对辣椒素通过MAP1S介导自噬减缓大鼠硬膜外纤维化的效果观察  

Effect observation of capsaicin mediates autophagy via MAP1S to decelerate rats epidural fibrosis

作　　者：娄林冰 戴纪杭 王静成[3] 许磊 Lou Linbing;Dai Jihang;Wang Jingcheng;Xu Lei(the Yangzhou School of Clinical Medicine of Dalian Medical University,Yangzhou 225001,China)

机构地区：[1]大连医科大学扬州临床医学院,扬州225001 [2]扬州大学附属苏北人民医院,225001 [3]江苏省苏北人民医院,扬州225001

出　　处：《骨科临床与研究杂志》2025年第1期44-51,共8页Journal Of Clinical Orthopedics And Research

基　　金：江苏省中医药科技发展计划项目(ZT202117);扬州市骨科学重点实验室(YZ2023249)。

摘　　要：目的探究局部应用辣椒素(Cap)对大鼠椎板切除术后硬膜外纤维化(EF)的效果。方法选取48只8周龄、体重260~320 g的雄性SD大鼠,随机分为假手术组、生理盐水对照组和辣椒素处理组(5 mg/ml和10 mg/ml)。对生理盐水对照组,辣椒素处理组大鼠行椎板切除术。假手术组仅暴露硬脊膜,不行椎板切除术。术前术后通过Basso-Beattie-Bresnahan(BBB)运动评分评估大鼠的关节活动、躯干运动及协调情况。术后1个月,通过大体观察、HE染色、免疫组织化(IHC)、Masson染色和天狼猩红染色评估EF程度。运用串联质谱标签(TMT)蛋白定量组学技术、qRT-PCR和组织免疫荧光染色检测微管相关蛋白1S(MAP1S)、自噬相关蛋白(ATG9A)、纤维连接蛋白(FN1)、肌动蛋白(ACTA2)、微管相关蛋白1轻链3B(LC3B)等基因表达水平。结果所有组别大鼠手术前后的BBB评分差异无统计学意义,表明手术和Cap无显著运动功能损害。宏观和组织学分析显示,Cap显著减缓了EF发生,抑制硬膜外细胞增殖,且效果与浓度相关。Masson和天狼猩红染色证实Cap减少了Ⅰ型胶原纤维沉积。TMT蛋白定量和qRTPCR分析揭示,Cap上调了MAP1S和ATG9A表达,下调了FN1和ACTA2表达。免疫荧光染色显示,Cap处理后MAP1S和LC3B表达显著增加。结论Cap通过激活MAP1S介导的自噬途径,有效抑制硬膜外区域细胞增殖和胶原纤维沉积,从而减缓EF进展。Objective To investigate the effects of local application of capsaicin(Cap)on epidural fibrosis(EF)in rats after laminectomy.Methods Forty-eight male SD rats aged 8 weeks and weighing 260-320 g were randomly divided into a sham operation group,a saline control group,and two capsaicin treatment groups(5 mg/ml and 10 mg/ml).Laminectomy was performed on rats in the saline control and capsaicin-treated groups.Sham operation group only exposed the dura mater without laminectomy.Basso-Beattie-Bresnahan(BBB)locomotor rating scale before and after operation was used to assess joint mobility,trunk movement and coordination in rats.One month postoperatively,EF was evaluated by gross observation,HE staining,immunohistochemistry(IHC),Masson staining,and Sirius red staining.Tandem mass tag(TMT)proteomics,qRT-PCR,and immunofluorescence were used to detect the gene expression levels of microtubule-associated protein 1S(MAP1S),autophagy-related protein 9A(ATG9A),fibronectin 1(FN1)and actin alpha 2(ACTA2),microtubule associated protein 1 light chain 3B(LC3B).Results BBB scores remained consistent pre-and post-surgery in all groups,indicating no significant motor function damage from the surgery or capsaicin.Gross and histological analysis revealed that capsaicin significantly slowed EF development and inhibited extramedullary proliferation,with an effect related to concentration.Masson and Sirius Red staining confirmed reduced deposition of type I collagen fibers following capsaicin treatment.TMT proteomics and qRTPCR showed upregulated expression of MAP1S and ATG9A,and downregulated expression of FN1 and ACTA2 in the capsaicin-treated groups.Immunofluorescence revealed significantly increased expression of MAP1S and LC3B after capsaicin treatment.Conclusion Cap effectively inhibits cell proliferation and collagen fiber deposition in the epidural space through activation of the MAP1S-mediated autophagic pathway,thus slowing the progression of EF.

关 键 词：局部应用 辣椒素 硬膜外纤维化 椎板切除术 

分 类 号：R683.2[医药卫生—骨科学]