黄芪注射液联合间质干细胞对大鼠脊髓损伤修复作用的实验研究

作者	单位
余勤	浙江中医药大学生物工程学院
白月双	浙江中医药大学第一临床医学院
林洁	浙江中医药大学生物工程学院
盛丽先	浙江中医药大学第一临床医学院
董勤	浙江中医药大学第一临床医学院
郭莹	浙江中医药大学生物工程学院
宣晓波	浙江中医药大学第一临床医学院

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中文摘要:

目的研究黄芪注射液与间质干细胞联合应用对大鼠脊髓损伤修复的影响。方法Wistar大鼠120只随机分成假手术组、模型组、PBS液对照组、黄芪注射液对照组、大鼠骨髓间质干细胞(rMSCs)移植治疗组和rMSCs移植加黄芪注射液治疗组。造模后3 d治疗组经脊髓局部注射rMSCs和脊髓局部注射rMSCs同时加用黄芪注射液腹腔注射,对照组则分别注射PBS液和黄芪注射液。移植后7、14、21、28天分别行(Basso Beattie Bresnahan,BBB)评分法检测大鼠神经功能恢复情况,进一步取脊髓组织作病理学检查,并应用双标免疫组化检测5-溴-2脱氧尿嘧啶核苷(BrdU)标记rMSCs的胶质纤维酸性蛋白(GFAP)和神经元中丝蛋白(NF-M)表达情况。结果治疗组较对照组损伤脊髓修复明显,神经功能评分显著提高(P<0.05)。rMSCs移植加黄芪注射液治疗组在不同时间点的神经功能评分都高于rMSCs移植治疗组。脊髓组织病理切片显示rMSCs移植加黄芪注射液治疗组较rMSCs移植治疗组组织水肿和炎性细胞浸润减轻更明显,胶质细胞增生更活跃。双标免疫组化显示移植的rMSCs在宿主脊髓中存活,从第7 d开始即有GFAP和NF-M表达并向损伤部位迁移。rMSCs加黄芪注射液治疗组中GFAP、NF-M阳性细胞数目均较rMSCs移植治疗组增多(P<0.05)。结论rMSCs移植是治疗脊髓损伤的有效方法;黄芪注射液在体内有诱导rMSCs向神经细胞分化的能力,并有协同rMSCs促进大鼠脊髓损伤修复的作用。

英文摘要:

Objective To observe the effect of Astragalus Injection(ASI) combined with rat′s mesenchymal stem cells transplantation(rMSCs) for repairing spinal cord injury in rats.Methods One hundred and twenty Wistar rats were randomly divided into 6 groups,named respectively by letters from A-F.they were treated respectively with none,PBS solution,ASI,rMSCs,and ASI+rMSCs,with the ASI administered via intraperitoneal injection and the rMSCs given by local injection to the spinal cord,on the 3rd day of operation.The condition of nerve function recovery was assessed on the 7th,14th,21st and 28th day after treatment by scoring according to the Basso-Beattie-Bresnahan(BBB) locomotor rating scale.Besides,the pathological change of the injured spinal cord was observed and expressions of glial fibriallary acidic protein(GFAP) and neurofilament-M(NF-M) in the BrdU-labeled rMSCs in the spinal cord tissue were examined by immuno histochemistry.Results The recovery of spinal cord in Group D,E and F was better than that in Group B and C,showing higher BBB scores.As compared with Group E,Group F showed a higher score of nerve function and a milder inflammatory cell infiltration with lessened tissue edema in the spinal cord and more active proliferation of gliacyte.Double-labelled immunohistochemical examination showed that the transplanted rMSCs were alive in the host′s spinal cord,revealing the expressions of GFAP and NF-M from the 7th day after transplantation,which were migrating to the injured site.The amount of GFAP and NF-M positive cells in Group F was much more than that in Group E(P<0.05).Conclusion Transplantation of rMSCs is an effective method in treatment of the spinal cord injury;ASI has the capacity for inducing rMSCs differentiated into neurons,and could synergize with rMSCs to promote the repairing of the spinal cord from injury.

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