首页|颈康合剂通过Akt/FoxO3a通路调控自噬对神经根型颈椎病根性神经痛的影响

颈康合剂通过Akt/FoxO3a通路调控自噬对神经根型颈椎病根性神经痛的影响

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目的 探讨颈康合剂通过蛋白激酶B(protein kinase B,Akt)/叉头框蛋白O3a(forkhead box O3a,FoxO3a)通路调控自噬影响神经根型颈椎病(cervical spondylotic radiculopathy,CSR)根性神经痛的作用机制.方法 将40只SD大鼠随机分为对照组(正常饲养、不作任何干预处理)、模型组(CSR造模+生理盐水1 ml灌胃)、颈康合剂组(CSR造模+颈康合剂浓缩药液1 ml灌胃)、3-甲基腺嘌呤(3-methyladenine,3-MA)组(CSR造模+3-MA自噬抑制剂1 ml灌胃),每组各10只.检测大鼠步态障碍评价评分、机械刺激缩足反应阈值(paw mechanical withdrawal threshold,PM-WT)与热刺激缩足反射潜伏期(paw withdraw thermal latency,PWTL).免疫组织化学法测定神经根组织微管相关蛋白1 轻链 3β(microtubule-associated proteins light chain 3β,LC3B)表达;苏木精-伊红(hematoxylin-eosin,HE)染色法观察大鼠神经根组织神经元;Western blot法检测神经根组织自噬相关基因Beclin1、自噬相关蛋白5(autophagy related protein 5,ATG5)及Akt、p-Akt、FoxO3a、p-FoxO3a的蛋白含量.结果 与对照组大鼠比较,模型组大鼠PMWT、PWTL明显降低(P均<0.01),神经细胞数量明显减少、体积缩小,分布疏散,神经根组织自噬蛋白Beclin1、ATG5、LC3B表达显著降低(P均<0.01);p-Akt/Akt、p-FoxO3a/FoxO3a表达水平显著升高(P均<0.01).与模型组比较,颈康合剂组大鼠PM-WT、PWTL明显提高(P均<0.01),神经细胞数量增多,神经根Beclin1、ATG5、LC3B表达显著提高(P均<0.01),p-Akt/Akt、p-FoxO3a/FoxO3a表达水平显著降低(P均<0.01);与模型组比较,3-MA组大鼠PMWT、PWTL明显降低(P均<0.01),神经细胞数量减少,神经根组织自噬蛋白Beclin1、ATG5、LC3B表达显著降低(P均<0.01).结论 颈康合剂可能通过调控Akt/FoxO3a信号通路,上调细胞自噬水平,减轻神经细胞损伤,进而提高CSR模型大鼠痛阈,缓解根性神经痛.
Effect of Jingkang Mixture on Radicular Pain in Cervical Spondylotic Radiculopathy by Regulating Autophagy Through the Akt/FoxO3a Pathway
Objective To investigate the mechanism of Jingkang mixture regulates autophagy through the the pro-tein kinase B(Akt)/forkhead box O3a(FoxO3a)signaling pathway to alleviate radicular pain in cervical spondylotic ra-diculopathy(CSR).Methods A total of 40 SD rats were randomly divided into control group(normal feeding without any intervention),model group(CSR molding+gavage of 1 ml normal saline),Jingkang mixture group(CSR molding+gavage of 1ml Jingkang mixture concentrated liquid)and 3-methyladenine(3-MA)group(CSR molding+gavage of 1 ml 3-MA autophagy inhibitor),with 10 rats in each.The evaluation score of gait disorder,paw mechanical withdraw thresh-old(PMWT)and paw withdraw thermal latency(PWTL)were measured.Immunohistochemistry was used to assess the expression of microtubule-associated proteins light chain 3β(LC3B)in the nerve root tissues;while hematoxylin-eosin(HE)staining was performed to observe nerve root tissue neurons;Western blot was used to detect the protein expres-sions of autophagy markers Beclin1,autophagy related protein 5(ATG5),Akt,p-Akt,FoxO3a and p-FoxO3a in the nerve root tissues.Results Compared with the control group,the PMWT and PWTL of rats in model group were significantly lower(all P<0.01);the number and volume of nerve cell were significantly reduced,while the distribution was dispread,and the expression of Beclin1,ATG5 and LC3B in nerve root tissues were significantly reduced(all P<0.01);the levels of p-Akt/Akt and p-FoxO3a/FoxO3a significantly increased(all P<0.01).Compared with the model group,the PMWT and PWTL of rats in the Jingkang mixture group significantly increased(all P<0.01),the number of nerve cell significantly in-creased,and the expression of Beclin1,ATG5 and LC3B in nerve root significantly increased(all P<0.01),while the levels of p-Akt/Akt and p-FoxO3a/FoxO3a were significantly reduced(all P<0.01);compared with the model group,the PMWT and PWTL of rats in the 3-MA group significantly decreased(all P<0.01),the number of nerve cell were significantly reduced,the expression of Beclin1,ATG5 and LC3B in nerve root tissues were significantly reduced(all P<0.01).Conclusion Jingkang mixture can alleviate nerve cell damage and increase pain thresholds in CSR rats and relieve radicular pain by upregulating autophagy via the Akt/FoxO3a signaling pathway.

Cervical spondylotic radiculopathyRadicular painJingkang mixtureProtein kinase B/forkhead box O3a signaling pathwayNerve cell damage

廖翼涛、肖清明、张贤、蔡建平、李超、董朋峻

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210038 江苏南京,南京中医药大学研究生院

南京中医药大学无锡附属医院骨伤科

江苏省名老中医药专家传承工作室

常州市武进区中医医院骨伤科

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神经根型颈椎病 根性神经痛 颈康合剂 蛋白激酶B/叉头框蛋白O3a通路 神经细胞损伤

2024

华南国防医学杂志
广州军区医学科学技术委员会

华南国防医学杂志

CSTPCD
影响因子:0.748
ISSN:1009-2595
年,卷(期):2024.38(12)