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100023 北京市2345信箱	世界华人消化杂志 2001年3月15日;9(3):279-283
Email: wcjd@public.bta.net.cn	世界华人消化杂志 ISSN 1009-3079 CN 14-1260/R
http:// www.wjgnet.com	版权归世界胃肠病学杂志社

⊙研究原著⊙

电针对胃经-脑肠肽-免疫网络的调控机制

高巍¹ 黄裕新¹ 陈洪² 赵宁侠³ 孙大勇⁴ 张洪新⁵ 王庆莉¹

中国人民解放军第四军医大学唐都医院 ¹消化内科 ²医教部 ³中医科 5介入科陕西省西安市 710038
⁴中国人民解放军广州军区总医院消化内科广东省广州市 510210
高巍，女，1971-02-03生，江苏省镇江人，汉族. 1994年西安医科大学毕业，2000年第四军医大学内科学消化专业毕业，医学硕士, 住院医师，助教. 发表论文6篇.
国家自然科学基金资助项目, No.39970888
项目负责人黄裕新，710038，陕西省西安市新寺路，中国人民解放军第四军医大学唐都医院消化内科.
¹Department of Gastroenterology， ²Department of Medical Affairs, 3Department of Traditional Chinese Medicine,⁵Department of Interventional Radiology, Tangdu Hospital, 4th Military Medical University, Xi＇an 710038, Shaanxi Province, China
⁴Department of Gastroenterology, Chinese PLA General Hospital of Guangzhou Command Area, Guangzhou 510210, Guangdong Province, China
Supported by the National Natural Science Foundation of China, No. 39970888
Correspondence to Dr. Wei Gao, Department of Gastroenterology,Tangdu Hospital, Fourth Military Medical University, Xi＇an 710038, Shaanxi Province, China
Tel. 0086-29-3577721, 3577128
Email. Chentx007@sohu.com
Received 2000-11-13 Accepted 2000-11-16

Regulatory mechanism of electroacupuncture on the stomach channel brain-gut peptide immune network

Wei Gao¹, Yu-Xin Huang¹, Hong Chen², Ning-Xia Zhao³, Da-Yong Sun⁴, Hong-Xin Zhang⁵ and Qing-Li Wang¹

Abstract

AIM To investigate regulatory effects of electro-acupuncture (EA) at Tsusanli (He-Sea Point, St 36) on changes in ir-SP and ir-VIP content in rats＇ pituitary gland and peripheral blood as well as on their cellular immunity and erythrocytic immunity; and to explore the mechanism, or the coordinating effects of “the stomach channel brain-gut immune network” and the possible inter relationship between the many factors involved in the coordination.

METHODS The rats were randomized into 5 groups, including the control, the Tsusanli, the immunosup-pressive model, the non-acupoint group and the Tsusanli+immunosuppressive groups. Changes in substance P (SP) and vasoactive intestinal peptide (VIP) contents in the pituitary gland and peripheral blood were determined by radioimmunoassay. With micro-whole-blood direct immunofluorescence staining and flow cytometry, T-lymphocytic subgroups were assayed to reflect the cellular immunity. Erythrocytic immunity was assayed by red blood cell C_3b receptor rosette (RBC-C_3bRR) and red blood cell immune complexes rosette (RBC-ICR).

RESULTS Ir-SP and ir-VIP content in the pituitary gland and peripheral blood and CD4+ and RBC-C_3bRR in the peripheral blood of the rats of the immunosuppressive group were significantly lower than that of the control group (P＜0.01, P＜0.01, P＜0.01 and P＜0.05 respectively). There was no significant change in CD8+ content (P＞0.05) of all the groups. There was no significant difference between the indexes of the Tsusanli+immunosuppressive group and that of the control group (P＞0.05). After EA at Tsusanli, a significant increase was observed in ir-SP and ir-VIP content in the pituitary gland and peripheral blood, CD4+, RBC-C_3bRR, RBC-ICR in the peripheral blood of the normal rats (P＜0.01) whereas CD8+ did not change significantly (P＞0.05). SP and VIP contents in the pituitary gland and the peripheral blood increased (P＜0.01). Besides, the change in CD4+ content and that in SP and VIP contents were positively correlated with each other (r=0.744, P＜0.05; r=0.738, P＜0.05;
r=0.822, P＜0.05; r=0.848, P＜0.05) and so were that between CD4+ and RBC-C_3bRR (r=0.719, P＜0.05). No significant difference was observed in the indexes between non-acupoint group and the control.

