scholarly journals Apatite Reduces Amelogenin Proteolysis by MMP-20 and KLK4 in vitro

2010 ◽  
Vol 89 (4) ◽  
pp. 344-348 ◽  
Author(s):  
Z. Sun ◽  
W. Carpiaux ◽  
D. Fan ◽  
Y. Fan ◽  
R. Lakshminarayanan ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Proteolytic Activity ◽  
Proteolytic Processing ◽  
Enamel Formation ◽  
Dependent Relationship ◽  
Kallikrein 4 ◽  
Synthetic Hydroxyapatite ◽  
Dose Dependent

Two enamel proteases, matrix metalloproteinase-20 (MMP-20) and kallikrein 4 (KLK4), are known to cleave amelogenin and are necessary for proper enamel formation. However, the effect of hydroxyapatite (HAP) on the proteolytic activity of these enzymes remains unclear. To investigate whether apatite affects normal amelogenin proteolysis, we used 2 different isoforms of amelogenin combined with the appropriate enzymes to analyze proteolytic processing rates in the presence or absence of synthetic hydroxyapatite (HAP) crystals (N = 3). We found a distinct dose-dependent relationship between the amount of HAP present in the proteolysis mixture and the rate of rP172 degradation by rpMMP-20, whereas the effect of HAP on proteolysis of either rP172 or rP148 by rhKLK4 was less prominent.

scholarly journals DPPI May Activate KLK4 during Enamel Formation

2009 ◽  
Vol 88 (4) ◽  
pp. 323-327 ◽  
Author(s):  
C.E. Tye ◽  
C.T. Pham ◽  
J.P. Simmer ◽  
J.D. Bartlett
Keyword(s):  
Serine Proteases ◽  
Immunohistochemical Staining ◽  
Proteolytic Processing ◽  
Enamel Organ ◽  
Enamel Matrix ◽  
Enamel Formation ◽  
Microhardness Testing ◽  
Kallikrein 4 ◽  
Fluorogenic Peptide

Kallikrein-4 (KLK4) is a serine protease expressed during enamel maturation, and proteolytic processing of the enamel matrix by KLK4 is critical for proper enamel formation. KLK4 is secreted as an inactive zymogen (pro-KLK4), and identification of its activator remains elusive. Dipeptidyl peptidase I (DPPI) is a cysteine aminopeptidase that can activate several serine proteases. In this study, we sought to examine DPPI expression in mouse enamel organ and determine if DPPI could activate KLK4. Real-time PCR showed DPPI expression throughout amelogenesis, with highest expression at maturation, and immunohistochemical staining of mouse incisors confirmed DPPI expression by ameloblasts. We demonstrate in vitro that DPPI activates pro-KLK4 to cleave a fluorogenic peptide containing a KLK4 cleavage site. Examination of mature enamel from DPPI null mice by FTIR showed no significant accumulation of protein; however, microhardness testing revealed that loss of DPPI expression significantly reduced enamel hardness.

scholarly journals Suppressive Activity of Vitamin D3 on Matrix Metalloproteinase Production From Cholesteatoma Keratinocytes In Vitro

2005 ◽  
Vol 2005 (4) ◽  
pp. 210-215 ◽  
Author(s):  
Hitome Kobayashi ◽  
Kazuhito Asano ◽  
Ken-ichi Kanai ◽  
Harumi Suzaki
Keyword(s):  
Extracellular Matrix ◽  
Matrix Metalloproteinase ◽  
Vitamin D3 ◽  
Biological Activities ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Suppressive Activity ◽  
Tissue Specimens ◽  
Culture Supernatants ◽  
Dose Dependent

