毛轲,高誉华,周松林,赵彦芬,张原源,杜思龙
MAO Ke,GAO Yuhua,ZHOU Songlin,ZHAO Yanfen,ZHANG Yuanyuan,DU Silong

• 1:河南省中医院麻醉科

摘要(Abstract)：

目的 探讨高迁移率族蛋白B1(high mobility group protein B1,HMGB1)加重神经性疼痛的作用机制。方法 在脂多糖(lipopolysaccharide, LPS)激活的BV-2细胞中干扰HMGB1表达，检测细胞凋亡、炎症因子分泌，以及JAK2和STAT3蛋白的磷酸化水平。检测LPS激活的BV-2细胞中JAK2和STAT3蛋白的磷酸化水平，并引入JAK2抑制剂探究JAK2/STAT3轴阻断对细胞的影响。建立HMGB1敲低的脊神经结扎(spinal nerve ligation, SNL)大鼠模型，检测脊髓组织中通路蛋白表达和炎症因子分泌，评估SNL大鼠的机械痛敏阈值和热痛敏阈值。结果 LPS处理的BV-2细胞中HMGB1表达上调，干扰HMGB1表达显著抑制细胞凋亡和炎症反应。LPS处理激活JAK2/STAT3轴，且JAK2/STAT3轴激活是由HMGB1表达上调介导的。在SNL大鼠模型中，脊髓组织中HMGB1和通路蛋白表达增加，炎症反应加重，机械痛敏阈值和热痛敏阈值均降低。通过敲低HMGB1表达，观察到HMGB1和通路蛋白的表达下调，炎症减轻，神经性疼痛缓解。结论 HMGB1通过激活JAK2/STAT3轴加重神经性疼痛。
Objective To explore the mechanism by which high mobility group protein B1(HMGB1)aggravates neuropathic pain.Methods The HMGB1 expression was interfered with in lipopolysaccharide(LPS)activated BV-2 cells, apoptosis, inflammatory factor secretion, and the phosphorylation levels of JAK2 and STAT3 proteins were examined.The phosphorylation levels of JAK2 and STAT3 proteins in LPS activated BV-2 cells were detected, and JAK2 inhibitors were introduced to explore the effects of JAK2/STAT3 axis blockade on the cells.A spinal nerve ligation(SNL)rat model with HMGB1 knockdown was established to detect pathway protein expression and inflammatory factor secretion in spinal cord tissue, and to evaluate the mechanical and thermal pain sensitivity thresholds in SNL rats.Results HMGB1 expression was upregulated in LPS treated BV-2 cells, and interfering with HMGB1 expression significantly inhibited the apoptosis and inflammation.LPS treatment activated the JAK2/STAT3 axis, and the activation of JAK2/STAT3 axis was mediated by HMGB1 upregulation.In the SNL rat model, HMGB1 and pathway protein expression in spinal cord tissue were increased, inflammatory responses were aggravated, and both mechanical and thermal pain sensitivity thresholds were decreased.Knockdown of HMGB1 expression downregulated the expressions of HMGB1 and pathway proteins, reduced inflammation, and alleviated neuropathic pain.Conclusion HMGB1 aggravates neuropathic pain by activating the JAK2/STAT3 axis.

关键词(KeyWords)： 神经性疼痛;高迁移率族蛋白B1;JAK2/STAT3轴
neuropathic pain;high mobility group protein B1;JAK2/STAT3 axis

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 毛轲,高誉华,周松林,赵彦芬,张原源,杜思龙
MAO Ke,GAO Yuhua,ZHOU Songlin,ZHAO Yanfen,ZHANG Yuanyuan,DU Silong

