Intervertebral disc degeneration(IVDD)is the primary contributor to a range of spinal diseases.Dynamin-related protein 1(Drp1)-mediated mitochondrial fission has recently been identified as a new cause of nucleus pulp...Intervertebral disc degeneration(IVDD)is the primary contributor to a range of spinal diseases.Dynamin-related protein 1(Drp1)-mediated mitochondrial fission has recently been identified as a new cause of nucleus pulposus cell(NPC)death and IVDD,but the underlying mechanisms remain unclear.Although the effects of Drp1 phosphorylation in IVDD have been studied,it is currently unknown if small ubiquitin-like modifications(SUMOylation)of Drp1 regulate IVDD.This study aimed to investigate the functions and mechanisms of mitochondria-anchored protein ligase(MAPL),a mitochondrial SUMO E3 ligase,during IVDD progression.The expression of genes related to SUMOylation and mitochondrial dynamics in TNF-α-stimulated NPCs was analysed via RNA sequencing.展开更多
基于天麻转录组数据,设计特异引物,克隆天麻DRP1E(dynamin-related protein 1E)基因,命名为GeDRP1E(GenBase登录号为C_AA018577.1),借助ExPASy、ClustalW、MEGA等软件对基因进行生物信息学分析利用拟南芥、马铃薯遗传转化体系获得转基...基于天麻转录组数据,设计特异引物,克隆天麻DRP1E(dynamin-related protein 1E)基因,命名为GeDRP1E(GenBase登录号为C_AA018577.1),借助ExPASy、ClustalW、MEGA等软件对基因进行生物信息学分析利用拟南芥、马铃薯遗传转化体系获得转基因植株,并对转基因拟南芥株高、结实率,转基因马铃薯微型薯的块茎长、宽、重量和淀粉含量等农艺性状进行检测和分析,初步解析GeDRP1E基因的功能。结果显示,GeDRP1E基因ORF全长1899 bp、编码632个氨基酸残基,相对分子质量为69.90 kDa,分子式为C3079H4973N883O933S19;理论等电点为7.27,不稳定系数为43.34,亲水性平均指数为-0.259,预测属于不稳定亲水性蛋白。GeDRP1E不存在跨膜结构,无信号肽,定位于细胞质。系统进化树显示,GeDRP1E与其他种类植物DRP1E蛋白之间具有较高的同源性,其中与铁皮石斛DcDRP1E(XP_020689662.1)同源性最高,为90.05%。运用双酶切法构建植物表达载体pCambia1300-35Spro-GeDRP1E,并通过农杆菌介导转化法获得了GeDRP1E基因的拟南芥互补突变体、马铃薯过表达株系。与拟南芥AtDRP1E基因突变体相比,GeDRP1E基因互补突变体株高、结实率表型得以恢复。与野生型马铃薯相比GeDRP1E基因过表达株系微型薯体积、重量显著增大,淀粉含量显著增高。初步推测GeDRP1E很可能参与调节线粒体的形态,从而影响拟南芥植株、马铃薯微型薯的生长发育。上述研究结果为深入阐明天麻块茎生长发育的分子机制奠定了基础。展开更多
目的本研究旨在探讨芒柄花黄素(formononetin,FN)通过干预ROS的生成抑制线粒体动力相关蛋白1(dynamic-related protein 1,DRP1)-NLRP3轴减轻过敏性气道炎症的有关机制。方法为建立过敏性哮喘小鼠模型,50只8周龄的BALB/c小鼠通过卵清蛋白...目的本研究旨在探讨芒柄花黄素(formononetin,FN)通过干预ROS的生成抑制线粒体动力相关蛋白1(dynamic-related protein 1,DRP1)-NLRP3轴减轻过敏性气道炎症的有关机制。方法为建立过敏性哮喘小鼠模型,50只8周龄的BALB/c小鼠通过卵清蛋白(ovalbumin,OVA)诱导后分为对照组、模型组、FN治疗组及地塞米松组。采用HE和Masson染色检测气道炎症和胶原沉积,ELISA测定支气管肺泡灌洗液(BALF)中Th2型细胞因子和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)以及IgE水平,DCFH-DA染色评估BEAS-2B细胞中ROS,免疫组织化学和免疫荧光检测肺组织和BEAS-2B细胞中DRP1表达,免疫印迹分析DRP1-NLRP3途径。结果FN治疗可有效改善哮喘小鼠模型的症状,包括减少嗜酸性粒细胞聚集、气道胶原沉积,降低Th2细胞因子和IgE水平,减少ROS和MDA生成,提高SOD和CAT活性,并调节DRP1-NLRP3途径相关蛋白表达,从而缓解炎症。结论FN通过调节DRP1-NLRP3途径改善哮喘气道炎症。展开更多
