| With increasing morbidity and predicted to be No.1 mortal disease in the 21 St century, Alzheimer抯 Disease (AD) is now holding the medical spotlight in this aging society. Modem western medicine has delved deep into the possible etiology and pathogeny, but made no breakthrough in therapy. Traditional Chinese medicine (TCM) has much originality concerning the understanding, prevention and treatment of senile dementia but shows inferiority in mechanism exploration. Acupuncture has affirmative effect on senile intelligence yet needs further research on acupoint specificity. Here we used Senescence Accelerated Mouse (SAM) PlO and the normal control SAMRI as models and applied mRNA-AFLP technology to study the therapeutic significance of different acupoints for senile dementia. In the light of TCM theory, we selected point Du26 to stimulate the brain-activity, point Pc6 to regulate the heart-activity, point Li3 to regulate the liver-Chi, point UB23 to tonify the kidney and point ST36 to reinforce the spleen and stomach. Differentially expressed bands were displayed through silver staining and then analyzed for research on the expression of encephalic senescence-associated genes and the interference effect of acupuncture.The main results were as follows:1. Blended by magnetic separation of mRNA, SMART-PCR cDNA synthesis and AFLP-silver staining, mRNA-AFLP technology showed high efficiency in displaying the differentially expressed genes. With just 13 primer pairs for selective AFLP amplification, 42 bands with evidently different expression level between the two murine strains (P vs R) were demonstrated and found to respond differently, to different acupoints. Therefore, mRNA-AFLP technology can greatly benefit the differential display of encephalic senescence-associated genes, the research on acupuncture interference and the exploration on the relative specificity of acupoints.2.Among the 42 differentially expressed bands in this experiment, 33 bands (78.6% of the total) showed relatively lower expression level in the P-strain of SAM than the R-strain, while the other 9 bands (21.4% of the total), relativelyhOhigher in the former strain. This phenomenon during the process of aging needs further study concerning its significance.3.The?expression level of the 42 differential bands could be regulated by acupuncture, indicating an interference effect on the senescence-associated abnormal change in the gene expression of brain. Those bands with lower expression level in the P-strain of SAM were upregulated while those higher, downregulated. Both showed a tendancy towards the expression level of the normal control, R-strain of SAM, suggesting a biphasic-regulation effect of acupuncture on the molecular level.4.The 42 differential bands responded differently to different acupoints, manifested by the number of bands and the extent to which the expression level was affected. There were respectively 34, 26, another26, 17 and 8 bands ( each accounting for 81.0%, 61.9%, 61.9%, 40.5% and 19.0% of the total) with expression level being remarkably regulated by points Du26, Pc6, Li3, ST36 and UB23. This reveals the relative specificity of acupoints in the treatment of senile dementia, with the former three points being more effective than the latter two.5.Among the aquired seven clones inserted with pure band of interest, four cDNA fragments were found to have high identity with known sequences in the GenBank while the other three, low identity. The essence of them needs further exploration.6.Through RT-PCR semi-quantification, a fragment of 231bp sharing 93% identity with human CD34 was preliminarily considered non-false positive. Its expression level in the 7-month-old SAMPIO brain was lower than that of SAMRI but could be upregulated by puncturing points Du26, Pc6 and Li3. This suggests that sustenance of the expression level of CD34 in the murine brain may have some relevance to the mechanism of acupuncture interference with encephalic senescence. However, the...
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