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Insight into vitronectin structural evolution on material surface chemistries: The mediation for cell adhesion

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Bioactive Materials 2020年第4期5卷 1044-1052页

作者： Tianjie Li Lijing Hao Jiangyu Li Chang Du Yingjun Wang Department of Biomedical Engineering School of Materials Science and EngineeringSouth China University of TechnologyGuangzhou510641PR China National Engineering Research Center for Tissue Restoration and Reconstruction PR China Key Laboratory of Biomedical Materials and Engineering Ministry of EducationSouth China University of TechnologyGuangzhou510006PR China Department of Mechanical Engineering University of WashingtonSeattle98195WashingtonUnited States Guangzhou Regenerative Medicine and Health Guangdong Laboratory 510005PR China

Biomaterial surface chemistry engenders profound consequences on cell adhesion and the ultimate tissue response by adsorbing proteins from extracellular matrix,where vitronectin(Vn)is involved as one of the crucial mediator *** the adsorption behaviors of Vn in molecular scale provides a useful account of how to design biomaterial *** the details of structural dynamics and consequential biological effect remain ***,both experimental and computational approaches were applied to delineate the conformational and orientational evolution of Vn during adsorption onto self-assembled monolayers(SAMs)terminating with-COOH,-NH2,-CH3 *** unravel the interplay between cell binding and the charge and wettability of material surface,somatomedin-B(SMB)domain of Vn holding the RGD cell-binding motif was employed in molecular dynamics(MD)simulations,with orientation initialized by Monte Carlo(MC)*** evidences including protein adsorption,cell adhesion and integrin gene expressions were thoroughly *** adsorption of Vn on different surface chemistries showed very complex *** adhesion was enabled on all Vn-adsorbed surfaces but with distinct mechanisms mostly determined by conformational change induced *** amount of Vn was observed on negatively charged surface(COOH)and hydrophobic surface(CH3).However,advantageous orientations defined by RGD loop conditions were only obtained on the charged surfaces(COOH and NH2).Specifically,COOH surface straightened up the Vn molecules and accumulated them into a higher density,whereas CH3 surface squashed Vn and stacked them into higher density multilayer by tracking adsorption but with the RGD loops *** findings may have a broad implication on the understanding of Vn functionality and would help develop new strategies for designing advanced biomaterials.

关键词： vitronectin Protein adsorption Surface chemistry Cell adhesion Molecular dynamics simulation

Improvement of human embryonic stem cell-derived retinal pigment epithelium cell adhesion, maturation, and function through coating with truncated recombinant human vitronectin

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International Journal of Ophthalmology(English edition) 2021年第8期14卷 1160-1167页

作者： Xin-Yue Zhu Yu-Hong Chen Ting Zhang Su-Jun Liu Xin-Yue Bai Xian-Yu Huang Mei Jiang Xiao-Dong Sun Department of Ophthalmology Shanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China National Clinical Research Center for Eye Diseases Shanghai 200080China Shanghai Key Laboratory of Fundus Diseases Shanghai 200080China Shanghai Engineering Center for Visual Science and Photomedicine Shanghai 200080China

AIM:To explore an xeno-free and defined coating substrate suitable for the culture of H9 human embryonic stem cell-derived retinal pigment epithelial(hES-RPE)cells in vitro,and compare the behaviors and functions of h ESRPE cells on two culture substrates,laminin521(LN-521)and truncated recombinant human vitronectin(VTN-N).METHODS:hES-RPE cells were used in the *** abilities of LN-521 and VTN-N at different concentrations to adhere to hES-RPE cells were compared with a high-content imaging *** real-time polymerase chain reaction was used to evaluate RPE-specific gene expression levels midway(day 10)and at the end(day 20)of the time *** polarity was observed by immunofluorescent staining for apical and basal markers of the *** phagocytic ability of hES-RPE cells was identified by flow cytometry and ***:The cell adhesion assay showed that the ability of LN-521 to adhere to hES-RPE cells was *** increasing coating concentration,an increasing number of cells attached to the surface of LN-521-coated *** contrast,VTN-N presented a strong adhesive ability even at a low *** optimal concentration of LN-521 and VTN-N required to coat and adhesion to hES-RPE cells were 2 and 0.25μg/cm^(2),***,both LN-521 and VTN-N could facilitate adoption of the desired cobblestone cellular morphology with tight junction and showed polarity by the hES-RPE ***,hES-RPE cells cultivated in VTN-N had a greater phagocytic ability,and it took less time for these hES-RPE cells to ***:VTN-N is a more suitable coating substrate for cultivating hES-RPE cells.

