Increased Levels Of MMP-9 Research Articles

Anxiogenic-like effect of chronic lipopolysaccharide is associated with increased expression of matrix metalloproteinase 9 in the rat amygdala

Pathways by which inflammatory stimuli influence behaviors can involve changes in neuronal plasticity, however, the evidence for this is still insufficient. This study aimed to evaluate the effects of chronic lipopolysaccharide (LPS) injected alone or together with tetracycline antibiotic doxycycline (Dox) on the levels of Iba-1, BDNF, Bcl-xL and MMP-9 in brain regions in relation to stress-induced behaviors in the elevated plus-maze (EPM). LPS injected to adult rats every 2 days for a total of 7 injections reduced body weight gain, increased spleen and adrenal weights, decreased locomotor activity, and increased anxiety-like behavior. These effects were associated with increased expression of Iba-1, a well-known marker for activated microglia, in most brain regions investigated. Co-treatment of LPS with Dox attenuated LPS-induced microglial activation and behavioral changes, supporting their relation to the neuroinflammation. LPS administration also produced pro-apoptotic changes in the brain. In the hypothalamus and striatum, the levels of anti-apoptotic protein Bcl-xL were decreased, whereas in the amygdala, a significant increase in MMP-9 protein levels was observed. The levels of Iba-1 as well as MMP-9 in the amygdala positively correlated with the numbers of defecation. The data suggest that mechanisms of anxiety associated with neuroinflammation may involve the increase in MMP-9 levels in the amygdala.

Neuregulin-1 protects cardiac electrical conduction through downregulating matrix metalloproteinase-9 and upregulating connexin 43 in a rat myocardial infarction model.

Aims: Neuregulin-1 (NRG-1) is a member of the epidermal growth factor family, and has an important role in cardiomyocyte development and myocardial regeneration. The aim of this study was to determine the protective effect of NRG-1 on cardiac electrical conduction in a rat myocardial infarction (MI) model. Methods: Thirty-three adult male SPF SD rats were randomized into three groups: sham-operated (n=9), acute myocardial infarction (AMI, n=12), and the NRG-1-treated (NRG-1, n=12) groups. All rats were sacrificed on day 8 after inducing MI. The 6-lead electrocardiograms (ECG) were recorded pre-operatively and eight days after operation, and analyzed. The expression levels of matrix metalloproteinase (MMP)-9 and gap junction protein connexin 43 (Cx43) in the infarcted myocardium were measured by Western blotting, and its in-situ distribution was evaluated using immunohistochemistry. Results: The PR, QRS and QT intervals were significantly prolonged in the AMI group compared to the sham operated animals (P<0.05, P<0.01 and P<0.01 respectively), and the PR and QRS intervals were partially restored in the NRG-1-treated rats (P<0.01 and P<0.01 compared to AMI group). Similarly, the increased levels of MMP-9 in the AMI group was restored upon NRG-1 treatment. The myocardial expression of Cx43 was decreased significantly in the AMI group, and was upregulated by NRG-1 treatment. Conclusions: NRG-1 attenuates MI-induced dysfunctional cardiac electrical conduction by downregulating MMP-9 and upregulating Cx43.

The Effect of Mesenchymal Stem Cell Wharton's Jelly on Matrix Metalloproteinase-1 and Interleukin-4 Levels in Osteoarthritis Rat Model.

BACKGROUND:Osteoarthritis (OA) is generally considered a degenerative joint disease caused by biomechanical changes and the ageing process. In OA pathogenesis, the development of OA is thought to be regulated largely by excess matrix metalloproteinase (MMP), which contributes to the degradation of extracellular matrices such as MMP-1 and Interleukin-4.AIM:This study aims to prove the influence of Mesenchymal Stem Cell Wharton Jelly on decreasing MMP-1 levels and increasing IL-4 which is a specific target as a target component in cases of osteoarthritis in vivo.MATERIAL AND METHODS:This research is an experimental study with the design of Post-Test-Only Control Group Design. The sample consisted of 16 OA rats as a control group and 16 OA rats treated with MSC-WJ as a treatment group. OA induction is done by injection of monosodium iodoacetate (MIA) into the intra-articular right knee. Giving MSC-WJ is done in the third week after MIA induction. The serum MMP-1 and IL-4 levels were measured after 3 weeks treated with MSC-WJ using the ELISA method. The statistical test used is an independent t-test. The value of p < 0.05 was said to be statistically significant.RESULTS:The result showed that serum MMP-1 levels were higher in the group treated with MSC-WJ than in the control group (p < 0.05). Serum IL-4 levels were higher in the group treated with MSC-WJ than in the control group (p < 0.05).CONCLUSION:This study concluded that MSC-WJ increased MMP-1 levels and IL-4 levels in serum OA rats. MSC-WJ showed a negative effect on MMP-1 in the serum of OA rats.

