Isolation And Characterization Of Novel Genes And Expression Of Relational Genes With Learn And Memory In The Hippcampus Of Vascular Dementia Rats After Acupuncture

Although more and more research papers mention the Vascular Dementia (VD), its mechanism has not been understood completedly. In recent years, studies of the acupuncture therapy of VD have shown that acupuncture could increase the blood flow of the brain microvascular vessel and improve learning and memory capability of VD patients. However, a few of reports covered the molecule mechanism of the acupuncture therapy of VD. Therefore, this study aims mainly to the molecule mechanism of VD and the molecule mechanism of the acupuncture therapy of VD. The study used the Vascular Dementia Model of rat (VDM) which was formed by 4-vessel occlusion method. According to the theories of Traditional Chinese Medicine and the papers of clinic studies, the study choosed three acupuncture points, baihi and shenyu and zusanli, to treat VDM. RNA fingerprinting, RT-PCR, immunohistochemistry, histochemistry, in situ hybridization, behaviour model such as Morris water maze were used in the study. The results are as following:First: The vascular dementia model of rat was performed by 4-vessel occlusion method. The learning and memory of VD rats were tested by Morris water maze. VDM was successful reproduced. It could copy the clinical feature of human VD and showed clearly the defection of learning and memory. There were selective neuronal damage in some regions of brain in VDM, such as hippcampus.Second: It is availably that acupuncture treat VDM on baihi and shenyu and zusanl points. The achievement of Morris water maze showed that the learning and memory of rats got well after acupuncture therapy. And the neuronal degeneration was reduced in hippocampus.Third: It is success to isolate differetially expressed genes associated with vascular dementia from hippcampus. The technique of RNA fingerprinting was used to investigate the changes of differentiallyexpressed cDNAs in the hippcampus of normal aging and VD rats. CDNAs expressing differently during the progression of VDM were isolated. Thirty-two candidate cDNA fragments were isolated with RNA fingerprinting. The two cDNA fragments which were only owned by VDM group were analysised by Northern blot. The results were compared with the databases in National Center of Biotechnology Information (NCBI) Genbank using the BLAST. Homology analysis through BLAST revealed that these genes were novel ones. Genbank accession numbers are BG937392^ BG937393. The study shows that RNA fingerprinting is a simple and quick technique for isolating differently expressed genes. The differentially expressed gene fragments may contribute significantly to the process of VD as lead-disease genes and protect genes.Forth: It is success to isolate differetially expressed cDNAs associated with vascular dementia rats from hippcampus after acupuncture treatment too. The study treated VDM of rats by acupuncture on Beihui and Shenyu and Zhusanli points. The learning and memory of VD rats who were treated by acupuncture were tested by Morris water maze. The technique of RNA fingerprinting was used to investigate the changes of differentially expressed cDNAs in the hippcampus of normal aging and VD and acupuncture treatment rats. CDNAs expressing differently during the progression of acupuncture treatment were isolated. Fifty-two candidate cDNAs fragments were isolated with RNA fingerprinting. The five cDNAs fragments which were only owned by acupuncture rats were analysised by Northern blot. Homology analysis through BLAST revealed that these genes were novel ones. Genbank accession numbers are BG937394> BG937395, BG937396> BG937397> BG937398. The study shows that the differentially expressed gene fragments may contribute significantly to the process of VD as protect genes or modificatory genes.Fifth: The study focuses on the probable molecule mechanism of therapy and the change of the brain-derived neurotrophic factor mRNA(BDNF mRNA) in hippocampus of rat after acupuncture. The level of BDNF mRNA in rat hippocampus was measured using reverse transcription PCR. The expression of the BDNF mRNA in...