米诺环素对脊神经结扎大鼠脊髓内M1/M2型小胶质细胞极化的影响

成志红; 冯松; 王霞; 倪锐男; 郭洋; 向宇; 杨正伟; 彭彬

doi:10.16016/j.2097-0927.202402020

您当前的位置：

首页 >

文章列表页 >

米诺环素对脊神经结扎大鼠脊髓内M1/M2型小胶质细胞极化的影响

基础医学 | 更新时间：2026-01-09

- 米诺环素对脊神经结扎大鼠脊髓内M1/M2型小胶质细胞极化的影响
- Effect of minocycline on polarization of types M1/M2 microglia in spinal cord in rats after spinal nerve ligation
- 陆军军医大学学报 2024年46卷第15期页码：1740-1750
- 作者机构：
  
  1.637000 四川南充，川北医学院基础医学与法医学研究所
  2.635000 四川达州，达州市第三人民医院神经外科
  3.637000 四川南充，川北医学院临床医学院
- 作者简介：
  
  [ 彭彬，E-mail: binpeng01@sina.com PENG Bin, E-mail: binpeng01@sina.com
- 基金信息：
  
  南充市科学技术局博士、硕士学位授权点和“一流学科”建设专项(20SXQT0304);四川省大学生创新创业训练计划项目(S202310634104)
- DOI：10.16016/j.2097-0927.202402020
  中图分类号： R322.81;R441.1;R978.14
- 收稿：2024-02-04，
  
  修回：2024-03-24，
  
  纸质出版：2024-08-15
- 稿件说明：
移动端阅览
成志红, 冯松, 王霞, 等. 米诺环素对脊神经结扎大鼠脊髓内M1/M2型小胶质细胞极化的影响[J]. 陆军军医大学学报, 2024,46(15):1740-1750.

Zhihong CHENG, Song FENG, Xia WANG, et al. Effect of minocycline on polarization of types M1/M2 microglia in spinal cord in rats after spinal nerve ligation[J]. Journal of Army Medical University, 2024, 46(15): 1740-1750.
成志红, 冯松, 王霞, 等. 米诺环素对脊神经结扎大鼠脊髓内M1/M2型小胶质细胞极化的影响[J]. 陆军军医大学学报, 2024,46(15):1740-1750. DOI： 10.16016/j.2097-0927.202402020.

Zhihong CHENG, Song FENG, Xia WANG, et al. Effect of minocycline on polarization of types M1/M2 microglia in spinal cord in rats after spinal nerve ligation[J]. Journal of Army Medical University, 2024, 46(15): 1740-1750. DOI： 10.16016/j.2097-0927.202402020.

摘要

目的

探讨米诺环素(minocycline，Mino)对脊神经结扎(spinal nerve ligation，SNL)致神经病理性疼痛(neuropathic pain，NP)大鼠脊髓内M1型(促炎型)和M2型(抗炎型)小胶质细胞极化的影响及其机制。

方法

36只成年雄性SD大鼠按体质量分层随机抽样分为Sham组、SNL组和M+SNL组(鞘内注射米诺环素)，于术前1 d和术后14 d内测定大鼠后足机械痛阈值和冷痛敏阈值。术后14 d于L5脊髓取材，采用免疫组化技术和体视学技术检测L5脊髓背角内小胶质细胞的总数以及M1/M2型小胶质细胞的数量；利用生物信息学技术筛选出脊髓小胶质细胞内的核心靶点

Cst

7，并采用Western blot检测小胶质细胞标志物Iba-1、M1型小胶质细胞标志物iNOS、M2型小胶质细胞标志物CD206、

Cst

7基因编码的蛋白质胱抑素F(cystatin F，CF)及CatS/CX3CL1/CX3CR1通路蛋白的表达水平；采用ELISA检测脊髓内TNF-α、IL-6和IL-10的蛋白表达量。

结果

M+SNL组大鼠术后7~14 d手术侧的机械痛和冷痛敏阈值均比SNL组显著升高(

0.01)。SNL组大鼠手术侧脊髓背角内小胶质细胞的总数，M1型、M2型小胶质细胞的数量以及脊髓CatS、CX3CL1、CX3CR1、TNF-α、IL-6和IL-10蛋白表达较Sham组均显著增加，而CF含量显著降低(

