The effect of thymosin β4 on articular cartilage chondrocyte matrix metalloproteinase expression

2002 ◽  
Vol 30 (6) ◽  
pp. 879-882 ◽  
Author(s):  
E. J. Blain ◽  
D. J. Mason ◽  
V. C. Duance
Keyword(s):  
Articular Cartilage ◽  
Matrix Metalloproteinases ◽  
Matrix Metalloproteinase ◽  
Mechanical Loading ◽  
Functional Role ◽  
Fold Increase ◽  
Thymosin Β4 ◽  
Cytoskeletal Organization ◽  
Primary Chondrocytes ◽  
The Matrix

Mechanical loading is paramount in regulating both the anabolic and catabolic activities of articular cartilage chondrocytes, essential for the matrix to retain its functional integrity. We have identified thymosin β4 as a putative mechanically regulated gene that may mediate load-enhanced synthesis and activation of matrix metalloproteinases (MMPs) 2 and 9 in articular cartilage. The objective of this study was to confirm the mechanical regulation of thymosin β4 and determine its effect on cartilage chondrocyte MMP production. Thymosin β4 mRNA expression, analysed by quantitative PCR, revealed a significant 20-fold increase in cartilage loaded for 10 min which was still evident after 30 min of loading. Treatment of primary chondrocytes with 2 and 4 μg · ml-1 thymosin β4 peptide for 4 h significantly increased pro-MMP 9 expression and activation. We postulate a functional role for load-induced thymosin β4 in modulating the cytoskeletal organization of articular cartilage chondrocytes to affect MMP expression.

scholarly journals Chondrocyte-mediated depletion of articular cartilage proteoglycans in vitro

1985 ◽  
Vol 225 (2) ◽  
pp. 493-507 ◽  
Author(s):  
J A Tyler
Keyword(s):  
Articular Cartilage ◽  
Core Protein ◽  
Fold Increase ◽  
Acid Proteinase ◽  
Binding Region ◽  
Guanidinium Chloride ◽  
The Core ◽  
The Matrix ◽  
Average Size

The degradation of proteoglycan was examined in cultured slices of pig articular cartilage. Pig leucocyte catabolin (10 ng/ml) was used to stimulate the chondrocytes and induce a 4-fold increase in the rate of proteoglycan loss from the matrix for 4 days. Material in the medium of both control and depleted cultures was mostly a degradation product of the aggregating proteoglycan. It was recovered as a very large molecule slightly smaller than the monomers extracted with 4M-guanidinium chloride and lacked a functional hyaluronate binding region. The size and charge were consistent with a very limited cleavage or conformational change of the core protein near the hyaluronate binding region releasing the C-terminal portion of the molecule intact from the aggregate. The ‘clipped’ monomer diffuses very rapidly through the matrix into the medium. The amount of proteoglycan extracted with 4M-guanidinium chloride decreased during culture from both the controls and depleted cartilage, and the average size of the molecules initially remained the same. However, the proportion of molecules with a smaller average size increased with time and was predominant in explants that had lost more than 70% of their proteoglycan. All of this material was able to form aggregates when mixed with hyaluronate, and glycosaminoglycans were the same size and charge as normal, indicating either that the core protein had been cleaved in many places or that larger molecules were preferentially released. A large proportion of the easily extracted and non-extractable proteoglycan remained in the partially depleted cartilage and the molecules were the same size and charge as those found in the controls. There was no evidence of detectable glycosidase activity and only very limited sulphatase activity. A similar rate of breakdown and final distribution pattern was found for newly synthesized proteoglycan. Increased amounts of latent neutral metalloproteinases and acid proteinase activities were present in the medium of depleted cartilage. These were not thought to be involved in the breakdown of proteoglycan. Increased release of proteoglycan ceased within 24h of removal of the catabolin, indicating that the effect was reversible and persisted only while the stimulus was present.

scholarly journals Recent insights into natural product inhibitors of matrix metalloproteinases

MedChemComm ◽  
2019 ◽  
Vol 10 (12) ◽  
pp. 2024-2037 ◽  
Author(s):  
Geetha B. Kumar ◽  
Bipin G. Nair ◽  
J. Jefferson P. Perry ◽  
David B. C. Martin
Keyword(s):  
Cardiovascular Disease ◽  
Matrix Metalloproteinases ◽  
Matrix Metalloproteinase ◽  
Natural Product ◽  
Human Health ◽  
Neurodegenerative Disorders ◽  
Biological Functions ◽  
Mmp Inhibitors ◽  
The Matrix ◽  
Health And Disease