There is much evidence that degradation of the extracellular matrix is essential for the development of cholesteatomas and that this is induced by activation of matrix metalloproteinases (MMPs). Vitamin D3 (VD3) has several well-recognised biological activities, including suppression of MMP production. The present study, therefore, was undertaken to examine whether VD3 could suppress MMP production from cholesteatoma keratinocytes in vitro. Keratinocytes (2.5×105cells/mL) induced from cholesteatoma tissue specimens were cultured with various concentrations of VD3. After one hour, lipopolysaccharide was added to the cell cultures at 100μg/mL. The culture supernatants were then collected and assayed for MMP-1 and MMP-3 by ELISA. We also used ELISA to measure the levels of both TIMP (tissue inhibitor of metalloproteinase)-1 and TIMP-2 in culture supernatants. Addition of VD3 into keratinocyte cultures caused the suppression of MMP and TIMP production, which was increased by LPS stimulation. This was dose-dependent. The present results showing the suppressive activity of VD3 on the production of MMPs, which are responsible for tissue remodeling, strongly suggest that VD3 would be a good candidate for an agent in the medical treatment of, or prophylaxis for, cholesteatomas.

CATECHOL OESTROGENS AND GONADOTROPHIN SECRETION IN THE EWE: AFFINITY FOR PITUITARY OESTROGEN RECEPTORS IN VITRO AND ACTION ON GONADOTROPHIN SECRETION IN VIVO

1980 ◽  
Vol 85 (3) ◽  
pp. 503-509 ◽  
Author(s):  
I. J. CLARKE ◽  
J. K. FINDLAY
Keyword(s):  
Negative Feedback ◽  
Oestrogen Receptor ◽  
Dose Response Relationship ◽  
Concurrent Administration ◽  
Feedback Effects ◽  
Dependent Relationship ◽  
Gonadotrophin Secretion ◽  
Dose Dependent

The binding of three catechol oestrogens, 2-OH-oestradiol-17β, 4-OH-oestrone and 2-OH-oestrone, to the ovine pituitary oestrogen receptor was measured in vitro to establish doses for the assessment of the effects of catechol oestrogens in vivo. Relative to oestradiol (100%) the compounds had receptor affinities of 30, 20 and 5% respectively. A dose of oestradiol sufficient to cause negative-feedback effects on the secretion of LH and FSH in ovariectomized ewes was established by intracarotid (i.c.) injections of 0·625–5·0 μg/dose (n = 3), and by measuring plasma levels of gonadotrophins in jugular venous samples taken at intervals of 20 min from 3 h before until 4 h after injection. A dose-dependent relationship (r = 0·88, P<0·001) was found for oestradiol and plasma LH levels. Plasma FSH was slightly (12–25%) but significantly (P<0·05) reduced by doses of 1·25–5·0 μg oestradiol, but no dose–response relationship was observed. Ovariectomized ewes (n = 4/group) were given 2·5 μg oestradiol (i.c.) simultaneously with 83 μg 2-OH-oestradiol, 125 μg 4-OH-oestrone or 500 μg 2-OH-oestrone. These doses of catechol oestrogens were chosen as being ten times that of oestradiol, with the relative affinities for oestrogen receptor taken into account. Concurrent administration of such doses of catechol oestrogens had no effect on the negative-feedback action of oestradiol in vivo. We have concluded that catechol oestrogens in the circulation probably do not modulate the action of oestradiol on release of LH or FSH; this does not preclude a possible role for them as locally produced regulators of oestrogen action.

scholarly journals Matrix metalloproteinase processing of monocyte chemoattractant proteins generates CC chemokine receptor antagonists with anti-inflammatory properties in vivo

Blood ◽  
2002 ◽  
Vol 100 (4) ◽  
pp. 1160-1167 ◽  
Author(s):  
G. Angus McQuibban ◽  
Jiang-Hong Gong ◽  
Julie P. Wong ◽  
John L. Wallace ◽  
Ian Clark-Lewis ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Chemokine Receptor ◽  
Chemokine Receptors ◽  
Leukemic Cell ◽  
Proteolytic Processing ◽  
Cc Chemokine ◽  
Monocyte Chemoattractant ◽  
The Matrix

