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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 study the effect of BMSC and BMNC on the content of brain-derived neurotrophic factor (BDNF) in rat brain after ischemia-reperfusion by subarachnoid transplantation and oral Chinese herbs. Methods: BMNCs and BMSCs were isolated and cultured. 140 Wistar rats were randomly divided into seven groups: ① normal group, ② sham operation group, ③ model group, ④ BMNC group, ⑤ BMNC combined with traditional Chinese medicine group, ⑥ BMSC group and ⑦ BMSC combined with traditional Chinese medicine group. Rats in group③④⑤⑥⑦ were subjected to MCAO (middle cerebral artery occlusion) and MCAO (middle cerebral artery occlusion) by thread occlusion method. After 24 h, 100 microliters of 0.01MPBS solution (group ③) and (4) Group, ⑤ group, 2.0 × 107) or BMSCs suspension (⑥ group, ⑦ group, 2.0 × 107), ⑤ group, ⑦ group on the day of gavage. The content of BDNF in the brain tissue was detected by quantitative ELISA at 4 days and 28 days after transplantation. Results: BMNC group and BMSC group doubled BDNF content in brain homogenate 4 d after transplantation, and remained higher than model group at 28 d after transplantation. There was no significant difference between the two groups in BMNC group and BMSC group. Compared with BMNC group or BMSC group, BDNF content in brain increased by 2 fold at 4 d and 28 d, and the level of BDNF in Chinese BMNC group was slightly higher than that of BMSC group at 28 d. CONCLUSIONS: BMSC and BMNC can significantly increase the level of BDNF in the brain tissue of MCAO rats after subarachnoid transplantation. The combination of BMNC and BMSC may double the level of BDNF, which is more superior than that of BMNC or BMSC alone.