Members of the matrix metalloproteinase (MMP) family have biological functions that are central to human health and disease, and MMP inhibitors have been investigated for the treatment of cardiovascular disease, cancer and neurodegenerative disorders.

scholarly journals Curcumin reduces prostaglandin E2, matrix metalloproteinase-3 and proteoglycan release in the secretome of interleukin 1β-treated articular cartilage

F1000Research ◽  
2013 ◽  
Vol 2 ◽  
pp. 147 ◽  
Author(s):  
Abigail L Clutterbuck ◽  
David Allaway ◽  
Pat Harris ◽  
Ali Mobasheri
Keyword(s):  
Articular Cartilage ◽  
Matrix Metalloproteinase ◽  
Therapeutic Potential ◽  
Inflammatory Diseases ◽  
Primary Cultures ◽  
Cartilage Explants ◽  
Prostaglandin E ◽  
Primary Chondrocytes ◽  
Chondrocyte Death ◽  
Anti Inflammatory

Objective: Curcumin (diferuloylmethane) is a phytochemical with potent anti-inflammatory and anti-oxidant properties, and has therapeutic potential for the treatment of a range of inflammatory diseases, including osteoarthritis (OA). The aim of this study was to determine whether non-toxic concentrations of curcumin can reduce interleukin-1beta (IL-1β)-stimulated inflammation and catabolism in an explant model of cartilage inflammation.Methods: Articular cartilage explants and primary chondrocytes were obtained from equine metacarpophalangeal joints. Curcumin was added to monolayer cultured primary chondrocytes and cartilage explants in concentrations ranging from 3μM-100μM. Prostaglandin E2 (PGE2) and matrix metalloproteinase (MMP)-3 release into the secretome of IL-1β-stimulated explants was measured using a competitive ELISA and western blotting respectively. Proteoglycan (PG) release in the secretome was measured using the 1,9-dimethylmethylene blue (DMMB) assay. Cytotoxicity was assessed with a live/dead assay in monolayer cultures after 24 hours, 48 hours and five days, and in explants after five days.Results: Curcumin induced chondrocyte death in primary cultures (50μM p<0.001 and 100μM p<0.001) after 24 hours. After 48 hours and five days, curcumin (≥25μM) significantly increased cell death (p<0.001 both time points). In explants, curcumin toxicity was not observed at concentrations up to and including 25μM after five days. Curcumin (≥3μM) significantly reduced IL-1β-stimulated PG (p<0.05) and PGE2 release (p<0.001) from explants, whilst curcumin (≥12μM) significantly reduced MMP-3 release (p<0.01).Conclusion: Non-cytotoxic concentrations of curcumin exert anti-catabolic and anti-inflammatory effects in cartilage explants.

scholarly journals Curcumin reduces prostaglandin E2, matrix metalloproteinase-3 and proteoglycan release in the secretome of interleukin 1β-treated articular cartilage

F1000Research ◽  
2013 ◽  
Vol 2 ◽  
pp. 147 ◽  
Author(s):  
Abigail L Clutterbuck ◽  
David Allaway ◽  
Pat Harris ◽  
Ali Mobasheri
Keyword(s):  
Articular Cartilage ◽  
Matrix Metalloproteinase ◽  
Therapeutic Potential ◽  
Inflammatory Diseases ◽  
Primary Cultures ◽  
Cartilage Explants ◽  
Prostaglandin E ◽  
Primary Chondrocytes ◽  
Chondrocyte Death ◽  
Anti Inflammatory

Objective: Curcumin (diferuloylmethane) is a phytochemical with potent anti-inflammatory and anti-oxidant properties, and has therapeutic potential for the treatment of a range of inflammatory diseases, including osteoarthritis (OA). The aim of this study was to determine whether non-toxic concentrations of curcumin can reduce interleukin-1beta (IL-1β)-stimulated inflammation and catabolism in an explant model of cartilage inflammation.Methods: Articular cartilage explants and primary chondrocytes were obtained from equine metacarpophalangeal joints. Curcumin was added to monolayer cultured primary chondrocytes and cartilage explants in concentrations ranging from 3μM-100μM. Prostaglandin E2 (PGE2) and matrix metalloproteinase (MMP)-3 release into the secretome of IL-1β-stimulated explants was measured using a competitive ELISA and western blotting respectively. Proteoglycan (PG) release in the secretome was measured using the 1,9-dimethylmethylene blue (DMMB) assay. Cytotoxicity was assessed with a live/dead assay in monolayer cultures after 24 hours, 48 hours and five days, and in explants after five days.Results: Curcumin induced chondrocyte death in primary cultures (50μM p<0.001 and 100μM p<0.001) after 24 hours. After 48 hours and five days, curcumin (≥25μM) significantly increased cell death (p<0.001 both time points). In explants, curcumin toxicity was not observed at concentrations up to and including 25μM after five days. Curcumin (≥3μM) significantly reduced IL-1β-stimulated PG (p<0.05) and PGE2 release (p<0.001) from explants, whilst curcumin (≥12μM) significantly reduced MMP-3 release (p<0.01).Conclusion: Non-cytotoxic concentrations of curcumin exert anti-catabolic and anti-inflammatory effects in cartilage explants.

