Bidirectional regulation of distal colon motility in rats with electroacupuncture of different intensities at "Tianshu" (ST25)

LI Tong; LIU Xiaoyu; WANG Xiaoyu; LUO Min; ZHANG Zhiyun; SU Yangshuai; JING Xianghong

doi:10.13703/j.0255-2930.20240409-k0001

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Bidirectional regulation of distal colon motility in rats with electroacupuncture of different intensities at "Tianshu" (ST25)

Study on Mechanism | 更新时间：2025-04-11

- Bidirectional regulation of distal colon motility in rats with electroacupuncture of different intensities at "Tianshu" (ST25)
- Chinese Acupuncture & Moxibustion Vol. 45, Issue 4, Pages: 460-472(2025)
- 作者机构：
  
  中国中医科学院针灸研究所，北京　100700
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20240409-k0001
  CLC：
- Received：09 April 2024，
  
  Online First：22 November 2024，
  
  Published：12 April 2025
- 稿件说明：
移动端阅览
LI Tong, LIU Xiaoyu, WANG Xiaoyu, et al. Bidirectional regulation of distal colon motility in rats with electroacupuncture of different intensities at "Tianshu" (ST25)[J]. Chinese Acupuncture & Moxibustion, 2025, 45(4): 460-472.
DOI：

LI Tong, LIU Xiaoyu, WANG Xiaoyu, et al. Bidirectional regulation of distal colon motility in rats with electroacupuncture of different intensities at "Tianshu" (ST25)[J]. Chinese Acupuncture & Moxibustion, 2025, 45(4): 460-472. DOI： 10.13703/j.0255-2930.20240409-k0001.

摘要

目的：

观察功能性便秘、腹泻模型大鼠体表敏化区的分布特点，探究不同强度电针“天枢”对大鼠远端结肠运动的调节规律，明确肠神经系统（ENS）内不同亚型神经元在该调节效应中发挥的作用。

方法：

选用90只SPF级雄性SD大鼠，将其中15只大鼠随机分为正常组、便秘组、腹泻组，每组5只，观察粪便形态与含水量，以及尾静脉注射伊文氏蓝（EB）后大鼠体表EB渗出点的分布规律；18只大鼠随机分为正常+2 mA组、正常+4 mA组和正常+6 mA组，每组6只，利用生理信号采集分析系统记录、分析大鼠结肠蠕动波曲线下面积和平均幅值，观察不同强度电针“天枢”对远端结肠蠕动的即刻效应；30只大鼠随机分为正常组、便秘组、腹泻组、便秘+2 mA组、腹泻+6 mA组，每组6只，观察电针“天枢”对便秘和腹泻模型大鼠结肠动力紊乱的累积效应；12只大鼠随机分为便秘+2 mA组、腹泻+6 mA组，每组6只，用于观察电针“天枢”对便秘和腹泻模型大鼠结肠动力紊乱的即刻效应；15只大鼠随机分为正常组、便秘组、腹泻组、便秘+2 mA组、腹泻+6 mA组，每组3只，采用whole-mount染色检测ENS内囊泡乙酰胆碱转运体（VAChT）阳性神经元和一氧化氮合酶（nNOS）阳性神经元表达情况。根据分组，通过盐酸洛哌丁胺灌胃（10 mg·kg

-1

·d

-1

，连续7 d）建立功能性便秘模型，番泻叶药液灌胃（10 mL/kg，每天1次，连续2 d）建立功能性腹泻模型；于双侧“天枢”行不同强度（电流2、4、6 mA）电针干预，连续波，频率10 Hz。

结果：

与正常组比较，便秘组大鼠粪便数量减少，粪便含水量降低（

0.001）；腹泻组大鼠出现便溏症状，粪便含水量升高（

0.001）。便秘组和腹泻组大鼠体表EB渗出点可见于T

-S

节段区域，且在T

-L

节段的下腹部和下背部较为集中。与电针前比较，正常+2 mA组、正常+4 mA组大鼠电针过程中远端结肠蠕动波曲线下面积和平均幅值升高（

0.01，

0.05），表现为促进性的即刻效应，2 mA电针具有后效应；

而正常+6 mA组大鼠电针过程中远端结肠蠕动波曲线下面积和平均幅值较电针前降低（

0.001），呈现出抑制性的即刻效应。多次电针干预结束后，与电针前比较，便秘+2 mA组大鼠电针过程中结肠蠕动波曲线下面积和平均幅值升高（

0.05，

0.01），腹泻+6 mA组大鼠电针过程中结肠蠕动波曲线下面积和平均幅值降低（

0.01，

0.05）；便秘+2 mA组大鼠结肠蠕动波曲线下面积高于便秘组（

0.001），腹泻+6 mA组大鼠结肠蠕动波曲线下面积低于腹泻组（

0.001）；电针对便秘+2 mA组大鼠结肠动力的促进效应强于正常+2 mA组（

0.05），电针的抑制效应在正常+6 mA组和腹泻+6 mA组大鼠间的差异无统计学意义（

0.05）。远端结肠局部ENS中，2 mA电针激活VAChT阳性神经元的比例高于其激活nNOS阳性神经元的比例（

0.001），而6 mA电针则以激活nNOS阳性神经元为主（

0.001）。

结论：

功能性便秘和腹泻模型状态下，大鼠体表敏化区集中分布于T

-L

节段的下腹部和下背部区域。电针“天枢”对远端结肠运动具有双向调节作用，该效应与电针强度相关，且可能由不同亚型的ENS神经元介导完成。

Abstract

Objective

To observe the distribution characteristics of sensitization areas on the body surface in the rat models with functional constipation and diarrhea