Hypoxic-ischemic injury is a common pathological dysfunction in clinical settings.Mitochondria are sensitive organelles that are readily damaged following ischemia and hypoxia.Dynamin-related protein 1(Drp1)regulates ...Hypoxic-ischemic injury is a common pathological dysfunction in clinical settings.Mitochondria are sensitive organelles that are readily damaged following ischemia and hypoxia.Dynamin-related protein 1(Drp1)regulates mitochondrial quality and cellular functions via its oligomeric changes and multiple modifications,which plays a role in mediating the induction of multiple organ damage during hypoxic-ischemic injury.However,there is active controversy and gaps in knowledge regarding the modification,protein interaction,and functions of Drp1,which both hinder and promote development of Drp1 as a novel therapeutic target.Here,we summarize recent findings on the oligomeric changes,modification types,and protein interactions of Drp1 in various hypoxic-ischemic diseases,as well as the Drp1-mediated regulation of mitochondrial quality and cell functions following ischemia and hypoxia.Additionally,potential clinical translation prospects for targeting Drp1 are discussed.This review provides new ideas and targets for proactive interventions on multiple organ damage induced by various hypoxic-ischemic diseases.展开更多
线粒体动力相关蛋白(dynamin-related protein 1,Drp1)是介导线粒体分裂的主要蛋白,Drp1表达增加,线粒体分裂增加,网状结构破坏,反之则有助线粒体融合,促进损伤线粒体修复。心肌缺血再灌注损伤与活性氧(ROS)的大量产生,线粒体通透性转换...线粒体动力相关蛋白(dynamin-related protein 1,Drp1)是介导线粒体分裂的主要蛋白,Drp1表达增加,线粒体分裂增加,网状结构破坏,反之则有助线粒体融合,促进损伤线粒体修复。心肌缺血再灌注损伤与活性氧(ROS)的大量产生,线粒体通透性转换孔(MPTP)的开放及细胞凋亡等密切相关。近年来大量研究发现Drp1介导的线粒体分裂参与心肌缺血再灌注损伤,本文就Drp1参与心肌缺血再灌注损伤的相关机制作一简要综述。展开更多
目的:探讨缺氧-复氧(hypoxia/reoxygenation,H/R)后人近曲肾小管上皮细胞(HK-2)内动力相关蛋白1(Dynamin related protein 1,DRP1)及视神经萎缩症蛋白1(optic atrophy,OPA1)表达的变化。方法:以人近曲肾小管上皮细胞株为研究对象,将培...目的:探讨缺氧-复氧(hypoxia/reoxygenation,H/R)后人近曲肾小管上皮细胞(HK-2)内动力相关蛋白1(Dynamin related protein 1,DRP1)及视神经萎缩症蛋白1(optic atrophy,OPA1)表达的变化。方法:以人近曲肾小管上皮细胞株为研究对象,将培养细胞随机分为正常对照组和H/R组。正常对照组常规培养,H/R组先缺氧24h,然后复氧培养6h。光镜观察细胞形态变化,CCK-8检测细胞活力,透射电子显微镜观察线粒体形态改变,免疫组织化学法检测细胞内DRP1和OPA1蛋白表达。结果:H/R组与对照组相比,细胞活力下降,线粒体出现凋亡相关改变,OPA1表达减少而DRP1的表达增加。结论:缺氧-复氧可导致HK-2细胞发生凋亡相关改变,其机制可能与诱导线粒体形态相关蛋白DRP1、OPA1的表达改变相关。展开更多
基金supported by National Natural Science Foundation of China(82272549,82472505,and 82472498)National key Research and Development plan,Ministry of Science and Technology of the People’s Republic of China(2022YFC2407203)+2 种基金the Young Health Talents of Shanghai Municipal Health Commission,China(2022YQ011)China Medical Education Association(3030537245)The Youth Talent Project of Huashan Hospital(30302164006).
文摘Intervertebral disc degeneration(IVDD)is the primary contributor to a range of spinal diseases.Dynamin-related protein 1(Drp1)-mediated mitochondrial fission has recently been identified as a new cause of nucleus pulposus cell(NPC)death and IVDD,but the underlying mechanisms remain unclear.Although the effects of Drp1 phosphorylation in IVDD have been studied,it is currently unknown if small ubiquitin-like modifications(SUMOylation)of Drp1 regulate IVDD.This study aimed to investigate the functions and mechanisms of mitochondria-anchored protein ligase(MAPL),a mitochondrial SUMO E3 ligase,during IVDD progression.The expression of genes related to SUMOylation and mitochondrial dynamics in TNF-α-stimulated NPCs was analysed via RNA sequencing.
