大黄苷元对骨髓间充质干细胞移植脑缺血大鼠神经细胞和神经营养因子的影响  

Influence of rhubarb aglycone on neurocytes and neurotrophic factors in rats with cerebral ischemia after BMSC transplantation

作　　者：李建生[1] 刘敬霞[1] 李宁[1] 赵跃武[2] 郭晓燕[1] 苏静[1] 孙捷[1] 

机构地区：[1]河南中医学院老年医学研究所,河南450008 [2]河南省人民医院

出　　处：《北京中医药大学学报》2008年第10期668-672,共5页Journal of Beijing University of Traditional Chinese Medicine

基　　金：河南省高校新世纪优秀人才支持计划资助项目(No.2006HANCET-05)

摘　　要：目的观察骨髓间充质干细胞(BMSCs)移植脑缺血大鼠神经细胞及神经营养因子表达的变化以及从大黄苷元对其变化的影响方面探讨二者联合的作用机制。方法体外全骨髓贴壁筛选培养法扩增BMSCs;线栓法制备局灶性脑缺血动物模型(MCAO)模型;术后24h将BMSCs由同侧颈内动脉移植脑内,大黄苷元灌胃用药;免疫组织化学法测定BMSCs移植后7d(早期)、14d(中期)和28d(后期)神经细胞特异性标志兔抗大鼠神经元特异性烯醇化酶抗体(NSE)和兔抗大鼠胶质纤维酸性蛋白抗体(GFAP)及神经营养因子兔抗大鼠神经生长因子抗体(NGF)和兔抗大鼠胶质源性神经营养因子抗体(GDNF)表达变化。结果假手术组大鼠NSE和GFAP表达显著,NGF和GDNF呈弱阳性。模型组7d的GFAP、NGF和GDNF表达增强,14d和28d的GFAP、NGF表达递减,14d的GDNF表达减弱、28d增强。大黄苷元组7d的NSE较模型组表达增强,GFAP减弱,28d的NGF表达增强,7d和14d的GDNF减弱。移植组28d的GFAP、NGF和GDNF表达均较模型组增强。联合组各时间点的NSE表达增强;7d的GFAP表达减弱,14d增强;联合组14d和28d的NGF表达均较大黄苷元组和移植组增强。结论脑缺血再灌注损伤后随时间延长神经元细胞呈递减反应,神经胶质细胞反应先增强再减弱;神经营养因子早期呈反应性增强,此后NGF递减,GDNF出现先减弱再增强的特点。BMSCs移植脑内中后期增强神经营养因子及神经细胞反应的作用明显;大黄苷元可使BMSCs移植后保护神经细胞的时间提前,并使其作用增强,其作用机制可能与上调早期NGF和GDNF及增强中后期NGF的表达有关。Objective To observe the expression changes of neurocytes and neurotrophic factors in rats with cerebral ischemia after of bone mesenchymal stem cells （BMSC） transplantation, and investigate the mechanisms of combination of rhubarb agiycone and BMSC transplantation based on the influence of rhubarb aglycone on these changes. Methods BMSC were purified and increased by using the methods of adherence and screening in vitro. The model of middle cerebral artery occlusion （MCAO） was established with suture method. After 24 hours BMSC were transplanted from internal carotid artery to the brain, and then rhubarb aglycone was used by intragastric administration. The expressions of the specific signs of neurocytes （NSE and GFAP ）, as well as neurotrophic factors （NGF and GDNF ）, weredetermined by using immunohistochemical method on 7^th（ earlier （ anaphase ） day respectively after BMSC transplantation. Results period）, 14^th（metaphase） and 28^th The expressions of NSE and GFAP were significant and expressions of NGF and GDNF showed weak positive in the sham-operation group. The expressions of GFAP, NGF and GDNF all increased on 7^th day, while those of GFAP and NGF decreased on 14^th and 28^th day, and GDNF expression reduced on 14^th day and increased on 28^th day in the model group. In rhubarb aglycone group, compared with the model group, NSE expression increased and GFAP expression decreased on 7^th day, and NGF expression increased on 28^th day, while GDNF expression decreased on 7^th and 14^th day. In he transplantation group, compared with the model group, the expressions of GFAP, NGF and GDNF increased all on 28^th day. In the combined group NSE expression increased at all time points, GFAP expression decreased on 7^th day and increased on 14^th day, and NGF expression, compared with the rhubarb aglylcone group and transplantation group, increased on 14^th and 28^th day. Conclusions Neuron showed a decreased response progressively following the prolongation of the lesion of cerebral is

关 键 词：脑缺血 大黄苷元 骨髓间充质干细胞 神经细胞 神经营养因子 大鼠 

分 类 号：R285.5[医药卫生—中药学]