explore the regulatory patterns of electroacupuncture (EA) of different intensities at "Tianshu" (ST25) on distal colon motility

and clarify the roles of the neurons of different subtypes in the enteric nervous system (ENS) displayed in the regulatory effect.

Methods

Of 90 SD male rats of SPF grade

15 rats were randomized into a normal group

a constipation group and a diarrhea group

5 rats in each one. The stool form and fecal water content

as well as the distribution of the Evans blue (EB) extravasation on the body surface after the intravenous injection with EB on the tails were observed. Eighteen rats were randomized into a normal +2 mA group

a normal +4 mA group and a normal + 6 mA group

6 rats in each one. Using physiological signal acquisition system

the area under the curve and the average amplitude of colon peristalsis were recorded and analyzed

and the immediate effect on distal colon peristalsis observed after EA with different intensities at "Tianshu" (ST25). Thirty rats were randomized into a normal group

a constipation group

a diarrhea group

a constipation +2 mA group

and a diarrhea +6 mA group

6 rats in each one

so as to observe the cumulative effect on colon motility disorder in the rat models of constipation and diarrhea after EA at "Tianshu" (ST25). Twelve rats were randomized into a constipation +2 mA group and a diarrhea +6 mA group

6 rats in each one

to observe the immediate effect on colon motility disorder in the rat models of constipation and diarrhea after EA at "Tianshu" (ST25). Fifteen rats were randomly divided into a normal group

a constipation group

a diarrhea group

a constipation +2 mA group

and a diarrhea + 6 mA group

3 rats in each one. Using the whole-mount staining technique

the expression of vesicular acetylcholine transporter (VAChT)-positive neurons and nitric oxide synthase (nNOS)-positive neurons in ENS was detected. According to the group divisions

the functional constipation models were established by intragastric administration of loperamide hydrochloride (10 mg/kg

once daily

for consecutive 7 days)

and the functional diarrhea models were prepared by intragastric administration of folium sennae decoction (10 mL/kg

once daily

for consecutive 2 days). The interventions were delivered with EA of different intensities (the electric current of 2

4 or 6 mA) at bilateral "Tianshu" (ST25)

separately

with the continuous wave and the frequency of 10 Hz used.

Results

Compared with the normal group

the fecal amount was decreased

and the fecal water content was reduced in the rats of the constipation group (

0.001); and loose stool was presented and the fecal water content increased in rats of the diarrhea group (

0.001). EB extravasation on the body surface happened in the region from T

to S

of the rats in the constipation and diarrhea groups

and it was more concentrated in the lower abdominal an

d the lower back regions from T

to L

. Compared with the indexes before EA

in the normal +2 mA group and the normal +4 mA group

the areas under the curve and the average amplitude of the distal colon peristalsis were higher during EA delivery (

0.01

0.05)

showing a stimulatory immediate effect; and the post-effect was obtained after EA at 2 mA. Whereas

these two indexes were declined during EA in the rats of the normal +6 mA group (

0.001)

showing an inhibitory immediate effect. After many interventions with EA

when compared with those before EA

the above two indexes rose in the constipation +2 mA group (

0.05

0.01)

and they were dropped in the diarrhea +6 mA group (

0.01

0.05). The area under the curve of the colon peristalsis in the constipation +2 mA group was higher than that of the constipation group (

0.001)

and that in the diarrhea +6 mA group was lower compared with that in the diarrhea group (

0.001). The stimulatory effect of EA on colon motility in the constipation +2 mA group was stronger than that of the normal + 2 mA group (

0.05)

and its inhibitory effect was not different statistically in comparison between the normal +6 mA group and the diarrhea +6 mA group (

0.05). In ENS of the distal colon

after EA at 2 mA

the proportion of VAChT-positive neurons was higher than that of the activated nNOS-positive neurons (

0.001); and after EA at 6 mA

the activated nNOS-positive neurons were dominant (

0.001).

Conclusion

In the functional constipation and diarrhea rat models

the sensitization areas on the body surface are centralized in the lower abdominal and the lower back regions of T

to L

. Electroacupuncture at "Tianshu" (ST25) has a bidir

ectional regulatory effect on distal colon motility

and this effect is coordinated with the intensity of electroacupuncture

and may be mediated by ENS neurons of different subtypes.

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