scholarly journals Quantification of a matrix metalloproteinase-generated aggrecan G1 fragment using monospecific anti-peptide serum

1995 ◽  
Vol 307 (1) ◽  
pp. 245-252 ◽  
Author(s):  
M W Lark ◽  
H Williams ◽  
L A Hoernner ◽  
J Weidner ◽  
J M Ayala ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Matrix Metalloproteinase ◽  
Limit Of Detection ◽  
Cartilage Destruction ◽  
Cathepsin G ◽  
Terminal Sequence ◽  
Leucocyte Elastase ◽  
The Matrix ◽  
Radio Immunoassay ◽  
Species Specific

Several members of the matrix metalloproteinase family have been reported to cleave aggrecan in the interglobular domain between Asn-341 and Phe-342. An antiserum was prepared against a peptide conjugate corresponding to the C-terminal sequence of the matrix metalloproteinase-generated aggrecan G1 fragment (Phe335-Val-Asp-Ile-Pro-Glu-Asn341). A quantitative radioimmunoassay, with a limit of detection of about 80 pM, was developed using this antiserum. This antiserum requires the free carboxyl group of the C-terminal asparagine for optimal recognition. If the C-terminal asparagine is excised from the sequence, replaced with closely related amino acids, or extended across the matrix metalloproteinase cleavage site, there is a 40-10,000-fold loss in detection. Using peptides cleaved from the N-terminus, it was determined that the antiserum requires the entire Phe-Val-Asp-Ile-Pro-Glu-Asn sequence for optimal recognition. The radioimmunoassay detects matrix metalloproteinase-generated G1 fragments with similar sensitivity to the Phe-Val-Asp-Ile-Pro-Glu-Asn peptide, but it does not recognize intact aggrecan. Immunoreactive aggrecan G1 fragments of molecular mass 50 kDa are generated by the matrix metalloproteinases stromelysin and gelatinase A. In contrast, under identical conditions, the closely related metalloproteinases, gelatinase B and collagenase, as well as cathepsin G, cathepsin B and human leucocyte elastase, did not generate a G1 fragment recognized by the antiserum. The anti-Phe-Val-Asp-Ile-Pro-Glu-Asn serum detects stromelysin-generated aggrecan G1 fragments from mouse, guinea pig, rabbit and human, indicating that the detection is not species-specific. This antiserum and radio-immunoassay should be useful for quantifying and characterizing matrix metalloproteinase-generated aggrecan G1 fragments in articular cartilage and synovial fluids from humans and various animal models of articular-cartilage destruction.

scholarly journals The history of matrix metalloproteinases: milestones, myths, and misperceptions

2012 ◽  
Vol 303 (8) ◽  
pp. H919-H930 ◽  
Author(s):  
Rugmani Padmanabhan Iyer ◽  
Nicolle L. Patterson ◽  
Gregg B. Fields ◽  
Merry L. Lindsey
Keyword(s):  
Matrix Metalloproteinases ◽  
Matrix Metalloproteinase ◽  
Cardiovascular System ◽  
Cdna Clone ◽  
Null Mouse ◽  
Mmp Inhibitor ◽  
The Matrix ◽  
History Of ◽  
Research Questions

Since the discovery of tadpole collagenase in 1962, the matrix metalloproteinase (MMP) family has emerged as a significant proteinase group with recognized effects on the cardiovascular system. Over the last 40 years, many milestones have been achieved, from the identification of the first MMP, to the generation of the first MMP cDNA clone and null mouse, to the clinical approval of the first MMP inhibitor. Over the years, a few myths and misunderstandings have interwoven into the truths. In this review, we will discuss the major milestones of MMP research, as well as review the misinterpretations and misperceptions that have evolved. Clarifying the confusions and dispelling the myths will both provide a better understanding of MMP properties and functions and focus the cardiovascular field on the outstanding research questions that need to be addressed.

scholarly journals Screening of the matrix metalloproteinase inhibitory activity on extracts from Cordyceps neovolkiana DL0004 and Cordyceps takaomontana DL0038A fungi

