葛根素通过抑制背根神经节Nav1.7上调治疗神经根性疼痛  

Puerarin alleviates radicular pain by inhibiting Nav1.7 upregulation in the dorsal root ganglia

作　　者：张玮丽 唐倩 邓镇 赵元淑 钟祎 ZHANG Weili;TANG Qian;DENG Zhen;ZHAO Yuanshu;ZHONG Yi(School of Basic Medical Sciences,Guangzhou Medical University,Guangzhou 511346,China)

机构地区：[1]广州医科大学基础医学院,广州511436

出　　处：《中国疼痛医学杂志》2023年第5期332-339,共8页Chinese Journal of Pain Medicine

基　　金：广东省普通高校特性创新项目(2019KTSCX145)。

摘　　要：目的:探究传统中药葛根素(puerarin,PU)对腰椎间盘突出症引起的神经根性疼痛的作用及机制。方法:采用自体髓核(neucleus pulposus,NP)移植的方法复制大鼠腰椎间盘突出症的模型。实验一:20只大鼠用完全随机化分组法分成假手术组(sham,n=4)和NP组(n=16),NP组采用免疫荧光染色方法检测背根神经节(dorsal root ganglia,DRG)中电压门控钠通道Nav1.7的表达,ELISA法检测DRG和脑脊液(cerebrospinal fluid,CSF)中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的含量;Western blot方法检测DRG组织TNF-α的表达。实验二:40只大鼠分为五组(n=8):sham组;NP移植组;NP+PU组;NP+Veh组;NP+PU+TNF-α组,对NP移植大鼠连续7天腹腔注射葛根素(100 mg/kg,每日1次)观察镇痛效果;在术后0、3、5、7、14天进行疼痛行为学检测,机械刺激缩足反射阈值(mechanical withdrawal threshold,MWT)和热缩足反射潜伏期(thermal withdrawal latency,TWL)作为痛觉阈值检测指标;Western blot方法检测DRG组织中TNF-α和磷酸化核转录因子-κB(phosphorylated-nucleus factor-κB,p-NF-κB)表达水平;免疫荧光染色方法检测Nav1.7表达情况。结果:NP移植组大鼠手术同侧DRG中Nav1.7表达上升,同侧DRG组织和CSF中TNF-α含量显著增加;葛根素明显缓解NP移植引起的单侧机械性痛觉过敏和热痛觉超敏;葛根素降低NP移植组大鼠同侧DRG中TNF-α、p-NF-κB(p-p65)和Nav1.7的表达;鞘内注射TNF-α(0.3 ng,每6小时1次,持续7天)阻断了葛根素的镇痛作用,并抑制了葛根素引起的Nav1.7的降低。结论:葛根素可通过抑制背根神经节Nav1.7上调治疗神经根性疼痛。Objective:To explore the role and mechanism of traditional Chinese medicine puerarin(PU)in radicular pain induced by lumbar disc herniation.Methods:Rat model of lumbar disc herniation was induced by autologous nucleus pulposus(NP)implantation.In experiment one,20 rats were completely randomized grouped into sham group(n=4)and NP group(n=16),voltage gated sodium channel Nav1.7 expression was detected by immunofluorescence;Tumor necrosis factor-α(TNF-α)level in the dorsal root ganglia(DRG)and cerebrospinal fluid(CSF)was detected by enzyme-linked immunosorbent assay(ELISA);The expression of TNF-αin DRG was measured by western blotting analysis.In experiment two,40 rats were divided into 5 groups(n=8):sham group,NP group,NP+PU group,NP+Veh group,NP+PU+TNF-αgroup,puerarin was successively delivered for 7 days(ip,100 mg/kg,qd);Pain thresholds were assessed by mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)on day 0,3,5,7,14 after surgery;The expression of TNF-αand p-NF-κB(p-p65)in DRG was measured by western blotting analysis;Nav1.7 expression was detected by immunofluorescence.Results:Nav1.7 expression in the DRG was increased in rats with NP implantation;TNF-αlevel in the DRG and CSF was increased in rats with NP implantation;Puerarin alleviated mechanical allodynia and thermal hyperalgesia induced by NP implantation;Puerarin reduced TNF-α,p-p65 and Nav1.7 expression in DRG of rats with NP;Intrathecal delivery of TNF-α(0.3 ng,q6 h,for 7 days)significantly obscured the effect of puerarin on Nav1.7 expression.Conclusion:Puerarin alleviates radicular pain by inhibiting Nav1.7 upregulation in the DRG.

关 键 词：葛根素 肿瘤坏死因子-α 背根神经节 电压依赖性钠通道 神经根性疼痛 腰椎间盘突出症 

分 类 号：R285[医药卫生—中药学]