基于TLR4/NF-κB信号通路探讨“气血并治”针法对神经根型颈椎病大鼠炎性损伤的影响

doi:10.20047/j.issn1673-7210.2023.23.01

摘要
图/表
参考文献(26)
相关文章 (15)

全文: PDF (976 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的探讨“气血并治”针法对神经根型颈椎病（CSR）大鼠炎性损伤的治疗效应及与调节Toll样受体4（TLR4）/核因子κB（NF-κB）信号通路的相关性。方法 24只5～7周龄（180～220 g）健康雄性SD大鼠，按照随机数字表法将其分为空白组、假手术组、模型组、“气血并治”针法组，每组6只。采用椎管插线法制备CSR模型，“气血并治”针法组于造模后第3天针刺双侧颈5和颈7夹脊穴、足三里和膈俞，颈夹脊穴予电针疏密波，2 Hz/50 Hz，强度1.5 mA，30 min/d，连续治疗14 d；空白组、假手术组、模型组束缚同“气血并治”针法组，不治疗。治疗结束后，比较各组步态评分，电镜观察脊髓各组超微结构，酶联免疫吸附试验法检测各组脊髓白细胞介素（IL）-1β和前列腺素E2（PGE2）含量，Western blot检测脊髓TLR4、NF-κB和环氧合酶2（COX-2）蛋白表达。结果治疗后，假手术组步态评分与空白组比较，差异无统计学意义（P＞0.05），模型组步态评分较空白组升高（P＜0.01），“气血并治”针法组步态评分较模型组降低（P＜0.05）。空白组和假手术组大鼠髓鞘外形规则，结构完整；模型组髓鞘结构不规则，部分髓鞘断裂消失，厚薄不一，板层结构松散、扭曲、破裂，线粒体肿胀，甚至部分溶解消失；“气血并治”针法组髓鞘板层松散程度有所减轻，结构较紧凑，可见清晰的线粒体。假手术组IL-1β和PGE2蛋白表达水平与空白组比较，差异无统计学意义（P＞0.05）；模型组IL-1β和PGE2蛋白表达水平高于空白组（P＜0.01）；“气血并治”针法组IL-1β和PGE2蛋白表达水平低于模型组（P＜0.01）。假手术组TLR4、NF-κB、COX-2蛋白相对表达量与空白组比较，差异无统计学意义（P＞0.05）；模型组TLR4、NF-κB、COX-2蛋白表达量较空白组升高（P＜0.01）；“气血并治”针法组TLR4、NF-κB、COX-2蛋白表达量低于模型组（P＜0.01）。结论 “气血并治”针法可以改善CSR大鼠的活动受限，其可能是通过阻抑TLR4/NF-κB信号通路变化，降低IL-1β和PGE2的表达，减轻炎性损伤发挥作用。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	王佳佳　　嵇波▲　　刘翼天　　石天宇　　葛云鹏　　谢亚娜　　方洋　　张玲