CONCLUSION EA at Tsusanli (He-Sea Point, St 36) can improve the cellular immunity and erythrocytic immuno-adhesiveness of normal rats and immunosuppressed rats. The changes in SP and VIP contents in the pituitary gland and peripheral blood indicate that EA can improve immunity of the body. The mechanism of this might be related to the increased synthesis and release of gastrointestinal hormones, which in turn affects the nerve-endocrine-immunoregulation network by means of these immuno-transmitters.

Subject headings electroacupuncture; stomach meridian; gastrointestinal hormones; T-lymphocyte subsets

Gao W, Huang YX, Chen H, Zhao NX, Sun DY, Zhang HX, Wang QL. Regulatory mechanism of electroacupuncture on the stomach channel brain-gut peptide immune network. Shijie Huaren Xiaohua Zazhi, 2001;9(3):279-283

摘要

目的探索“胃经-脑肠肽-免疫调节网络”在针刺调控胃肠功能中的多因素协调作用及可能的内在相互联系.

方法将大鼠随机分成正常对照组、非经非穴组、足三里组、免疫抑制组、足三里+免疫抑制组，同步观察电针后某些脑肠肽及免疫功能的变化. 应用放射免疫法测定脑垂体和外周血中P物质（SP）、血管活性肠肽（VIP）的含量；应用流式细胞仪技术，通过微量全血直接免疫荧光染色法测定外周血T细胞亚群以反映细胞免疫功能；应用红细胞C_3b受体-酵母菌花环试验和红细胞-IC花环试验检测红细胞免疫功能.

结果电针刺激足三里穴后正常大鼠外周血的CD4+, RBC-C_3bRR, RBC-ICR均明显升高（P均＜0.01），CD8+无显著变化（P＞0.05）；脑垂体和外周血中SP, VIP的含量增加（P均＜0.01）；且CD4+与SP, VIP含量的变化（r=0.744，P＜0.05；r=0.738，P＜0.05；r=0.822，P＜0.05；r=0.848，P＜0.05）、CD4+和RBC-C_3bRR呈显著正相关（r=0.719，P＜0.05）. 免疫抑制组大鼠CD4+, RBC-C_3bRR明显低于正常对照组（P均＜0.05），CD8+无显著变化（P＞0.05）；脑垂体和外周血中SP, VIP的含量减少（P均＜0.01）.足三里+免疫抑制组CD4+（P＜0.01）, RBC-C_3bRR（P＜0.01）以及脑垂体和外周血中SP, VIP的含量（P均＜0.01）与免疫抑制组有差异，与正常对照组相比无显著差异（P＞0.05）. 非经非穴组的各项指标与正常对照组相比无显著差异（P＞0.05）.

结论电针刺激足三里穴在提高正常大鼠及免疫抑制模型大鼠的细胞免疫功能和红细胞免疫粘附功能的同时可引起多种脑肠肽的变化，提示针刺胃经对神经-内分泌-免疫网络具有调节作用，这些脑肠肽又作为调节因子，对全身免疫系统发生影响，同时免疫网络之间、调节因子之间也存在着相互调节和制约，形成一个复杂、有机的网络体系，从而实现针刺对机体的双向性良性调衡作用.

主题词 电针；穴，足三里/针灸效应；胃经/针灸效应；胃肠激素类/针灸效应；T淋巴细胞亚群

高巍, 黄裕新, 陈洪, 赵宁侠, 孙大勇, 张洪新, 王庆莉. 电针对胃经-脑肠肽-免疫网络的调控机制. 世界华人消化杂志，2001；9(3)：279-283

0 引言

针灸对机体各系统、器官的生理功能和代谢活动具有相对穴位特异性的调衡作用.这种调衡作用是针灸治疗各系统多种疾病的基础^［¹^］. 研究显示，胃经上相应穴位的电针治疗对胃肠功能确有良好的调整作用^［^2-6^］, 其部分机制是通过对脑肠肽的影响而实现对胃肠功能的调节作用^［^7-15^］. 同时，电针足三里等穴位可直接或间接对免疫系统产生影响^［^16-26^］，而许多胃肠激素对免疫系统具有调节作用^［^27-34^］，据此，不能排除胃肠激素作为一种针刺调节免疫反应的调节因子而影响全身免疫系统. 那么，针刺调节免疫反应的同时对神经-内分泌-免疫调节网络有无影响它是通过什么途径如何影响这一网络的其主要递质是什么它们相互之间有无联系目前国内外的研究多为单因素、分散的研究，缺乏系统、同步的实验研究资料. 本研究从“胃经-脑肠肽-免疫网络调控”同步研究入手，探索这一网络在针刺调控胃肠功能中的多因素协调作用及可能的内在相互联系，力求为针刺治疗相关疾病提供进一步的科学理论依据.