Serum of Patients with Psoriasis Modulates the Production of MMP-9 and TIMP-1 in Cells of Monocytic Lineage

ABSTRACTPsoriasis is triggered by several stimuli that share a systemic production of interferon (IFN)-γ and other inflammatory mediators, which are key to regulate the production of matrix metalloproteinase (MMP)-9 and its inhibitor (TIMP)-1 by cells of monocytic lineage. This study evaluates the effect of the sera of 55 patients with psoriasis and 41 non-psoriatic individuals on the production of MMP-9 and TIMP-1 in cultured monocytes from a single healthy blood donor and in U937 cells. The effect of IFN-γ stimulation was also evaluated. Serum and supernatant concentrations of IFN-γ, MMP-9, and TIMP-1 were measured by enzyme-linked immunoassays, and the MMP-9/TIMP-1 ratios were calculated. In monocytes, incubation with psoriasis’ sera increased the production of MMP-9 and TIMP-1 in comparison with both baseline and monocytes incubated with non-psoriatic sera. Although the MMP-9/TIMP-1 ratio was significantly higher compared to the baseline, no differences between groups were observed. In contrast, IFN-γ stimulation in monocytes previously exposed to psoriasis’ sera increased MMP-9 levels and decreased TIMP-1 levels, whereas stimulation in monocytes exposed to non-psoriatic sera did not further modify the levels of MMP-9 or TIMP-1. Consequently, the MMP-9/TIMP-1 ratio in cells exposed to psoriatic serum was significantly higher than in cells exposed to non-psoriatic sera (24.5 versus 16.7; P < 0.05). Similar results were observed in U937 cells. Therefore, our results suggest that soluble mediators in psoriatic sera may enhance the proteolytic phenotype of monocytes when stimulated with IFN-γ, which supports the existence of a primed state in the inflammatory cells of patients with psoriasis.

Anti-inflammatory and ECM gene expression modulations of \u03b2-eudesmol via NF-\u03baB signaling pathway in normal human dermal fibroblasts

Backgroundβ-eudesmol is a kind of aromatic compound belonging to sesquiterpenoid which exists within not only the bark of magnolia but also Nardostachys jatamansi, Atractylodes lancea, Pterocarpus santalinus, Ginkgo bilobal, Cryptomeria japonica, etc., and there has been progress in medical and pharmaceutic researches on antitumor, anticancer, and anti-inflammatory effects; nervous system stabilization; and vasodilator effects, etc., but not in researches on skin cares and cosmetics at all. Therefore, this study pretreated β-eudesmol with human dermal fibroblasts (HDFs) and then gave oxidative stresses with H2O2 to examine antioxidation, anti-inflammatory, and cell preservation effects. Through this process, it proves the possibility of β-eudesmol as cosmetic materials.MethodsThis study verified the effectiveness of β-eudesmol through cell viability analysis, reactive oxygen estimation, associated β-galactosidase assay, nuclear factor-kappa B (NF-κB) luciferase assay, and quantitative real-time polymerase chain reaction (qRT-PCR).ResultsThe cell viability which decreased due to H2O2 increased as per dose-dependent manners of β-eudesmol. Also, at the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity assay, intracellular reactive oxygen species (ROS) quantitative analysis and glutathione (GSH) estimation, the relative levels which were changed by H2O2 treatment, showed attenuated or protective transition forms, depending on the concentration of β-eudesmol. Additionally, reduced superoxide dismutase 1 (SOD1) and catalase (CAT) gene expression by H2O2 were increased by β-eudesmol. As the result of the promoter activity analysis of NF-κB which has a key role in inflammation and skin aging, NF-κB activity decreases as β-eudesmol concentration increases, and also this study proves that gene expression of interleukin 1 beta (IL-1β) which is a downstream gene of NF-κB related to inflammatory response decreases as well as tumor necrosis factor-alpha (TNF-α) gene expression depending on the concentration of β-eudesmol.ConclusionsThrough these results, this study suggests there are anti-inflammatory effects by shutting out the NF-κB pathway. Following the results of the extracellular matrix (ECM) regulating gene expression analysis, this study proves that oxidative stress-induced increased MMP1 levels were decreased depending on the concentration of β-eudesmol and verifies that it hinders the collapse of collagen through inhibition of transcriptional activity of NF-κB. From the result of β-eudesmol regulating tissue inhibitors of metalloproteinase (TIMP)-1 gene expression which hinders matrix metalloproteinase (MMP), activation and alteration of gene expression of collagen type I alpha 1 (COL1A1) underpins the above consequences. Through this research, it is considered that β-eudesmol as one of natural cosmetic materials with such effects as antioxidation, anti-inflammation, and cell preservation is worthy of notice.

