Pharmacological Studies for animal model of Binswanger's disease (encephalopathy)

本研究采用永久性阻断大鼠双侧颈总动脉（2 VO）的方法建立慢性脑低灌注模型。2 VO大鼠表现出学习记忆障碍、白色物质稀疏、胶质标记物增多等特点，提示该模型有可能成为Binswannger病（脑病）模型。本研究应用差异显示RT-PCR技术，对2 VO大鼠脑缺血再灌注损伤的内在因素进行了初步研究。分别在2 VO治疗后4天和4个月从大鼠脑中分离到11和12的表达增加的克隆。对所有这些分离的克隆进行序列分析，其仍然是部分序列，并通过半定量PCR方法定量表达水平。2 VO治疗4天和4个月后表达明显增强的因子分别命名为vof-21和vof-16。vof-21基因全长1,600 bp，vof-16基因全长780 bp。这些是小说，尽管编码位点序列仍然未知。vof-21的表达水平在2 VO后7天达到最高，14天恢复到对照水平。vof-16在海马、顶盖带、梨状皮质和主动脉周围区域表达丰富。用RT-PCR方法鉴定NG 108 -15细胞中vof-16和vof-21的存在。用反义寡核苷酸（asODN）方法研究这两种因子的细胞功能是有用的。对asODN递送的基本检查显示pH敏感脂质体是有用的。我们正在研究vof-16和vof-21的全序列及其细胞功能。asODN脑内转运的基础研究也在进行中。

We have established a chronic cerebral hypoperfusion model that is produced by permanent occlusion of bilateral common carotide arteries (2VO) in rats. The 2VO rats exhibited impairment of learning and memory, rarefaction in the white matter and increase of the glial makers, suggesting that this 2VO model has possibility to be a model of Binswannger's disease (encephalopathy). In this study, we tried to identify the intrinsic factors related with the disease in the 2VO rats using the differentianl display RT-PCR methods. The increased expression clones of 11 and of 12 were isolated form the rat brain 4 days and 4 months after 2VO treatment, respectively. All of these isolated clones were clarified the sequences, which were still partial sequences, and quantified the expression levels by semi-quantitative PCR methods. The factors of which expressions were markedly enhanced by 2VO for 4 days and 4 months were named vof -21 and vof-16, respectively. The sequences of 1,600 bp of vof-21 and 780 bp of vof-16 were clarified. These were novels, although the coding site sequences are still unknown. The expression levels of vof-21 reached maximum at 7 days after 2VO and returned to the control level at 14 days. The expression of vof-16 was abundant in the hippocampus, the tenia tecta, the piriform cortex and the area around the aorta. The existance of both vof-16 and vof-21 in NG108-15 cells were identified by RT-PCR methods. The cells seems to be useful to clarify the cellular functions of both factors by antisenseoligodeoxynucleotides (asODN) methods. The basic examinations for delivery of asODN revealed that pH-sensitive liposomes were useful. We are now studing to clarify the whole sequence and cellular functions of vof-16 and vof-21. Basic examination of delivery of asODN to rat brain are also studying.