Bone marrow mesenchymal stem cells improve learning ability of the aging rat

doi:10.3969/j.issn.2095-4344.0506

Abstract

Abstract:

BACKGROUND: At present, studies have shown that bone marrow mesenchymal stem cells (BMSCs) have self-renewal ability, which can be used as ideal seed cells for repairing tissue and organ damages caused by aging and lesions.
OBJECTIVE: To study the changes in the levels of oxidation, inflammatory factors and neurotrophic factors (BDNF) in the brain of aging rats undergoing BMSCs transplantation, and to analyze the mechanism underlying the repair of learning and memory ability in the aging rats.
METHODS: A total of 30 clean Sprague-Dawley rats were randomly divided into control group, model group and BMSCs group, 10 rats in each group. Aging models were made in the rats by 3-month subcutaneous injection of D-galactose. After modeling, BMSCs treatment was performed via tail vein injection in the BMSCs group. The injection was performed once a week, for 8 continuous weeks. Morris water maze was used to detect the learning and memory abilities of the rats in each group after the final injection of BMSCs. Superoxide dismutase activity in the brain tissue of rats was detected by xanthine oxidase method. Level of malondialdehyde in the rat brain tissue was detected by thiobarbituric acid method. Total antioxidant capacity of the brain tissue was detected by Fe3+ reduction method. Real-time PCR and western blot assay were used to detect the expression of brain-derived neurotrophic factor mRNA and protein in the brain tissue of the aging rat, respectively.
RESULTS AND CONCLUSION: Compared with the model group, the BMSCs group exhibited significantly higher activity of superoxide dismutase, stronger total antioxidant capacity, and higher levels of brain-derived neurotrophic factor mRNA and protein (P < 0.05), but the lower malondialdehyde level in the brain (P < 0.05). Compared with the model group, there was less time and higher frequency for passing through the platform in the BMSCs group (P < 0.05). Our findings further indicate that BMSCs can improve the abilities of learning and memory in aging rats, and the underlying mechanism is likely to improve antioxidant capacity and to regulate the level of brain-derived neurotrophic factors.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cell Transplantation, Aging, Brain-Derived Neurotrophic Factor, Tissue Engineering

CLC Number:

R394.2

Liu Yang, Wang Fei-qing, Liu Yan-qing, Li Hong-ri, Zhang Bo, Li Yan-ju. Bone marrow mesenchymal stem cells improve learning ability of the aging rat[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(13): 1999-2004.

Figures/Tables 1

2.1 骨髓间充质干细胞的培养形态以及鉴定结果刚接种时，于显微镜下观察可见悬浮于其中的大量骨髓细胞，无贴壁细胞；第3天换液，大量的造血干血细胞被除去，于显微镜下可观察到培养瓶底壁贴壁的细胞分布不均，部分呈巢状生长；第6天可见形成多个成纤维样细胞集落，10 d左右细胞融合80%-90%。传代培养后细胞形态基本上趋于一致，光镜下见细胞呈星形，梭形，见图1。经流式细胞仪检测，第3代骨髓间充质干细胞的特异性表面标志物CD105，CD90，CD73呈阳性表达，阳性率分别为99.81%，99.62%，96.25%；CD11b，CD45，HLA-DR，CD19，CD34呈阴性表达，阴性率分别为0.11%，0.22%，0.08%，0.07%，0.03%，提示第3代贴壁细胞为骨髓间充质干细胞，见图2。 2.2 实验动物数量分析 30只SD大鼠均进入结果分析，无脱失。 2.3 水迷宫检测大鼠学习记忆能力变化通过水迷宫空间探索实验，模型组和细胞治疗组大鼠首次穿越平台时间明显大于对照组，差异有显著性意义(P < 0.05)。与模型组比较，细胞治疗组大鼠穿越平台时间减少，差异有显著性意义(P < 0.05)。穿越平台次数对照组明显多于模型组和细胞治疗组，差异有显著性意义(P < 0.05)。与模型组比较，细胞治疗组大鼠穿越平台次数明显多于模型组，差异有显著性意义(P < 0.05)，见表2。 2.4 光镜观察大鼠脑组织病理形态 200倍光镜下观察，对照组可见形态规则的尼氏小体、星形细胞，核仁清晰，未见衰老现象(图3A)。模型组大鼠脑组织尼氏小体明显减少，部分核仁固缩边缘化，细胞出现肿胀，呈现明显衰老的现象(图3B)。细胞治疗组大鼠脑组织尼氏小体增加，核仁清晰，脑组织未见明显肿胀衰老迹象，与模型组比较明显改善(图3C)。 2.5 衰老大鼠脑组织超氧化物歧化酶、总抗氧化能力和丙二醛水平模型组和细胞治疗组大鼠脑组织超氧化物歧化酶活性低于对照组，模型组与对照组比较差异有显著性意义(P < 0.05)。细胞治疗组大鼠脑组织超氧化物歧化酶活性与模型组比较，差异无显著性意义(P > 0.05)。模型组和细胞治疗组大鼠脑组织丙二醛水平高于对照组 (P < 0.05)，细胞治疗组大鼠脑组织丙二醛水平低于模型组(P < 0.05)。模型组和细胞治疗组大鼠脑组织总抗氧化能力水平低于对照组(P < 0.05)，细胞治疗组大鼠脑组织总抗氧化能力水平高于模型组(P < 0.05)，见表3。 2.6 Real-time PCR测定脑组织中BDNF mRNA表达与对照组比较，模型组和细胞治疗组大鼠脑组织BDNF mRNA的表达降低，差异有显著性意义(P < 0.05)。与模型组比较，细胞治疗组大鼠脑组织BDNF mRNA升高，差异有显著性意义(P < 0.05)，见表4。 2.7 Western Blot检测脑组织BDNF蛋白表达与对照组比较，模型组和细胞治疗组大鼠脑组织BDNF蛋白表达明显降低，差异有显著性意义(P < 0.05)。与模型组比较，细胞治疗组大鼠脑组织BDNF蛋白表达升高，差异有显著性意义(P < 0.05)，见表5。"

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