Objective To observe the effect of needling "Danzhong"(CV17)

"Zhongwan"(CV12)

"Qihai"(CV6)

"Xuehai"(SP10)and"Zusanli"(ST36)(Triple Energizer Acupuncture Method)on the learning-memory ability and profile of hippocampal differentially-expressed genes and proteins of SAMP8 mice(rapid aging mice)

so as to explore its underlying mechanisms in improving Alzheimer's disease(AD).Methods A total of 60 SAMP8 were used as the dementia model and randomly divided into blank control

acupoint and non-acupoint groups(n=20 per group).The 5 acupuncture points and non-acupoints(subcostal region)on the bilateral sides were needled with filiform needles and manipulated manually for 30 sat each one

once daily

6 times a week for 4 weeks.The Morris water maze tests(location navigation tests and space probing trials)were used to evaluate the mouse's learning-memory ability.The hippocampal tissue was extracted to detect differentially expressed genes and proteins related to acupuncture intervention by gene chip and isobaric tags for relative and absolute quantitation(iTRAQ)techniques as well as bioinformatic analysis

separately.The information is analyzed through bioinformatics database tools.Finally

immunofluorescence staining was used to verify the results of microarray analysis.Results Compared with mice of the control and non-acupoint groups

the escape latency of location navigation task of Morris water maze tests on 4 th and 5 th day of training was significantly shortened in mice of the acupoint group

and the duration of stay in the original safe-platform quadrant was significantly increased in the acupoint group(P <0.05).Gene microarray displayed that in comparison with the control group

898 differentially expressed genes were up-regulated

418 genes were down-regulated in the hippocampus of acupoint group.The iTRAQ analysis indicated that in the acupoint and non-acupoint groups

286 and 299 differentially expressed proteins were up-regulated

319 and 179 proteins down-regulated

respectively.Of the 34 terms containing 47 proteins up-regulated by acupoint needling

including intermediate filament

keratin filament

myelin sheath

postsynaptic density

neuron projection were related with neurite and cytoskeleton.While in the non-acupoint group

of the 24 terms were listed by the system

only the myelin sheath involving 11 differentially expressed proteins functions in activities of neurite and cytoskeleton.Immunofluorescence staining of the hippocampal tissue showed that the high-density distribution areas of neurons and neurite fibers were characterized by decentralization and disordering

with the highlighted areas being mainly near the cell body parts in control mice

but in mice of the acupoint group

the highlighted areas at the neurite were relatively dense

the morphology of hippocampal cells was complete

the fiber structure was clear

dense and orderly

and the neurites were closely arranged and in order

indicating an improvement of the distribution and arrangement of nerve fibers after acupuncture.The height of neurite highlight area of the acupoint group was significantly higher than that of the non-acupoint group(P<0.05).Conclusion The "Triple Energizer Acupuncture"of acupoints is able to improve the learning-memory ability in SAMP8

which may be related to its effects in regulating the expression and function of hippocampal genes and proteins related to neurite and cytoskeleton.