Monocyte chemoattractant protein (MCP)–3 is inactivated upon cleavage by the matrix metalloproteinase (MMP) gelatinase A (MMP-2). We investigated the susceptibility to proteolytic processing of the 4 human MCPs by 8 recombinant MMPs to determine whether MCP-3 is an isolated example or represents a general susceptibility of chemokines to proteolytic inactivation by these important inflammatory proteases. In addition to MMP-2, MCP-3 is efficiently cleaved by membrane type 1 (MT1)–MMP, the cellular activator of MMP-2, and by collagenase-1 and collagenase-3 (MMP-1, MMP-13) and stromelysin-1 (MMP-3). Specificity was shown by absence of cleavage by matrilysin (MMP-7) and the leukocytic MMPs neutrophil collagenase (MMP-8) and gelatinase B (MMP-9). The closely related chemokines MCP-1, MCP-2, and MCP-4 were not cleaved by MMP-2 or MT1-MMP, but were cleaved by MMP-1 and MMP-3 with varying efficiency. MCPs were typically cleaved between residues 4 and 5, but MCP-4 was further processed at Val7-Pro8. Synthetic MCP analogs corresponding to the MMP-cleaved forms bound CC chemokine receptor (CCR)–2 and CCR-3, but lacked chemoattractant activity in pre-B cells transfected with CCR-2 and CCR-3 or in THP-1 monocytic cells, a transformed leukemic cell line. Moreover, the truncated products of MCP-2 and MCP-4, like MCP-3, were potent antagonists of their cognate CC chemokine receptors in transwell cell migration assays in vitro. When they were injected 24 hours after the initiation of carrageenan-induced inflammation in rat paws, their in vivo antagonist activities were revealed by a greater than 66% reduction in inflammatory edema progression after 12 hours. We propose that MMPs have an important role in modulating inflammatory and immune responses by processing chemokines in wound healing and in disease.

scholarly journals SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Zhen-Zhen Guo ◽  
Qun-An Cao ◽  
Zong-Zhuang Li ◽  
Li-Ping Liu ◽  
Zhi Zhang ◽  
...  
Keyword(s):  
Aortic Aneurysm ◽  
Matrix Metalloproteinase ◽  
Molecular Mechanisms ◽  
Macrophage Migration ◽  
Dependent Manner ◽  
Monocyte Chemoattractant ◽  
Abdominal Aortic ◽  
Major Chemical ◽  
Dose Dependent

Nicotine, a major chemical component of cigarettes, plays a pivotal role in the development of abdominal aortic aneurysm (AAA). c-Jun N-terminal kinase (JNK) has been demonstrated to participate in elastase-induced AAA. This study aimed to elucidate whether the JNK inhibitor SP600125 can attenuate nicotine plus angiotensin II- (AngII-) induced AAA formation and to assess the underlying molecular mechanisms. SP600125 significantly attenuated nicotine plus AngII-induced AAA formation. The expression of matrix metalloproteinase- (MMP-) 2, MMP-9, monocyte chemoattractant protein- (MCP-) 1, and regulated-on-activation, normal T-cells expressed and secreted (RANTES) was significantly upregulated in aortic aneurysm lesions but inhibited by SP600125.In vitro, nicotine induced the expression of MCP-1 and RANTES in both RAW264.7 (mouse macrophage) and MOVAS (mouse vascular smooth muscle) cells in a dose-dependent manner; expression was upregulated by 0.5 ng/mL nicotine but strongly downregulated by 500 ng/mL nicotine. SP600125 attenuated the upregulation of MCP-1 and RANTES expression and subsequent macrophage migration. In conclusion, SP600125 attenuates nicotine plus AngII-induced AAA formation likely by inhibiting MMP-2, MMP-9, MCP-1, and RANTES. The expression of chemokines in MOVAS cells induced by nicotine has an effect on RAW264.7 migration, which is likely to contribute to the development of nicotine-related AAA.

scholarly journals Cleavage of HMGB1 by Proteolytic Enzymes Associated with Inflammatory Conditions

