| Aim:This study explored the part of mechanism that acupuncture’s anti anxiety regulated T cell subsets, in order to provide the basis of preliminary study on the mechanism of acupuncture’s anti anxiety, also provide scientific experimental basis for new clinical application.Methods:Clean grade male SD rats were selected and randomly divided into blank group, model group and electric acupuncture (EA) group. The model group and the EA group were both stimulated by chronic unpredictable stress (CUS) method to duplicate model of anxiety. EA was used in electric acupuncture group to penetrate "Neiguan" and "Shenmen" point. EA parameters:dilatational wave, frequency 15/25Hz, the intensity of stimulation in rats with EA was based on the slight shaking of limbs, and each treatment lasted for 15min. Treatment started on the modeling day, every other day for 1 times, left right alternation, which lasted 15 days. The elevated plus maze test was used to test the anxiety rats’ behavior, HE staining was used to observe the pathologic change of thymus tissue, the plasma concentrations of CD4+, CD8+ and CORT were measured by enzyme linked immunosorbent assay and the expression of mature thymocytes’ CD4+, CD8+ and GR were observed by immunohistochemistry.Results:1. Results of behavior:Compared with the blank group, OE% in model group were decreased significantly (P<0.05); while compared with the model group, OE% in EA group’s were increased significantly (P<0.05); OE% in EA group and control group were no significant difference (P>0.05). OT %in Each group was no significant difference (P>0.05),which meant the model animals were in anxiety state. In other words, the experimental model was successful established; and EA could be used to reduce the anxiety of model animals.2. Results of the plasma levels of T cell subsets:Compared with the blank control group, the concentration of CD4+ of model group was decreased significantly (P＜0.05), while the concentration of CD8+ was increased significantly (P＜0.05), and CD4+/ CD8+ ratio was showed a decreasing trend (P=0.054); compared with the model group, CD4+ concentration and CD4+/CD8+ ratio of EA group were increased significantly (P＜0.05, P<0.01),the concentration of CD8+ were decreased significantly (P<0.05). It suggested that CUS induced the decrease of CD4+ levels and CD4+/CD8+ ratio and the increase of CD8+ in plasma, while EA could up-regulate the excessively low CD4+ levels and CD4+/CD8+ ratio and down-regulate excessively high CD8+ levels in anxiety rats.3. Results of thymus morphology and T cell subsets:(1) Thymus tissue HE staining:The thymus tissue structures of the blank group rats were normal, the boundaries between cortex and medulla were clear, the number of lymphocytes in the cortex was dense,while in the medulla, those were rare and the thymus corpuscle’s volume did not increase obviously. Compared with the blank group, the thymus tissue morphologies of the model group rats atrophied severely, both the structures of thymus lobules and the boundaries between cortex and medulla were not clear, the number of lymphocytes in both cortex and medulla were loose and the thymus corpuscle’s volume were increased obviously. However, the thymus of electric acupuncture group rats atrophied mildly, the thymus lobules’ structures were basic integrity, the boundaries between cortex and medulla were clear, the number of lymphocytes in the cortex was dense, the medulla was expanded and the thymus lobules’ structures were normal. It suggested that EA could inhibit the pathological injury in thymus tissue of anxiety rats.(2) The expression CD4+ and CD8+ in thymus cell of medulla:Compared with the blank group, the expression of CD4+ and CD8+ in thymocyte in the model group were decreased significantly (P<0.01); while compared with the model group, that of the electric acupuncture group were increased significantly (P<0.01), suggesting that the CD4+and CD8+ level of thymus in anxiety rats were decreased, in other words, EA could up- regulate the levels of CD4+ and CD8+ in thymus.4. Results of plasma CORT and thymus GR:(1)Plasma CORT concentration:Compared with the blank group, the plasma CORT concentration of the model group increased; compared with the model group, that of EA group were decreased significantly (P<0.05). It suggested that EA had an effect on down-regulating the excessively high CORT concentration in plasma of anxiety rats.(2) The expression of thymus GR:Compared with the blank group, the expression of thymus GR of model group excessively increased significantly (P <0.01); while compared with the model group, that of EA group were decreased significantly (P<0.01). It suggested that EA could down-regulate the excessively overexpression of thymus GR of anxiety rats.Conclusions:1. Electric acupuncture could not only be used to reduce the anxiety of rats, but also could inhibit the impaired morphology of thymus and benignly regulate T cells subsets level in peripheral plasma and thymus.2. The effect of regulating immune function of electric acupuncture in anxiety rats might be through reducing the CORT concentration of plasma and the expression level of GR of thymus, and might be related to HPA axis. |
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