Wang Lu, Chen Ying, Xu Ming-Min, Cao Wei, Zheng Qian-Hua, Zhou Si-Yuan, Yao Jun-Peng, Xi Meng-Han, Qin Hai-Yan, Li Ying, Zhang Wei
Acupuncture and Tuina School, Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China.
School of Traditional Chinese Medicine, Jinan University, Guangzhou, 510632, China.
Chin J Integr Med. 2023 May;29(5):459-469. doi: 10.1007/s11655-023-3594-3. Epub 2023 Mar 28.
To investigate autophagy-related mechanisms of electroacupuncture (EA) action in improving gastrointestinal motility in mice with functional constipation (FC).
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.
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.
EA treatment can inhibit PI3K/AKT/mTOR signaling in the colonic tissues of FC mice, thereby promoting EGCs autophagy to improve intestinal motility.
探讨电针（EA）改善功能性便秘（FC）小鼠胃肠动力的自噬相关机制。
在实验I中，根据随机数字表将昆明小鼠分为正常对照组、FC组和EA组。在实验II中使用自噬抑制剂3-甲基腺嘌呤（3-MA）观察其是否拮抗EA的作用。通过灌胃地芬诺酯建立FC模型。然后对小鼠进行天枢（ST 25）和上巨虚（ST 37）穴位的EA刺激。采用首次排黑便时间、8小时粪便数量、重量和含水量以及肠道传输率来评估肠道传输情况。对结肠组织进行组织病理学评估，并通过免疫组织化学染色检测自噬标志物微管相关蛋白1轻链3（LC3）和Beclin-1的表达。分别通过蛋白质免疫印迹法和定量逆转录-聚合酶链反应研究磷酸肌醇3-激酶（PI3K）-蛋白激酶B（AKT）-雷帕霉素靶蛋白（mTOR）信号通路成员的表达。通过共聚焦免疫荧光显微镜、定位分析和电子显微镜观察肠胶质细胞（EGC）与自噬之间的关系。
EA治疗缩短了FC小鼠的首次排黑便时间，增加了8小时粪便的数量、重量和含水量，并提高了肠道传输率（P<0.01）。在假定的自噬机制方面，EA治疗促进了FC小鼠结肠组织中LC3和Beclin-1蛋白的表达（P<0.05），胶质纤维酸性蛋白（GFAP）和LC3显著共定位。此外，EA通过抑制PI3K/AKT/mTOR信号通路促进FC小鼠结肠自噬（P<0.05或P<0.01）。3-MA阻断了EA对FC小鼠肠道动力的积极作用。
EA治疗可抑制FC小鼠结肠组织中的PI3K/AKT/mTOR信号通路，从而促进EGC自噬以改善肠道动力。
