Experimental Research On Aging Related Genes And Acupuncture Intervention Effect

With the world population aging, senescence-related cardiovascular diseases and cerebrovascular diseases have become the primary cause of death while senile dementia turned out to be the fourth mortal disease. More and more people and families are falling victim to it, which causes heavy burden to the society. Therefore, the research on aging and anti-aging gets awfully urgent and important for current medical and biological sciences. Nematodes, fruit flies, mice, cultured cells and transgenic animals are all useful models and have contributed to the discovery of some aging-related genes. However. there is still a long way to go in this field of study for being short of a senescence-naturalized animal model. SAM(Senescence Accelerated Mouse), whose process of aging accelerates but resembles that of human being, just fits our needs and has been internationally acknowledged to be an ideal model. SAM-P/lO, a substrain characterized by encephalatrophy, was selected for our study here. The homologous strain Rh, which has regular process of aging,was used as control. mRNA differential display technique was applied for the screening of senescence-associated genes and for the study on acupuncture interference with aging. The main conclusions drawn from our work are as follows: I. Two annealing temperature were performed in differential display:first,a lower temperature, 370, was adopted to assure the pairing between primers and templates: then, the temperature was raised to 500 for purpose of specific amplification. The differentially expressed cDNA fragments shown by silver staining were directly cut from the gel, eluted and then amplified. The manipulation was convenient and safe and the results showed good specificity and reproducibility. 2. Forty-two primers were used altogether, from which two were randomly selccted for each PCR reaction.Some differentially expressed cDNA fragments showed regular changes which could be repeated by amplifications using different primer pairs. This implies that the accelerated cerebral senescence of SAM-P/I 0 is probably related to the abnormal expression of aging-associated genes. 3. The expression amount of the cDNA fragments changed obviously after acupuncture, suggesting that the acupuncture method we used, 揟iao Shen Yi Zhi, Ping Gan Tong Luo? could defer the progress of senescence through interfering with the expression of aging-associated genes. 4.lt is for the first time that acupuncture exhibits its beneficial regulation effect at the molecular level, down-regulating the overexpression of some genes while up-regulating the low expression of others. Both showed a tendancy towards that of the young group. This confirms from the molecular angle that the core of acupuncture lies in its regulation effect. S. The results show that puncturing acupoints has more apparent effect on the expression level of genes than puncturing non-points, therefore verifying the relative specificity of acupoints at the molecular level. 6. Some differentially displayed cDNA fragments were cloned, sequenced and homology analysized, three of which are probably new discoveries since they have no homology with the already known genes. There is a cDNA fragment of 5 lObp length sharing high homology with eukaryotic translation initiation factor 3(eIF3), 98% identity with that of mouse and 89% with that of human. In addition,there is another cDNA fragment of 1 l6bp length sharing 93% homology with the nuclear gene encoding mitochondrial protein, a subunit of eIF3. Northern blot analysis reveals that...