0.01)；与SNL组比较，M+SNL组小胶质细胞的总数、M1型小胶质细胞的数量以及CatS、CX3CL1、CX3CR1、TNF-α、IL-6蛋白表达均显著减少，而M2型小胶质细胞的数量以及CF、IL-10的蛋白表达均显著增加(

0.05)。

结论

米诺环素改善SNL致神经病理性疼痛的机制可能是通过上调小胶质细胞内CF蛋白表达，进而抑制CatS/CX3CL1/CX3CR1信号通路，促进小胶质细胞由M1型向M2型转化，以平衡M1/M2小胶质细胞极化失衡、减轻神经炎症。

Abstract

Objective

To investigate the effect of minocycline (Mino) on the polarization of types M1/M2 microglia (pro- and anti-inflammatory type) in the spinal dorsal horn of rats with neuropathic pain (NP) induced by spinal nerve ligation (SNL) and its underlying mechanism.

Methods

A total of 36 adult male SD rats were randomly stratified into Sham-operation (Sham) group

SNL group and Mino+SNL group by stratified random sampling based on body weight. Mechanical pain threshold and cold nociceptive thresholds of rat hind paw were measured in 1 d before and 14 d after modelling. Spinal cord tissue at the lumbar 5 (L5) segment was taken at 14 d after modelling

and the total number of microglia as well as the numbers of M1 and M2 microglia in the spinal dorsal horn were measured with immunohistochemistry and stereology. With aid of bioinformatics techniques

the core target in the spinal cord

Cst

was selected. Then

the protein levels of microglia marker Iba-1

M1 microglia marker iNOS

M2 microglia marker CD206

Cst

7 encoded protein cystatin F (CF) and pathway CatS/CX3CL1/CX3CR1 were detected with Western blotting. The expression levels of TNF-α

IL-6 and IL-10 in the spinal cord tissues were measured with ELISA.

Results

The mechanical pain and cold nociceptive thresholds were both significantly higher in the M+SNL group than the SNL group at 7~14 d after modelling (

0.01). The total number of microglia and the numbers of M1/M2 microglia in the spinal dorsal horn as well as the expression levels of CatS

CX3CL1

CX3CR1

TNF-α

IL-6

and IL-10 in the spinal cord tissues were obviously increased

and the expression level of CF was notably decreased in the SNL model group than the Sham group (

0.01). While

Mino treatment remarkably reversed above phenomena

with decreased total number of microglia and number of M1 microglia as well as expression levels of CatS

CX3CL1

CX3CR1

TNF-α and IL-6

and increased number of M2 microglia as well as CF and IL-10 levels when compared with the SNL group (

0.05).

Conclusion

Mino alleviates SNL induced neuropathic pain

probably through up-regulating CF in the microglia

and thus inhibiting the CatS/CX3CL1/CX3CR1 signaling pathway

promoting the conversion of microglia from type M1 to M2 to balance the imbalance in the M1/M2 polarization

and thus reducing neuroinflammation.

关键词

Keywords

references

LORIO M, KIM C, ARAGHI A, et al . International Society for the Advancement of Spine Surgery Policy 2019—surgical treatment of lumbar disc herniation with radiculopathy[J ] . Int J Spine Surg, 2020, 14(1): 1-17. DOI: 10.14444/7001.

殷利军, 陈辉, 熊利泽. 神经病理性疼痛动物模型的选择与建立[J]. 中国疼痛医学杂志, 2023, 29(10): 725-730. DOI: 10.3969/j.issn.1006-9852.2023.10.002.

YIN L J, CHEN H, XIONG L Z. Selection and establishment of animal models of neuropathic pain[J]. Chin J Pain Med, 2023, 29(10): 725-730. DOI: 10.3969/j.issn.1006-9852.2023.10.002.

XU F T, HUANG J, HE Z H, et al . Microglial polarization dynamics in dorsal spinal cord in the early stages following chronic sciatic nerve damage[J ] . Neurosci Lett, 2016, 617: 6-13. DOI: 10.1016/j.neulet.2016.01.038.

方茜, 吴文, 唐丹喆. 脊髓小胶质细胞的极化对神经病理性疼痛的影响[J]. 中国疼痛医学杂志, 2022, 28(4): 285-289. DOI: 10.3969/j.issn.1006-9852.2022.04.008.

