Neuromuscular electrical stimulation promoting the recovery of motor function in mice after spinal cord injury by regulating interleukin-6/signal transducer and activator of transcription-3 signaling pathway

doi:10.16098/j.issn.0529-1356.2024.03.002

Abstract

Abstract:

Objective To observe the effect of neuromuscular electrical stimulation (NMES) on interleukin-6 (IL-6)/STAT3 signaling pathway in mice after spinal cord injury, and to explore the mechanism of its effect on motor function recovery. Methods Seventy-two SPF grade mice were randomly divided into sham operation group, spinal cord injury (SCI) group and NMES group. BMS score, inclined plane test and neuromuscular electrophysiology (EMG) were used to evaluate the recovery of spinal cord injury in mice. Western blotting and Real-time PCR were used to detect the expression of inflammatory factors，IL-6/STAT3, glial fibrillary acidic protein (GFAP)and brain-derived neurotrophic factor (BDNF) in spinal cord tissues of three groups of mice. HE staining was used to observe the pathological changes of spinal cord injury. Results BMS scores and the inclined plane test of mice in the NMES group were higher than those in SCI group (P<0.05). The maximum amplitude of motor evoked potential in NMES group was higher than that in SCI group (P<0.05). The expressions of TNF-α, IL-12A and GFAP in the spinal cord of NMES group were lower than that of SCI group (P<0.05), while the expressions of TGF-β, IL-10 and BDNF were higher than that of SCI group (P<0.05). The protein expressions of IL-6/STAT3 signaling pathway of NMES group were lower than that of SCI group(P<0.05). Conclusion Neuromuscular electrical stimulation plays an anti-inflammatory role by inhibiting the IL-6/STAT3 signaling pathway, thereby promoting the recovery of hind limb motor function in mice after spinal cord injury.

Key words: Spinal cord injury, Inflammation, Interleukin-6/signal transducer and activator of transcription-3 signaling pathway, Neuromuscular electrical stimulation, Western blotting, Mouse

CLC Number:

R322.81

QIN Jian-feng SONG Hai-wang SUN Bao-fei JI Yang-dan LONG Si-fang YANG Dan. Neuromuscular electrical stimulation promoting the recovery of motor function in mice after spinal cord injury by regulating interleukin-6/signal transducer and activator of transcription-3 signaling pathway[J]. Acta Anatomica Sinica, 2024, 55(3): 260-267.

References

［1］ Hamid R, Averbeck M A, Chiang H, et al. Epidemiology and pathophysiology of neurogenic bladder after spinal cord injury［J］. World J Urol, 2018,36(10):1517-1527.

［2］ Karsy M, Hawryluk G. Modern medical management of spinal cord injury［J］. Curr Neurol Neurosci Rep, 2019,19(9):65-71.

［3］ Hachem LD, Fehlings MG. Pathophysiology of spinal cord injury［J］. Neurosurg Clin N Am, 2021,32(3):305-313.

［4］ Hellenbrand DJ, Quinn CM, Piper ZJ, et al. Inflammation after spinal cord injury: a review of the critical timeline of signaling cues and cellular infiltration［J］. J Neuroinflammation, 2021,18(1):284-299.

［5］ Pang QM, Chen SY, Xu QJ, et al. Neuroinflammation and scarring after spinal cord injury: therapeutic roles of MSCs on inflammation and glial scar［J］. Front Immunol, 2021,12：751021.

［6］ Deng WW，Min LX，Feng Zh，et al. A preliminary study on effects of optogenetic stimulation of glutamate neurons in the M1 region on the functional recovery of rats with spinal cord injury［J］.Chinese Journal of Rehabilitation Medicine, 2021, 36(5):520-525. (in Chinese)

邓维维, 闵令霞, 冯州, 等. 光遗传学靶向激活M1区谷氨酸能神经元促进脊髓损伤大鼠运动功能恢复的初步研究［J］. 中国康复医学杂志, 2021,36(5):520-525.

［7］ Huang L, Deng Y, Zheng X, et al. Transcranial direct current stimulation with halo sport enhances repeated sprint cycling and cognitive performance［J］. Front Physiol, 2019, 10:118-124.

［8］ Cebeci GC, Cebeci H, Kucuk MP, et al. Neuromuscular Electrical Stimulator as a Protective Treatment against Intensive Care Unit Muscle Wasting in Sepsis/Septic Shock Patients ［J］. J Coll Physicians Surg Pak, 2022,32(10):1300-1307.