2020 ◽  
Vol 11 ◽  
Author(s):  
Agnieszka Sowinska ◽  
Merlin Rensing ◽  
Lena Klevenvall ◽  
Manoj Neog ◽  
Peter Lundbäck ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Matrix Metalloproteinase ◽  
Neutrophil Elastase ◽  
Human Neutrophil ◽  
Proteolytic Enzymes ◽  
Proteolytic Processing ◽  
Cathepsin G ◽  
Human Neutrophil Elastase ◽  
Matrix Metalloproteinase 3

Extracellular HMGB1 acts as an alarmin in multiple autoimmune diseases. While its release and functions have been extensively studied, there is a substantial lack of knowledge regarding HMGB1 regulation at the site of inflammation. Herein we show that enzymes present in arthritis-affected joints process HMGB1 into smaller peptides in vitro. Gel electrophoresis, Western blotting and mass spectrometry analyses indicate cleavage sites for human neutrophil elastase, cathepsin G, and matrix metalloproteinase 3 within the HMGB1 structure. While human neutrophil elastase and matrix metalloproteinase 3 might alter the affinity of HMGB1 to its receptors by cleaving the acidic C-terminal tail, cathepsin G rapidly and completely degraded the alarmin. Contrary to a previous report we demonstrate that HMGB1 is not a substrate for dipeptidyl peptidase IV. We also provide novel information regarding the presence of these proteases in synovial fluid of juvenile idiopathic arthritis patients. Correlation analysis of protease levels and HMGB1 levels in synovial fluid samples did not, however, reveal any direct relationship between the recorded levels. This study provides knowledge of proteolytic processing of HMGB1 relevant for the regulation of HMGB1 during inflammatory disease.

scholarly journals The correlation between IL-33 and inflammation factors in rheumatoid arthritis patients in vivo and in fibroblast-like synoviocytes in vitro

2020 ◽  
Author(s):  
Jing Wu ◽  
Qiang Li ◽  
Jia-Xin Deng ◽  
Jin-Jun Zhao ◽  
Qing-Hong Yu
Keyword(s):  
Rheumatoid Arthritis ◽  
Synovial Fluid ◽  
Interleukin 33 ◽  
Cytokines And Chemokines ◽  
Dependent Relationship ◽  
The Relationship ◽  
Dose Dependent ◽  
Fibroblast Like Synoviocytes

Abstract Background: Interleukin-33 (IL-33) is a member of the IL-1 family of cytokines whose role remains controversial in rheumatoid arthritis (RA), because no clear conclusion has been established regarding the relationship between IL-33 and other cytokines and chemokines. The present study was conducted to evaluate the correlation of IL-33 with other cytokines and chemokines in serum and the synovia, and to explore the nature of the relationship.Results: IL-33 was found to exhibit an inverted-U-shaped correlation with multiple cytokines and chemokines in synovial fluid, including IL-6, IL-1β, CXCL8 (IL-8), CXCL9 (MIG) and CXCL10 (IP-10), but not in serum. Moreover, in vitro experiments confirmed that IL-33 also exhibits U-type dose-dependent regulation of FLS function.Conclusions: IL-33 exhibit an inverted-U-shaped correlation with multiple cytokines and chemokines in synovial fluid of RA patients. IL-33 affects the secretion of cytokines and chemokines in the synovium in a U-type dose-dependent relationship.

scholarly journals Captopril as an Adjuvant to Temozolomide Prolongs Survival in a Rat Intracranial Gliosarcoma Model via Downregulation of Matrix Metalloproteinase-2

Neurosurgery ◽  
2019 ◽  
Vol 66 (Supplement_1) ◽  
Author(s):  
Leon Pinheiro ◽  
Alexander Perdomo-Pantoja ◽  
Joshua Casaos ◽  
Iddo Paldor ◽  
Veronica Vigilar ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Matrix Metalloproteinase 2 ◽  
Treated Animal ◽  
Western Blots ◽  
Significant Survival ◽  
Dose Dependent Decrease ◽  
Dose Dependent ◽  
F344 Rats

