。脑梗死后6h经尾静脉注射BMMNCs或PBS,脑梗后1,4,7d采用神经功能损害评分表(mNSS)、免疫荧光法、Fluoro—JadeB(FJB)染色进行神经功能、小胶质细胞活性及神经元变性死亡数目的评估。脑梗死后7d,TTC染色评价脑梗死体积。结果(1)BMMNCs在常氧或低氧条件下培养24h后,低氧环境下BMMNCs分泌细胞因子(IL-10、VEGF和IGF-1)的能力明显强于常氧环境,差异有统计学意义(P〈0.05);(2)与PBS治疗组相比,BMMNCs治疗组小鼠在脑梗死后第4,7天的mNSS评分、小胶质细胞的活性、神经元变性死亡数量均显著降低,差异有统计学意义(P〈0.05);(3)BMMNCs治疗组小鼠脑梗死体积[(27.8±7.5)%]明显小于PBS治疗组[(37.1±6.9)%],差异有统计学意义0P〈0.05)。结论骨髓单个核细胞可通过分泌细胞因子IL-10、VEGF和IGF-1抑制小胶质细胞的活性,明显抑制脑梗死后神经元的变性死亡,减少脑梗死体积,促进神经功能恢复。
Objective To investigate the effects of bone marrow mononuclear cells (BMMNCs) on neuro- degeneration after cerebral ischemia in mice.Methods Adult male mice were randomly divided into PBS treated group and BMMNCs treated group. The model was established with suture emboli method;BMMNCs were isolated with density gradient centrifugation method. The production of cytokines from BMMNCs including interleukin-10 ( IL-10), vascular endothelial growth factor (VEGF) and insuliulike growth factor-I ( IGF-1 ) were tested by en- zyme-linked immunosorbent assay (ELISA) under normoxic or hypoxic conditions.Mice were injected with PBS or BMMNCs via tail vein 6 h after ischemia reperfusion.Neurological functional deficits, the activity of microglial cell and neurodegenration were assessed on day 1,4,7 after MCAO,and infarct volume was assessed on day 7.Results ( 1 ) Compared with BMMNCs under normoxic conditions, BMMNCs under hypoxic conditions secreted more IL-10, VEGF, IGF-1 after 24h of cuhivation(P〈0.05). (2) BMMNCs transplantation significantly reduced neurologic func- tional deficits and the number of activated microglia and degenerated neurons on days 4,7 compared with PBS trea- ted groups (P〈0.05). ( 3 ) Compared with PBS treated group ( ( 37.1 ± 6.9 ) % ), BMMNCs transplantation signifi- cantly reduced infarct volume((27.8±7.5)%) (P〈0.05).Conclusion Bone marrow mononuclear cells can signifi- cantly inhibit neurodegeneration, promote neurological recover and reduce infarct volume through inhibition of mi- croglial cell activation by secretion of cytokines including IL-10,VEGF and IGF-1.
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2013年第11期961-964,共4页
Chinese Journal of Behavioral Medicine and Brain Science
基金
基金项目:国家自然科学基金项目(81271284)