Ming-Chung Lin, Ming-Wei Lin, Erna Sulistyowati, Ching-Chieh Kao, Chung-Jung Liu, Shu-Ping Huang, S. C. Hsu, Bin Huang
{"title":"比较不同涂层基质对剪切流下内皮细胞中细胞结合和一氧化氮介导的蛋白 S-亚硝基化的影响","authors":"Ming-Chung Lin, Ming-Wei Lin, Erna Sulistyowati, Ching-Chieh Kao, Chung-Jung Liu, Shu-Ping Huang, S. C. Hsu, Bin Huang","doi":"10.2174/0115701646300960240606093535","DOIUrl":null,"url":null,"abstract":"\n\nShear flow is a mechanical signal regulating the function of Endothelial\nCells (ECs). The present study aimed to investigate the effects of different matrices on cell binding,\nNitric Oxide (NO) production, protein S-nitrosylation, expression of adhesion proteins, ROS\ngeneration, and cell viability in ECs under shear flow.\n\n\n\nThe ECs growing on glass slides separately coated with poly-L-lysine (p-Lys), collagen\n(Colla), fibronectin (Fibro), and a combined matrix (Colla+Fibro) were exposed to shear flow (25\ndyne/cm2) for 0, 1, 4, 8 h. The number of ECs remaining attached on the glass slide was calculated.\nThe expressions of endothelial Nitric Oxide Synthase (eNOS), peNOSS1177, VE-cadherin, FAK,\nand S-nitrosylated proteins were investigated by western blotting. The production of Nitric Oxide\n(NO) was measured by a specific reagent. Finally, the levels of ROS and cell viability were monitored.\n\n\n\nUnder a constant shear flow for 1 h, the physiological responses of ECs were similar between\nthese four matrices. When shear flow was extended to 4 and 8 h, higher cell binding, elevated\nNO production, increased S-nitrosylated proteins, enhanced expressions of FAK and VE-cadherin,\nmildly accumulated ROS, and cell death were observed in the matrix of Fibro and Colla+Fibro.\n\n\n\nWe have concluded fibronectin to be the optimal matrix facilitating NO-mediated Snitrosylation\nthat might be essential for superior binding efficiency, thereby preventing the stripping\nof ECs under shear flow. The results can be broadly applied to diverse biomechanical\nstudies.\n","PeriodicalId":50601,"journal":{"name":"Current Proteomics","volume":null,"pages":null},"PeriodicalIF":0.5000,"publicationDate":"2024-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Comparison of the Effects of Different Coating Matrices on Cell Binding and Nitric Oxide-Mediated Protein S-Nitrosylation in Endothelial Cells\\nunder Shear Flow\",\"authors\":\"Ming-Chung Lin, Ming-Wei Lin, Erna Sulistyowati, Ching-Chieh Kao, Chung-Jung Liu, Shu-Ping Huang, S. C. Hsu, Bin Huang\",\"doi\":\"10.2174/0115701646300960240606093535\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"\\n\\nShear flow is a mechanical signal regulating the function of Endothelial\\nCells (ECs). The present study aimed to investigate the effects of different matrices on cell binding,\\nNitric Oxide (NO) production, protein S-nitrosylation, expression of adhesion proteins, ROS\\ngeneration, and cell viability in ECs under shear flow.\\n\\n\\n\\nThe ECs growing on glass slides separately coated with poly-L-lysine (p-Lys), collagen\\n(Colla), fibronectin (Fibro), and a combined matrix (Colla+Fibro) were exposed to shear flow (25\\ndyne/cm2) for 0, 1, 4, 8 h. The number of ECs remaining attached on the glass slide was calculated.\\nThe expressions of endothelial Nitric Oxide Synthase (eNOS), peNOSS1177, VE-cadherin, FAK,\\nand S-nitrosylated proteins were investigated by western blotting. The production of Nitric Oxide\\n(NO) was measured by a specific reagent. Finally, the levels of ROS and cell viability were monitored.\\n\\n\\n\\nUnder a constant shear flow for 1 h, the physiological responses of ECs were similar between\\nthese four matrices. 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Comparison of the Effects of Different Coating Matrices on Cell Binding and Nitric Oxide-Mediated Protein S-Nitrosylation in Endothelial Cells
under Shear Flow
Shear flow is a mechanical signal regulating the function of Endothelial
Cells (ECs). The present study aimed to investigate the effects of different matrices on cell binding,
Nitric Oxide (NO) production, protein S-nitrosylation, expression of adhesion proteins, ROS
generation, and cell viability in ECs under shear flow.
The ECs growing on glass slides separately coated with poly-L-lysine (p-Lys), collagen
(Colla), fibronectin (Fibro), and a combined matrix (Colla+Fibro) were exposed to shear flow (25
dyne/cm2) for 0, 1, 4, 8 h. The number of ECs remaining attached on the glass slide was calculated.
The expressions of endothelial Nitric Oxide Synthase (eNOS), peNOSS1177, VE-cadherin, FAK,
and S-nitrosylated proteins were investigated by western blotting. The production of Nitric Oxide
(NO) was measured by a specific reagent. Finally, the levels of ROS and cell viability were monitored.
Under a constant shear flow for 1 h, the physiological responses of ECs were similar between
these four matrices. When shear flow was extended to 4 and 8 h, higher cell binding, elevated
NO production, increased S-nitrosylated proteins, enhanced expressions of FAK and VE-cadherin,
mildly accumulated ROS, and cell death were observed in the matrix of Fibro and Colla+Fibro.
We have concluded fibronectin to be the optimal matrix facilitating NO-mediated Snitrosylation
that might be essential for superior binding efficiency, thereby preventing the stripping
of ECs under shear flow. The results can be broadly applied to diverse biomechanical
studies.
Current ProteomicsBIOCHEMICAL RESEARCH METHODS-BIOCHEMISTRY & MOLECULAR BIOLOGY
CiteScore
1.60
自引率
0.00%
发文量
25
审稿时长
>0 weeks
期刊介绍:
Research in the emerging field of proteomics is growing at an extremely rapid rate. The principal aim of Current Proteomics is to publish well-timed in-depth/mini review articles in this fast-expanding area on topics relevant and significant to the development of proteomics. Current Proteomics is an essential journal for everyone involved in proteomics and related fields in both academia and industry.
Current Proteomics publishes in-depth/mini review articles in all aspects of the fast-expanding field of proteomics. All areas of proteomics are covered together with the methodology, software, databases, technological advances and applications of proteomics, including functional proteomics. Diverse technologies covered include but are not limited to:
Protein separation and characterization techniques
2-D gel electrophoresis and image analysis
Techniques for protein expression profiling including mass spectrometry-based methods and algorithms for correlative database searching
Determination of co-translational and post- translational modification of proteins
Protein/peptide microarrays
Biomolecular interaction analysis
Analysis of protein complexes
Yeast two-hybrid projects
Protein-protein interaction (protein interactome) pathways and cell signaling networks
Systems biology
Proteome informatics (bioinformatics)
Knowledge integration and management tools
High-throughput protein structural studies (using mass spectrometry, nuclear magnetic resonance and X-ray crystallography)
High-throughput computational methods for protein 3-D structure as well as function determination
Robotics, nanotechnology, and microfluidics.