Department of Massage, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193.
National Clinical Research Center of Chinese Medicine Acupuncture and Moxibustion, Tianjin 300193, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2022 Apr 28;47(4):488-496. doi: 10.11817/j.issn.1672-7347.2022.210260.
Electroacupuncture can enhance autophagic flow, promote neuronal regeneration, axonal and myelin remodeling to achieve the protection of spinal cord injury, but its role in neurogenic urine retention is not completely clear. This study aims to investigate whether the mechanism of electroacupuncture in the treatment of neurogenic urine retention is through autophagy mediated by adenosine monophosphate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway.
A rat model of neurogenic urine retention after sacral spinal cord injury was established. The rats with successful model were randomly divided into a model group, an electroacupuncture group (electro-acupuncture for Ciliao, Zhongji, and Sanyinjiao by electronic stimulation, once a day, 20 min each time for 7 days), and an electroacupuncture+AMP-activated protein kinase (AMPK) inhibitor group (on the basis of the treatment of electroacupuncture group, 100 μg of AMPK inhibitor compound C was injected intramuscularly around the L intervertebral space on the 1st and 4th day). The normal group did not receive any treatment. The maximum bladder volume, bladder basal pressure, leak point pressure, and bladder compliance were recorded by multi-channel physiological recorder; the morphology of bladder tissue was observed by HE staining; autophagy was observed under transmission electron microscope; the expressions of LC3II and Beclin1 protein were observed by immunofluorescence staining; the protein levels of AMPK, phosphorylated-AMPK (p-AMPK), mTOR, phosphorylated-mTOR (p-mTOR), microtubule associated protein 1 light chain 3 (LC3) II and Beclin1 in bladder tissue were detected by Western blotting.
Compared with the normal group, the maximum bladder capacity, leak point pressure, bladder compliance, p-AMPK, LC3II, Beclin1 protein expressions in the bladder tissue of the model group increased, and the p-mTOR protein expressions were decreased (all <0.05); compared with the model group, the maximum bladder capacity, bladder compliance, p-mTOR protein expression in the bladder tissue of the electroacupuncture group were decreased, and the p-AMPK, LC3II, and Beclin1 protein expressions were increased (all <0.05); compared with the electroacupuncture group, the maximum bladder capacity, bladder compliance, p-mTOR protein expression in the bladder tissue of the electroacupuncture+AMPK inhibitor group were increased, the p-AMPK, LC3II, and Beclin1 protein expressions were decreased (all <0.05). In the model group, the bladder became larger, with unclear and varying degrees of degeneration, severe tissue damage and autophagosome appeared; the bladder of the electroacupuncture group was smaller than that of the model group, and all levels were clearly visible with autophagy bodies; the layers were slightly disordered and damaged in the electroacupuncture + AMPK inhibitor group.
Electroacupuncture can activate autophagy through AMPK/mTOR pathway, thereby reducing neurogenic urine retention caused by spinal cord injury.
电针对脊髓损伤后神经源性尿潴留具有治疗作用，其机制可能与促进自噬、促进神经元再生、轴突和髓鞘重塑有关，但其对神经源性尿潴留的作用机制尚不完全清楚。本研究旨在探讨电针对神经源性尿潴留的治疗作用是否通过 AMPK/mTOR 通路介导的自噬实现。
采用大鼠骶髓损伤后神经源性尿潴留模型，成功建模的大鼠随机分为模型组、电针组（电子刺激 Ciliao、Zhongji 和 Sanyinjiao，每日 1 次，每次 20 min，共 7 天）和电针+AMPK 抑制剂组（在电针组治疗的基础上，于第 1、4 天在 L 椎间肌周围肌肉注射 100 μg AMPK 抑制剂化合物 C）。正常组不做任何处理。多道生理记录仪记录最大膀胱容量、膀胱基础压、漏点压和膀胱顺应性；HE 染色观察膀胱组织形态；透射电镜观察自噬体；免疫荧光染色观察 LC3II 和 Beclin1 蛋白表达；Western blot 检测膀胱组织中 AMPK、磷酸化 AMPK（p-AMPK）、mTOR、磷酸化 mTOR（p-mTOR）、微管相关蛋白 1 轻链 3（LC3）II 和 Beclin1 蛋白水平。
与正常组比较，模型组大鼠最大膀胱容量、漏点压、膀胱顺应性、膀胱组织 p-AMPK、LC3II、Beclin1 蛋白表达升高，p-mTOR 蛋白表达降低（均<0.05）；与模型组比较，电针组大鼠最大膀胱容量、膀胱顺应性、膀胱组织 p-mTOR 蛋白表达降低，p-AMPK、LC3II、Beclin1 蛋白表达升高（均<0.05）；与电针组比较，电针+AMPK 抑制剂组大鼠最大膀胱容量、膀胱顺应性、膀胱组织 p-mTOR 蛋白表达升高，p-AMPK、LC3II、Beclin1 蛋白表达降低（均<0.05）。模型组大鼠膀胱体积增大，形态不一，有不同程度的退行性变，组织损伤严重，自噬体出现；电针组大鼠膀胱体积小于模型组，各层均可见，可见自噬体；电针+AMPK 抑制剂组大鼠膀胱组织层次稍紊乱，有损伤。
电针可能通过 AMPK/mTOR 通路激活自噬，从而减轻脊髓损伤引起的神经源性尿潴留。
