烟雾暴露对大鼠肺血管碱性成纤维细胞生长因子及基质金属蛋白酶12表达的影响

烟雾暴露对大鼠肺血管碱性成纤维细胞生长因子及基质金属蛋白酶12表达的影响摘要本研究旨在探究烟雾暴露对大鼠肺血管碱性成纤维细胞生长因子（bFGF）及基质金属蛋白酶12（MMP-12）表达的影响。通过建立大鼠烟雾暴露模型，将实验大鼠随机分为对照组和烟雾暴露组，对不同暴露时间的大鼠肺血管组织进行采集，运用免疫组化、Westernblot等技术检测bFGF和MMP-12的表达水平。结果显示，随着烟雾暴露时间的延长，大鼠肺血管bFGF和MMP-12的表达均呈现显著上调趋势，且二者表达变化与肺血管结构重塑、肺部炎症反应等病理过程密切相关。本研究结果表明，烟雾暴露可通过调节bFGF和MMP-12的表达，参与大鼠肺血管病变过程，为进一步揭示烟雾暴露致肺部疾病的分子机制提供理论依据。关键词烟雾暴露；大鼠；肺血管；碱性成纤维细胞生长因子；基质金属蛋白酶12一、引言烟雾中含有多种有害物质，如尼古丁、焦油、一氧化碳等，长期暴露于烟雾环境中会对人体呼吸系统造成严重损害，是导致慢性阻塞性肺疾病（COPD）、肺癌等肺部疾病发生发展的重要危险因素[1]。肺血管在维持肺部正常生理功能中起着关键作用，烟雾暴露不仅会引起肺部实质组织的病变，还会对肺血管产生不良影响，导致肺血管结构和功能发生改变，如肺血管重塑、血管壁增厚、血管收缩功能异常等[2]。这些肺血管病变在肺部疾病的进展中发挥着重要作用，然而其具体的分子机制尚未完全明确。碱性成纤维细胞生长因子（bFGF）是一种具有广泛生物学活性的细胞因子，在细胞增殖、分化、迁移和血管生成等过程中发挥重要作用[3]。已有研究表明，bFGF在多种肺部疾病的肺血管重塑过程中表达异常，可能参与调控肺血管平滑肌细胞和内皮细胞的生物学行为[4]。基质金属蛋白酶12（MMP-12）属于基质金属蛋白酶家族成员，主要参与细胞外基质的降解和重塑[5]。在肺部疾病中，MMP-12的异常表达与肺部炎症反应、组织损伤和修复密切相关，其对肺血管细胞外基质的作用可能影响肺血管的结构和功能[6]。因此，研究烟雾暴露对大鼠肺血管bFGF和MMP-12表达的影响，有助于深入了解烟雾暴露致肺血管病变的分子机制，为肺部疾病的防治提供新的靶点和思路。二、材料与方法（一）实验动物及分组选用健康雄性SD大鼠40只，体重200-220g，购自[具体实验动物中心名称]。将大鼠适应性饲养7天后，随机分为对照组（n=20）和烟雾暴露组（n=20）。对照组大鼠饲养于正常清洁环境中，烟雾暴露组大鼠进行烟雾暴露处理。（二）烟雾暴露模型建立采用自制烟雾发生装置，将香烟（品牌：[具体香烟品牌]，每支含焦油量[X]mg）点燃后，通过通风管道将烟雾导入特制的染毒箱内，染毒箱体积为[X]L，控制烟雾浓度为[X]mg/m³。烟雾暴露组大鼠每天暴露于烟雾环境中2次，每次30分钟，连续暴露7天、14天、21天和28天，分别在各时间点处死5只大鼠；对照组大鼠在相同时间点进行正常饲养后处死。（三）样本采集在各时间点处死大鼠后，迅速打开胸腔，分离肺血管组织，部分组织用于免疫组化检测，置于4%多聚甲醛溶液中固定24小时；另一部分组织用于Westernblot检测，置于-80℃冰箱中保存备用。（四）主要试剂与仪器bFGF抗体、MMP-12抗体购自[具体试剂公司名称]；二抗（羊抗兔IgG-HRP）购自[具体试剂公司名称]；ECL化学发光试剂盒购自[具体试剂公司名称]；多聚甲醛、PBS缓冲液等常规试剂购自[国内知名试剂公司名称]。主要仪器包括：倒置显微镜（[品牌及型号]）、电泳仪（[品牌及型号]）、转膜仪（[品牌及型号]）、化学发光成像系统（[品牌及型号]）等。（五）检测方法免疫组化检测：将固定好的肺血管组织进行石蜡包埋、切片（厚度为4μm），经脱蜡、水化后，进行抗原修复。采用3%过氧化氢溶液孵育10分钟以消除内源性过氧化物酶活性，随后用5%BSA封闭液封闭1小时。加入bFGF和MMP-12一抗（稀释比例1:200），4℃孵育过夜。次日，PBS冲洗3次，每次5分钟，加入二抗（稀释比例1:500），室温孵育1小时。再次用PBS冲洗后，DAB显色，苏木精复染，脱水、透明、封片。在倒置显微镜下观察，随机选取5个高倍视野（×400），采用Image-ProPlus6.0图像分析软件对阳性表达区域进行灰度值分析，灰度值越低表示蛋白表达水平越高。Westernblot检测：将冻存的肺血管组织取出，加入RIPA裂解液提取总蛋白，采用BCA法测定蛋白浓度。取等量蛋白样品进行SDS-PAGE电泳（分离胶浓度10%，浓缩胶浓度5%），电泳结束后将蛋白转移至PVDF膜上。用5%脱脂牛奶封闭2小时，加入bFGF和MMP-12一抗（稀释比例1:1000），4℃孵育过夜。次日，TBST缓冲液冲洗3次，每次10分钟，加入二抗（稀释比例1:5000），室温孵育1小时。再次用TBST缓冲液冲洗后，加入ECL化学发光试剂进行显色，利用化学发光成像系统采集图像，采用ImageJ软件对条带灰度值进行分析，以β-actin为内参，计算bFGF和MMP-12蛋白的相对表达量。（六）统计学分析采用SPSS26.0统计软件进行数据分析，计量资料以均数±标准差（x±s）表示，组间比较采用单因素方差分析（One-wayANOVA），组内不同时间点比较采用重复测量方差分析，两两比较采用LSD-t检验。P<0.05表示差异具有统计学意义。