1 材料和方法

1.1 材料上海产SD健康大白鼠（动物由本校实验动物研究中心提供），体质量150g～180g，12wk～14wk. 应用分段随机分组法（blocked randomization）将大鼠分为5组（每组8只）：正常对照组、足三里组、非经非穴组、免疫抑制组、足三里+免疫抑制组. G6805-A型电针治疗仪由上海医疗仪器厂生产，放免试剂盒由北京海科锐生物制品有限公司提供，Epics-profileⅡ型流式细胞仪由美国Coulter公司产.

1.2 方法 ① 免疫抑制模型的制作：于d1大鼠腹腔注射（ip）新鲜配制的环磷酰胺（CY）溶液，剂量为100mg·kg^-1体重，d4加强注射1次，制成免疫抑制模型. 其余各组大鼠给腹腔注射同剂量的生理盐水. ② 针刺方法：所有实验大鼠均在安静环境下统一喂养. 穴位：双侧足三里穴^［^1,35^］，非经非穴点选足三里穴旁0.5cm处. 参数为断续波，频率2Hz，强度3V，以大鼠后肢轻微抖动为宜. 足三里组大鼠固定后每天辰时于双侧“足三里”连续刺激30min， 7d；正常对照组大鼠每天辰时以同样方法固定30min，但不行电针刺激. ③ 标本采集及测定：所有大鼠于电针d7 ip 10g·L^-1戊巴比妥钠，30mg·kg^-1，麻醉后断头采血，125KU·L^-1肝素抗凝. 应用放射免疫分析法（Radioimmunoassay, RIA）测定脑垂体和外周血中ir-SP, ir-VIP含量，按试剂盒说明书进行. 微量全血直接免疫荧光染色法检测T淋巴细胞亚群，抗大鼠CD4FITC/CD8PE单克隆抗体由英国Serotec公司提供. 应用花环法^［³⁶^］测定大鼠的C_3b受体花环率（RBC-C_3bRR）、红细胞免疫复合物花环率（RBC-ICR）.

统计学处理采用SPSS 8.0软件进行数据分析，所用统计学方法为方差分析、直线相关.

2 结果

T淋巴细胞亚群和红细胞免疫功能（表1）、外周血及脑垂体中P物质、血管活性肠肽含量（表2）均有改变. 足三里组T细胞亚群和红细胞免疫粘附功能增高，同时，脑垂体和外周血中ir-SP, ir-VIP的含量也明显升高，且CD4+与外周血和脑垂体中SP, VIP含量的变化, CD4+与C_3b受体花环率（RBC-C_3bRR）呈显著正相关. 非经非穴组大鼠各项指标与正常对照组相比无显著差异.

表1 T细胞亚群和红细胞免疫功能的变化（n=8，x±s）

组别	CD4+	CD8+	RBC-C_3bRR	RBC-ICR
正常对照组	43.1±3.1	23.6±2.6	8.6±2.1	6.8±2.3
非经非穴组	44.7±4.7	21.0±4.3	11.1±1.2	8.9±2.9
免疫抑制组	34.5±2.5^b	26.2±8.4	5.3±1.2^a	9.8±4.4
足三里组	65.5±8.4^b	29.5±8.2	15.9±3.0^b	12.1±1.2^b
足三里+
免疫抑制组	48.8±6.0^d	22.6±7.0	16.4±4.1^d	15.0±5.2^bd

^aP＜0.05,^bP＜0.01,vs 正常对照组; ^dP＜0.01,vs 免疫抑制组.

表2 外周血和脑垂体中P物质、血管活性肠肽含量的变化（ng·L^-1，n=8，x±s）

组别	P物质		血管活性肠肽
	脑垂体	外周血	脑垂体	外周血
正常对照组	569