Research Of The Effect About "Double-reinforcing And One-unblocking" Electro-acupuncture Therapy On Aging Rats And The Mechanism Of The T Cell In Immunization

ObjectiveAccording to the theory of preventive treatment of disease in Chinese medicine, and the previous studies, this research try to observe and discuss the effect and mechanism of Double-reinforcing and One-unblocking Electro-acupuncture therapy on aging model rats induced by D-galactose, to provide the evidence for clinical study researches, and imorove the application of in the Double-reinforcing and One-unblocking Electro-acupuncture therapy in the control aging field.Methods40SD rats (female),5-month old, weight200～220g. All rats were randomly divided into youth control group, aging model group (model), Double-reinforcing and One-unblocking electro-acupuncture therapy group (EA) and a control electro-acupuncture group (EAc),10rats in each group. The aging model rats induced by D-galactose, except the youth control group. Double-reinforcing and One-unblocking group (EA) used electro-acupuncture therapy on "GuanYuan"(GV4),"HouSanli"(ST36)and "BaiHui"(GV20); And control electro-acupuncture group (EAc) used electro-acupuncture therapy on " ZhongJi "(GV3),"YinLingquan"(SP9) and "YinTang"(GV29). All points treated with the electrically of2Hz and1mA,15min per day,6-time per week, and last for4weeks. Morris water maze experiment to test the ability of learning and memory of all rats, detect serum SOD activity, the MDA content in serum, the spleen index and thymus index, and to observe the change of ultra-microstructure of thymocyte in aging model rats, to discuss the effect of Double-reinforcing and One-unblocking electro-acupuncture therapy, to test the Positive expression of T cells subgroup CD4+, CD4+CD45R0+, CD8+,CD8+CD45RO+,CD45RA+, CD45RO+, CD45RA+/CD45R0+; And to explore the mechanism of anti-aging by Double-reinforcing and One-unblocking electro-acupuncture therapy.Results1. Morris water maze testing, compared with the normal group, in aging model group, the incubation period extended (P<0.01), more errors appeared (P<0.01); Compared with aging model group, in two electro-acupuncture therapy groups used less time in finding platform(P<0.05) and made fewer mistakes (P<0.05); In two electro-acupuncture therapy groups,compared with EAc group, the effect of EA group was much better (P<0.05).2.Compared with the normal control group, the serum SOD activity in aging model group reduced significantly (P<0.001), and the serum MDA content increased significantly (P<0.01); Compared with the model group, the SOD activity of two electro-acupuncture therapy groups improved (P<0.01), and the MDA content reduced (P<0.05); the two electro-acupuncture therapy groups compared between each other, the SOD activity of the EA group was highter (P<0.05), and the MDA content was lower (P<0.05).3. The changes of spleen and thymus index. Compared with normal control group, the spleen index and the thymus index in model group were lower (P<0.05); Compared with aging model group, the spleen index and thymus index in two electro-acupuncture therapy groups were increased (P<0.05), and the effect of the EA group was better than the control group (P<0.05).4.The changes of the thymocyte ultrastructure, compared with the normal control group, the thymocyte of the model rats became pyknotic, nuclear membrane retracted, cytoplasm marked edema, mitochondria crest fractured, and degeneration appeared, swelling and emptybubble, lipofuscins accumulated, and there were obviously apoptotic cells, euchromatin in edge of nuclear membrane; Compared with the model group, the thymus cell apoptosis and the cell membrane became destructive, nucleus and organelles in the two electro-acupuncture therapy groups were lighter than the model group, and the effect of the EA group was much better.3.Adoption of the fluorescent monoclonal antibody three notation method and FCM to detect the positive expression of T cells subsets.①Compared with the normal control group, in model group, the positive expression of CD4+cells reduced (P<0.01), the positive expression of CD4+CD45RO+cells increased (P<0.01); Compared with the model group; The positive expression rate of CD4+increased both in the EA group (P+<0.01)and EAc group(P+<0.05), the positive expression rate of CD4+CD45R0+cells reduced both in the EA group (P+<0.01) and EAc group (P+<0.05); Compared with the EA group, there were remarkably difference between the two electro-acupuncture therapy groups, the effect of EA group was better。②Compared with the normal control group, in the model group, the positive expression of CD8+(P<0.01), CD8+CD45RO+(P<0.05) cells increased; Compared with the model group, the positive expression rate of CD8+(P<0.01) and CD8+CD45RO+(P<0.05)cells in Double-reinforcing and One-unblocking electro-acupuncture therapy group reduced obviously; the positive expression of CD8+T cells in the control group reduced (P<0.05), but the expression of CD8+CD45RO+cells had no obviously difference (P>0.05); Compared with the EA group, the positive expression rate in EAc group was higher, CD8+T cells (P<0.05) and CD8+CD45R0+T cells (P<0.05).③Compared with the normal control group, in the modelgroup, the positive expression of CD45RA+cells reduced (P<0.01), the positive expression of CD45R0+cells rose (P<0.01), the specific value of CD45RA+/CD45R0+reduced(P<0.05); Compared with the model group, positive expression of CD45RA+of the EA group increased (P<0.05), the positive expression of CD45R0+reduced (P<0.01), the specific value of CD45RA+/CD45R0+increased (P<0.01); the positive expression of CD45RA+in the EAc group also increased (P<0.05), the positive expression of CD45R0+reduced(P<0.01) the specific value of CD45RA+/CD45R0+increased (P<0.01); compared of the EA group, the positive expression of EAc group, CD45RA+Tcell s reduced (P<0.05), CD45R0+Tcells increased (P<0.05), CD45RA+/CD45R0+reduced (P<0.05)。 Conclusion1. Double-reinforcing and One-unblocking electro-acupuncture therapy can enhance the ability of study and spatial memory effectively; regulate the free radical metabolism and the immune related index, and improve the thymus ultrastructural structure.2. the Double-reinforcing and One-unblocking electro-acupuncture therapy can increase the positive expression of CD4+; CD45RA+and increase the specific value of CD45RA+/CD45R0+; moreover, the Double-reinforcing and One-unblocking electro-acupuncture therapy can reduce he positive expression of CD4+CD45RO+; CD8+; CD8+CD45R0+andCD45R0+, thus, this treatment can regulate the immune system in vivo, and adjust the cellular immunity.3. The effect of Double-reinforcing and One-unblocking electro-acupuncture therapy group is better than ordinary acupuncture method group.