2020 ◽  
Vol 10 (1) ◽  
pp. 65-71
Author(s):  
Nguyen Nguyet Hong ◽  
Cao Ha Tim ◽  
Nguyen Chi Dung ◽  
Dinh Minh Hiep ◽  
Ngo Ke Suong
Keyword(s):  
Matrix Metalloproteinases ◽  
Matrix Metalloproteinase ◽  
Tumor Growth ◽  
Inhibitory Activity ◽  
Jurkat Cell ◽  
Fruit Body ◽  
Inhibition Assay ◽  
Invasion And Metastasis ◽  
The Matrix ◽  
Mcf 7

Matrix metalloproteinases (MMPs) are endopeptidases, they are involved in tumor growth, and the processes of invasion and metastasis. Cordyceps spp. were recorded to have the anticancer potential. In previous studies, extracts from Cordyceps neovolkiana strain DL0004 and Cordyceps takaomontana strain DL0038A were capable of cytotoxic activity against MCF-7 and Jurkat cell lines. In this study, 26 extracts were prepared from biomasses and fruit bodies of C. neovolkiana DL0004 and C. takaomontana DL0038A, then proceed to screen for MMP inhibition assay by concentration ranges of 2000 µg/mL, 200 µg/mL, 20 µg/mL. The results showed that the CPS extract of the fruit body of the C. neovolkiana DL0004 had the highest activity in MMP inhibition with 84.27 ± 4.59% at 2000 µg/mL. The results were achievement for further studies of metastatic inhibition activities of Cordyceps extracts.

scholarly journals Matrix metalloproteinase degradation of elastin, type IV collagen and proteoglycan. A quantitative comparison of the activities of 95 kDa and 72 kDa gelatinases, stromelysins-1 and -2 and punctuated metalloproteinase (PUMP)

1991 ◽  
Vol 277 (1) ◽  
pp. 277-279 ◽  
Author(s):  
G Murphy ◽  
M I Cockett ◽  
R V Ward ◽  
A J P Docherty
Keyword(s):  
Matrix Metalloproteinases ◽  
Connective Tissue ◽  
Matrix Metalloproteinase ◽  
Quantitative Data ◽  
Type Iv Collagen ◽  
Quantitative Comparison ◽  
Degrading Enzymes ◽  
Type Iv ◽  
The Matrix

The abilities of the matrix metalloproteinases 95 kDa and 72 kDa gelatinases (type IV collagenases), stromelysins-1 and -2 and punctuated metalloproteinase (PUMP) to degrade insoluble elastin, type IV collagen films and proteoglycan have been compared. The gelatinases and PUMP were markedly more active in the degradation of elastin than were the stromelysins. PUMP and the stromelysins were more potent proteoglycan-degrading enzymes. All of the enzymes studied degraded soluble native type IV collagen, but the gelatinases were more effective at higher temperatures. These quantitative data allow an analysis of the potential relative roles of these metalloproteinases in the breakdown of the key components of connective tissue matrices.

scholarly journals Activation profiles of human kallikrein-related peptidases by matrix metalloproteinases

2013 ◽  
Vol 394 (1) ◽  
pp. 137-147 ◽  
Author(s):  
Hyesook Yoon ◽  
Sachiko I. Blaber ◽  
Wu Li ◽  
Isobel A. Scarisbrick ◽  
Michael Blaber
Keyword(s):  
Matrix Metalloproteinases ◽  
Matrix Metalloproteinase ◽  
Neurodegenerative Disease ◽  
Recent Work ◽  
Present Report ◽  
Therapeutic Targets ◽  
Amino Terminal ◽  
The Matrix ◽  
Regulatory Cascades ◽  
Functional Regulation

Abstract The 15 human kallikrein-related peptidases (KLKs) are clinically important biomarkers and therapeutic targets of interest in inflammation, cancer, and neurodegenerative disease. KLKs are secreted as inactive pro-forms (pro-KLKs) that are activated extracellularly by specific proteolytic release of their amino-terminal pro-peptide, and this is a key step in their functional regulation. Physiologically relevant KLK regulatory cascades of activation have been described in skin desquamation and semen liquefaction, and work by a large number of investigators has elucidated pairwise and autolytic activation relationships among the KLKs with the potential for more extensive activation cascades. More recent work has asked whether functional intersection of KLKs with other types of regulatory proteases exists. Such studies show a capacity for members of the thrombostasis axis to act as broad activators of pro-KLKs. In the present report, we ask whether such functional intersection is possible between the KLKs and the members of the matrix metalloproteinase (MMP) family by evaluating the ability of the MMPs to activate pro-KLKs. The results identify MMP-20 as a broad activator of pro-KLKs, suggesting the potential for intersection of the KLK and MMP axes under pathological dysregulation of MMP-20 expression.

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