Down-Regulation of Neuropathy Target Esterase in Preeclampsia Placenta Inhibits Human Trophoblast Cell Invasion via Modulating MMP-9 Levels

Background/Aims: Neuropathy target esterase (NTE, also known as neurotoxic esterase) is proven to deacylate phosphatidylcholine (PC) to glycerophosphocholine as a phospholipase B. Recently; studies showed that artificial phosphatidylserine/PC microvesicles can induce preeclampsia (PE)-like changes in pregnant mice. However, it is unclear whether NTE plays a key role in the pathology of PE, a pregnancy-related disease, which was characterized by deficient trophoblast invasion and reduced trophoblast-mediated remodeling of spiral arteries. The aim of this study was to investigate the expression pattern of NTE in the placenta from women with PE and normal pregnancy, and the molecular mechanism of NTE involved in the development of PE. Methods: NTE expression levels in placentas from 20 pregnant women with PE and 20 healthy pregnant women were detected using quantitative PCR and immunohistochemistry staining. The effect of NTE on trophoblast migration and invasion and the underlying mechanisms were examined in HTR-8/SVneo cell lines by transfection method. Results: NTE mRNA and protein expression levels were significantly decreased in preeclamptic placentas than normal control. Over-expression of NTE in HTR-8/SVneo cells significantly promoted trophoblast cells migration and invasion and was associated with increased MMP-9 levels. Conversely, shRNA-mediated down-regulation of NTE markedly inhibited the cell migration and invasion. In addition, silencing NTE reduced the MMP-9 activity and phosphorylated Erk1/2 and AKT levels. Conclusions: Our results suggest that the decreased NTE may contribute to the development of PE through impairing trophoblast invasion by down-regulating MMP-9 via the Erk1/2 and AKT signaling pathway.

Poorly controlled diabetes mellitus type 2 is associated with increased mmp-9 levels and occurance of myocardial infarction-a pilot study

Poorly controlled diabetes mellitus type 2 is associated with increased mmp-9 levels and occurance of myocardial infarction-a pilot study

Pam3CSK4 induces MMP-9 expression in human monocytic THP-1 cells

Abstract Background Matrix metalloproteinase (MMP)-9 is known to degrade the extracellular matrix and increased MMP-9 levels are related with the pathogenesis of many inflammatory conditions including obesity. Pam3CSK4 is a synthetic triacylated lipopeptide (LP) which is a potent activator of immune cells and induces cytokine production. However, it is unclear whether Pam3CSK4 is able to induce MMP-9 expression in monocytic cells. We, therefore, determined MMP-9 production by Pam3CSK4-treated THP-1 cells and also investigated the signal transduction pathway(s) involved. Methods MMP-9 expression was determined by real-time qPCR and ELISA and MMP-9 activity was assessed by zymography. THP-1 cells, THP1-XBlue™ cells, THP1-XBlue™-defMyD cells, anti-TLR2 mAb and selective pharmacological inhibitors were used to study signaling pathways involved. Phospho and total proteins were detected by western blotting. Results Pam3CSK4 induced MMP-9 expression (P&amp;lt;0.05) at both mRNA and protein levels in human monocytic THP-1 cells. Increased NF-κB/AP-1 activity was detected in Pam3CSK4-treated THP-1 cells and MMP-9 production in these cells was significantly suppressed by pre-treatment with anti-TLR2 neutralizing antibody or by inhibition of clathrin-dependent endocytosis. Also, MyD88−/− THP-1 cells did not express MMP-9 following treatment with Pam3CSK4. Inhibition of JNK, MEK/ERK, p38 MAPK and NF-κB significantly suppressed MMP-9 gene expression (P&amp;lt;0.05). Conclusion Pam3CSK4 induces MMP-9 production in THP-1 cells through the TLR-2/MyD88-dependent mechanism involving ERK, JNK, p38 MAPK and NF-κB/AP-1 activation.