关键词： retinal pigment epithelial stem cell surface coating laminin vitronectin cell adhesion phagocytosis

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vitronectin的止血效应

国外医学（生理病理科学与临床分册） 1993年第2期13卷 83-85页

作者：周金成

The cellular microenvironment modulates the role of PAI-1 and vitronectin in mediating cell-matrix interactions

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Advances in Biological Chemistry 2013年第1期3卷 114-132页

作者： Sumit Goswami Lawrence C. Thompson Lindsay Wickman Cynthia B. Peterson The Department of Biochemistry and Cellular and Molecular Biology The University of Tennessee Knoxville USA

Plasminogen activator inhibitor-1 (PAI-1), a member of the serine protease inhibitor (serpin) superfamily of proteins, circulates in a complex with vitronectin. Furthermore, these two proteins are co-localized in the extracellular matrix (ECM) in many different pathophysiological conditions. Though PAI-1 is a well-characterized inhibitor of serine proteases, recent emphasis has also focused on its protease-independent functions. vitronectin, a multi-domain protein that binds a wide variety of ligands and proteins, exists in the circulation in a preferred monomeric state, while in the extracellular matrix it exists as a multimer resulting from an altered conformation. Though the mechanism for the conformational alterations and compartmentalization in tissues is unknown, there are a number of biomolecules including PAI-1 that appear to cause such changes. Experimental analysis has established that PAI-1 induces association of vitronectin to higher-order species in a concentration-dependent fashion [1]. This report extends our investigations into the mechanism of the interaction between vitronectin and PAI-1 to explore the physiological relevance of these higher-order complexes for cellular adhesion and migration. In this study, we evaluate the effects of the pericellular microenvironment on the functions of the multimeric complexes in a variety of relevant biological settings. Our findings underscore the importance of the variability of components within this microenvironment, including different receptors and ECM components, in governing the way in which the vitronectin/PAI-1 complex mediates cell-matrix interactions.

关键词： PAI-1 vitronectin Extracellular Matrix Receptors

鸭疫里默杆菌通过外膜蛋白Omp45结合鸭补体调节因子vitronectin

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中国兽医学报 2023年第12期43卷 2472-2479页

作者：何忆箫秦豆豆彭兰青程中林高继业李继祥西南大学动物医学院重庆402460

旨在筛选和鉴定与鸭补体调节因子vitronectin(Vn)结合的鸭疫里默杆菌(Riemerella anatipestifer,***)外膜蛋白。在酵母表达鸭Vn及间接免疫荧光和免疫斑点杂交试验检测鸭疫里默杆菌结合Vn的基础上,以重组Vn为诱饵蛋白进行His pull-down及... 详细信息

旨在筛选和鉴定与鸭补体调节因子vitronectin(Vn)结合的鸭疫里默杆菌(Riemerella anatipestifer,***)外膜蛋白。在酵母表达鸭Vn及间接免疫荧光和免疫斑点杂交试验检测鸭疫里默杆菌结合Vn的基础上,以重组Vn为诱饵蛋白进行His pull-down及LC-MS/MS质谱鉴定,筛选外膜蛋白并进行生物信息学分析;原核表达候选外膜蛋白及截短片段,制备多克隆抗体,利用Far-Western blot验证与鸭Vn结合,并用间接ELISA测定肝素和氯化钠对结合的影响;测定Vn及抗外膜蛋白抗体对血清杀菌活性的影响。结果显示,鸭疫里默杆菌能在体外结合鸭Vn,经His-pull down、质谱鉴定及蛋白质结构分析,筛选出的外膜蛋白Omp45具有β桶状结构及7个暴露于细胞膜外的多肽环;成功原核表达Omp45及截短片段,制备的抗Omp45多克隆抗体间接ELISA效价超过1∶12800,Western blot检测多克隆抗体可以与重组蛋白发生特异性反应;Far-Western blot及间接ELISA结果显示,Omp45、包含2个及以上细胞膜外多肽环的截短片段同时以肝素和氯化钠敏感方式结合Vn;血清杀菌试验结果显示,鸭疫里默杆菌对缺乏Vn的健康鸭血清的抵抗力降低,在抑制补体经典激活途径条件下抗Omp45抗体也能极显著降低细菌在鸭血清中的存活率(P<0.01)。结果表明,成功筛选并鉴定了与鸭Vn结合的鸭疫里默杆菌外膜蛋白Omp45,为进一步阐明鸭疫里默杆菌致病机制奠定了基础。

关键词：鸭疫里默杆菌补体调节因子 vitronectin 外膜蛋白

PAI-1和vitronectin在慢性移植物肾损伤中的表达及致纤维化作用的初步探讨

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PAI-1和vitronectin在慢性移植物肾损伤中的表达及致纤维化作用的...