文摘基于天麻转录组数据,设计特异引物,克隆天麻DRP1E(dynamin-related protein 1E)基因,命名为GeDRP1E(GenBase登录号为C_AA018577.1),借助ExPASy、ClustalW、MEGA等软件对基因进行生物信息学分析利用拟南芥、马铃薯遗传转化体系获得转基因植株,并对转基因拟南芥株高、结实率,转基因马铃薯微型薯的块茎长、宽、重量和淀粉含量等农艺性状进行检测和分析,初步解析GeDRP1E基因的功能。结果显示,GeDRP1E基因ORF全长1899 bp、编码632个氨基酸残基,相对分子质量为69.90 kDa,分子式为C3079H4973N883O933S19;理论等电点为7.27,不稳定系数为43.34,亲水性平均指数为-0.259,预测属于不稳定亲水性蛋白。GeDRP1E不存在跨膜结构,无信号肽,定位于细胞质。系统进化树显示,GeDRP1E与其他种类植物DRP1E蛋白之间具有较高的同源性,其中与铁皮石斛DcDRP1E(XP_020689662.1)同源性最高,为90.05%。运用双酶切法构建植物表达载体pCambia1300-35Spro-GeDRP1E,并通过农杆菌介导转化法获得了GeDRP1E基因的拟南芥互补突变体、马铃薯过表达株系。与拟南芥AtDRP1E基因突变体相比,GeDRP1E基因互补突变体株高、结实率表型得以恢复。与野生型马铃薯相比GeDRP1E基因过表达株系微型薯体积、重量显著增大,淀粉含量显著增高。初步推测GeDRP1E很可能参与调节线粒体的形态,从而影响拟南芥植株、马铃薯微型薯的生长发育。上述研究结果为深入阐明天麻块茎生长发育的分子机制奠定了基础。
文摘目的本研究旨在探讨芒柄花黄素(formononetin,FN)通过干预ROS的生成抑制线粒体动力相关蛋白1(dynamic-related protein 1,DRP1)-NLRP3轴减轻过敏性气道炎症的有关机制。方法为建立过敏性哮喘小鼠模型,50只8周龄的BALB/c小鼠通过卵清蛋白(ovalbumin,OVA)诱导后分为对照组、模型组、FN治疗组及地塞米松组。采用HE和Masson染色检测气道炎症和胶原沉积,ELISA测定支气管肺泡灌洗液(BALF)中Th2型细胞因子和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)以及IgE水平,DCFH-DA染色评估BEAS-2B细胞中ROS,免疫组织化学和免疫荧光检测肺组织和BEAS-2B细胞中DRP1表达,免疫印迹分析DRP1-NLRP3途径。结果FN治疗可有效改善哮喘小鼠模型的症状,包括减少嗜酸性粒细胞聚集、气道胶原沉积,降低Th2细胞因子和IgE水平,减少ROS和MDA生成,提高SOD和CAT活性,并调节DRP1-NLRP3途径相关蛋白表达,从而缓解炎症。结论FN通过调节DRP1-NLRP3途径改善哮喘气道炎症。
基金This work was supported by the National Natural Science Foundation of China(82272252,82270378)the Senior Medical Talents Program of Chongqing for Young and Middle-agedthe Kuanren Talents Program of the Second Affiliated Hospital of Chongqing Medical University.
文摘Hypoxic-ischemic injury is a common pathological dysfunction in clinical settings.Mitochondria are sensitive organelles that are readily damaged following ischemia and hypoxia.Dynamin-related protein 1(Drp1)regulates mitochondrial quality and cellular functions via its oligomeric changes and multiple modifications,which plays a role in mediating the induction of multiple organ damage during hypoxic-ischemic injury.However,there is active controversy and gaps in knowledge regarding the modification,protein interaction,and functions of Drp1,which both hinder and promote development of Drp1 as a novel therapeutic target.Here,we summarize recent findings on the oligomeric changes,modification types,and protein interactions of Drp1 in various hypoxic-ischemic diseases,as well as the Drp1-mediated regulation of mitochondrial quality and cell functions following ischemia and hypoxia.Additionally,potential clinical translation prospects for targeting Drp1 are discussed.This review provides new ideas and targets for proactive interventions on multiple organ damage induced by various hypoxic-ischemic diseases.
文摘线粒体动力相关蛋白(dynamin-related protein 1,Drp1)是介导线粒体分裂的主要蛋白,Drp1表达增加,线粒体分裂增加,网状结构破坏,反之则有助线粒体融合,促进损伤线粒体修复。心肌缺血再灌注损伤与活性氧(ROS)的大量产生,线粒体通透性转换孔(MPTP)的开放及细胞凋亡等密切相关。近年来大量研究发现Drp1介导的线粒体分裂参与心肌缺血再灌注损伤,本文就Drp1参与心肌缺血再灌注损伤的相关机制作一简要综述。
文摘目的:探讨缺氧-复氧(hypoxia/reoxygenation,H/R)后人近曲肾小管上皮细胞(HK-2)内动力相关蛋白1(Dynamin related protein 1,DRP1)及视神经萎缩症蛋白1(optic atrophy,OPA1)表达的变化。方法:以人近曲肾小管上皮细胞株为研究对象,将培养细胞随机分为正常对照组和H/R组。正常对照组常规培养,H/R组先缺氧24h,然后复氧培养6h。光镜观察细胞形态变化,CCK-8检测细胞活力,透射电子显微镜观察线粒体形态改变,免疫组织化学法检测细胞内DRP1和OPA1蛋白表达。结果:H/R组与对照组相比,细胞活力下降,线粒体出现凋亡相关改变,OPA1表达减少而DRP1的表达增加。结论:缺氧-复氧可导致HK-2细胞发生凋亡相关改变,其机制可能与诱导线粒体形态相关蛋白DRP1、OPA1的表达改变相关。