FANG Q, WU W, TANG D Z. Effect of polarization of spinal microglia on neuropathic pain[J]. Chin J Pain Med, 2022, 28(4): 285-289. DOI: 10.3969/j.issn.1006-9852.2022.04.008.

KARAVIS M Y, SIAFAKA I, VADALOUCA A, et al . Role of microglia in neuropathic pain[J ] . Cureus, 2023, 15(8): e43555. DOI: 10.7759/cureus.43555.

黄小丽, 谭川, 林菁艳, 等. 体视学研究SNL模型大鼠脊髓背角胶质细胞数量的动态变化[J]. 神经解剖学杂志, 2016, 32(5): 629-634. DOI: 10.16557/j.cnki.1000-7547.2016.05.0014.

HUANG X L, TAN C, LIN J Y, et al . Stereological study on the dynamic changes in the number of glial cells in the spinal dorsal horn of rats with spinal nerve ligation[J ] . Chin J Neuroanat, 2016, 32(5): 629-634. DOI: 10.16557/j.cnki.1000-7547.2016.05.0014.

ZHOU X X, TAO L, ZHAO M R, et al . Wnt/β-catenin signaling regulates brain-derived neurotrophic factor release from spinal microglia to mediate HIV 1 gp120-induced neuropathic pain[J ] . Mol Pain, 2020, 16: 1744806920922100. DOI: 10.1177/1744806920922100..

SHINODA M, FUJITA S, SUGAWARA S, et al . Suppression of superficial microglial activation by spinal cord stimulation attenuates neuropathic pain following sciatic nerve injury in rats[J ] . Int J Mol Sci, 2020, 21(7): 2390. DOI: 10.3390/ijms21072390.

ASANO S, HAYASHI Y, IWATA K, et al . Microglia-astrocyte communication via C1q contributes to orofacial neuropathic pain associated with infraorbital nerve injury[J ] . Int J Mol Sci, 2020, 21(18): 6834. DOI: 10.3390/ijms21186834..

BURKE N N, KERR D M, MORIARTY O, et al . Minocycline modulates neuropathic pain behaviour and cortical M1-M2 microglial gene expression in a rat model of depression[J ] . Brain Behav Immun, 2014, 42: 147-156. DOI: 10.1016/j.bbi.2014.06.015.

SHIN D A, KIM T U, CHANG M C. Minocycline for controlling neuropathic pain: a systematic narrative review of studies in humans[J]. J Pain Res, 2021, 14: 139-145. DOI: 10.2147/JPR.S292824.

KONG G G, HUANG Z P, ZHU Q G, et al . Comparison of two modified methods of intrathecal catheterization in rats[J ] . Exp Anim, 2020, 69(2): 219-223. DOI: 10.1538/expanim.19-0108.

杨林凤, 黄静, 杨雁琪, 等. 神经生长因子对坐骨神经损伤大鼠脊髓内神经纤维的影响[J]. 神经解剖学杂志, 2022, 38(6): 677-682. DOI: 10.16557/j.cnki.1000-7547.2022.06.011.

YANG L F, HUANG J, YANG Y Q, et al . The effects of nerve growth factor on nerve fibers within spinal cord in rats with sciatic nerve injury[J ] . Chin J Neuroanat, 2022, 38(6): 677-682. DOI: 10.16557/j.cnki.1000-7547 .2022.06.011.

LIN J Y, HE Y N, ZHU N, et al . Metformin attenuates increase of synaptic number in the rat spinal dorsal horn with painful diabetic neuropathy induced by type 2 diabetes: a stereological study[J ] . Neurochem Res, 2018, 43(12): 2232-2239. DOI: 10.1007/s11064-018-2642-4.

KIM S H, CHUNG J M. An experimental model for peripheral neuropathy produced by segmental spinal nerve ligation in the rat[J]. Pain, 1992, 50(3): 355-363. DOI: 10.1016/0304-3959(92)90041-9.

MARTIN P Y, DOLY S, HAMIEH A M, et al . mTOR activation by constitutively active serotonin6 receptors as new paradigm in neuropathic pain and its treatment[J ] . Prog Neurobiol, 2020, 193: 101846. DOI: 10.1016/j.pneurobio.2020.101846.

薛松, 孙涛. 小胶质细胞介导脊髓损伤后神经病理性疼痛机制研究进展[J]. 中国疼痛医学杂志, 2022, 28(3): 165-172. DOI: 10.3969/j.issn.1006-9852.2022.03.002.

