摘要
目的探究针刺对腰椎间盘突出症(LDH)模型大鼠疼痛反应及CXC趋化因子配体12(CXCL12)/CXC趋化因子受体4(CXCR4)信号通路的影响。方法采用自体髓核移植法复制LDH大鼠模型,将造模成功大鼠随机分为LDH组、针刺组、AMD3100(CXCL12/CXCR4信号通路抑制剂)组、针刺+AMD3100组,每组12只,另选12只为正常组。针刺组大鼠予针刺干预,AMD3100组予20μg的AMD3100鞘内注射,针刺+AMD3100组予针刺+AMD3100鞘内注射,LDH组和正常组予等量0.9%氯化钠注射液鞘内注射,每日1次,持续21 d。比较各组大鼠造模前1 d和干预后3、7、14、21 d机械缩足反射阈值(MWT)和热刺激缩足反射潜伏期(TWL);苏木精-伊红(HE)染色观察各组大鼠背根神经节形态改变;酶联免疫吸附法(ELISA)检测大鼠血清肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)及P物质(SP)、5-羟色胺(5-HT)水平;实时荧光定量聚合酶链反应和蛋白免疫印迹法分别检测各组大鼠背根神经节CXCL12、CXCR4 mRNA及蛋白表达。结果与造模前比较,干预后3、7、14、21 d LDH组、针刺组、AMD3100组、针刺+AMD3100组MWT、TWL均降低(P<0.05);与正常组比较,干预后3、7、14、21 d LDH组MWT、TWL均降低(P<0.05);与LDH组比较,干预后3、7、14、21 d针刺组、AMD3100组、针刺+AMD3100组MWT、TWL均升高(P<0.05),且针刺+AMD3100组最高(P<0.05)。与正常组比较,LDH组TNF-α、IL-1β、SP、5-HT水平和CXCL12、CXCR4 mRNA及蛋白表达升高(P<0.05);与LDH组比较,针刺组、AMD3100组、针刺+AMD3100组TNF-α、IL-1β、SP、5-HT水平和CXCL12、CXCR4 mRNA及蛋白表达降低(P<0.05),且针刺+AMD3100组最低(P<0.05)。结论针刺能抑制炎症反应,改善LDH大鼠疼痛反应,其作用机制可能与抑制CXCL12/CXCR4信号传导有关。
Objective To explore the impacts of acupuncture and moxibustion(AM)on pain response and CXC chemokine ligand 12(CXCL12)/CXC chemokine receptor 4(CXCR4)signaling pathway in lumbar disc herniation(LDH)model in rats.Methods The LDH rat model was reproduced by autologous nucleus pulposus transplantation.The successful rats were randomly separated into LDH model group(Vaseline external coating intrathecal injection of 0.9% sodium chloride,qd),AM group(AM,qb),AMD3100 group(intrathecal injection of 20μg AMD3100 which was an inhibitor of CXCL12/CXCR4 signaling pathway,qb),AM+AMD310 group(the combination of AM+intrathecal injection of 20μg AMD3100,12 rats in each group);Another 12 rats were selected as Sham group(intrathecal injection of 0.9% sodium chloride,qb).After 21days of continuous treatment,the mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were detected at one day before modeling and 3th,7th,14th,21th days after intervention.The Morphological changes in the dorsal root ganglion of rats were assessed by hematoxylin and eosin(H&E)staining.The Serum levels of tumor necrosis factor(TNF)-α,interleukin-1β(IL-1β),substance P(SP),and serotonin(5-HT)were determined by Enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expression of CXCL12 and CXCR4 were detected by quantitative reverse-transcription polymerase chain reaction(qRT-PCR)and Western blot,respectively.Results Compared with before modeling,MWT and TWL at 3th,7th,14th,21th days after intervention of the LDH group,AM group,AMD3100 group,and AM+AMD310 group were significantly decreased(P<0.05).Compared with Sham group,MWT and TWL at 3th,7th,14th,21th days after intervention of the LDH group were significantly decreased(P<0.05).Compared with LDH group,MWT and TWL at 3th,7th,14th,21th days after intervention of the AM group,AMD3100 group,and AM+AMD310 group were significantly increased(P<0.05),especially in the AM+AMD3100 group(P<0.05).Compared with Sham group,the mRNA and protein expression of TNF-α,IL-1β,SP,5-HT,CXCL12,and CXCR4 of LDH group were significantly increased(P<0.05).Compared with LDH group,the mRNA and protein expression of TNF-α,IL-1β,SP,5-HT,CXCL12,and CXCR4 of the AM group,AMD3100 group,and AM+AMD310 group were significantly decreased(P<0.05),the above changes were most pronounced in the AM+AMD3100 group(P<0.05).Conclusion AM can inhibit inflammatory response and improve pain response in LDH rats by the inhibition of CXCL12/CXCR4 signal transduction.
作者
徐军
黄移生
范舟云
张玉丰
XU Jun;HUANG Yisheng;FAN Zhouyun;ZHANG Yufeng(Department of Acupuncture and Moxibustion,Huangshi Hospital of Traditional Chinese Medicine,Huangshi,Hubei 435000)
出处
《河北中医》
2025年第12期2042-2047,共6页
Hebei Journal of Traditional Chinese Medicine
基金
湖北省中医药管理局2023~2024年度中医药科研立项项目(编号:ZY2023M034)。
