Regulation of matrix metalloproteinase-3 gene expression in inflammation: A molecular study
Introduction: Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways. Materials and Methods: Lipopolysaccharide-indu...
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Published in | Journal of conservative dentistry Vol. 21; no. 6; pp. 592 - 596 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
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India
Wolters Kluwer India Pvt. Ltd
01.11.2018
Medknow Publications and Media Pvt. Ltd Medknow Publications & Media Pvt. Ltd Medknow Publications & Media Pvt Ltd |
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Abstract | Introduction: Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways.
Materials and Methods: Lipopolysaccharide-induced murine macrophage-like RAW 264.7 cell lines were obtained and maintained in Dulbecco's modified Eagle's medium plus 10% fetal bovine serum under optimal temperatures. Primers used were MMP-3 forward primer, MMP-3 reverse primer, and glyceraldehyde-3-phosphate dehydrogenase forward primer and glyceraldehyde-3-phosphate reverse primer. Total RNA was isolated, the sample was prepared, and electrophoresis was performed. The first strand of cDNA was synthesized and amplification of specific isolated gene using polymerase chain reactor (PCR). The amplified products were then separated on a 1.0% agarose gel in 1XTBE at 75 V for 3 h. The gel was stained with ethidium bromide, and the amplified product was visualized and photographed on Gel Doc system.
Results: Real-time PCR showed only bands at expected size of 595 bp for internal control amplification of glyceraldehyde-3-dehydrogenase gene. Analysis was done with densitometry, and these values are compared with the negative control. Results showed a statistically significant rise in the relative levels of MMP-3-mRNA when compared with negative control at 1, 2, and 3 h.
Conclusion: This study proved the significantly increased levels of MMP gene at different period, thereby it can be concluded that MMP-3 levels are higher in inflammatory conditions. |
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AbstractList | INTRODUCTIONMatrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways. MATERIALS AND METHODSLipopolysaccharide-induced murine macrophage-like RAW 264.7 cell lines were obtained and maintained in Dulbecco's modified Eagle's medium plus 10% fetal bovine serum under optimal temperatures. Primers used were MMP-3 forward primer, MMP-3 reverse primer, and glyceraldehyde-3-phosphate dehydrogenase forward primer and glyceraldehyde-3-phosphate reverse primer. Total RNA was isolated, the sample was prepared, and electrophoresis was performed. The first strand of cDNA was synthesized and amplification of specific isolated gene using polymerase chain reactor (PCR). The amplified products were then separated on a 1.0% agarose gel in 1XTBE at 75 V for 3 h. The gel was stained with ethidium bromide, and the amplified product was visualized and photographed on Gel Doc system. RESULTSReal-time PCR showed only bands at expected size of 595 bp for internal control amplification of glyceraldehyde-3-dehydrogenase gene. Analysis was done with densitometry, and these values are compared with the negative control. Results showed a statistically significant rise in the relative levels of MMP-3-mRNA when compared with negative control at 1, 2, and 3 h. CONCLUSIONThis study proved the significantly increased levels of MMP gene at different period, thereby it can be concluded that MMP-3 levels are higher in inflammatory conditions. Introduction: Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways. Materials and Methods: Lipopolysaccharide-induced murine macrophage-like RAW 264.7 cell lines were obtained and maintained in Dulbecco's modified Eagle's medium plus 10% fetal bovine serum under optimal temperatures. Primers used were MMP-3 forward primer, MMP-3 reverse primer, and glyceraldehyde-3-phosphate dehydrogenase forward primer and glyceraldehyde-3-phosphate reverse primer. Total RNA was isolated, the sample was prepared, and electrophoresis was performed. The first strand of cDNA was synthesized and amplification of specific isolated gene using polymerase chain reactor (PCR). The amplified products were then separated on a 1.0% agarose gel in 1XTBE at 75 V for 3 h. The gel was stained with ethidium bromide, and the amplified product was visualized and photographed on Gel Doc system. Results: Real-time PCR showed only bands at expected size of 595 bp for internal control amplification of glyceraldehyde-3-dehydrogenase gene. Analysis was done with densitometry, and these values are compared with the negative control. Results showed a statistically significant rise in the relative levels of MMP-3-mRNA when compared with negative control at 1, 2, and 3 h. Conclusion: This study proved the significantly increased levels of MMP gene at different period, thereby it can be concluded that MMP-3 levels are higher in inflammatory conditions. Introduction: Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways. Materials and Methods: Lipopolysaccharide-induced murine macrophage-like RAW 264.7 cell lines were obtained and maintained in Dulbecco's modified Eagle's medium plus 10% fetal bovine serum under optimal temperatures. Primers used were MMP-3 forward primer, MMP-3 reverse primer, and glyceraldehyde-3-phosphate dehydrogenase forward primer