蛛网膜下腔移植骨髓源干细胞与中药联合治疗对缺血再灌大鼠脑内BDNF的影响

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

目的:研究骨髓间充质干细胞BMSC和骨髓单个核细胞BMNC经蛛网膜下腔移植与口服中药联合应用对缺血再灌注脑组织中脑源性神经营养因子(BDNF)含量的影响。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。③④⑤⑥⑦组应用线栓法建立大鼠大脑中动脉缺血再灌2 h模型(MCAO),24 h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 d、28 d检测脑组织)内BDNF的含量。结果:BMNC组和BMSC组在移植后4 d即可成倍提高脑匀浆内BDNF含量,至28 d时仍较模型组含量高,两组间比较无差异;中药联合BMNC组和中药联合BMSC组在4 d、28 d时均较单纯BMNC或BMSC移植组脑内BDNF含量成倍增高,且中药联合BMNC组在28 d时较中药联合BMSC组水平略高。结论:BMSC和BMNC经蛛网膜下腔移植可明显上调MCAO大鼠脑组织中BDNF水平,中药联合BMNC或BMSC治疗可使BDNF水平成倍提高,比单纯BMNC或BMSC移植更显优势。

Objective：To investigate the influence of traditional Chinese medicine（TCM） on the brain derived neurotrophic factor（BDNF） after treatment stroke rats with bone mesenchymal stem cells（BMSCs） and bone marrow mononuclear cells（BMNCs）.Methods：BMSCs and BMNCs were separated and cultivated.140 adult Wistar rats were randomly divided into 7 groups（n=20）：group1：normal group,group 2：pretent surgery group,group 3：model group,group 4：BMNC group,group 5： BMNCTCM group,group 6：BMSC group,group 7： BMSCTCM group.Group 3,4,5,6,7 were performed（2 hours） middle cerebral arterial occlusion（MCAO） models.100 μL 0.01 MPBS（group 3）,BMNCS（group 4 and 5,2×107）or BMSCS（group 6 and 7,2×107） were injected into rats＇ subarachnoid cavity after 24 h of MCAO corresponded.Group 5 and 7 were treated united with TCM for gavage.BDNF in brain was measured by ELISA at the 4th day and 28th day after transplantation.Result：The BDNF level of model group was increased at the 4th and 28th day.The BDNF levels of group 4,5,6,7 were higher than the model group＇s（P〈0.05） at the 4th and 28th day.The BDNF level between group 4 and 6 was not different.The BDNF levels of group 5 and 7 were significantly higher than that in the group 4 and 6（P〈0.05）.At the 28th day,the BDNF levels of group 7 were higher than that in the group 5（P〈0.05）.Conclusion：Cerebral arterial occlusion can increase the BDNF level in brain of rats.Transplantation of BMNCs/ BMSCs will increase the level further and has obvious advantages than single BMNC or BMSC.