［9］ Silva PE, de Cássia Marqueti R, LivinodeCarvalho K, et al. Neuromuscular electrical stimulation in critically ill traumatic brain injury patients attenuates muscle atrophy, neurophysiological disorders, and weakness: a randomized controlled trial［J］. J Intensive Care, 2019,7:59-71.

［10］ Wu WD, Wang LH, Wei NX, et al. MicroRNA-15a inhibits inflammatory response and apoptosis after spinal cord injury via targeting STAT3［J］. Eur Rev Med Pharmacol Sci, 2019,23(21):9189-9198.

［11］ Wang Y, Sang X, Shao R, et al. Xuanfei Baidu Decoction protects against macrophages induced inflammation and pulmonary fibrosis via inhibiting IL-6/STAT3 signaling pathway［J］. J Ethnopharmacol, 2022,283(30):114701.

［12］ Ren Q, Tao S, Guo F, et al. Natural flavonol fisetin attenuated hyperuricemic nephropathy via inhibiting IL-6/JAK2/STAT3 and TGF-β/SMAD3 signaling［J］. Phytomedicine, 2021,87:153552.

［13］ Starsichova A, Lincova E, Pernicova Z, et al. TGF-beta1 suppresses IL-6-induced STAT3 activation through regulation of Jak2 expression in prostate epithelial cells［J］. Cell Signal, 2010,22(11):1734-1744.

［14］ Ho CH, Triolo RJ, Elias AL, et al. Functional electrical stimulation and spinal cord injury［J］. Phys Med Rehabil Clin N Am, 2014,25(3):631-654.

［15］ Doucet BM, Lam A, Griffin L. Neuromuscular electrical stimulation for skeletal muscle function［J］. Yale J Biol Med, 2012,85(2):201-215.

［16］ Eli I, Lerner DP, Ghogawala Z. Acute traumatic spinal cord injury［J］. Neurol Clin, 2021,39(2):471-488.

［17］ Wu X, Wei S, Chen M, et al. P2RY13 Exacerbates Intestinal Inflammation by Damaging the Intestinal Mucosal Barrier via Activating IL-6/STAT3 Pathway［J］. Int J Biol Sci, 2022,18(13):5056-5069.

［18］ Candido FD, Fiorino G, Spadaccini M, et al. Are surgical rates decreasing in the biological era in IBD ［J］ ? Curr Drug Targets, 2019,20(13):1356-1362.

［19］ Li Y, de Haar C, Chen M, et al. Disease-related expression of the IL6/STAT3/SOCS3 signalling pathway in ulcerative colitis and ulcerative colitis-related carcinogenesis［J］. Gut, 2010,59(2):227-235.

［20］ Zhao W, Chai Y, Fang QM, et al. Effect of small interferon RNA combined with electro-acupuncture on the expression of connective tissue growth factor after spinal cord injury in rats［J］. Acta Anatomia Sinica, 2016, 47(2):185-190. (in Chinese)

赵伟, 柴勇, 房清敏, 等. 小RNA干扰联合电针治疗对脊髓损伤后结缔组织生长因子的沉默作用［J］. 解剖学报, 2016,47(2):185-190.

［21］ Kisucka A, Bimbova K, Bacova M, et al. Activation of neuroprotective microglia and astrocytes at the lesion site and in the adjacent segments is crucial for spontaneous locomotor recovery after spinal cord injury［J］. Cells, 2021,10(8):1692-1943.

［22］ Chen S, Wu C, Hwang W, et al. More Insight into BDNF against neurodegeneration: anti-apoptosis, anti-oxidation, and suppression of autophagy［J］. Int J Mol Sci, 2017,18(3):545-564.

［23］ Gao P, Wang Z, Lei M, et al. Daphnetin ameliorates Aβ pathogenesis via STAT3/GFAP signaling in an APP/PS1 double-transgenic mouse model of Alzheimer’s disease［J］. Pharmacol Res, 2022,180:106227.

［24］ Yao Y, Du JY, Zhou WJ. Protective effect and mechanism of ginsenoside Rb1 on repairing sciatic nerve injury in mice［J］. Acta Anatomia Sinica,2022,53(1):19-27.(in Chinese)

姚远,杜静怡,周文娟.人参皂苷Rb1促进小鼠坐骨神经损伤修复的作用及机制［J］.解剖学报,2022,53(1):19-27.

