从NF-κB和Notch-1通路探索紫杉醇-重组水蛭素介入涂层抗再狭窄的细胞学机制  

Investigating the cytological mechanisms of paclitaxel-recombinant hirudin interventional coating to prevent restenosis via NF-κB and Notch-1 pathways

作　　者：王婷 徐元珊 李红梅 WANG Ting;XU Yuan-shan;LI Hong-mei(School of Life Science,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学生命科学学院,北京100029

出　　处：《中国介入心脏病学杂志》2024年第10期576-587,共12页Chinese Journal of Interventional Cardiology

基　　金：北京市自然科学基金面上项目(7222279)。

摘　　要：目的 探索紫杉醇-重组水蛭素介入涂层复合物(LFN)调控Notch-1和NF-κB通路间交互作用关键位点抗再狭窄的微观机制。方法 利用网络药理学技术初筛LFN调控双信号通路交互作用的预测靶点,构建LPS+Jagged-1诱导人冠状动脉平滑肌细胞(HCASMCs)增殖迁移模型,通过敲减或过表达双通路间枢纽基因,明确其交互作用与HCASMCs增殖迁移的相关性。在此基础上,对LFN初筛靶点进行精筛,进而在HCASMCs模型上予以LFN干预,从LFN双向调控Notch-1和NF-κB通路切入,深入阐释LFN防治介入术后再狭窄的细胞学机制。结果 网络药理学筛选结果表明,LFN对再狭窄的调控包含多种生物学模块和过程。联合LPS和Jagged-1以1μg/ml的浓度24 h持续刺激HCASMCs可诱导建立双通路活化细胞模型。与模型组比较,最佳浓度下的LFN(1μmol/L的紫杉醇配比0.2 mg/ml比伐芦定)对造模活化后的HCASMCs迁移和增殖变化具有显著抑制作用,可下调HCASMCs中NF-κB和Notch-1基因表达、上调IκBα基因表达,降低NICD、VEGF、MMP2、MMP9和Bcl-xL基因表达,同时下调OPN、PCNA、Notch-1和NF-κB(细胞核)蛋白表达、上调NF-κB(细胞质)蛋白表达(P<0.05)。其中,Notch-1与NICD表达的变化可直接影响LFN的作用效果。结论 LFN的抗再狭窄效应是通过调节Notch-1和NF-κB双通路间交互作用、阻断HCASMCs从收缩型向分泌型转变而实现。Objective Investigating how the intervention coating composite of paclitaxel-recombinant hirudin(LFN)regulates the connection between Notch-1 and NF-κB pathways to avoid restenosis.Methods Targets and enrichment analysis of LFN anti-ISR were determined using a network-based technique.Human coronary artery smooth muscle cells(HCASMCs)were employed as a model.Lipopolysaccharide(LPS)and Jagged-1 were utilized to activate the Notch-1 and NF-κB pathways in HCASMCs.It is possible to provide light on the relationship between pathway interactions and the growth and migration of HCASMCs by regulating dual pathways.The mechanism of LFN in preventing and curing postoperative restenosis was explained molecularly.Results Computational results revealed that LFN therapy of in-stent restenosis regulated many biological modules.The ideal modeling setting for infl ammatory activation of HCASMCs was found to be 1μg/ml of LPS and Jagged-1 for 24 hours.When compared to the model group,the migration and proliferation changes of modeling-activated HCASMCs were significantly inhibited by LFN(1μmol/L paclitaxel with 0.2 mg/ml bivalirudin)at the optimal concentration.This resulted in a down-regulation of NF-κB and Notch-1,an up-regulation of IκBα,and a decrease in the expression of NICD,VEGF,MMP2,MMP9,and Bcl-xL.It also down-regulated the expression of OPN,PCNA,Notch-1,and NF-κB(nucleus)and up-regulated the expression of NF-κB(cytoplasm)(P<0.05).Among them,the impact of LFN may be directly impacted by changes in Notch-1 and NICD expression.Conclusions LFN prevents restenosis by modulating the Notch-1 and NF-κB pathways and inhibiting the shift of HCASMCs from contractile to synthetic phenotype.

关 键 词：紫杉醇-重组水蛭素介入涂层 抗再狭窄 NF-κB和Notch-1通路 细胞学机制 微观调控 

分 类 号：R54[医药卫生—心血管疾病]