Effect of “Lumbar Three Needles” on expression of oxidative stress factors in rats with multifidus muscles injury

doi:10.3969/j.issn.2095-4344.2863

Abstract

Abstract:

BACKGROUND: Posterior lumbar spine surgery is currently the main surgical approach for many surgical operations such as discectomy and spinal canal decompression. However, 10%-40% of patients with posterior lumbar spine surgery can have low back pain and related dysfunctions soon after surgery, which is related to excessive stretching or blunt injury to the paraspinal muscles such as the multifidus muscle during the operation.

OBJECTIVE: To explore the effect of “Lumbar Three Needles” on the expression of oxidative stress factors, including malondialdehyde, reactive oxygen species, superoxide dismutase, glutathione peroxidase and phosphorylated protein kinase B, in rats with multifidus muscles injury.

METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into control group, model group and “Lumbar Three Needles” group, with 8 rats in each group. An animal model of multifidus muscles injury was made by intramuscular injection of 0.5% bupivacaine hydrochloride in the model and treatment groups in control and “Lumbar Three Needles” group, while normal saline was injected in the control group. No acupuncture intervention was given in the control group and model group, while Dachangshu, Shenshu and Weizhong acupoints were electroacupunctured in the “Lumbar Three Needles” group. Needles were then stimulated electrically using a Han's Acupoint Nerve Stimulator with density wave, frequency of 2 Hz/10 Hz and continuous current of 1 mA for 20 minutes, once daily for 7 days in total. The multifidus muscle was stained with hematoxylin-eosin staining to observe morphological changes, and kits were used to observe the expression of malondialdehyde, reactive oxygen species, superoxide dismutase, and glutathione peroxidase, while western blot was applied to observe the expression of phosphorylated protein kinase B. The study protocol was approved by the Animal Ethic Committee of Guangdong Second Traditional Chinese Medicine Hospital with an approval No. 048617.

RESULTS AND CONCLUSION: Hematoxylin-eosin staining results indicated that the muscle fibers were partially fixed in the model group and there were still a lot of macrophages after 7 days of intervention. Compared with the model group, more newborn muscle fibers and fewer macrophages could be seen in the “Lumbar Three Needles” group. After intervention, the cross-sectional area of the multifidus muscles in the “Lumbar Three Needles” group was significantly bigger than that in the model group (P < 0.01). The expression levels of malondialdehyde and reactive oxygen species in the model group were significantly increased compared with the control group (P < 0.01) and “Lumbar Three Needles” group (P < 0.01). The expression of superoxide dismutase, glutathione peroxidase and phosphorylated protein kinase B in the “Lumbar Three Needles” group was significantly higher than that in the model group (P < 0.01 or P < 0.05). Therefore, “Lumbar Three Needles” can significantly decrease the oxidative stress level after multifidus muscle injury, which may be related to the improvement of phosphorylated protein kinase B.

Key words: experiment, rat, Lumbar Three Needles, multifidus muscle, oxidative stress, malondialdehyde, reactive oxygen species, superoxide dismutase, oxidative stress factors

CLC Number:

Bai Zhenjun, Liu Tong, Wang Zirun, Zhang Huiyu, Xu Peng. Effect of “Lumbar Three Needles” on expression of oxidative stress factors in rats with multifidus muscles injury[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(32): 5145-5150.

Figures/Tables 4

References

[1] SNIDER RK. A prospective, randomized study of lumbar fusion: preliminary results.Spine.1994;19(1):109.

[2] AGHION D, CHOPRA P, OYELESE A A. Failed back syndrome.Med Health R I. 2012;95(12):391-393.

[3] DZIK K, SKROBOT W, FLIS DJ, et al. Vitamin D supplementation attenuates oxidative stress in paraspinal skeletal muscles in patients with low back pain. Eur J Appl Physiol. 2018;118(1):143-151.

[4] 刘通,于佳妮,邹德辉,等.电针血清对多裂肌卫星细胞增殖及Pax-7､成肌分化抗原､磷酸化蛋白激酶B表达的影响[J].针刺研究, 2016,41(5):402-409.

[5] 刘通,于佳妮,邹德辉,等.电针调控miRNA206/HDAC4轴促进损伤多裂肌修复过程中成肌细胞分化的机制[J].中国组织工程研究,2019,23(33):5341-5346.

[6] TAGUCHI T, HOHEISEL U, MENSE S.Dorsal horn neurons having input from low back structures in rats. Pain. 2008; 138:119–129.

