人参皂甙Rg1促进嗅鞘细胞迁移及修复脊髓损伤的实验研究

发布时间：2018-05-31 08:26

  本文选题：嗅鞘细胞 + 人参皂甙Rg1　； 参考：《苏州大学》2015年硕士论文

【摘要】：第一部分:人参皂甙Rg1对嗅鞘细胞迁移能力的影响目的:探讨人参皂甙Rg1对嗅鞘细胞(OECs)迁移能力及迁移相关因子基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)及神经细胞黏附分子1(NCAM1)表达的影响。方法:从新生3~5d的SD大鼠嗅球取材获得嗅鞘细胞体外培养、纯化后,进行人参皂甙Rg1干预(40μg/ml浓度下干预72 h)并设置对照(加等量DMEM/F12培养基),通过划痕实验和Transwell小室实验检测人参皂甙Rg1对嗅鞘细胞迁移能力的影响。反转录-聚合酶链反应(RT-PCR)分别测定两组细胞迁移相关因子MMP-2、MMP-9和NCAM1的m RNA表达。通过Western-blot研究Rg1对嗅鞘细胞PI3K/Akt通路的影响。结果:Rg1组划痕愈合面积明显高于对照组(P0.05);Rg1组嗅鞘细胞穿过隔离膜的细胞数量较对照组明显增多(P0.05);RT-PCR结果表明:人参皂甙Rg1显著上调嗅鞘细胞MMP-2、MMP-9和NCAM1的m RNA表达(P0.05)。Rg1能够促进嗅鞘细胞PI3K/Akt通路的活化。结论:人参皂甙Rg1通过上调嗅鞘细胞迁移相关因子MMP-2、MMP-9和NCAM1的表达,提高细胞的迁移能力,这一作用可能通过PI3K/Akt通路完成。第二部分:人参皂甙Rg1促进嗅鞘细胞修复脊髓损伤的实验研究目的:研究人参皂甙Rg1对嗅鞘细胞(olfactory ensheathing cells,OECs)移植修复脊髓损伤能力的影响。方法:新生SD大鼠嗅球组织体外培养、纯化嗅鞘细胞,人参皂甙Rg1干预并设置对照,Allen打击建立大鼠脊髓损伤模型,随机分成三组,对照组(control)、嗅鞘细胞移植组(OECs组)和Rg1干预嗅鞘细胞移植组(Rg1+OECs组),于移植后第1、3、7、14、28天分别进行肢体活动BBB评分,并于第28d取损伤部位脊髓组织制作石蜡切片进行免疫组化检测;RT-PCR分别测定OECs组和Rg1+OECs组脊髓组织中CNTF和VEGF的m RNA表达。结果:Rg1+OECs组后肢运动恢复情况优于OECs组和对照组,组织切片示Rg1+OECs组神经元数量明显高于OECs组和对照组(P0.01),GFAP表达量显著低于OECs组和对照组(P0.01),HE染色示Rg1+OECs组脊髓组织结构清晰,囊性病变及细胞水肿坏死数量明显少于其它各组;RT-PCR结果显示人参皂甙Rg1显著上调嗅鞘细胞在移植宿主体内CNTF、VEGF的m RNA表达(P0.05)。结论:人参皂甙Rg1能够通过上调嗅鞘细胞在宿主体内CNTF、VEGF的表达,提高嗅鞘细胞移植修复脊髓损伤的能力。
[Abstract]:Part one: effects of ginsenoside Rg1 on the migration of olfactory ensheathing cells objective: to investigate the migration ability of ginsenoside Rg1 on olfactory ensheathing cells (OECs) and the migration related factors matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and nerve cells. The effect of NCAM1) on the expression of NCAM1. Methods: olfactory ensheathing cells were cultured from olfactory bulb of SD rats for 3 days. Ginsenoside Rg1 was used to intervene at 40 渭 g/ml for 72 h. The effect of ginsenoside Rg1 on the migration of olfactory ensheathing cells was detected by scratch test and Transwell chamber experiment. Reverse transcription-polymerase chain reaction (RT PCR) was used to detect the expression of MMP-2, MMP-9 and NCAM1, respectively. The effect of Rg1 on the PI3K/Akt pathway of olfactory ensheathing cells was studied by Western-blot. Results compared with control group, the number of olfactory ensheathing cells passing through the isolation membrane was significantly higher in group 1 than that in group Rg1. The RT-PCR results showed that ginsenoside Rg1 could significantly up-regulate the expression of m RNA of MMP-2, MMP-9 and NCAM1 in olfactory ensheathing cells. It can promote the activation of PI3K/Akt pathway in olfactory ensheathing cells. Conclusion: ginsenoside Rg1 can increase the migration ability of olfactory ensheathing cells by up-regulating the expression of MMP-2, MMP-9 and NCAM1, which may be accomplished by PI3K/Akt pathway. Part two: experimental study of ginsenoside Rg1 promoting olfactory ensheathing cells to repair spinal cord injury objective: to study the effect of ginsenoside Rg1 on the ability of olfactory ensheathing cells to repair spinal cord injury. Methods: the olfactory bulb tissue of newborn SD rats was cultured in vitro, the olfactory ensheathing cells were purified, ginsenoside Rg1 was used to intervene and the control group Rg1 was used to set up the spinal cord injury model of rats, and they were randomly divided into three groups. Control group, olfactory ensheathing cell transplantation group (OECs group) and Rg1 intervention olfactory ensheathing cell transplantation group (Rg1 OECs group) were evaluated for limb activity BBB scores on day 1,1428 after transplantation. On the 28th day, paraffin sections were taken from the injured spinal cord to detect the m RNA expression of CNTF and VEGF in the spinal cord tissues of OECs group and Rg1 OECs group, respectively. Results the recovery of hind limb movement in the OECs group was better than that in the OECs group and the control group. The number of neurons in the Rg1 OECs group was significantly higher than that in the OECs group and the control group. The expression of GFAP in the Rg1 OECs group was significantly lower than that in the OECs group and the control group. The results of HE staining showed that the structure of the spinal cord in the Rg1 OECs group was clear. The results of RT-PCR showed that ginsenoside Rg1 upregulated the expression of m RNA in olfactory ensheathing cells. Conclusion: ginsenoside Rg1 can improve the ability of olfactory ensheathing cell transplantation to repair spinal cord injury by upregulating the expression of CNTF-VEGF in olfactory ensheathing cells in host.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2

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