Abstract INTRODUCTION Evidence suggests that captopril might inhibit matrix metalloproteinase-2 (MMP-2), an endopeptidase that selectively breaks down elements of the extracellular matrix and that is associated with glioma cell migration. We analyzed the effects of captopril on MMP-2 expression in gliosarcoma cells in Vitro and on survival in an in Vivo murine intracranial gliosarcoma model. METHODS For survival, F344 rats were implanted intracranially with 9 L gliosarcoma tumor fragments, and were then divided into groups: (1) control, receiving no treatment; (2) oral captopril, 30 mg/kg/day on days 5 to 14; (3) oral temozolomide (TMZ), 50 mg/kg on days 5 to 9; and (4) oral TMZ and oral captopril. For immunohistochemistry, on day 0, additional F344 rats were implanted intracranially with 9 L gliosarcoma cells and divided into groups: (1) control; and (2) oral captopril, 30 mg/kg/day. Rats from each group were sacrificed on days 6, 8, and 11. To assess the MMP-2 protein level, cultured 9 L gliosarcoma cells were treated with either 100, 150, or 200 μg/mL of captopril or ultrapure water as the vehicle, and protein expression was assessed via Western blots at 24, 48, 72, and 96 h. RESULTS Captopril in Vivo extended survival in a murine intracranial gliosarcoma model significantly. Within a period of 30 d, adjuvant captopril presented a statistically significant survival advantage compared to the control, captopril-only, and TMZ-only groups (27.5 d versus 14, 16, and 23 d, P < .001, < .001, and .02, respectively). Immunohistochemistry analysis of treated animal brains demonstrated a dose-dependent decrease in MMP-2 expression with captopril, while Western blots of treated 9 L cells in Vitro demonstrated a decreased MMP-2 expression. CONCLUSION Captopril as adjuvant therapy to TMZ increased survival in a rat intracranial gliosarcoma model. A dose-dependent reduction in MMP-2 expression with captopril was demonstrated in Vitro. Captopril may serve as a potential adjuvant to TMZ therapy and deserves further investigation.

scholarly journals Activities of the Matrix Metalloproteinase Stromelysin-2 (MMP-10) in Matrix Degradation and Keratinocyte Organization in Wounded Skin

2004 ◽  
Vol 15 (12) ◽  
pp. 5242-5254 ◽  
Author(s):  
Monika Krampert ◽  
Wilhelm Bloch ◽  
Takako Sasaki ◽  
Philippe Bugnon ◽  
Thomas Rülicke ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Proteolytic Processing ◽  
Expression Level ◽  
Skin Wounds ◽  
Matrix Degradation ◽  
Wound Site ◽  
Laminin 5 ◽  
Keratinocyte Migration ◽  
The Matrix

The matrix metalloproteinase stromelysin-2 is expressed in keratinocytes of the epithelial tongue of skin wounds, suggesting a role in keratinocyte migration. Here, we show that stromelysin-2 enhances migration of cultured keratinocytes. To gain insight into the in vivo activities of stromelysin-2 in epithelial repair, we generated transgenic mice expressing a constitutively active stromelysin-2 mutant in keratinocytes. These animals had no alterations in skin architecture, and the healing rate of skin wounds was normal. Histologically, however, we found abnormalities in the organization of the wound epithelium. Keratinocytes at the migrating epidermal tip were scattered in most sections of mice with high expression level, and there was a reduced deposition of new matrix. In particular, the staining pattern of laminin-5 at the wound site was altered. This may be due to proteolytic processing of laminin-5 by stromelysin-2, because degradation of laminin-5 by this enzyme was observed in vitro. The inappropriate matrix contact of keratinocytes was accompanied by aberrant localization of β1-integrins and phosphorylated focal adhesion kinase, as well as by increased apoptosis of wound keratinocytes. These results suggest that a tightly regulated expression level of stromelysin-2 is required for limited matrix degradation at the wound site, thereby controlling keratinocyte migration.

Sign in / Sign up

Export Citation Format

Share Document