| 1 BackgroundAlzheimer’s disease(AD),which often occurs undetectively and progresses chronically,is a neurodegenerative disease of central nervous system.It can cause progressive cognitive and memory deterioration at the beginning which can be followed by syndromes of personality change,aphasia,disability,cognitive impairment,and so on.It is also the main cause of Senile Dementia.AD is very complex pathogenesis,the main hypothesis is the Aβprotein theory,gene mutation theory,tau protein theory,cholinology theory,etc.,all kinds of theories are intricate,but these hypotheses can not fully explain the pathogenesis of AD.Therefore,the study of the mechanism of neuritic plaque has gradually becoming the hot spot in recent years,and currently it is believed that the inflammatory response caused by microglia stimulated by the formation of neuritic plaque which is composed mainly of extracellularβamyloid deposion.As the deepening acknowledgment of the whole process of microglia joining the occurrence and development of AD,targeting at microglia activation inhibition using the traditional Chinese acupuncture combining with the modern music electro-acupuncture can also be an important treatment strategy.2 ObjectiveIn this study,7 months of APP/PS1 double transgenic mice were used as subjects,and inhibition of microglia activation and the release of the associated inflammatory factor TNF-αwas treated as the starting point.The study is to observe the differences between the influence of different kinds of electro-acupuncture therapies on the spatial learning and memory ability,based on "Tongdu Qishen" principle;and eventually to provide the objective experimental proof for music-electro-acupuncture clinical practice,and further the supplement of "Tongdu Qishen" academic thought.3 MethodThirty 7-month-old APP/PS1 transgenic male mice were equally and randomly divided into model control group,EA treatment group and music EA group;and 10 C57BL/6 mice were used as the normal control group.The total number of mice is forty.Normal control group:In the same feeding conditions,and do nothing but to catch twice when treating EA treatment group and music EA group,using the same mouse tied for 20min.Model group was dealt with as the normal control group.In electro-acupuncture group,the mice were placed in self-made rat sleeves and treated by electro-acupuncture.First we chose Renzhong using pricking needling,and then chose Baihui and Yintang using horizontal needling with the direction downward and upward respectively.The puncture depth is 0.5cm.The needle handles were connected with HANS-LH202 electro-acupuncture device,2Hz of frequency,2V of the voltage,and 0.1 mA of the intensity with mice heads needles slightly shaking for 20min.The treatments were done once a day for 15 days.In the music electro-acupuncture group,acupuncture points and methods selection were just as those of the electro-acupuncture group,with needle handles connected with ZJ-12H Musical electro-acupuncture device.Rhythm clear,moderate speed and intensity of the music prescription was selected,and adjusted the intensity so that the animals kept quiet during the course of treatment.The treatment lasted for 20min,for in total 15 days.After 15-day’s treatment,morris water maze test was used to observe the ethology changes of mice in each group.The expression levels of MG in cerebral cortex was assayed by immunohistochemistry.Western blotting was used to observe TNF-a in cerebral cortex of mice.4 Results4.1 MWM Result4.1.1 Result of pace navigationCompared with model control group,the normal group mice escape latency were significantly lower than the model control group from first day to fifth day,and the difference were statistically significant(P<0.01).Compared with the model group mice,the music-EA group have a downward trend in the third day(P>0.05).Until the forth day,the EA group have statistical difference in the forth day(P>0.05).On the fourth day and fifth day,compared with model group,the normal group,the EA group and music-EA group have a downward trend,the difference were statistically significant(P<0.01)(P<0.05)(P<0.01);With the time of the normal group is the shortest,and next are those of EA group and music-EA group,and the time of the model group is the longest.After 15 days treatment of EA and music-EA,the APP/PS1 mice had a significantly improvement of the ability of spatial location learning compared with the model group,but there are no statistical difference between EA group and music-EA group(P>0.05).4.1.2 Result of spatial probeThe times of crossing platform,compared with the model group,the normal group,EA group and music-EA group are of obvious advantages,the difference were statistically significant(P<0.01)(P<0.01)(P<0.01).According to the results,the times of platform-cross of the normal control group is the most,and the times of platform-cross of the model group is the least,while the compared with EA group,the times of the platform-cross of music-EA group is more but has no statistical difference(P>0.05).The normal group,the EA group and the music-EA group compared with the model group,had an increasing trend of the ratio of swimming distance in the target quadrant to the total swimming distance,and the difference were statistically significant(P<0.01)(P<0.01)(P<0.01),with the music-EA group performed better than the EA group(P<0.01).4.2 Results of immunohistochemistryThe comparison of activation state of microglia in frontal lobe of each group is as follows.Microglia Iba-1 immunohistochemical staining positive expression of cytoplasm,cell membrane dark brown dye.The positive expression of Iba-1 in the normal control group was the lowest,the microglia was branched,the cell body was small and the cell protrusion was elongated.The positive expression of Iba-1 in the microglia of the model group was significantly increased and the number of positive cells was significantly increased,microglia cells become larger,oval,thickened cells,showing "bush" like a high branch,significantly compared with other groups,the maximum amount of microglia activation.Compare with the model group,the expression of IBA-1-immunore activity and the number of the IBA-1-immunopositive cells were obviously decreased in the EA group and music EA group(P<0.05).And the music-EA group performed better than EA group(P<0.05).4.3 Results of Western blotting detectionWestern blotting detection displayed that the TNF-α expression in cerebral cortex of the model group was markedly higher than that of the normal control group(P<0.01).After the treatment of EA group and music-EA group,TNF-a expression level was moderately down-regulated(P<0.05)(P<0.01);the strip of music-EA group is significantly grew wider compared with EA group,and the protein expression levels decreased a lot(P<0.05).Western blotting detection displayed that the IL-12B expression in cerebral cortex of the model group was markedly higher than that of the normal control group(P<0.01).After the treatment of EA group and music-EA group,IL-12B expression level was moderately down-regulated(P<0.01);the strip of music-EA group is significantly grew wider compared with EA group,and the protein expression levels decreased a lot(P<0.05).5 Conclusions(1)The 7-month-old APP/PS1 mice had an obvious cognitive impairment of learning and memory and the high expression of the relevant inflammatory cytokines,which is one of the ideal models of AD.(2)The therapies of pulse-EA and music-EA based on "Tongdu Qishen" principle can improve the learning-memory ability of the 7-month-old APP/PS1 double transgenic mice;while the music-EA on learning memory keep better than the pulse-EA.(3)The therapy of music-EA based on "Tongdu Qishen" principle can inhibit of microglia activation and significantly reduce the release of relevant inflammatory cytokines,which has better treatment effect of AD than pulse electro-acupuncture. |
PDF Full Text Request