| Purpose:Under the guidance of the theory "brain and kidney with Leniency", to observe the effect of Bushen Yijing fill pulp electro acupuncture therapy of "Tiangui dried up" the peri-menopausal depression, and to explore its mechanism of action, for the clinical treatment of peri Menopause Depression to provide guidance and experimental basis.Material and method:From 17 week old female SD 120 rats were randomly divided into normal group, model group, drug group and electroacupuncture group and s-ham operation group, 24 rats in each group. 2 rats in a normal feeding cage.1 Behavioral science indexes: rats in modeling before the start of the first behavioral indicators of observation and behavioral indicators in the open field test, the total mov-ing distance and open field in the central region of the residence time; sucrose prefer-ence test in the consumption of sucrose and total fluid consumption amount and suga-r consumption rate; forced swimming test in rats and the rest time in water.2 preparation of perimenopausal depression model rats in the normal group does not a-ccept the model, normal feeding. Model group, drug group and electroacupuncture gro-up rats were bilaterally ovariectomized and sham operation group without excision of ovarian resection, only in ovarian and ovarian big fat near. Postoperative recovery of the rats for two weeks, after 5 consecutive days of vaginal smear, determine the ope-ration group rats had no estrous cycle. That was ovariectomized successfully. Rats in the normal group were fed with no stimulation. The rats in model group, drug group and sham operation group received 21 days of chronic non mild stimulation(CUMS) for a period of days. Stimulation includes: ice water swimming, hot water swimming, no water fasting, clip tail, restraint action, tilt the cage and day and night upside down. Each stimulus appeared randomly, and each stimulus was not less than 3 times in 21 days. At the end of the 21 days after the stimulus, the model of the depression model of the rat model was successfully induced by the change of behavior index.3 electroacupuncture treatment: normal rats normal feeding, does not receive the treatm-ent. electroacupuncture group and sham operation group rats received electroacupunctu-re treatment for 28 days, according to the "brain and kidney with Leniency" theory, select Baihui, Shenshu, Sanyinjiao, daily electroacupuncture treatment 1 and continued for 20 minutes. Rats in drug group selected tricyclic antidepressant clomipramine hydrochloride as the control, with clomipramine hydrochloride orally daily by gavage 1 times. After the end of treatment through behavioral index change evaluation of electroacupuncture on perimenopausal depression treatment effects in rats.4. The behavior observed after anesthesia, killed all the rats were killed, rat brain paraffin sections were used for rat hippocampal structure and neural cell morphology changes were observed; serum and by enzyme linked immunosorbent assay for the detecti-on of serum Remove the rat hippocampus and hippocampal tissue were detected by midbrain derived neural neu-rotrophic factor(BDNF) by enzyme- linked immunosorbe nt assay and receptor tyrosi-ne kinase B(Trk B) and c AMP response element binding protein(CREB), B cell lym-phoma 2(Bcl-2) and Bcl-2 associated X protein(Bax) co ntent, and real-time fluoresc-ence quantitative PCR method for detection of hippocamp al BDNF, Trk B, CREB, Bcl-2 and Bax m RNA expression.Results:1. before the start of the model rats in the open field test in the total moving distanc-e and center area of residence time had no difference(P>0.05). Experiment in syrup, sugar consumption amount, total fluid consumption amount and sugar consumption w as no difference(P>0.05); forced swimming test in immobility time no difference(P>0.05).2. in addition to the normal group and sham operated group, the vaginal smears of th-e rats in each group had no cycle.3. After 21 days chronic unpredictable mild stress. The results of the open field: compared with normal group compared, rats in other groups were total movement distance of each group were lower than the normal group, the difference is statistically signifi-cant(P<0.05). The remaining rats in open field center area of residence time were le-ss than the normal group, the difference was statistically significant(P<0.05).4. After 21 days chronic unpredictable mild stress, sucrose test results: group and normal compared the rats in the other groups, sugar consumption rate were lower than th-e normal group, the difference was statistically significant(P<0.05). Compared with the model group, sham operation group rats sugar consumption rate is higher than th-e model group, the difference was statistically significant(P<0.05).5.after 21 days of chronic non mild stimulation, forced swimming test results: compared with the normal group, the rest of the rats in the water at rest time is greater tha n the normal group, the difference was statistically significant(P<0.05).6. after 28 days of acupuncture and drug treatment, the results of the open field: com-pared with the normal group, model group rats of total moving distance is less than the normal group, central region of residence time is less than the normal group, the difference is statistically significant(P<0.05). Compared with the model group, drug gr-oup and acupuncture group and sham operation group of total moving distance is g-r eater than that of model group, Kuang center of the field area residence time is larg-e r than that of the model group, the difference was statistically significant(P<0.05).7. After 28 days of electroacupuncture and medication, sugar on experimental results: group and normal compared the rats in the other groups, sugar consumption rate was higher than that in normal group, the difference was statistically significant(P<0.05). Compared with model group, sham operation group, electroacupuncture group and a m--edication group of sugar consumption was higher than that in the model group, the difference was statistically significant(P<0.05). Compared with the electroacupuncture gr-oup, sham operation group, sugar consumption rate is higher than the acupuncture g-roup, the difference was statistically significant(P<0.05).8. after 28 days of electro acupuncture and medicine treatment, forced swimming test results: compared with the normal group, model group rats were still higher than the normal group, the difference was statistically significant(P<0.05). Compared with mod-el group, the rest time of drug group, electro acupuncture group and sham operation g-roup were less than model group, and the difference was statistically significant(P <0.05).9. After 28 days of electroacupuncture and medication, under the light microscope obs-ervation of rats hippocampus paraffin section, the normal control group rat hippocamp-al CA3 pyramidal cell layer and dentate to granular zone over the number of cell layers of cells, arranged in neat rows, the structural integrity of neurons, the cell nuclea-r and cytoplasmic staining of uniform, clear. It