关键词 ：神经根型颈椎病, 气血并治, 炎症, Toll样受体4, 核因子&kappa, 4, 环氧合酶2

Abstract：Objective To investigate the therapeutic effect of qi and blood combined treatment acupuncture on inflammatory injury in rats with cervical spondylotic radiculopathy (CSR) and the correlation between toll-like receptor 4 (TLR4)/nuclear factor κB (NF-κB) signaling pathway. Methods Twenty-four 5-7-week-old healthy male SD rats (180-220 g) were divided into blank group, sham surgery group, model group, and “qi and blood combined treatment” acupuncture group by random number table method, with six rats in each group. The CSR model was prepared by spinal canal insertion method. On the 3rd day after the model was made, the “qi and blood combined treatment” acupuncture group was treated by acupuncture at ji?觃jǐ points of cervical 5 and cervical 7, zus?觃nlǐ and géshù, and the cervical ji?觃jǐ points were treated by electro-acupuncture with density wave at 2 Hz/50 Hz, with an intensity of 1.5 mA for 30 min/d for 14 days. The restrain of blank group, sham operation group, and model group were the same as those of “qi and blood combined treatment” acupuncture group, no treatment. The restrain of the gait score of all groups was compared, the ultrastructure of the spinal cord was observed by electron microscope, the contents of interleukin(IL)-1β and prostaglandin E2(PGE2) in the spinal cord were detected by enzyme-linked immunosorbent assay, and the expression of TLR4, NF-κB, and cyclocycesterase 2 (COX-2) in the spinal cord were detected by Western blot. Results After treatment, there was no significant difference in gait score between sham operation group and blank group (P＞0.05), the gait score of model group was higher than that of blank group (P＜0.01), and the gait score of “qi and blood combined treatment” acupuncture group was lower than that of model group (P＜0.05). The appearance of myelin sheath of rats in blank group and sham operation group was regular and the structure was complete. In the model group, the myelin structure was irregular, some myelin was broken and disappeared, the thickness was different, the lamellar structure was loose, twisted and broken, and the mitochondria was swollen or even partially dissolved and disappeared; in the “qi and blood combined treatment” acupuncture group, the myelin lamella was less loose, the structure was more compact, and clear mitochondria could be seen. There was no significant difference in IL-1β and PGE2 protein expression between sham operation group and blank group (P＞0.05). The expression levels of IL-1β and PGE2 in model group were higher than those in blank group (P＜0.01). The expression levels of IL-1β and PGE2 protein in “qi and blood combined treatment” acupuncture group were lower than those in model group (P＜0.01). There was no significant difference in the relative expressions of TLR4, NF-κB, and COX-2 protein between the sham operation group and the blank group (P＞0.05). The expressions of TLR4, NF-κB, and COX-2 in model group were higher than those in blank group (P＜0.01). The protein expressions of TLR4, NF-κB, and COX-2 in “qi and blood combined treatment” acupuncture group were lower than those in model group (P＜0.01). Conclusion The “qi and blood combined treatment” acupuncture method can improve the limited activity of CSR rats, probably by blocking changes in the TLR4/NF-κB signaling pathway, reducing the expression of IL-1β and PGE2, and reducing inflammatory damage.

Key words： Cervical spondylosis radiculopathy Qi and blood combined treatment Inflammatory Toll-like receptor 4 Nuclear factor kappa-B Cycloxygenase-2

基金资助:国家自然科学基金资助项目（82174505）。

通讯作者: ▲通讯作者

引用本文:

王佳佳　　嵇波▲　　刘翼天　　石天宇　　葛云鹏　　谢亚娜　　方洋　　张玲 . 基于TLR4/NF-κB信号通路探讨“气血并治”针法对神经根型颈椎病大鼠炎性损伤的影响[J]. 中国医药导报, 2023, 20(23): 4-7,22.
WANG Jiajia JI Bo▲ LIU Yitian SHI Tianyu GE Yunpeng XIE Yana FANG Yang ZHANG Ling . Investigating the effect of “qi and blood combined treatment” acupuncture on inflammatory injury of cervical spondylotic radiculopathy rats based on TLR4/NF-κB signaling pathway. 中国医药导报, 2023, 20(23): 4-7,22.

链接本文:

https://www.yiyaodaobao.com.cn/CN/10.20047/j.issn1673-7210.2023.23.01 或 https://www.yiyaodaobao.com.cn/CN/Y2023/V20/I23/4