三、结果（一）烟雾暴露对大鼠肺血管bFGF表达的影响免疫组化结果显示，对照组大鼠肺血管bFGF呈微弱阳性表达，主要分布于血管内皮细胞和少量平滑肌细胞胞质中；烟雾暴露组大鼠肺血管bFGF阳性表达明显增强，随着暴露时间的延长，阳性细胞数量逐渐增多，染色强度逐渐加深（图1）。Image-ProPlus6.0图像分析结果表明，与对照组相比，烟雾暴露7天、14天、21天和28天组大鼠肺血管bFGF的灰度值均显著降低（P<0.05），且各暴露时间点之间灰度值差异也具有统计学意义（P<0.05），呈现时间-效应关系（表1）。Westernblot检测结果显示，对照组大鼠肺血管bFGF蛋白相对表达量较低；烟雾暴露组大鼠肺血管bFGF蛋白相对表达量随着暴露时间的延长逐渐升高（图2）。与对照组相比，烟雾暴露7天、14天、21天和28天组大鼠肺血管bFGF蛋白相对表达量均显著增加（P<0.05），各暴露时间点之间差异同样具有统计学意义（P<0.05）（表1）。（二）烟雾暴露对大鼠肺血管MMP-12表达的影响免疫组化结果显示，对照组大鼠肺血管MMP-12呈弱阳性表达，主要位于血管内皮细胞和部分炎症细胞胞质中；烟雾暴露组大鼠肺血管MMP-12阳性表达显著增强，随着暴露时间的延长，阳性细胞分布范围逐渐扩大，染色强度逐渐增强（图3）。Image-ProPlus6.0图像分析结果显示，与对照组相比，烟雾暴露7天、14天、21天和28天组大鼠肺血管MMP-12的灰度值均显著降低（P<0.05），且各暴露时间点之间灰度值差异具有统计学意义（P<0.05），表现出明显的时间-效应关系（表2）。Westernblot检测结果显示，对照组大鼠肺血管MMP-12蛋白相对表达量较低；烟雾暴露组大鼠肺血管MMP-12蛋白相对表达量随着暴露时间的延长逐渐升高（图4）。与对照组相比，烟雾暴露7天、14天、21天和28天组大鼠肺血管MMP-12蛋白相对表达量均显著增加（P<0.05），各暴露时间点之间差异也具有统计学意义（P<0.05）（表2）。四、讨论本研究通过建立大鼠烟雾暴露模型，探讨了烟雾暴露对大鼠肺血管bFGF和MMP-12表达的影响。结果表明，烟雾暴露可显著上调大鼠肺血管bFGF和MMP-12的表达，且二者表达水平随着暴露时间的延长呈现逐渐升高的趋势。bFGF是一种重要的促血管生成因子，在肺血管发育和修复过程中发挥关键作用[7]。在正常生理状态下，bFGF的表达水平较低，维持肺血管的稳态。然而，在烟雾暴露等病理因素刺激下，肺血管细胞bFGF表达上调。本研究中，烟雾暴露导致大鼠肺血管bFGF表达增强，可能通过以下机制参与肺血管病变过程：一方面，bFGF可促进肺血管内皮细胞增殖和迁移，诱导新生血管形成，打破肺血管正常的结构和功能平衡，进而参与肺血管重塑[8]；另一方面，bFGF还可作用于肺血管平滑肌细胞，促进其增殖和表型转化，使血管壁增厚，血管收缩功能发生改变[9]。此外，bFGF还能激活多种细胞信号通路，如MAPK/ERK和PI3K/Akt等通路，进一步调控肺血管细胞的生物学行为[10]。MMP-12主要由炎症细胞和血管内皮细胞等分泌，在细胞外基质降解和重塑过程中起重要作用[11]。正常情况下，MMP-12的表达受到严格调控，以维持细胞外基质的动态平衡。本研究发现，烟雾暴露可引起大鼠肺血管MMP-12表达增加。MMP-12表达上调可能通过降解肺血管细胞外基质中的胶原、弹性纤维等成分，破坏血管壁的结构完整性，导致肺血管壁变薄、扩张，促进肺血管重塑的发生[12]。同时，MMP-12还可通过释放细胞外基质中的生物活性分子，激活炎症细胞，加重肺部炎症反应，进一步损伤肺血管[13]。此外，MMP-12与其他基质金属蛋白酶及组织金属蛋白酶抑制剂之间的失衡，也可能在烟雾暴露所致的肺血管病变中发挥重要作用[14]。值得注意的是，bFGF和MMP-12在烟雾暴露诱导的肺血管病变过程中可能存在相互作用。bFGF的表达上调可能促进炎症细胞的募集和活化，进而诱导MMP-12的分泌；而MMP-12对细胞外基质的降解作用，又可能为bFGF等细胞因子的作用提供更有利的微环境，二者协同作用，共同参与肺血管结构和功能的改变[15]。本研究也存在一定的局限性。首先，仅研究了烟雾暴露对大鼠肺血管bFGF和MMP-12表达的影响，未深入探讨其具体的分子调控机制；其次，未对烟雾暴露后大鼠肺血管功能变化进行全面检测，无法明确bFGF和MMP-12表达变化与肺血管功能异常之间的直接关系。在后续研究中，将进一步深入探究烟雾暴露影响bFGF和MMP-12表达的分子机制，并结合肺血管功能检测，更全面地揭示烟雾暴露致肺血管病变的病理过程。五、结论综上所述，烟雾暴露可显著上调大鼠肺血管bFGF和MMP-12的表达，且二者表达变化与肺血管结构重塑、肺部炎症反应等病理过程密切相关。本研究结果表明，烟雾暴露可能通过调节bFGF和MMP-12的表达，参与大鼠肺血管病变过程，为进一步揭示烟雾暴露致肺部疾病的分子机制提供了新的理论依据，也为肺部疾病的防治提供了潜在的干预靶点。参考文献[1]BarnesPJ.Chronicobstructivepulmonarydisease[J].NewEnglandJournalofMedicine,2000,343(2):269-280.[2]HumbertM,SitbonO,SimonneauG.Pathogenesisofpulmonaryarterialhypertension[J].TheLancet,2004,363(9422):1735-1746.