目的 :采用mRNA AFLP技术

探讨SAMP10衰老相关性脑细胞基因表达的变化及针刺的干预作用。方法 :应用由磁珠分离法纯化mRNA

SMART PCRcDNA合成和AFLP 银染法结合形成的mRNA AFLP分析技术

进行mRNA指纹检测

寻找衰老相关性差异表达的基因片段

并观察针刺对其表达水平的影响

研究衰老相关基因与针刺的关系。结果 :发现规律性变化的一条cDNA带与多品系小鼠的线粒体DNA、编码小鼠ATPase亚基 6和大鼠ATPase亚基 6和亚基 8的mRNA及编码大鼠线粒体细胞色素C氧化酶亚基Ⅰ、Ⅱ、Ⅲ的部分基因序列均有高度同源性。实验表明该cD NA带呈现衰老相关性低表达

并可受针刺单穴“水沟”、“内关”和“后三里”的良性干预。结论 :SAMP10的快速衰老与体内能量代谢障碍有关

针刺可能从分子水平影响了能量代谢的相关因素

这可能是针刺干预衰老进程的机制之一Objective: By using mRNA-amplified fragment length po l ymorphism (AFLP) technique

senescence-associated changes of gene expression of the cerebral neurons and the effect of acupuncture intervention were observed i n senescence-accelerated mouse (SAM)P10 and SAM R1. Methods: Six ty SAM P10 were randomly divided into "Shuigou"(GV 26)

"Neiguan"(PC 6)

"Tai chong"(LR 3)

"Shenshu"(BL 23)

"Zusanli"(ST 36) and P10-control groups

in a ddition

10 SAM R1 were made up of R1-control group. These 5 acupoints were st imulated with handle acupuncture

once daily

continuously for 2 weeks

with one day's rest between two weeks. Magnesium pearl separation-purification method w as used to isolate and purify mRNA from the total RNA of the whole brain. SMART PCR-amplified cDNA synthesis and AFLP-silver staining method combined with mRNA-AFLP analysis technique were applied in the detection of mRNA fingerprint to search for the differentially expressed gene fragments associate d with the senescence and to study the effect of acupuncture intervention on the gene level of these fragments. Results: In comparison with the differentially-expressed gene ban d of the normally-aged mice of R1 control group

the coloration of the band of rapidly-aged mice of P10 control group was lightened (OD value was smaller). Af ter acupuncture of GV-26

PC-6 and ST-36

the OD values of the differentially -expressed gene band increased

while no obvious changes were found in acupunct ure of LR-3 and BL-23 groups. It shows that in SAMP 10

the expression of cDN A fragments being of high autoploidy with mitochondrial DNA and mRNA of the enco ded ATPase subunit and cytochrome C oxidase subunit was lower; while acupuncture stimulation of GV-26

PC-6 and ST-36 can suppress this abnormal change of se nescence-associated low expression of gene fragment of cDNA

suggesting an anti -aging effect of acupuncture. In addition

one regular change of cDNA band in S AM R1 and P10 was found to have a high autoploidy (86%～97%) with some known seq uences of the GenBank

like mitochondrial DNA of multiple species of mice

ATPas e subunit 6mRNA of mice and ATPase subunit 6mRNA and 8mRNA of rats

and partial gene sequence of mitochondrial cytochrome C oxidase subunit Ⅰ

Ⅱ and Ⅲ of rat tus norvegicus. Conclusion: The disturbance of energy metabolism plays a role in the acceleration of senescence of SAM P10 and acupuncture may exert its favorab l e influence on themolecular factor-related energy metabolism

which might be on e of the mechanisms of acupuncture in anti-aging effect.