[1] Descalzi G，Mitsi V，Purushothaman I，et al. Neuropathic pain promotes adaptive changes in gene expression in brain networks involved in stress and depression [J]. Sci Signal，2017， 10（471）：eaaj1549.
[2] Woods BI，Hilibrand AS. Cervical radiculopathy：epidemiology，etiology，diagnosis，and treatment [J]. J Spinal Disord Tech， 2015，28（5）：E251-E259.
[3] 魏戌，高云，张兴平，等.基于临床科研信息共享系统的神经根型颈椎病发病特征分析[J].中医杂志，2015，56（19）：1663-1666.
[4] 谢炜，赵伟宏，于林，等.川芎提取物对神经根型颈椎病模型大鼠根性疼痛的保护作用研究[J].广东药学院学报，2008，24（5）：496-499.
[5] Kawakami M，Weinstein JN，Spratt KF，et al. Experimental lumbar radiculopathy. Immunohistochemical and quantitative demonstrations of pain induced by lumbar nerve root irritation of the rat [J]. Spine （Phila Pa 1976），1994，19（16）：1780- 1794.
[6] 华兴邦，周浩良.大鼠穴位图谱的研制[J].实验动物与动物实验，1991（1）：1-5.
[7] 朱立国，于杰，高景华，等.神经根型颈椎病患者的疼痛和麻木观测[J].中国中医骨伤科杂志，2009，17（4）：1-3.
[8] 杨雨婷，段航，吴嘉萍，等.基于数据挖掘探讨电针治疗颈椎病参数应用规律[J].河南中医，2021，41（7）：1087- 1091.
[9] 王春英，孙忠人，王琳晶，等.电针颈夹脊穴治疗神经根型颈椎病40例临床观察[J].湖南中医杂志，2021，37（10）：74-76.
[10] 张鸥，李燕.夹脊穴与脊髓神经节段支配及其经络脏腑效应[J].中国中医基础医学杂志，2007（9）：701-702.
[11] 崔承斌，王京京，吴中朝.从背俞穴与夹脊穴的关系论背俞功能带[J].中国针灸，2005（7）：483-486.
[12] 王健，张卫华.张卫华分期辨治神经根型颈椎病撷英[J].中国中医骨伤科杂志，2023，31（3）： 86-88.
[13] 覃美相，粟胜勇，张熙，等.温和灸对神经根型颈椎病大鼠脊髓组织LC3/Bax表达的影响[J].针刺研究，2022，47（3）：244-249.
[14] 林媛媛，粟胜勇，许铱杨，等.麦粒灸对神经根型颈椎病大鼠脊髓背角Beclin-1/GRP78表达的影响[J].中国针灸，2022，42（5）： 533-539.
[15] 朱栋，许金海，莫文.基于PI3K/Akt信号通路探究鹿灵活络颗粒对神经根型颈椎病大鼠炎症反应的影响[J].长春中医药大学学报，2022，38（5）： 506-509.
[16] 曾欢欢，黄英如，李子健，等.大黄素对大鼠急性脊髓损伤后氧化应激和炎症反应的影响研究[J].中国中药杂志，2018，43（9）：1886-1893.
[17] 吴子健.通腑逐瘀法指导下抵当汤加减调控AQP4表达治疗大鼠急性脊髓损伤的作用机制研究[D].武汉：湖北中医药大学，2021.
[18] 高永辉，王俊英，韩焱晶，等.脊髓Toll样受体4和热休克蛋白90参与电针缓解大鼠慢性神经病理性疼痛机制研究[J].针刺研究，2021，46（9）： 735-741.
[19] Wang B，Shen PF，Qu YX，et al. miR-940 promotes spinal cord injury recovery by inhibiting TLR4/NF-κB pathway- mediated inflammation [J]. Eur Rev Med Pharmacol Sci，2019，23（8）：3190-3197.
[20] 蒋学余.电针颈夹脊穴对神经根型颈椎病神经病理性疼痛模型大鼠中枢神经元-胶质细胞-趋化因子镇痛机制的研究[D].长沙：湖南中医药大学，2021.
[21] Ma Y，Song X，Ma T，et al. Aminoguanidine inhibits IL-1β- induced protein expression of iNOS and COX-2 by blocking the NF-κB signaling pathway in rat articular chondrocytes [J]. Exp Ther Med，2020，20（3）：2623-2630.
[22] Jin GJ，Peng X，Chen ZG，et al. Celastrol attenuates chronic constrictive injury-induced neuropathic pain and inhibits the TLR4/NF-κB signaling pathway in the spinal cord [J]. J Nat Med，2022，76（1）：268-275.
[23] Sun WH，He F，Zhang NN，et al. Time dependent neuroprotection of dexamethasone in experimental focal cerebral ischemia：The involvement of NF-κB pathways [J]. Brain Res，2018，1701：237-245.
[24] Nagano T，Tsuda N，Fujimura K，et al. Prostaglandin E2 increases the expression of cyclooxygenase-2 in cultured rat microglia [J]. J Neuroimmunol，2021，361：577724.
[25] Pei W，Zou Y，Wang W，et al. Tizanidine exerts anti-nociceptive effects in spared nerve injury model of neuropathic pain through inhibition of TLR4/NF-κB pathway [J]. Int J Mol Med，2018，42（6）：3209-3219.
[26] Wang YW，Zhang X，Chen CL，et al. Protective effects of Garcinol against neuropathic pain-Evidence from in vivo and in vitro studies [J]. Neurosci Lett，2017，647：85-90.