and glyceraldehyde-3-phosphate reverse primer. Total RNA was isolated, the sample was prepared, and electrophoresis was performed. The first strand of cDNA was synthesized and amplification of specific isolated gene using polymerase chain reactor (PCR). The amplified products were then separated on a 1.0% agarose gel in 1XTBE at 75 V for 3 h. The gel was stained with ethidium bromide, and the amplified product was visualized and photographed on Gel Doc system. Results: Real-time PCR showed only bands at expected size of 595 bp for internal control amplification of glyceraldehyde-3-dehydrogenase gene. Analysis was done with densitometry, and these values are compared with the negative control. Results showed a statistically significant rise in the relative levels of MMP-3-mRNA when compared with negative control at 1, 2, and 3 h. Conclusion: This study proved the significantly increased levels of MMP gene at different period, thereby it can be concluded that MMP-3 levels are higher in inflammatory conditions. Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways. Lipopolysaccharide-induced murine macrophage-like RAW 264.7 cell lines were obtained and maintained in Dulbecco's modified Eagle's medium plus 10% fetal bovine serum under optimal temperatures. Primers used were MMP-3 forward primer, MMP-3 reverse primer, and glyceraldehyde-3-phosphate dehydrogenase forward primer and glyceraldehyde-3-phosphate reverse primer. Total RNA was isolated, the sample was prepared, and electrophoresis was performed. The first strand of cDNA was synthesized and amplification of specific isolated gene using polymerase chain reactor (PCR). The amplified products were then separated on a 1.0% agarose gel in 1XTBE at 75 V for 3 h. The gel was stained with ethidium bromide, and the amplified product was visualized and photographed on Gel Doc system. Real-time PCR showed only bands at expected size of 595 bp for internal control amplification of glyceraldehyde-3-dehydrogenase gene. Analysis was done with densitometry, and these values are compared with the negative control. Results showed a statistically significant rise in the relative levels of MMP-3-mRNA when compared with negative control at 1, 2, and 3 h. This study proved the significantly increased levels of MMP gene at different period, thereby it can be concluded that MMP-3 levels are higher in inflammatory conditions. |
Audience | Academic |
Author | Ramesh, Sindhu Priya, Vishnu Teja, Kavalipurapu |
AuthorAffiliation | Department of Conservative Dentistry and Endodontics, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India 1 Department of Biochemistry, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India |
AuthorAffiliation_xml | – name: 1 Department of Biochemistry, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India – name: Department of Conservative Dentistry and Endodontics, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India |
Author_xml | – sequence: 1 givenname: Kavalipurapu surname: Teja fullname: Teja, Kavalipurapu organization: Department of Conservative Dentistry and Endodontics, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu – sequence: 2 givenname: Sindhu surname: Ramesh fullname: Ramesh, Sindhu organization: Department of Conservative Dentistry and Endodontics, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu – sequence: 3 givenname: Vishnu surname: Priya fullname: Priya, Vishnu organization: Department of Biochemistry, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu |
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CitedBy_id | crossref_primary_10_1016_j_injury_2021_11_065 crossref_primary_10_4103_JCDE_JCDE_289_23 crossref_primary_10_1016_j_jddst_2023_104528 crossref_primary_10_1016_j_prp_2024_155130 crossref_primary_10_1021_acs_bioconjchem_3c00266 crossref_primary_10_1016_j_ijbiomac_2021_07_165 crossref_primary_10_1016_j_jddst_2022_103600 |
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Keywords | real-time-polymerase chain reactor pulpitis periapical abscess RAW 264-7 cell lines Matrix metalloproteinase-3 gene |
Language | English |
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Snippet | Introduction: Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of... Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by... INTRODUCTIONMatrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of... |
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SubjectTerms | Biochemistry Cell lines Cellular signal transduction Densitometry Dentin Electrophoresis Enzymes Ethidium bromide Extracellular matrix Gene expression Genes Glyceraldehyde-3-phosphate dehydrogenase Inflammation Inflammatory diseases Lipopolysaccharides Macrophages Matrix metalloproteinase Metalloproteinase Metalloproteins Mitogens Monosaccharides Original Pathology Phosphates Physiology Polymerase chain reaction Primers Protein kinase Protein kinases RNA Statistical analysis Stromelysin 1 |
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Title | Regulation of matrix metalloproteinase-3 gene expression in inflammation: A molecular study |
URI | http://www.jcd.org.in/article.asp?issn=0972-0707;year=2018;volume=21;issue=6;spage=592;epage=596;aulast=Teja;type=0 https://www.ncbi.nlm.nih.gov/pubmed/30546201 https://www.proquest.com/docview/2132199692 https://search.proquest.com/docview/2157661279 https://pubmed.ncbi.nlm.nih.gov/PMC6249951 |
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