补阳还五汤联合鞘内注射间充质干细胞治疗脊髓损伤的研究  

Bu yang Huan wu Decoction Combined with Mesenchymal Stem Cell Intrathecal Inject Transplantation for Treating Spinal Cord Injury in Rats

作　　者：徐志华[1] 余勤[2] 白月双[1] 宣晓波[1] 盛丽先[1] 李佩佩[1] 王标[2] 单威[2] 周丽萍[2] 刘伟[2] 

机构地区：[1]浙江中医药大学第一临床医学院,浙江杭州310053 [2]浙江中医药大学生物工程学院,浙江杭州310053

出　　处：《中华中医药学刊》2012年第12期2726-2729,I0029,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省中医药科技计划项目(2009CA006)

摘　　要：目的:探讨补阳还五汤联合鞘内注射大鼠间充质干细胞(rMSCs)对大鼠脊髓损伤(SCI)的修复作用。方法:将66只Wistar大鼠,随机分成模型组、假手术组、PBS对照组(简称对照组)、补阳还五汤组(简称中药组)、鞘内注射rMSCs组(简称移植组)和补阳还五汤联合鞘内注射rMSCs组(简称联合组)。造模后3天,对照组鞘内注射PBS液,中药组予以补阳还五汤灌胃30天,移植组鞘内注射rMSCs,联合组鞘内注射rMSCs同时给予补阳还五汤灌胃30天。治疗后第7、14、21、28天分别行BBB分级法检测神经功能恢复情况,HE染色法观察脊髓损伤病理改变和修复情况,应用免疫组化技术检测BrdU标记的rMSCs胶质纤维酸性蛋白(GFAP)和神经元特异烯醇化酶(NSE)表达情况。结果:成功制作了大鼠脊髓半横断损伤模型,移植组和联合组BBB分级法检测平均得分较模型组、对照组和中药组有显著提高(P<0.05),联合组在不同时间点的BBB平均得分都高于移植组。脊髓组织病理切片显示联合组较移植组组织水肿和炎性细胞浸润减轻更明显,胶质细胞增生更加活跃。双标免疫组化显示移植的rMSCs可迁移到宿主脊髓损伤处并存活,从第7天开始即有NSE和GFAP表达。联合组中GFAP、NSE阳性细胞数目均较移植组多(P<0.05)。结论:鞘内注射移植rMSCs对大鼠脊髓损伤具有修复作用,联合应用补阳还五汤有协同增效的效果,有利于大鼠脊髓功能的恢复。Objective ： To observe the effect of Bu yang Huan wu decoction combined with rMSCs transplanted intrathe cally on the recovery of motor function of rats with spinal cord injury（ SCI）, and analyze its possibly mechanism. Meth ods ：66 Wistar rats were randomly divided into model group（ Group A）, sham -operated group （Group B）, PBS solution control group （Group C）, Bu yang Huan wu decoction treated group （Group D）, rMSCs treated group （Group E） and Bu yang Huan wu decoction combined with rMSCs treated group （ Group F）. 3 days after the models were madeed, Group C was intrathecally injected with PBS solution, Group D was fed with Bu yang Huan wu decoction daily for 30 days, Group E was transplanted rMSCs by intrathecal injection, and Group F was treated with both rMSCs and Bu yang Huan wu decoction. On the 7 th 14th 21th and 28th day after treated, nerve function was tested by BBB locomotor rating scale （BBB） , The pathological change and tissue recovery of the SCI were observed by HE staining, expression of neu- ron - specific enolase （NSE） or glial fibers acid protein （GFAP） on rMSCs were showed by double - labeled immunohis- tochemistry （IHC） . Results ： Hemisection spinal cord injury model in rats had been made successfully , The average BBB scores of the Group E and Group F had been increased significantly（P 〈 0.05 ）Compared with the Group A Group B and Group D . The average scores of the Group F were significantly higher than the Group E at every time point . The patho- logical of spinal cord showed that . Compared with the Group E, edema and infiltration of inflammatory cells were de- creased significantly and hyperplasia of glial cells were more active in Group F. The double - labeled IHC showed rMSCstransplanted could migrated to the injury site and survived, from the 7th day on, they were found some of the Brdu -la- beled cells were reactive for NSE or GFAP. There were more GFAP and NSE positive cells in the Group F than in the Group E（P 〈 O. 05）

关 键 词：脊髓损伤 间充质干细胞 移植 补阳还五汤 

分 类 号：R285.5[医药卫生—中药学]