[7] 陈玉佩,刘通,邹德辉,等.局部注射布比卡因建立大鼠骨骼肌损伤模型的组织形态学评价[J].中国组织工程研究,2016,20(18): 2615-2621.

[8] 李忠仁. 实验针灸学[M].北京:中国中医药出版社, 2003:316.

[9] HUNG CW, WU MF, HONG RT, et al. Comparison of multifidus muscle atrophy afterposterior lumbar interbody fusion with conventional and cortical bone trajectory. Clin Neurol Neurosurg. 2016;145:41-45.

[10] CHA JR, KIM YC, JANG C, et al. Pedicle screw fixation and posterior fusion for lumbar degenerative diseases: effects on individual paraspinal muscles and lower back pain; a single-center, prospective study. BMC Musculoskelet Disord. 2016;17:63.

[11] GILBERT F, HEINTEL TM, JAKUBIETZ MG, et al. Quantitative MRI comparison of multifidus muscle degeneration in thoracolumbar fractures treated with open and minimally invasive approach.BMC Musculoskelet Disord. 2018;19(1):75.

[12] YARJANIAN JA, FETZER A, YAMAKAWA KS, et al. Correlation of Paraspinal Atrophy and Denervation in Back Pain and Spinal Stenosis Relative to Asymptomatic Controls. PM&R.2013;5(1):39-44.

[13] WASCHKE A, HARTMANN C, WALTER J, et al. Denervation and atrophy of paraspinal muscles after open lumbar interbody fusion is associated with clinical outcome-electromyographic and CT-volumetric investigation of 30 patients. ActaNeurochirurgica.2014;156(2):235-244.

[14] GILLE O, JOLIVET E, DOUSSET V, et al. Erector Spinae Muscle Changes on Magnetic Resonance Imaging Following Lumbar Surgery Through a Posterior Approach. Spine. 2007; 32(11):1236-1241.

[15] 季传婷,徐平,谢龙.电针对Cardiotoxin肌肉损伤小鼠MyoD表达的影响[J].南京中医药大学学报, 2012,28(2):136-138.

[16] WATANABE K, MATSUMOTO M, IKEGAMI T, et al. Reduced postoperative wound pain after lumbar spinous process–splitting laminectomy for lumbar canal stenosis: a randomized controlled study. J Neurosurg Spine. 2011;14(1): 51-58.

[17] 房芳,魏艳霞.腰三针治疗腰椎间盘突出症的临床观察及影像学变化[J].世界中医药,2018,13(5):1250-1253+1257.

[18] 曲淑婕,张帅,戈磊,等.腰三针配合穴位推拿对LDH患者治疗效果及超氧化物歧化酶和α1-AGP水平影响[J].辽宁中医药大学学报,2019,21(5):211-214.

[19] 张洁.腰三针温针灸治疗退行性腰椎管狭窄症的临床效果观察[J].临床医药文献电子杂志,2018,5(90):45+47.

[20] HALLIWELL B. Biochemistry of oxidative stress. Biochem Soc Trans. 2007;35(Pt 5):1147-1150.

[21] LU K, LIANG CL, CHO CL, et al. Oxidative stress and heat shock protein response in human paraspinal muscles during retraction.J Neurosurg. 2002;97(1 Suppl):75-81.

[22] LU K, LIANG CL, CHEN HJ, et al. Nuclear factor-kappaB- regulated cyclooxygenase-2 expression in surgery-associated paraspinal muscle injury in rats.J Neurosurg. 2003 Mar;98(2 Suppl):181-187.

[23] 胡志军. 腰椎后路手术原性多裂肌损伤和萎缩的相关研究:解剖基础和因素分析[D].杭州:浙江大学, 2012.

[24] REDDY NM, POTTETI HR, VEGIRAJU S, et al. PI3K-AKT Signaling via Nrf2 Protects against Hyperoxia-Induced Acute Lung Injury, but Promotes Inflammation Post-Injury Independent of Nrf2 in Mice. Plos One. 2015;10(6):e0129676.

[25] LIEN EC, LYSSIOTIS CA, JUVEKAR A, et al. Glutathione biosynthesis is a metabolic vulnerability in PI(3)K/Akt-driven breast cancer. Nat Cell Biol. 2016;18(5):572-578.

[26] 陈伟巍,王敏,吴明辉.Trx-1过表达通过NF-κB信号通路减轻MPP~+所致PC12细胞的氧化应激损伤[J].中国病理生理杂志, 2019,35(07):1206-1212.

[27] 王宇霆. 乌司他丁通过Akt/mTOR通路保护受氧化应激的人脐静脉血管内皮细胞[D].杭州:浙江大学,2013.