[1]	XIA Yu, REN Shu-yu, LI Tao, MEI Feng. Function and distribution of growth hormone receptor positive neurons in neonatal mouse brain [J]. Acta Anatomica Sinica, 2024, 55(4): 422-429.
[2]	LI Hai-yan, LI Zhong-da, HE Li-juan. Effects of conditional knockout ceruloplasmin in astrocytes on brain iron metabolism of mice [J]. Acta Anatomica Sinica, 2024, 55(4): 475-481.
[3]	GONG Chuan-hui QIU Jia-yi YIN Ke-xin ZHANG Ji-ru HE Cheng YUAN Ye LÜ Guang-ming. Optimisation of CUBIC tissue clearing technology based on perfusion methods [J]. Acta Anatomica Sinica, 2024, 55(3): 363-370.
[4]	CAO Cong HUANG Qin-wen WANG Hong XU Ze-ting ZHANG Chan3 SHAN Yi-wen FAN Xiao-xiao LIAO Min . Exercise and complex environment inhibiting lipopolysaccharide-induced dopaminergic neuron damage in substantia nigra [J]. Acta Anatomica Sinica, 2024, 55(3): 253-259.
[5]	CHEN Lin YAN Li-min XING Huai-jie CHEN Min LI Xiao-yan ZENG Chao-sheng . Protective effect of Xuebijing on hippocampal neurons in mice with anti-N-methyl-D-aspartate receptor encephalitis and its mechanism#br# [J]. Acta Anatomica Sinica, 2024, 55(3): 268-275.
[6]	LU Jia-yin YAO Zhi-chao HAO Xiao-jing YAN Yi MA Pei ZHANG Hui-ling WANG Hai-dong. College of Veterinary Medicine, Shanxi Agricultural University, Shanxi Taigu 030801, China [J]. Acta Anatomica Sinica, 2024, 55(3): 285-294.
[7]	XU Lin XIONG Xi-wen LI Zun HUANG Rong MA Hong-hui MA Jie. Roles of hepatic Dishevelled/Egl-10/pleckstrin domain-containing protein 5/mammalian target of rapamycin complex 1 signaling axis on the development of non-alcoholic fatty liver disease [J]. Acta Anatomica Sinica, 2024, 55(3): 295-301.
[8]	LIN Wei LI Chao-yi TANG Jie ZHANG Pi-jun. Osteoblast-derived exosome mediating the effect of microRNA-494 on bone metabolism and bone remodeling balance in osteoporotic rats [J]. Acta Anatomica Sinica, 2024, 55(3): 302-310.
[9]	CHEN Pei CHEN Xiao-ju DU Zhu-man WANG Cao-hui . Mechanism of peimine improving chronic obstructive pulmonary disease induced by lipopolysaccharide combined with cigarette smoke in mice [J]. Acta Anatomica Sinica, 2024, 55(2): 215-221.
[10]	WU Yue-wu HU Bin GUO Xiao-dong FU Qin ZOU Zhi-jia . Effect of microRNA-30a regulation of mitogen-activated protein kinase pathway on aortic coarctation in rats [J]. Acta Anatomica Sinica, 2024, 55(2): 222-228.
[11]	YANG Shan-shan PAN Yu-xiang ZHENG Wan WANG Zheng . Exendin-4 inhibiting cyclophilin A reducing the pathological phenotype of atherosclerotic mice [J]. Acta Anatomica Sinica, 2024, 55(2): 229-236.
[12]	CHENG Quan-cheng DING Hui-ru WANG Zi-yuan FANG Jin-yu ZHANG Xiao-li ZHANG Wei-guang. Analysis of recognition sites and application for commercial and homemade antibodies to aquaporin 9 [J]. Acta Anatomica Sinica, 2024, 55(2): 237-240.
[13]	WANG Yang WU Zhen-hua Lü Hong-bo LUO Dong-bo . Epithelial transformation sequence 2 affecting the in vitro metastatic activity of esophageal squamous carcinoma cells by regulating the expression of p33 inhibitor growth-1 [J]. Acta Anatomica Sinica, 2024, 55(2): 203-209.
[14]	HUANG Jie-he WANG Qian JIA Shun-jie YANG Sheng. Effects of microRNA-103a-3p on osteoporosis through tumor protein 53-regulated inhibitor of apoptosis 1/P53 [J]. Acta Anatomica Sinica, 2024, 55(2): 174-180.
[15]	REN Jian CHEN Xiao-yan. Lentiviral vector mediated steroidogenic factor-1 silencing regulating decidualization and autophagy of endometrial stromal cells [J]. Acta Anatomica Sinica, 2024, 55(2): 188-195.