Pam3CSK4 Induces MMP-9 Expression in Human Monocytic THP-1 Cells

Background: Matrix metalloproteinase (MMP)-9 is known to degrade the extracellular matrix and increased MMP-9 levels are related with the pathogenesis of many inflammatory conditions including obesity. Pam3CSK4 is a synthetic triacylated lipopeptide (LP) which is a potent activator of immune cells and induces cytokine production. However, it is unclear whether Pam3CSK4 is able to induce MMP-9 expression in monocytic cells. We, therefore, determined MMP-9 production by Pam3CSK4-treated THP-1 cells and also investigated the signal transduction pathway(s) involved. Methods: MMP-9 expression was determined by real-time qPCR and ELISA. MMP-9 activity was assessed by zymography. THP-1 cells, THP1-XBlue^{^TM} cells, THP1-XBlue^{^TM}-defMyD cells, anti-TLR2 mAb and selective pharmacological inhibitors were used to study signaling pathways involved. Phosphorylated and total proteins were detected by western blotting. Results: Pam3CSK4 induced MMP-9 expression (P<0.05) at both mRNA and protein levels in human monocytic THP-1 cells. Increased NF-κB/AP-1 activity was detected in Pam3CSK4-treated THP-1 cells and MMP-9 production in these cells was significantly suppressed by pre-treatment with anti-TLR2 neutralizing antibody or by inhibition of clathrin-dependent endocytosis. Also, MyD88-/- THP-1 cells did not express MMP-9 following treatment with Pam3CSK4. Inhibition of JNK, MEK/ERK, p38 MAPK and NF-κB significantly suppressed MMP-9 gene expression (P<0.05). Conclusion: Pam3CSK4 induces MMP-9 production in THP-1 cells through the TLR-2/MyD88-dependent mechanism involving MEK/ERK, JNK, p38 MAPK and NF-κB/AP-1 activation.

Matrix metalloproteinase-9, its inhibitor-1 and interleukines in experimental traumatic brain injury

Traumatic brain injury (TBI) is accompanied by high rates of morbidity and mortality in both developed and undeveloped countries that makes it one of the most actual medical and social problems. In recent years matrix metalloproteinases are in increasing interest while studying TBI pathogenesis because of their ability to increase permeability of the blood-brain barrier and to cause nervous tissue matrix reorganization.The goal of given study was to investigate the role of matrix metalloproteinase MMP-9 and its inhibitor TIMP-1 in pathogenesis of TBI.Methods: The study was performed on 98 mature white rats. Moderate severity TBI was modeled with one blow on the cranial vault by means of free-falling plummet. Control group included 30 rats. Cytokines (IL-1b, IL-6, IL-8, TNF-a), MMP-9 and TIMP-1 levels were investigated in animals blood by means of ELISA on the 1st, 3rd, 7th, 14th and 21st days after trauma.Results and discussion: MMP-9 levels increased by only 38,2% on the 1st day, but on the 3rd day there was its marked increase to 538%. It is known that metalloproteinases are released from the cells under the influence of various factors, including cytokines. On the 1st day after trauma it was IL-1β which increased by 705% showing the highest rise among other cytokines and exceeding increase in MMP-9 levels. This might indicate regulatory role of IL-1β. A marked increase in MMP-9 levels in its turn led to TIMP-1 activation. Significant increase in TIMP-1 levels was determined on the 3rd day after trauma. On the 7th day there was a critical period with the highest levels of IL-1β (2147,2%), MMP-9 (720,3%) and TIMR-1 (339,3%). Then all research indicators were decreasing with the most pronounced decrease in IL-1β and MMP-9.Conclusion: MMP-9 levels began to increase on the 1st day after trauma due to influence of mainly IL-1β. An abrupt increase in MMP-9 in its turn caused an increase in TIMR-1 levels. Identified changes in IL-1β, ММР-9 and ТІМР-1 levels after TBI indicate complex relationships between cytokines and intercellular matrix reorganization regulators in formation of intercellular cooperation and neuroinflammation development.

