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. 2023 Aug 1;130(2):380-391.
doi: 10.1152/jn.00156.2023. Epub 2023 Jul 12.

Electroacupuncture in the treatment of IBS in rats: investigation of the mechanisms of CRH+ neurons in the paraventricular nucleus

Fang Gao  1 , Wei-Hua Yuan  1 , Sheng-Bing Wu  1 , Zi-Bei Wang  2 , Guo-Qi Zhu  1 , Mei-Qi Zhou  1
Affiliations

Electroacupuncture in the treatment of IBS in rats: investigation of the mechanisms of CRH+ neurons in the paraventricular nucleus

Fang Gao et al. J Neurophysiol. .

Abstract

Electroacupuncture (EA) is well documented to treat irritable bowel syndrome (IBS). However, the mechanism of the central nervous system related to IBS and acupuncture stimulation is still not well known. In this study, a rat model of IBS was established by cold-restraint comprehensive stresses for 15 days, and it was found that the levels of corticotropin-releasing hormone (CRH), corticosterone (CORT), and adrenocorticotropic hormone (ACTH) in the peripheral serum were increased; the visceral sensitivity was enhanced; and the intestinal motility was accelerated, specifically, there was an enhancement in the discharge frequency of neurons in the paraventricular nucleus (PVN). EA treatment for 3 days, 20 min/day, alleviated the increase in the levels of CRH, CORT, and ACTH in the peripheral serum of rats, reduced the visceral sensitivity of IBS rats, and inhibited colon movement and discharge frequency of the neurons in the PVN. In addition, EA could reduce the excitability of CRH neurons and the expression of corticotropin-releasing hormone receptor 1 (CRHR1) and corticotropin-releasing hormone receptor 2 (CRHR2) in PVN. At the same time, the expression of CRH, CRHR1, and CRHR2 in the peripheral colon was decreased. Taken together, EA appears to regulate intestinal functional activity through the central CRH nervous system, revealing the central regulation mechanism of EA in IBS rats, and providing a scientific research basis for the correlation among the meridians, viscera, and brain.NEW & NOTEWORTHY The purpose of this research was to determine the central regulatory mechanism of electroacupuncture (EA) in rats with irritable bowel syndrome (IBS). Our results showed that combined with the serum changes in corticotropin-releasing hormone (CRH), corticosterone (CORT), and adrenocorticotropic hormone (ACTH), the improvement of IBS by EA was related to them. Furthermore, EA could regulate intestinal functional activity through the central CRH+ nervous system.

Keywords: corticotropin-releasing hormone; corticotropin-releasing hormone receptor 1; corticotropin-releasing hormone receptor 2; electroacupuncture; irritable bowel syndrome.

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Conflict of interest statement

No conflicts of interest, financial or otherwise, are declared by the authors.

Figures

None
Graphical abstract
Figure 1.
Figure 1.
Electroacupuncture (EA) increased visceral pain threshold and inhibited intestinal motility. A: experimental process diagram. After 7 days of adaptation, except for the control group rats, the other groups rats were treated with chronic allotype stress for 15 days. Except for the control group and irritable bowel syndrome (IBS) group, the other groups were treated with acupuncture for 3 days. Finally, visceral pain threshold, electrophysiological test, and intestinal propulsion rate were detected, and abdominal aortic blood was collected. B: the body weight of rats in each group was recorded every other day for 17 days. C: statistical graphics of visceral pain threshold values of each group. D: statistical graphics of intestinal propulsion rate values of each group. The data were analyzed using repeated-measures one-way ANOVA and t tests. (n = 8 rats/group). *P < 0.05 vs. IBS group; **P < 0.05 vs. EA group. Values are presented as means ± SD.
Figure 2.
Figure 2.
Electroacupuncture (EA) decreased the serum concentrations of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and corticosterone (CORT) of irritable bowel syndrome (IBS) rats. A: statistical graphics of the serum concentration of CRH of each group. B: statistical graphics of the serum concentration of CORT of each group. C: statistical graphics of the serum concentration of ACTH of each group. The data were analyzed using repeated-measures one-way ANOVA and t tests. (n = 8 rats/group). *P < 0.05 vs. IBS group; **P < 0.05 vs. EA group. Values are presented as means ± SD.
Figure 3.
Figure 3.
Analysis of paraventricular nucleus (PVN) discharge frequency in each group. A: cluster analysis of the discharge frequency of PVN neurons in irritable bowel syndrome (IBS) rats undergoing electroacupuncture (EA). B: discharge frequency of the PVN in IBS rats undergoing EA (statistical graphics of rate, time = 300 s). The data were analyzed using repeated-measures one-way ANOVA and t tests. (n = 5 rats/group). *P < 0.05 vs. IBS group; **P < 0.05 vs. EA group. Values are presented as means ± SD. C: real-time spectrum analysis of PVN neurons discharge. The four groups were sequenced as follows: IBS group > nonacupoint > control group > EA group.
Figure 4.
Figure 4.
Electroacupuncture (EA) regulated the excitatory and inhibitory effects of paraventricular nucleus (PVN) corticotropin-releasing hormone (CRH) neurons. A: experimental process diagram. After 5 days of adaptation, except for the EA group, the PVN of rats in other groups was injected with virus mixture, then each group was treated with chronic allotype stress for 15 days. Except for the hm4D(Gi) group and hm3D(Gq) group rats, the other groups were treated with acupuncture for 3 days, at the same time. Except for the EA group rats, the other groups were intraperitoneally injected with clozapine N-oxide (CNO). B: virus mixture injection diagram. C: PVN (bregma: −1.8 mm, LR: 0.4 mm, and H: 8 mm) was accurately localized by hematoxylin-eosin (H&E) (100 µm). D1–F5: immunofluorescence sections of PVN, DAPI, and CRH-C-FOS of each group (20 µm). G1–G5: CRH-C-FOS co-labeled scatterplot by Scatter J of each group.
Figure 5.
Figure 5.
Electroacupuncture (EA) decreased the expression of corticotropin-releasing hormone receptor 1 (CRHR1) and corticotropin-releasing hormone receptor 2 (CRHR2) in the paraventricular nucleus (PVN) of experimental rats. A1–F5: immunofluorescence sections of PVN, CRHR1, CRHR2, and diamidino-phenyl indole (DAPI) of each group (20 µm). G and H: mean fluorescence intensity values in statistical graphics of CRHR1 and CRHR2 of each group. The data were analyzed using repeated-measures one-way ANOVA and t tests. (n = 5 rats/group). *P < 0.05, the hm4D(Gi) group vs. the EA + hm4D(Gi) group and the hm3D(Gq) group vs. the EA + hm3D(Gq); **P < 0.05 vs. EA group. Values are presented as means ± SD.
Figure 6.
Figure 6.
Electroacupuncture (EA) reduced the expression of corticotropin-releasing hormone (CRH), corticotropin-releasing hormone receptor 1 (CRHR1), and corticotropin-releasing hormone receptor 2 (CRHR2) in the colon. A–C: immunofluorescence sections of CRH, CRHR1, and CRHR2 in the colon from the same site (50 µm). D–F: Western blot representative stripes CRH, CRHR1, and CRHR2 of the colon and CRH/GAPDH, CRHR1/GAPDH, and CRHR2/GAPDH statistical graphics analysis of each group. The data were analyzed using repeated-measures one-way ANOVA and t tests. (n = 5 rats/group). *P < 0.05, the hm4D(Gi) group vs. the EA + hm4D(Gi) group and the hm3D(Gq) group vs. the EA + hm3D(Gq); **P < 0.05 vs. EA group. Values are presented as means ± SD.
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