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目的:研究黄芪注射液联合鞘内移植大鼠骨髓间充质干细胞(rMSCs)对大鼠缺氧缺血性脑损伤(HIBD)修复的影响。方法:115只Wistar大鼠随机分成假手术组、模型组、PBS对照组、黄芪注射液对照组、rMSCs治疗组和rMSCs加黄芪注射液治疗组。造模后3天治疗组鞘内注射rMSCs,或同时腹腔注射黄芪注射液,对照组鞘内注射PBS或腹腔注射黄芪注射液,造模后24天行Morris水迷宫实验检测大鼠学习、记忆能力,移植后1、4、7、14、21、28天分别取脑组织作病理学检查,并应用双标免疫组化检测经5-溴-2-脱氧嘧啶(BrdU)标记rMSCs的神经元特异性烯醇化酶(NSE)和胶质纤维酸性蛋白(GFAP)表达情况。结果:治疗组学习、记忆能力较对照组有显著改善(P<0.05),rMSCs加黄芪注射液治疗组第3天平均逃避潜伏期显著低于rMSCs治疗组(P<0.05)。脑组织病理切片显示rMSCs加黄芪治疗组较rMSCs治疗组脑组织水肿、神经细胞变性和坏死程度减轻更明显。双标免疫组化显示rMSCs移植后1天就出现在脑组织,并表达NSE与GFAP。rMSCs加黄芪治疗组BrdU阳性细胞数除移植后14天外均显著多于rMSCs治疗组(P<0.05),NSE阳性率移植后1、7、14天显著高于rMSCs治疗组(P<0.05),GFAP阳性率除移植后14天外均显著高于rMSCs治疗组(P<0.05)。结论:鞘内移植rMSCs是治疗大鼠HIBD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HIBD修复的作用。
Objective: To investigate the effects of Astragalus injection combined with intrathecal transplantation of rat bone marrow mesenchymal stem cells (rMSCs) on the repair of hypoxic-ischemic brain damage (HIBD) in rats. Methods: One hundred and fifteen Wistar rats were randomly divided into sham operation group, model group, PBS control group, astragalus injection control group, rMSCs treatment group and rMSCs plus Astragalus injection group. Three days after modeling, intrathecal rMSCs were injected into the treatment group, or astragalus injection was given intraperitoneally. In the control group, intrathecal injection of PBS or intraperitoneal injection of astragalus injection was performed. Morris water maze test was performed 24 days after the model was established to study the learning and memory abilities At 1, 4, 7, 14, 21 and 28 days after transplantation, the brain tissue was taken for pathological examination, and the neuron specificities of rMSCs labeled with 5-bromo-2-deoxy pyrimidine (BrdU) Enolase (NSE) and glial fibrillary acidic protein (GFAP) expression. Results: The learning and memory abilities of the treatment group were significantly improved compared with the control group (P <0.05). The mean escape latency of rMSCs plus astragalus injection group on the third day was significantly lower than that of the rMSCs treatment group (P <0.05). Pathological sections of brain tissue showed rMSCs plus astragalus treatment group than rMSCs treatment group brain edema, nerve cell degeneration and necrosis reduced more significantly. Double-labeled immunohistochemistry showed that rMSCs appeared in brain tissue one day after transplantation and expressed NSE and GFAP. The number of BrdU positive cells in rMSCs plus astragalus group was significantly higher than those in rMSCs treated group (P <0.05) 14 days after transplantation. The positive rate of NSE in rMSCs plus astragalus group was significantly higher than that in rMSCs treated group (P <0.05) The positive rate of GFAP was significantly higher than that of rMSCs treatment group (P <0.05) except 14 days after transplantation. Conclusion: Intrathecal transplantation of rMSCs is an effective method for the treatment of HIBD in rats. Astragalus injection induces the differentiation of rMSCs into neural cells in vivo and has synergistic effect of rMSCs on the repair of HIBD in rats.