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  

Effect of,Jjingkang Mixture on Radicular Pain in Cervical Spondylotic Radiculopathy by Regulating Autophagy Through the Akt/FoxO3a Pathway

作　　者：廖翼涛 肖清明[2] 张贤[2] 蔡建平 李超[2] 董朋峻 LIAO Yitao;XIAO Qingming;ZHANG Xian;CAI Jianping;LI Chao;DONG Pengjun(Department of GraduateSchool,Nanjing University of Chinese Medicine,Nanjing Jiangsu 210038,China)

机构地区：[1]南京中医药大学研究生院,江苏南京210038 [2]南京中医药大学无锡附属医院骨伤科 [3]蔡建平江苏省名老中医药专家传承工作室 [4]常州市武进区中医医院骨伤科

出　　处：《联勤军事医学》2024年第12期1016-1021,共6页Military Medicine of Joint Logistics

基　　金：蔡建平江苏省名老中医药专家传承工作室建设项目(苏中医科教[2021]7号);无锡市中医药管理局科技项目(ZYYB04)。

摘　　要：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,PMWT)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PMWT、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。Ohjective To investigate the mechanism of Jingkang mixture regulates autophagy through the the pro-tein kinase B(Akt)/forkhead box O3a(Fox03a)signaling pathway to alleviate radicular pain in cervical spondylotic rardiculopathy(CSR),Methods A total of 40 SD rats were randomly divided into control group(normal feeding withoutany intervention),model group(CSR molding+gavage of 1 ml normal saline),Jingkang mixture group(CSR molding+gavage of 1 ml Jingkang mixture concentrated liquid)and 3-methyladenine(3-MA)group(CSR molding+gavage of 1 ml3-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 theexpression of microtubule-associated proteins light chain 33(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 groupwere significantly lower(all P<0.01);the number andvolume of nerve cell were significantly reduced,while thedistribution was dispread,and the expression of Beclin1,ATG5 and LC3B in nerve root tissues were significantlyreduced(all P0.01);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 thelevels of p-Akt/Akt and p-Fox03a/Fox03a 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 weresignificantly reduced,the expression of

关 键 词：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 

分 类 号：R289[医药卫生—方剂学] R274[医药卫生—中药学]