MMP1-1607 polymorphism increases the risk for periapical lesion development through the upregulation MMP-1 expression in association with pro-inflammatory milieu elements.

ABSTRACTIncreased matrix metalloproteinases (MMPs) activity is a hallmark of periapical granulomas. However, the factors underlying the MMPs expression modulation in healthy and diseased periapical tissues remains to be determined. Objective In this study, we evaluated the association between the MMP1-1607 polymorphism (rs1799750) and pro-inflammatory milieu elements with MMP-1 mRNA levels in vivo.Material and Methods MMP1-1607 SNP and the mRNA levels of MMP-1, TNF-a, IFN-g, IL-17A, IL-21, IL-10, IL-4, IL-9, and FOXp3 were determined via RealTimePCR in DNA/RNA samples from patients presenting periapical granulomas (N=111, for both genotyping and expression analysis) and control subjects (N=214 for genotyping and N=26 for expression analysis). The Shapiro-Wilk, Fisher, Pearson, Chi-square ordinal least squares regression tests were used for data analysis (p<0.05 was considered statistically significant).Results The MMP1-1607 1G/2G and 1G/2G+2G/2G genotypes were significantly more prevalent in the patients than in controls, comprising a risk factor for periapical lesions development. MMP-1 mRNA levels were higher in periapical lesions than in healthy periodontal ligament samples, as well as higher in active than in inactive lesions. The polymorphic allele 2G carriers presented a significantly higher MMP-1 mRNA expression when compared with the 1G/1G genotype group. The ordered logistic regression demonstrated a significant correlation between the genetic polymorphism and the expression levels of MMP-1. Additionally, the pro- and anti-inflammatory cytokines IL-17A, IFN-g, TNF-a, IL-21, IL-10, IL-9, and IL-4 were significant as complementary explanatory variables of MMP-1 expression.Conclusion The MMP1-1607 SNP was identified as a risk factor for periapical lesions development, possibly due to its association with increased MMP-1 mRNA levels in periapical lesions. The MMP-1 expression is also under the control of the inflammatory milieu elements, being the cytokines TNF-a, IL-21, IL-17A, and IFN-g associated with increased MMP-1 levels in periapical lesions, while IL-10, IL-9, or IL-4 presented an inverse association.

Association of Matrix Metalloproteinase-2 Gene Promoter Polymorphism and the Associated Phenotype Variation with Myocardial Infarction

In many developing countries Myocardial infarction has Becoming a major problem in public health 1,2. MI is a multifactorial Disease caused by genetic and environmental factors.The major cause of Death in the world is Myocardial infarction 3. The high plasma lipid Levels, high plasma glucose levels,high blood pressure, obesity, smoking, And family history of cardiac disease are the most important risk factors For MI. MI is mainly due to atherosclerosis of the coronary arteries . The structural changes, which permits the accumulation of cells, Extracellular matrix and lipids in the intimate layer of the diseased artery And allows the growth of atherosclerotic plaque. The fibrous cap lining Atheromatous plaque gets ruptured, gives rise to thrombosis, and its Complications4. Pathophysiology of MI involves a wide variety of proteins, Including the matrix metalloproteinases (mmps).Atheromatous plaque Formation is facilitated by the action of mmps. Major extra cellular Components of the basal lamina around blood vessels such as type 1V Collagen, laminin, and fibronectin are degraded by MMP-2. Mmps Also weakens the arterial wall, resulting destabilizing of atheromatous Plaque and dissolution of fibrous cap leading to MI5. Matrix metalloproteinases are zinc dependent endopeptidases That degrade components of the extracellular matrix (ECM). 72kda type IV collagenase also known as MMP-2, is an ubiquitous Metalloproteinase involved in various functions such as vascular Remodeling, atheromatous plaque rupture, and degrading matrix Proteins. MMP-2, also known as gelatinase A . In human, it is Encoded by the MMP-2 gene6. MMP-2 is produced as zymogen, after the production pro MMP- 2 is thought to be bound to its specific inhibitor, called as tissue Inhibitor of matrix metalloproteinases-2 . There are several pathways for Activation of the proenzyme, but mainly the most important pathway is Activation by membrane type metalloproteinases-1 (MT1-MMP)7. MTIMMP Binds to TIMP-2,this complexed structure comes near to the Active site of the MT-MMP enzyme. This results to removal of two Specific pro peptides from pro MMP-2 and the production of an Active 72 kda MMP-2 enzyme8. The gene for human MMP-2 contains 27,862 basepair genomic DNA and is composed of 13 exons. It has been localized on chromosome 16q219. Several common restriction fragment length polymorphisms (rflps) have been reported in the MMP-2 gene locus. MMP-2 gene – 1306C>T promoter region is linked with development of MI10. This Base transition is situated in CCACC box of the sp1 binding site. Increased MMP-2 levels have been found in the plasma of Patients with MI11. Elevated MMP-2 has also been found in Atherosclerotic plaques of coronary arteries12. In view of this we have evaluated the distribution of MMP-2 Promoter gene polymorphism by PCR- RFLP and the concerned Phenotype (MMP-2) was analyzed by using ELISA.

