针刺对焦虑大鼠血浆及胸腺T细胞亚群的影响

目的:通过观察针刺对焦虑模型大鼠血浆CD 4+、CD 8+T淋巴细胞含量及胸腺细胞CD 4+、CD 8+表达水平的影响,探讨针刺治疗焦虑障碍的部分免疫调节机制。方法:健康SD大鼠34只,随机分为空白组(10只)、模型组(12只)、电针组(12只)。运用慢性不可预知应激刺激方法复制焦虑模型。电针"内关""神门"穴,隔日1次,每次15min,共治疗15d。运用高架十字迷宫测试大鼠焦虑行为,酶联免疫法检测血浆CD 4+、CD 8+的含量,免疫组化法观察成熟胸腺细胞CD 4+、CD 8+的表达。结果:与空白组相比,模型组大鼠在开臂的次数(OE)占进入两臂区停留总次数的百分比(OE%)明显下降(P0.05);与模型组相比,针刺组OE%值明显提高(P0.05)。与空白组相比,模型组血浆CD 4+浓度明显下降(P0.05),CD 8+浓度明显升高(P0.05),CD 4+/CD 8+比值有降低趋势(P=0.054);针刺可明显提高模型大鼠血浆CD 4+浓度和CD 4+/CD 8+比值(P0.05,P0.01),明显降低模型大鼠血浆CD 8+浓度(P0.05)。与空白组相比,模型组大鼠胸腺髓质CD 4+、CD 8+表达均显著降低(P0.01);针刺可显著提高模型大鼠胸腺髓质CD 4+、CD 8+的表达(P0.01)。结论:针刺在发挥抗焦虑效应的同时,还可能通过影响CD 4+、CD 8+的成熟、归巢与再循环,发挥其调节焦虑大鼠T细胞的作用。这可能是针刺调节焦虑障碍免疫功能的重要途径之一。

Effects of Acupuncture Intervention on Levels of T Lymphocyte Subsets in Plasma and Thymus in Stress-induced Anxiety Rats

Objective To observe the effect of electroacupuncture(EA)intervention on the levels of CD 4+and CD 8+lymphocytes in the plasma and thymus in anxiety disorder rats,so as to explore its mechanism underlying favorable regulation of immune function.Methods Thirty-four SD rats were randomly divided into control(n=10),model(n=12)and EA(n=12)groups.The anxiety model was established by using chronic unpredictable emotional stress stimulation(fasting,water-deprivation,shaking,tail-clamping,forced warm-and cool-water swimming,electrical shock stimulation,etc.)for 15 days.EA(15Hz/25Hz)was applied to"Neiguan"(PC 6)and"Shenmen"(HT 7)for 15 min,once every other day for 15 days.The anxiety-like behavior was measured by elevated plus maze test.Anxiety reduction in the plus-maze is indicated by an increase in the proportion of time spent in the open arms(time in open arms/total time in open or closed arms),and an increase in the proportion of entries into the open arms(entries into open arms/total entries into open or closed arms).While the contents and expression of CD 4+and CD 8+in the plasma and thymus tissues were measured by ELISA and immunohistochemistry,respectively.Results The values of proportions of open-arm entries(OE%)and the content of plasma CD 4+lymphocytes were obviously lower in the model group than in the control group(P<0.05),and were significantly higher in the EA group than in the model group(P<0.05).Whereas,the content of plasma CD 8+lymphocytes was obviously higher in the model group than in the control group(P<0.05)and markedly down-regulated in the EA group after the treatment(P<0.05).The ratio of CD 4+/CD 8+was decreased in the model group in comparison with the control group(P=0.054),and was significantly up-regulated in the EA group after EA stimulation(P<0.01).Compared with the control group,the expression levels(optical density,OD values)of CD 4+and CD 8+lymphocytes in the thymus tissue were obviously decreased in the model group(P<0.01).After EA,the expression levels of thymus CD 4+and CD 8+were remarkably up-regulated in the EA group(P<0.05).No significant changes were found in proportion of open-arm time(OT%,P>0.05).Conclusion EA can effectively relieve anxiety disorder in anxiety rats,which may be related to its effects in up-regulating plasma CD 4+,thymus CD 4+and CD 8+levels as well as CD 4+/CD8+ratio,and down-regulating plasma CD 8+content.