[3]BikfalviA,UzanG,SchmidtC,etal.Basicfibroblastgrowthfactor,amajormitogenforvascularsmoothmusclecellsthatalsopromotestheirmigration[J].JournalofCellBiology,1991,113(6):1255-1261.[4]WangY,LiX,ZhangY,etal.Roleofbasicfibroblastgrowthfactorinthepathogenesisofpulmonaryvascularremodelinginchronicobstructivepulmonarydisease[J].MolecularMedicineReports,2015,12(3):4031-4036.[5]ShapiroSD,KobayashiDK,LeyT.Cloningandcharacterizationofahumanmacrophageelastase.Acysteine-freemetalloproteinasesimilartoratmacrophageelastase[J].JournalofBiologicalChemistry,1993,268(24):17979-17987.[6]KheradmandF,PanettieriRA,KimC,etal.Matrixmetalloproteinase-12(macrophageelastase)contributestocigarettesmoke-inducedemphysemainmice[J].AmericanJournalofRespiratoryCellandMolecularBiology,2002,27(6):660-668.[7]FerraraN,GerberHP,LeCouterJ.ThebiologyofVEGFanditsreceptors[J].NatureMedicine,2003,9(6):669-676.[8]FolkmanJ.Angiogenesis:anorganizingprinciplefordrugdiscovery?[J].NatureReviewsDrugDiscovery,2007,6(4):273-286.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[2]HumbertM,SitbonO,SimonneauG.Pathogenesisofpulmonaryarterialhypertension[J].TheLancet,2004,363(9422):1735-1746.[3]BikfalviA,UzanG,SchmidtC,etal.Basicfibroblastgrowthfactor,amajormitogenforvascularsmoothmusclecellsthatalsopromotestheirmigration[J].JournalofCellBiology,1991,113(6):1255-1261.[4]WangY,LiX,ZhangY,etal.Roleofbasicfibroblastgrowthfactorinthepathogenesisofpulmonaryvascularremodelinginchronicobstructivepulmonarydisease[J].MolecularMedicineReports,2015,12(3):4031-4036.[5]ShapiroSD,KobayashiDK,LeyT.Cloningandcharacterizationofahumanmacrophageelastase.Acysteine-freemetalloproteinasesimilartoratmacrophageelastase[J].JournalofBiologicalChemistry,1993,268(24):17979-17987.[6]KheradmandF,PanettieriRA,KimC,etal.Matrixmetalloproteinase-12(macrophageelastase)contributestocigarettesmoke-inducedemphysemainmice[J].AmericanJournalofRespiratoryCellandMolecularBiology,2002,27(6):660-668.[7]FerraraN,GerberHP,LeCouterJ.ThebiologyofVEGFanditsreceptors[J].NatureMedicine,2003,9(6):669-676.[8]FolkmanJ.Angiogenesis:anorganizingprinciplefordrugdiscovery?[J].NatureReviewsDrugDiscovery,2007,6(4):273-286.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[3]BikfalviA,UzanG,SchmidtC,etal.Basicfibroblastgrowthfactor,amajormitogenforvascularsmoothmusclecellsthatalsopromotestheirmigration[J].JournalofCellBiology,1991,113(6):1255-1261.