经鼻通路给予bFGF诱导在体成年大鼠SVZ区
南京军区南京总医院神经内科，南京大学医学院临床学院 南京 (210002)
【摘要】
目的：bFGF，即FGF-2 或者heparin-bingding growth factor 2 (hbgf-2), prostatropin, 是一种具有多潜能的多肽类营养因子，它对于血管生成及干细胞分化诱导都具有一定的作用。由于血脑屏障的存在，使具有生物效应的蛋白分子经静脉途径给药后不能到达中枢神经系统内环境。本实验运用不同的方法观察比较用静脉，经鼻等不同途径给药后bFGF在大鼠不同脑区的分布，并检测进入脑区的bFGF对正常大鼠及MCAO成年缺血大鼠SVZ区干细胞分化及迁移的影响。
方法：首先，bFGF以经鼻途径，静脉给药等在体方式给予正常及MCAO成年大鼠1 h后，分别用放免标记、免疫组化及蛋白印迹等不同方法检测并比较嗅球及SVZ区bFGF的存在及其含量；然后，将大鼠随机分为正常对照组，正常大鼠给药组，MCAO组，及MCAO后给药组，在给药或MCAO术后不同时相点(1，4，7，14 d)当天予以BrdU腹腔注射(100mg/kg)，9-10 h后灌注处死，取嗅球横切及SVZ区冠状切面，用免疫荧光及免疫共聚焦法观察以BrdU及干细胞特异性抗体双标法标记的干细胞及其分化和分布。
, 百拇医药
结果：我们发现不同在体方式给药1 h后，放免标记、免疫组化及蛋白印迹实验均证实，经鼻给药后1 h，嗅球及SVZ区bFGF浓度均高于静脉给药组及对照组。同时，在不同处理组中，与正常对照组相比，MCAO组，MCAO后给药组，及正常大鼠给药组均有不同程度的干细胞分化和迁移。其中，MCAO后给药组大鼠干细胞分化及迁移率显著高于其他分组(P<0.01)。
结论：我们的结果证实了以经鼻方式给予bFGF可以到达嗅球及SVZ区，与经静脉方式相比，有更高的局部血药浓度；因此，经鼻方式给药可以降低药物用量并得到理想的治疗效果，同时，可以降低因药物用量过大而在外周引起的毒付作用。另外，我们的实验证实以经鼻给药方式进入嗅球及SVZ区的bFGF可以诱导正常大鼠及MCAO大鼠局部成体静止干细胞分化并发生相应的迁移。
【关键词】 MCAO bFGF 经鼻给药
, 百拇医药
Intranasal delivery of inductive agents to rats leads to in vivo
proliferation and differentiating of adult neural
progenitor cells in subventricular zone in rats
MA Haihan , LIU Xinfeng , GUO Ruibing , ZHANG Renliang , WU Wentao, MAO Xiaowei
Department of Neurology, Jinling Hospital, Medical college of Nanjing University, Nanjing, China, 210002
, http://www.100md.com
【Abstract】
Objective: Basic fibroblast growth factor is a polypeptide not only with potent multi-potential trophic effects but also stem cell inducing effects. This study was carried out to investigate the distribution of bFGF by several administration methods and the general pharmacological effects of bFGF on proliferation and differentiating of adult neural progenitor cells in vivo in sub-ventricular zone.
Methods: Firstly the bFGF was delivered to the rats by several in vivo methods including intravenous administration, intranasal delivery, the radio-immunity methods, immunohistochemistry technique and Western blotting methods were used to detect the distribution of the delivered agents; then mature SD rats received, with or without, intranasal administration of bFGF, previously or immediately after focal ischemia infarcts were made in the right lateral cerebral cortex by permanent distal middle cerebral artery occlusion. Intraoperative measurements of core temperature, arterial blood pressure and blood geses, blood glucose concentration, and hematocrit and postoperative measurements of temperature revealed no differences among bFGF-treated and MCAO animals. 1, 4, 7, 14 days later, animals were killed 8.5-10 hours after BrdU injection.
, http://www.100md.com
Results: Using several different in vivo assays, we found that the bFGF concentrations in olfactory bulb and sub-ventricular zone are higher by intranasal than intravenous. And using anti-BrdU and different special stem cell antibodys by the immno-fluorescence double-staining methods we assess that the intranasal group showed more double-stained neural progenitor cells than the intravenous group; Also we found the several migration stem cells from the sub-ventricular zone.
Conclusion: Our study showed that intranasal bFGF in vivo could induce the proliferation and differentiating and even migration of the adult progenitor cells in adult sub-ventricular zone.
