Preelinical analyses of the therapeutic potential of allopregnanolone to promote neurogenesis in vitro and in vivo in transgenic mouse model of Alzheimer's disease
Preelinical analyses of the therapeutic potential of allopregnanolone to promote neurogenesis in vitro and in vivo in transgenic mouse model of Alzheimer's disease
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DOI：
10.2174/156720506775697160
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发表时间：
2006-02-01
影响因子：
2.1
通讯作者：
Wang, Jun Ming
中科院分区：
文献类型：
--
作者：
Brinton, Roberta Diaz;Wang, Jun Ming
在这里，我们提供的数据支持临床前的概念证明，即别孕酮促进神经发生的治疗潜力。我们最近的工作表明，神经活性孕酮代谢产物别孕酮(3α-羟基-5α-孕酮-20-酮)(APα)以剂量依赖的方式诱导大鼠海马神经前体细胞(NPC)和大脑皮层神经干细胞(HNSM)的增殖显著增加[1]。通过掺入BrdU和H-3-胸腺嘧啶核苷、流式细胞仪分析MuLV-GFP标记的有丝分裂神经前体细胞和细胞总数来确定细胞的增殖效率。别孕酮诱导的细胞增殖是异构体和类固醇特异性的，立体异构体3β-羟基-5β-孕烷-20-酮和相关类固醇不增加H-3-胸腺嘧啶核苷摄取。对鼻咽癌标志物Nestin和Tuj1的免疫荧光分析表明，新形成的细胞属于神经元谱系。此外，细胞周期基因的微阵列分析、实时RT-PCR和Western印迹验证表明，别孕酮增加了促进有丝分裂的基因的表达，抑制了抑制细胞增殖的基因的表达。电压门控型L钙通道阻滞剂硝苯地平可拮抗别孕酮诱导的海马神经元细胞增殖，这与别孕酮通过GABA型受体激活的L型钙通道引起海马神经元细胞内钙快速升高的发现一致。初步的吗？活体数据显示，通过对3月龄雄性三重转基因阿尔茨海默病小鼠的BrdU阳性细胞的无偏向体视学分析，24小时的APA显著增加了齿状回的神经发生。APα在体外和体内的成神经性，加上低分子量、易穿透血脑屏障和无毒性，是开发APα作为神经发生/再生疗法用于阿尔茨海默病患者神经元修复所需的关键因素。
Herein, we present data to support a preclinical proof of concept for the therapeutic potential of allopregnanolone to promote neurogenesis. Our recent work has demonstrated that the neuroactive progesterone metabolite, allopregnanolone (3 alpha-hydroxy-5 alpha-pregnan-20-one), (AP alpha) induced, in a dose dependent manner, a significant increase in proliferation of neuroprogenitor cells (NPCs) derived from tile rat hippocampus and human neural stein cells (hNSM) derived from, the cerebral cortex [1]. Proliferative efficacy was determined by incorporation of BrdU and H-3-thymidine, FACS analysis of MuLV-GFP-labeled mitotic NPCs and quantification of total Cell number. Allopregnanolone-induced proliferation was isomer and steroid specific, in that the stereoisomer 3 beta-hydroxy-5 beta-pregnan-20-one and related steroids did not increase H-3-thymidine uptake. Immunofluorescent analyses for the NPC markers, nestin and Tuj1, indicated that newly formed cells were of neuronal lineage. Furthermore, microarray analysis of cell cycle genes and real time RT-PCR and western blot validation revealed that allopregnanolone increased tile expression of genes which promote mitosis and inhibited the expression of genes that repress cell proliferation. Allopregnanolone-induced proliferation was antagonized by the voltage gated L-type calcium channel blocker nifedipine consistent with the finding that allopregnanolone induces a rapid increase in intracellular calcium in hippocampal neurons via a GABA type A receptor activated L-type calcium channel. Preliminary it? vivo data indicate that APa for 24 hrs significantly increased neurogenesis in dentate gyros, as determined by unbiased stereological analysis of BrdU positive cells, of 3-month-old male triple transgenic Alzheimer's disease mice. The in vitro and in vivo neurogenic properties of AP alpha coupled with a low molecular weight, easy penetration of the blood brain barrier and lack of toxicity, are key elements required for developing AP alpha as a neurogenic/regenerative therapeutic for restoration of neurons in victims of Alzheimer's disease.