XUE S, SUN T. Research progress on the mechanism of microglia in neuropathic pain after spinal cord injury[J]. Chin J Pain Med, 2022, 28(3): 165-172. DOI: 10.3969/j.issn.1006-9852.2022.03.002.

ATTA A A, IBRAHIM W W, M OHAMED A F, et al . Microglia polarization in nociplastic pain: mechanisms and perspectives[J ] . Inflammopharmacology, 2023, 31(3): 1053-1067. DOI: 10.1007/s10787-023-01216-x.

ZHANG L Q, GAO S J, SUN J, et al . DKK3 ameliorates neuropathic pain via inhibiting ASK-1/JNK/p-38-mediated microglia polarization and neuroinflammation[J ] . J Neuroinflammation, 2022, 19(1): 129. DOI: 10.1186/s12974-022-02495-x..

ANDERSEN AM, KAALUND S S, MARNER L, et al . Quantitative cellular changes in multiple system atrophy brains[J ] . Neuropathol Appl Neurobiol, 2023, 49(6): e12941. DOI: 10.1111/nan.12941.

MVHLFELD C, SCHIPKE J. Methodological progress of stereology in cardiac research and its application to normal and pathological heart development[J]. Cells, 2022, 11(13): 2032. DOI: 10.3390/cells11132032.

LIN J Y, ZHU N, HE Y N, et al . Stereological study on the numerical plasticity of myelinated fibers and oligodendrocytes in the rat spinal cord with painful diabetic neuropathy[J ] . Neuroreport, 2020, 31(4): 319-324. DOI: 10.1097/WNR.0000000000001407.

LI Z L , WEI H, PⅡRAINEN S, et al . Spinal versus brain microglial and macrophage activation traits determine the differential neuroinflammatory responses and analgesic effect of minocycline in chronic neuropathic pain[J ] . Brain Behav Immun, 2016, 58: 107-117. DOI: 10.1016/j.bbi.2016.05.021.

DENK F, CROW M, DIDANGELOS A, et al . Persistent alterations in microglial enhancers in a model of chronic pain[J ] . Cell Rep, 2016, 15(8): 1771-1781. DOI: 10.1016/j.celrep.2016.04.063.

WATTS C. Lysosomes and lysosome-related organelles in immune responses[J]. FEBS Open Bio, 2022, 12(4): 678-693. DOI: 10.1002/2211-5463.13388.

MA J M, TANAKA K F, SHIMIZU T, et al . Microglial cystatin F expression is a sensitive indicator for ongoing demyelination with concurrent remyelination[J ] . J Neurosci Res, 2011, 89(5): 639-649. DOI: 10.1002/jnr.22567.

MAGISTER S, OBERMAJER N, MIRKOVIC B, et al . Regulation of cathepsins S and L by cystatin F during maturation of dendritic cells[J ] . Eur J Cell Biol, 2012, 91(5): 391-401. DOI: 10.1016/j.ejcb.2012.01.001.

CLARK A K, MALCANGIO M. Microglial signalling mechanisms: Cathepsin S and Fractalkine[J]. Exp Neurol, 2012, 234(2): 283-292. DOI: 10.1016/j.expneurol.2011.09.012.

SILVA R, MALCANGIO M. Fractalkine/CX 3 CR 1 pathway in neuropathic pain: an update[J ] . Front Pain Res, 2021, 2: 684684. DOI: 10.3389/fpain.2021.684684..

DUAN W, WANG H X, FAN Q L, et al . Cystatin F involvement in adenosine A 2A receptor-mediated neuroinflammation in BV2 microglial cells[J ] . Sci Rep, 2018, 8(1): 6820. DOI: 10.1038/s41598-018-25031-5.00000000000000000.

浏览量

149

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

抑郁症模型大鼠海马内小胶质细胞数量和炎症因子表达增加

脊髓背角P2X₇受体激活引起神经病理性疼痛的发生

短期丰富生存环境干预下中老年大鼠海马结构有髓神经纤维的性别差异

运用新的体视学方法研究大鼠胼胝体及胼胝体额区有髓神经纤维

甲基莲心碱通过抑制ROS/NLRP3/Caspase-1通路减少小胶质细胞焦亡并改善帕金森病神经炎症