作者：赵霞娟复旦大学

学位级别：硕士

背景慢性移植物肾损伤(CAI)是造成移植物失功最主要的原因。肾脏的慢性纤维化是以小球及小管间质中细胞外基质(ECM)的过度积聚为典型表现。在一些慢性肾脏病中发现,1型纤溶酶原激活剂抑制物(PAI-1)和玻璃粘连蛋白(vitronectin)可能通过... 详细信息

背景慢性移植物肾损伤(CAI)是造成移植物失功最主要的原因。肾脏的慢性纤维化是以小球及小管间质中细胞外基质(ECM)的过度积聚为典型表现。在一些慢性肾脏病中发现,1型纤溶酶原激活剂抑制物(PAI-1)和玻璃粘连蛋白(vitronectin)可能通过相互结合的方式参与了基质的降解,促进小球的硬化和间质的纤维化。而在人慢性移植物肾损害中,国内外尚未有证实PAI-1通过与vitronectin相互结合的方式促进非特异性的纤维化的报道。本文旨在通过免疫组化的方法,进一步观察在人慢性移植物肾损害的标本中,是否存在PAI-1与vitronectin表达的共同上调,并通过相互结合的方式发挥其致纤维化的机制。本研究可为近一步寻找阻断移植肾纤维化过程的治疗途径提供理论依据。方法入选24例。肾移植术后患者,分为正常对照组(8例)、急性排异组(AR组)(8例)、CAI组(8例)。免疫组化法检测PAI-1,vitronectin以及PAI-1和vitronectin的双重染色。对正常对照组和CAI组的标本都进行PAS及masson染色,观察小球硬化、基质增生、小管萎缩、间质纤维化的情况,收集正常对照组及CAI组的基本临床资料,包括年龄、体重、肾穿时肌酐情况,根据CG公式计算其Ccr。随访两组肾穿后至少9个月的肌酐情况。结果采用半定量面积计数法。所有数据采用直线相关分析、t检验等进行统计学检验。P＜0.05认为具有统计学意义。结果PAI-1在正常对照组中仅极少量的表达(小球6.35±2.90,间质3.53±2.20),与正常对照组相比,PAI-1在AR组、CAI组都有表达上调(小球中分别为28.66±4.70,49.10±6.51;间质中分别是8.81±2.20,15.13±1.45),具有统计学意义(P＜0.05)。CAI组中的表达显著高于AR组,具有统计学的差异(P＜0.05)。vitronectin在正常对照组中已有大量表达(小球21.14±1.80,间质3.95±1.46),与AR组(小球28.44±9.46,间质5.53±1.83)相比,无统计学差异(P＞0.05)。CAI组(小球42.30±8.23,间质12.29±2.36)显著高于其他两组,有统计学差异(P＜0.05)。PAS染色结果,CAI组肾小球硬化指数(2.80±0.49)与正常对照组(0.91±0.23)相比,具有统计学差异(P＜0.01)。CAI组肾小球ECM面积/肾小球面积值(0.50±0.07)与正常对照组(0.14±0.03)相比,具有统计学差异(P＜0.01)。PAI-1半定量计数值与肾小球ECM面积/肾小球面积值呈直线正相关(R=0.944,P＜0.01),具有统计学意义。vitronectin半定量计数值与肾小球ECM面积/肾小球面积值呈直线正相关(R=0.815,P＜0.01),具有统计学意义。Masson染色结果,CAI组间质纤维化评分(2.50±0.27)与正常对照组(0.44±0.22)相比,具有统计学差异(P＜0.01)。CAI组小管损伤评分(2.49±0.29)与正常对照组(0.65±0.32)相比,具有统计学差异(P＜0.01)。PAI-1半定量计数值与vitronectin半定量计数值两者呈直线正相关,有统计学意义(R=0.826,P＜0.01)。双重染色结果显示,在CAI组标本中,观察到PAI-1与vitronectin两者在小球系膜区及小管间质处广泛共表达。肾脏局部PAI-1、vitronectin与患者的肌酐值呈线性正相关,具有统计学差异(R=0.858,P＜0.01及R=0.860,P＜0.01);两者与患者的Ccr呈线性负相关,具有统计学差异(R=-0.730,P＜0.01及R=-0.708,P＜0.05)。在肾穿后9个月的随访中,PAI-1高表达组Scr都显著高于PAI-1低表达组,有统计学差异(P＜0.01),且在随访第6及第9个月时,PAI-1高表达组分别发生1例因移植肾不可逆的纤维化导致移植物失功的不良事件。结论PAI-1及vitronectin在慢性移植物肾损伤纤维化的过程中表达上调,与基质的增生相关,PAI-1与vitronectin在基质积聚处广泛共表达。推论两者参与了移植肾纤维化的过程,PAI-1作为致纤维化的分子,与vitronectin相结合,通过依赖或非依赖纤溶蛋白酶的途径减少细胞外基质的降解,促进ECM的积聚,导致纤维化的发生及发展。

关键词：慢性移植物肾损伤 PAI-1 vitronectin ECM

VTN Silencing Inhibit Hepatocellular Carcinoma in vitro and in vivo

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VTN Silencing Inhibit Hepatocellular Carcinoma in vitro and ...