【Key words】bFGF intranasal middle cerebral artery occlusion, http://www.100md.com(马海涵 刘新峰 郭芮兵 张仁良 吴文涛 毛晓薇)
【摘要】
目的：bFGF，即FGF-2 或者heparin-bingding growth factor 2 (hbgf-2), prostatropin, 是一种具有多潜能的多肽类营养因子，它对于血管生成及干细胞分化诱导都具有一定的作用。由于血脑屏障的存在，使具有生物效应的蛋白分子经静脉途径给药后不能到达中枢神经系统内环境。本实验运用不同的方法观察比较用静脉，经鼻等不同途径给药后bFGF在大鼠不同脑区的分布，并检测进入脑区的bFGF对正常大鼠及MCAO成年缺血大鼠SVZ区干细胞分化及迁移的影响。
方法：首先，bFGF以经鼻途径，静脉给药等在体方式给予正常及MCAO成年大鼠1 h后，分别用放免标记、免疫组化及蛋白印迹等不同方法检测并比较嗅球及SVZ区bFGF的存在及其含量；然后，将大鼠随机分为正常对照组，正常大鼠给药组，MCAO组，及MCAO后给药组，在给药或MCAO术后不同时相点(1，4，7，14 d)当天予以BrdU腹腔注射(100mg/kg)，9-10 h后灌注处死，取嗅球横切及SVZ区冠状切面，用免疫荧光及免疫共聚焦法观察以BrdU及干细胞特异性抗体双标法标记的干细胞及其分化和分布。
, 百拇医药
结果：我们发现不同在体方式给药1 h后，放免标记、免疫组化及蛋白印迹实验均证实，经鼻给药后1 h，嗅球及SVZ区bFGF浓度均高于静脉给药组及对照组。同时，在不同处理组中，与正常对照组相比，MCAO组，MCAO后给药组，及正常大鼠给药组均有不同程度的干细胞分化和迁移。其中，MCAO后给药组大鼠干细胞分化及迁移率显著高于其他分组(P<0.01)。
结论：我们的结果证实了以经鼻方式给予bFGF可以到达嗅球及SVZ区，与经静脉方式相比，有更高的局部血药浓度；因此，经鼻方式给药可以降低药物用量并得到理想的治疗效果，同时，可以降低因药物用量过大而在外周引起的毒付作用。另外，我们的实验证实以经鼻给药方式进入嗅球及SVZ区的bFGF可以诱导正常大鼠及MCAO大鼠局部成体静止干细胞分化并发生相应的迁移。
【关键词】 MCAO bFGF 经鼻给药
, 百拇医药
Intranasal delivery of inductive agents to rats leads to in vivo
proliferation and differentiating of adult neural
progenitor cells in subventricular zone in rats
MA Haihan , LIU Xinfeng , GUO Ruibing , ZHANG Renliang , WU Wentao, MAO Xiaowei
Department of Neurology, Jinling Hospital, Medical college of Nanjing University, Nanjing, China, 210002
, http://www.100md.com
【Abstract】
Objective: Basic fibroblast growth factor is a polypeptide not only with potent multi-potential trophic effects but also stem cell inducing effects. This study was carried out to investigate the distribution of bFGF by several administration methods and the general pharmacological effects of bFGF on proliferation and differentiating of adult neural progenitor cells in vivo in sub-ventricular zone.
Methods: Firstly the bFGF was delivered to the rats by several in vivo methods including intravenous administration, intranasal delivery, the radio-immunity methods, immunohistochemistry technique and Western blotting methods were used to detect the distribution of the delivered agents; then mature SD rats received, with or without, intranasal administration of bFGF, previously or immediately after focal ischemia infarcts were made in the right lateral cerebral cortex by permanent distal middle cerebral artery occlusion. Intraoperative measurements of core temperature, arterial blood pressure and blood geses, blood glucose concentration, and hematocrit and postoperative measurements of temperature revealed no differences among bFGF-treated and MCAO animals. 1, 4, 7, 14 days later, animals were killed 8.5-10 hours after BrdU injection.
, http://www.100md.com
Results: Using several different in vivo assays, we found that the bFGF concentrations in olfactory bulb and sub-ventricular zone are higher by intranasal than intravenous. And using anti-BrdU and different special stem cell antibodys by the immno-fluorescence double-staining methods we assess that the intranasal group showed more double-stained neural progenitor cells than the intravenous group; Also we found the several migration stem cells from the sub-ventricular zone.
Conclusion: Our study showed that intranasal bFGF in vivo could induce the proliferation and differentiating and even migration of the adult progenitor cells in adult sub-ventricular zone.
【Key words】bFGF intranasal middle cerebral artery occlusion, http://www.100md.com(马海涵 刘新峰 郭芮兵 张仁良 吴文涛 毛晓薇)