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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 investigate the effect of Buyang Huanwu Decoction combined with intrathecal injection of rat mesenchymal stem cells (rMSCs) on the repair of spinal cord injury (SCI) in rats. Methods: Sixty-six Wistar rats were randomly divided into three groups: model group, sham operation group, PBS control group (abbreviated as control group), Buyang Huanwu Decoction group (abbreviated as TCM group), intrathecal rMSCs group Buyang Huanwu Decoction combined with intrathecal rMSCs group (referred to as the combination group). Three days after model establishment, the control group received intrathecal injection of PBS solution. The Chinese herb group was treated with Buyang Huanwu Decoction for 30 days. The rMSCs in the transplantation group were intrathecally injected with rMSCs. day. At 7, 14, 21 and 28 days after treatment, the recovery of neurological function was detected by BBB grading, the pathological changes and repair of spinal cord injury were observed by HE staining, and the BrdU labeled rMSCs glial fibrillary acidic protein GFAP) and neuron specific enolase (NSE) expression. Results: The rat model of spinal cord transection was successfully established. The mean score of BBB grading test in the transplantation group and the combined group was significantly higher than that in the model group, the control group and the traditional Chinese medicine group (P <0.05). The BBB The average score was higher than the transplantation group. The pathological sections of spinal cord showed that the edema and inflammatory cell infiltration in the combined group were more obvious than those in the transplantation group, and glial cell proliferation was more active. Double-labeled immunohistochemistry showed that the transplanted rMSCs could migrate to the host spinal cord injury site and survived, and the expression of NSE and GFAP began from the 7th day. The number of GFAP and NSE positive cells in the combined group was more than that in the transplantation group (P <0.05). Conclusion: Intrathecal injection of rMSCs can repair spinal cord injury in rats. Combined with Buyang Huanwu decoction has the synergistic effect, which is beneficial to the recovery of spinal cord function in rats.