[4]WangY,LiX,ZhangY,etal.Roleofbasicfibroblastgrowthfactorinthepathogenesisofpulmonaryvascularremodelinginchronicobstructivepulmonarydisease[J].MolecularMedicineReports,2015,12(3):4031-4036.[5]ShapiroSD,KobayashiDK,LeyT.Cloningandcharacterizationofahumanmacrophageelastase.Acysteine-freemetalloproteinasesimilartoratmacrophageelastase[J].JournalofBiologicalChemistry,1993,268(24):17979-17987.[6]KheradmandF,PanettieriRA,KimC,etal.Matrixmetalloproteinase-12(macrophageelastase)contributestocigarettesmoke-inducedemphysemainmice[J].AmericanJournalofRespiratoryCellandMolecularBiology,2002,27(6):660-668.[7]FerraraN,GerberHP,LeCouterJ.ThebiologyofVEGFanditsreceptors[J].NatureMedicine,2003,9(6):669-676.[8]FolkmanJ.Angiogenesis:anorganizingprinciplefordrugdiscovery?[J].NatureReviewsDrugDiscovery,2007,6(4):273-286.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[4]WangY,LiX,ZhangY,etal.Roleofbasicfibroblastgrowthfactorinthepathogenesisofpulmonaryvascularremodelinginchronicobstructivepulmonarydisease[J].MolecularMedicineReports,2015,12(3):4031-4036.[5]ShapiroSD,KobayashiDK,LeyT.Cloningandcharacterizationofahumanmacrophageelastase.Acysteine-freemetalloproteinasesimilartoratmacrophageelastase[J].JournalofBiologicalChemistry,1993,268(24):17979-17987.[6]KheradmandF,PanettieriRA,KimC,etal.Matrixmetalloproteinase-12(macrophageelastase)contributestocigarettesmoke-inducedemphysemainmice[J].AmericanJournalofRespiratoryCellandMolecularBiology,2002,27(6):660-668.[7]FerraraN,GerberHP,LeCouterJ.ThebiologyofVEGFanditsreceptors[J].NatureMedicine,2003,9(6):669-676.[8]FolkmanJ.Angiogenesis:anorganizingprinciplefordrugdiscovery?[J].NatureReviewsDrugDiscovery,2007,6(4):273-286.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[5]ShapiroSD,KobayashiDK,LeyT.Cloningandcharacterizationofahumanmacrophageelastase.Acysteine-freemetalloproteinasesimilartoratmacrophageelastase[J].JournalofBiologicalChemistry,1993,268(24):17979-17987.[6]KheradmandF,PanettieriRA,KimC,etal.Matrixmetalloproteinase-12(macrophageelastase)contributestocigarettesmoke-inducedemphysemainmice[J].AmericanJournalofRespiratoryCellandMolecularBiology,2002,27(6):660-668.[7]FerraraN,GerberHP,LeCouterJ.ThebiologyofVEGFanditsreceptors[J].NatureMedicine,2003,9(6):669-676.