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Electroacupuncture Alleviates Functional Constipation in Mice by Activating Enteric Glial Cell Autophagy via PI3K/AKT/mTOR Signaling

  • Acupuncture Research
  • Published:
Chinese Journal of Integrative Medicine Aims and scope Submit manuscript

Abstract

Objective

To investigate autophagy-related mechanisms of electroacupuncture (EA) action in improving gastrointestinal motility in mice with functional constipation (FC).

Methods

According to a random number table, the Kunming mice were divided into the normal control, FC and EA groups in Experiment I. The autophagy inhibitor 3-methyladenine (3-MA) was used to observe whether it antagonized the effects of EA in Experiment II. An FC model was established by diphenoxylate gavage. Then the mice were treated with EA stimulation at Tianshu (ST 25) and Shangjuxu (ST 37) acupoints. The first black stool defecation time, the number, weight, and water content of 8-h feces, and intestinal transit rate were used to assess intestinal transit. Colonic tissues underwent histopathological assessment, and the expressions of autophagy markers microtubule-associated protein 1 light chain 3 (LC3) and Beclin-1 were detected by immunohistochemical staining. The expressions of phosphoinositide 3-kinases (PI3K)-protein kinase B (AKT)-mammalian target of rapamycin (mTOR) signaling pathway members were investigated by Western blot and quantitative reverse transcription-polymerase chain reaction, respectively. The relationship between enteric glial cells (EGCs) and autophagy was observed by confocal immunofluorescence microscopy, localization analysis, and electron microscopy.

Results

EA treatment shortened the first black stool defecation time, increased the number, weight, and water content of 8-h feces, and improved the intestinal transit rate in FC mice (P<0.01). In terms of a putative autophagy mechanism, EA treatment promoted the expressions of LC3 and Beclin-1 proteins in the colonic tissue of FC mice (P<0.05), with glial fibrillary acidic protein (GFAP) and LC3 significantly colocalized. Furthermore, EA promoted colonic autophagy in FC mice by inhibiting PI3K/AKT/mTOR signaling (P<0.05 or P<0.01). The positive effect of EA on intestinal motility in FC mice was blocked by 3-MA.

Conclusion

EA treatment can inhibit PI3K/AKT/mTOR signaling in the colonic tissues of FC mice, thereby promoting EGCs autophagy to improve intestinal motility.

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Authors and Affiliations

  1. Acupuncture and Tuina School, Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China

    Lu Wang, Ying Chen, Wei Cao, Qian-hua Zheng, Si-yuan Zhou, Jun-peng Yao, Meng-han Xi, Hai-yan Qin, Ying Li &  Wei Zhang

  2. School of Traditional Chinese Medicine, Jinan University, Guangzhou, 510632, China

    Ming-min Xu

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  1. Lu Wang
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Contributions

Wang L, Chen Y, Xu MM and Zhang W were responsible for design of the study and drafted manuscript; Cao W, Yao JP, Xi MH and Qin HY performed the experiments; Zheng QH and Zhou SY were responsible for data analysis; Wang L, Zhang W and Li Y revised the manuscript. All authors read and approved the final manuscript for publication.

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Correspondence to Wei Zhang.

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Additional information

Supported by the National Natural Science Foundation of China (No. 82074554), the Foundation of the Science and Technology Department of Sichuan Province, China (No. 2021YJ0197), and Chengdu University of Traditional Chinese Medicine Xinglin Scholars Program (No. YXRC2018007)

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Wang, L., Chen, Y., Xu, Mm. et al. Electroacupuncture Alleviates Functional Constipation in Mice by Activating Enteric Glial Cell Autophagy via PI3K/AKT/mTOR Signaling. Chin. J. Integr. Med. 29, 459–469 (2023). https://doi.org/10.1007/s11655-023-3594-3

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