DOI: 10.14066/j.cnki.cn21-1349/r.2021.0463

参考文献(References)：

• [1] CHENG J W,JING J H,DU B R,et al.Early clinical diagnostic value of HMGB1 in type 2 diabetes mellitus with retinopathy[J].Journal of Shenyang Pharmaceutical University(沈阳药科大学学报),2021,38(S2):24-25.
• [2] SHANG J,LIU W,YIN C,et al.Cucurbitacin E ameliorates lipopolysaccharide-evoked injury,inflammation and MUC5AC expression in bronchial epithelial cells by restraining the HMGB1-TLR4-NF-κB signaling[J].Mol Immunol,2019,114:571-577.
• [3] WANG X,GUO Y,WANG C,et al.MicroRNA-142-3p inhibits chondrocyte apoptosis and inflammation in osteoarthritis by targeting HMGB1[J].Inflammation,2016,39(5):1718-1728.
• [4] DENG M,TANG Y,LI W,et al.The endotoxin delivery protein HMGB1 mediates caspase-11-dependent lethality in sepsis[J].Immunity,2018,49(4):740-753.e7.
• [5] ZHOU H,DENG M,LIU Y,et al.Platelet HMGB1 is required for efficient bacterial clearance in intra-abdominal bacterial sepsis in mice[J].Blood Adv,2018,2(6):638-648.
• [6] ZHU J R,LU H D,GUO C,et al.Berberine attenuates ischemia-reperfusion injury through inhibiting HMGB1 release and NF-κB nuclear translocation[J].Acta Pharmacol Sin,2018,39(11):1706-1715.
• [7] LIU L,ZHAO Z,LU L,et al.Icariin and icaritin ameliorated hippocampus neuroinflammation via inhibiting HMGB1-related pro-inflammatory signals in lipopolysaccharide-induced inflammation model in C57BL/6J mice[J].Int Immunopharmacol,2019,68:95-105.
• [8] MAKINDE H M,JUST T B,GADHVI G T,et al.Microglia adopt longitudinal transcriptional changes after traumatic brain injury[J].J Surg Res,2020,246:113-122.
• [9] WEST P K,VIENGKHOU B,CAMPBELL I L,et al.Microglia responses to interleukin-6 and type I interferons in neuroinflammatory disease[J].Glia,2019,67(10):1821-1841.
• [10] ZHAO F,FAN S Q,CHENG X Y,et al.Kaempferol alleviates neuropathic pain in rats with spinal nerve ligation by upregulating miR-340-5p[J].Journal of Shenyang Pharmaceutical University(沈阳药科大学学报),2023,40(1):75-82.
• [11] SEN TA NA H,NUO M,MENG Q T,et al.The pathway of let-7a-1/2-3p and HMGB1 mediated dexmedetomidine inhibiting microglia activation in spinal cord ischemia-reperfusion injury mice[J].J Mol Neurosci,2019,69(1):106-114.
• [12] GAO T,CHEN Z,CHEN H,et al.Inhibition of HMGB1 mediates neuroprotection of traumatic brain injury by modulating the microglia/macrophage polarization[J].Biochem Biophys Res Commun,2018,497(1):430-436.
• [13] FAN H,TANG H B,CHEN Z,et al.Inhibiting HMGB1-RAGE axis prevents pro-inflammatory macrophages/microglia polarization and affords neuroprotection after spinal cord injury[J].J Neuroinflammation,2020,17(1):295.
• [14] XIN P,XU X,DENG C,et al.The role of JAK/STAT signaling pathway and its inhibitors in diseases[J].Int Immunopharmacol,2020,80:106210.
• [15] WANG L,ZHANG H B,ZHANG P,et al.Recombinant human p43 protein inhibits angiogenesis through JAK-STAT pathway[J].Journal of Shenyang Pharmaceutical University(沈阳药科大学学报),2020,37(11):1043-1049.
• [16] LIU Q,XIE W,WANG Y,et al.JAK2/STAT1-mediated HMGB1 translocation increases inflammation and cell death in a ventilator-induced lung injury model[J].Lab Invest,2019,99(12):1810-1821.
• [17] WU Y,XU J,XU J,et al.Study on the mechanism of JAK2/STAT3 signaling pathway-mediated inflammatory reaction after cerebral ischemia[J].Mol Med Rep,2018,17(4):5007-5012.