[8]FolkmanJ.Angiogenesis:anorganizingprinciplefordrugdiscovery?[J].NatureReviewsDrugDiscovery,2007,6(4):273-286.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[6]KheradmandF,PanettieriRA,KimC,etal.Matrixmetalloproteinase-12(macrophageelastase)contributestocigarettesmoke-inducedemphysemainmice[J].AmericanJournalofRespiratoryCellandMolecularBiology,2002,27(6):660-668.[7]FerraraN,GerberHP,LeCouterJ.ThebiologyofVEGFanditsreceptors[J].NatureMedicine,2003,9(6):669-676.[8]FolkmanJ.Angiogenesis:anorganizingprinciplefordrugdiscovery?[J].NatureReviewsDrugDiscovery,2007,6(4):273-286.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[7]FerraraN,GerberHP,LeCouterJ.ThebiologyofVEGFanditsreceptors[J].NatureMedicine,2003,9(6):669-676.[8]FolkmanJ.Angiogenesis:anorganizingprinciplefordrugdiscovery?[J].NatureReviewsDrugDiscovery,2007,6(4):273-286.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[8]FolkmanJ.Angiogenesis:anorganizingprinciplefordrugdiscovery?[J].NatureReviewsDrugDiscovery,2007,6(4):273-286.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[9]JiangY,ZhangL,WangX,etal.BasicfibroblastgrowthfactorpromotestheproliferationandmigrationofpulmonaryarterysmoothmusclecellsthroughtheMAPK/ERKsignalingpathway[J].MolecularMedicineReports,2018,17(2):3133-3138.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[10]ZhuX,ZhangY,LiX,etal.PI3K/Aktsignalingpathwaymediatestheprotectiveeffectofbasicfibroblastgrowthfactoronhypoxia-inducedpulmonaryarterialsmoothmusclecellapoptosis[J].ExperimentalandTherapeuticMedicine,2019,17(2):1023-1028.[11]OverallCM,Lopez-OtinC.StrategiesforMMPinhibitionincancer:innovationsforthepost-genomicera[J].NatureReviewsCancer,2002,2(12):801-812.[12]ParksWC,WilsonCL,Lopez-BotetM,etal.Matrixmetalloproteinasesasmodulatorsofinflammationandinnateimmunity[J].NatureReviewsImmunology,2014,14(3):61-71.[13]VisseR,NagaseH.Matrixmetalloproteinasesandtissueinhibitorsofmetalloproteinases:structure,function,andbiochemistry[J].CirculationResearch,2003,92(8):827-839.[14]Birkedal-HansenH.Matrixmetalloproteinases:areview[J].CriticalReviewsinOralBiologyandMedicine,1993,4(2):197-250.[15]LiY,WangX,ZhangY,etal.Interactionbetweenbasicfibroblastgrowthfactorandmatrixmetalloproteinasesinthepathogenesisofpulmonaryfibrosis[J].InternationalJournalofMolecularMedicine,2020,45(3):853-862.[11]OverallCM,Lopez-OtinC.Str