was found that the cytoplasm of the neurons was abundant in the cytoplasm of the cells, which was purple blue, and the morphology and structure of the axon dendrites of neurons were complete. Model group rats hippocampus CA3 cells that are small in number, the intercellular space increased, malalignment of the loose, even disrupting layer of cells, Nissl staining showed the neuronal nucleus disappear, cytoplasmic atrophy, empty, axon dendritic structure is not complete, even disappearance of cell structure. The situation of drug group, electr oacupuncture group and sham operation group were between the two.10. After 28 days of electroacupuncture and medication, hippocampal tissue enzyme linked immunosorbent adsorption method results: compared with the normal group, mod-el group rats hippocampal BDNF protein content decreased significantly, the differenc-e was statistically significant(P<0.05); compared with the model group, drug group a nd acupuncture group and sham operation group were higher than that of the model g roup, the difference was statistically significant(P<0.05); compared with the electroacu-puncture group, sham operation group, the numerical value is higher than that of the normal group, the difference is statistically significant(P<0.05). Compared with the no-rmal group, model group rats hippocampal Trk B protein content decreased significantl y, the difference is statistically significant(P<0.05); compared with the model group, drug group and acupuncture group and sham operation group were higher than that of the model group, the difference is statistically significant(P<0.05); compared with EA group, sham operation group of numerical lower than the normal group, the differe-nces were statistically significant(P<0.05). Compared with the normal group, model gr-oup rats hippocampal CREB protein content decreased, the difference is statistically significant(P<0.05); compared with the model group, drug group, EA group and sham operation group, the content of higher than the model group, the difference is statisti-cally significant(P<0.05); compared with the electroacupuncture group and sham oper-ation group, the content of higher than electro acupuncture group, the difference is st-atistically significant(P<0.05). Compared with the normal group, Bcl-2 protein conten-t in the hippocampus of the model group rats were lower than normal group. Bax w-as higher than that in the normal group, the difference is statistically significant(P<0.05); compared with the model group, drug group, EA group and sham operation group content of Bcl-2 is higher than that of model group. Bax content lower than model group, the difference was statistically significant(P<0.05). And parity in the normal group, rats in the model group, the ratio of Bcl-2/Bax ratio is larger than that of the normal group, the difference was statistically significant(P<0.05); compared with the model group, drug group and acupuncture group and sham operation group Bcl-2/Bax ratio between is larger than that of the model group, the difference was statistically si-gnificant(P<0.05); compared with the electroacupuncture group and sham operation gr-oup Bcl-2/Bax ratio greater than EA group, the difference was statistically significant(P<0.05).11. After 28 days of electroacupuncture and medication, hippocampal tissue by real-time fluorescence quantitative PCR results showed that: compared with the normal group, model group rats hippocampal BDNF m RNA relative expression was lower than that of the normal group, the difference was statistically significant(P < 0.05); compared with the model group, drug group and acupuncture group and sham operation group relative expression quantity is higher than that of model group, the difference was statistically significant(P < 0.05); compared with the electroacupuncture group and sham operation group, BDNF m RNA relative expression was higher than that in EA group, the difference is statistically significant(P < 0.05). Compared with the normal group, the relative expression of m RNA Trk B in hippo-campus of model group was higher than that in normal group, and the difference was statistically significant(P < 0.05). Compared with the normal group, model group rats hippocampal CREB m RNA relative expression quantity of low and normal group, the difference is statistically significant(P < 0.05); compared with the model group, drug group and acupuncture group and sham operation group relative expression quantity is higher than that of model group, the difference is statistically significant(P < 0.05); compared with the electroacupuncture group and sham operation group of CREB m RNA relative expression was higher than that in EA group, the difference is statistically significant(P < 0.05). Compared with the normal group, model group rats hippocampus bcl-2 m RNA relative expression was lower than that of normal group. Bax m RNA relative expression were significantly higher than those in normal group, the difference was statistically significant(P < 0.05); compared with the model group, drug group and acupuncture group and sham operation group, bcl-2 m RNA relative expression quantity is higher than that of model group, Bax m RNA relative expression was lower than that in the model group, the difference is statistically significant(P < 0.05); compared with the electroacupuncture group and sham group, bcl-2 m RNA relative expression was higher than that in EA group. Bax m RNA relative expression amount less than that in EA group, the difference is statistically significant(P < 0.05).Conclusion:1 the establishment of a model of depression in the peri menopausal period with the combination of the model of chronic and non predictable mild stimulation.2 electroacupuncture and clomipramine can improve depressive symptoms in perimenop-ausal rats.3. In the sham operation group and electroacupuncture group behavior index results in a slightly different, indicating that "Tiangui dried up" in the peri menopausal depressi-on, the incidence and prognosis of have an important role.4. Peri menopausal depression rats "emptiness of the sea of marrow" corresponds to t he biological basis may be to reduce the number of hippocampal neurons in the brain and cellular structure damage, which may associated with increased cell apoptosis in hippocampal neurons, hippocampal neurogenesis occurs is related to the decrease of.5. Electroacupuncture can improve the content of rat hippocampus BDNF and TrkB and CREB, promote the neurogenesis in the hippocampus, can increase the ratio of Bcl-2 and Bax, and inhibit neuronal apoptosis and Electroacupuncture of "Bushen Yijing fill marrow" is through the promotion of neurogenesis and inhibiting cell apoptosis.6. Model group rats hippocampus Trk B m RNA relative expression of high volume but Trk B protein content is low, illustrates the peri menopausal depression rats certain fa-ctors hinder the Trk B gene expression, which may is chronic unpredictable mild stim ulation causes reduced hippocampal Trk B and BDNF levels.
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