中药联合干细胞蛛网膜下腔移植对缺血再灌注大鼠脑脊液NGF的影响

Effect of Transplantation Bone Marrow Mesenchymal Stem Cell Through Subarachnoid Associated with TCM on the Neural Growth Factor in CSF of Cerebal Ischemai Rats

目的:研究中药对蛛网膜下腔移植骨髓单个核细胞(BMNC)或骨髓间充质干细胞(BMSC)治疗缺血性脑卒中大鼠干细胞因子的影响。方法:分离培养BMNCs和BMSCs,140只Wistar大鼠随机分为7组:1正常组,2假手术组,3模型组,4BMNC组,5BMNC联合中药组,6BMSC组,7BMSC联合中药组。3、4、5、6、7组应用线栓法建立大鼠大脑中动脉缺血再灌2h模型(MCAO),24h后通过蛛网膜下腔注射法移植100微升0.01MPBS溶液(3组)、BMNCS悬液(4组、5组,2.0×107个)或BMSCs悬液(6组、7组,2.0×107个),5组、7组当日进行中药灌胃。应用定量ELISA法于移植后4天、28天检测脑脊液(CSF)内Neural growth factor(NGF)含量。结果:模型组CSF中NGF水平在移植后28天有所升高,4、5、6、7组在移植后4dNGF水平均较模型组明显升高(P<0.05),5、6、7组在移植后28天NGF水平均较模型组明显升高。BMNC组和BMSC组间比较没有明显差异。中药联合BMNC组在治疗28天时较单纯BMNC组明显升高,中药联合BMSC组在治疗4天、28天时均较单纯BMSC组增高。两组中药联合组间比较,治疗4天时中药联合BMSC组NGF含量更高。两中药联合治疗组在治疗28天时均较4天水平升高。结论:脑动脉阻塞本身就可以使大鼠脑脊液中NGF水平升高,BMNC和BMSC移植可进一步提高MCAO大鼠CSF中NGF水平,中药联合BMNC或BMSC移植治疗对NGF水平的升高具有协同增效作用。

Objective：Investigate the influence of traditional Chinese medicine（TCM） on the stem cell factor after treatment stroke rats with bone mesenchymal stem cells（BMSCs） and Bone marrow mononuclear cells（BMNCs）.ethods BMSCs and BMNCs were separate and cultivated.140 adult Wistar rats were divided randomly into 7 groups（n=20）：group1：normal group,group2：pretent surgery group,group3：model group,group4：BMNC group,group5： BMNCTCM group,group6：BMSC group,group7： BMSCTCM group.Group 3,4,5,6,7 were performed transmit（2hours） middle cerebral arterial occlusion（MCAO）.100μl 0.01M PBS（group 3）、BMNCS（group 4、5,2×107）or BMSCS（group 6、7,2×107） were injected into rats＇ subarachnoid after 24 h of MCAO corresponded.Group 5 and 7 were treated united with TCM the transplantantion day（po.qd.）.Neural growth factor（NGF） in cerebrospinal fluid（CSF） Elisa analyzed were performed at the 4th day and 28th day after transplantation.Result The NGF level of model group is increased at the 28th day.The NGF levels of group 4,5,6,7 are higher significantly than the model group＇s（P0.05） at the 4th day.At the 28th day,the NGF levels of group 5,6,7 are higher than the model＇s.The NGF level between group 4 and 6 isn＇t different.The level of group 5 is higher than group 4 at the 28th day.The level of group 7 is higher than group 6 at the 4th and 28th day.The level of group 7 is higher than group 5 at the 4th day.At the 28th day the levels of group 5 and 7 are increased in contrast to the 4th day＇s.Conclusion Cerebral arterial occlusion can increase the NGF level in CSF of rats.Transplantantion of BMNCs/ BMSCs will increase the level further.The transplantantion united with TCM inprove the capability of the enhance.