| Objective:In our prophase research, anti-anxiety and down-regulation expression level of central C-type natriuretic peptide receptor (natriuretic peptide receptor B, NPR-B) of the acupuncture were confirmed. And on the basis of it, neurochemistry agonist was applied to activate C type natriuretic peptide receptor of center paraventricular nucleus of hypothalamus (paraventricular nucleus of hypothalamus, PVN) to observe whether the anti-anxiety effect and regulation of HPA axis of acupuncture on anxiety model rats can be blocked by activation of NPR-B. Then whether NPR-B of the hypothalamus is involved in the anxiolytic effect of acupuncture can be disproved to provide some scientific experimental basis for the acupuncture in the treatment of anxiety disorders in clinical application.Methods:Healthy and clean SD (Sprague Dawley,SD) male rats were domesticated for a week, and then undesirable weight of rats were eliminated. The filtered rats were randomly divided into six groups according to their behavioral science by using SPSS 17.0 statistical analysis software.Three of them were combined to be operation/ surgery group, in other words, the rats were grouped as follows, blank (1 group), model (1 group), acupuncture (1 group), surgery (3 groups) groups. Then blank, model and acupuncture groups were domesticated for another week, the operation/ surgery group was performed in lateral ventricle buried tube operation at the same time. The injection catheter was implanted and fixed into the rat brain's specific position by stereotaxic apparatus according to the PaXinos and Watson rat brain stereotaxic atlas. And then the operation/surgery group was divided into 3 groups according to their behavioral science after postoperative recovery for one week, namely, intracerebroventricular injection of physiological saline group (sham injection group), intracerebroventricular injection of NPR-B excited agent group (model drug group), lateral ventricle injection of npr-b excited agent in the acupuncture group (acupuncture& drug group).Neurochemical receptor activation technique was used in the model drug group and acupuncture & drug group. Namely, NPR-B excited agent CNP22 (2?g/2?l) was microinjected into intracerebroventricular of acupuncture & drug group, and 0.9% saline(2?l) of sham injection group.7 kinds of stimulation methods were randomly arranged in the 15 days based on the improved reference to establish an unpredictable chronic emotional stress (CUS) anxiety model. In the acupuncture group, "Neiguan" and "Shenmen" points of one side were choosed to treat anxiety rats by using hand acupuncture, alternating left and right, once every other day. Each point was twirled by reinforcing-reducing method for 0.5min, once every 5 minutes, lasting 30 min, and acupuncture treatment lasted from beginning to end of modeling.At the end of the modeling, the EPM (elevated plus maze test, EPM) was used to test the behavior of rats in each group. Method of immunohistochemistry (immunohistochemistry,IHC) was used to detect the expression of C-type natriuretic peptide (C-type natriuretic peptide,CNP), its receptor NPR-B and corticotropin-releasing hormone (corticotropin-releasing hormone, CRH) in paraventricular nucleus of rats at the same time. Enzymelin-ked immunosorbent assay (Enzymelin-ked immunosorbent assay, Elisa) was used to detect corticosterone (corticosterone, CORT) levels in plasma.Result:1?The results of behavioral test:(1) OT% values:The OT% values of model group was significantly reduced in the blank group (P< 0.01), but the OT% values of acupuncture group was obviously higher than that of model group (P<0.01).The OT% value in false drug injection group, the model+drug group, acupuncture +drug group were lower than that in group of acupuncture group (P< 0.05 or P< 0.01),there was no statistical difference in the model+drug group, acupuncture +drug group (P> 0.05). (2) OE% values:acupuncture group was increased significantly (P<0.05) compared with model group, but no statistical difference in other groups.2?The CNP, NPR-B expression of hypothalamus:(1) CNP:compared with the blank group, the CNP average optical density of model group, false drug injection group acupuncture+drug group, mold +drug group in hypothalamus, were obviously increased (P<0.01);Compared with model group, the CNP values of acupuncture group was dramatically reduced (P<0.01), there was a higher trend in acupuncture+drug group and mold +drug group than false drug injection group, but no difference between in acupuncture+drug group and mold +drug group (P> 0.05).(2) NPR-B:compared with the normal animals, NPR-B expression of hypothalamus in all groups but acupuncture groups were significantly higher (P<0.01);Acupuncture group had a significantly lower NPR-B levels compared with model group (P< 0.01);Than to the false injection group, there was a higher tend about NPR-B content in acupuncture+drug group, mold +drug group,which compared to false drug injection group, the NPR-B level of acupuncture+drug group was lower than, mold +drug group, but no significant statistical difference in the two groups (P> 0.05).3% Related parameters of HPA axis test results:(1) The hypothalamus CRH:the CRH levels of model group, the false drug injection group, the model +drug group, acupuncture+drug group all were significantly higher in the blank group (P< 0.05), the acupuncture group significantly was lower than the model group (P< 0.05);Mold medicine group, the needle medicine group a false CRH injection group were higher, and the needle medicine group increased significantly (P< 0.05).(2) The plasmaCORT: the contents of plasmaCORT in model group and model +drug group were obviously higher than in the blank group (P<0.01), acupuncture group was significantly decreased compared with model group (P<0.01);although CORT of acupuncture+drug group had a lower tend, but no statistical difference in the two groups (P> 0.05).Conclusion:1?The anxiolytic effect of acupuncture is distinct,it may be related to regulating CNP and its receptors in hypothalamus,inhibiting the actvity of HPA axis.CNP-HPA axis can be another important role of acupuncture in the treatment of anxiety disorders.2?The hypothalamus paraventricular nucleus NPR-B agonists can weaken the anti-anxiety effect of acupuncture, at the same time, the influence of acupuncture on the HPA axis adjustment function, and then confirmed the central hypothalamic paraventricular nucleus NPR-B could be one of the key links in acupuncture play anti-anxiety effect. |
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