Serum Matrix Metalloproteinases as Quantitative Biomarkers for Myocardial Fibrosis and Sudden Cardiac Death Risk Stratification in Patients With Hypertrophic Cardiomyopathy

BackgroundHypertrophic cardiomyopathy (HCM) is associated with an increased risk of sudden cardiac death due to ventricular tachycardia (VT), and myocardial fibrosis reflects an important risk factor. Several matrix metalloproteinases (MMPs) and procollagen N-terminal propeptides (PNPs) are involved in collagen turnover and discussed as fibrosis biomarkers. We aimed to identify biomarkers that correlate with myocardial fibrosis in late-gadolinium-enhancement cardiac magnetic resonance imaging (LGE-CMR) and/or cardiac events (syncope, VT) in HCM patients. Methods and ResultsIn 54 HCM patients (age 55.9 ± 14.3 y, 50% female) fibrosis was quantified by LGE-CMR. Serum concentrations of MMP-1, -2, -3, -9, and tissue inhibitor of MMP (TIMP) 1 were analyzed by means of enzyme-linked immunosorbent assay and PINP, PIIINP, and type I collagen C-terminal telopeptide (ICTP) concentrations by radioimmunoassay. MMP-9 was associated with fibrosis in LGE-CMR (mean increase 0.66 g/unit MMP9 [95% confidence interval [CI] 0.50–0.82]; P < .001) and with cardiac events in women (odds ratio [OR] 1.07 [1.01–1.12], P = .01) but not in men. Increased MMP-2 levels in women were associated with lower fibrosis (0.05 [−0.09 to −0.01]; P = .015). MMP-3 levels were positively associated with cardiac events (OR 1.13 [1.05–1.22]; P = .001) independently from fibrosis and sex. No association was detected for MMP-1, TIMP-1, PNPs, and ICTP. ConclusionsThese data suggest that MMP-9 is a useful biomarker for fibrosis and cardiac events in female HCM patients, whereas MMP-3 is associated with a higher event rate independent from fibrosis and sex.

Mesenchymal stem cells and serelaxin synergistically abrogate established airway fibrosis in an experimental model of chronic allergic airways disease

This study determined if the anti-fibrotic drug, serelaxin (RLN), could augment human bone marrow-derived mesenchymal stem cell (MSC)-mediated reversal of airway remodeling and airway hyperresponsiveness (AHR) associated with chronic allergic airways disease (AAD/asthma). Female Balb/c mice subjected to the 9-week model of ovalbumin (OVA)-induced chronic AAD were either untreated or treated with MSCs alone, RLN alone or both combined from weeks 9–11. Changes in airway inflammation (AI), epithelial thickness, goblet cell metaplasia, transforming growth factor (TGF)-β1 expression, myofibroblast differentiation, subepithelial and total lung collagen deposition, matrix metalloproteinase (MMP) expression, and AHR were then assessed. MSCs alone modestly reversed OVA-induced subepithelial and total collagen deposition, and increased MMP-9 levels above that induced by OVA alone (all p<0.05 vs OVA group). RLN alone more broadly reversed OVA-induced epithelial thickening, TGF-β1 expression, myofibroblast differentiation, airway fibrosis and AHR (all p<0.05 vs OVA group). Combination treatment further reversed OVA-induced AI and airway/lung fibrosis compared to either treatment alone (all p<0.05 vs either treatment alone), and further increased MMP-9 levels. RLN appeared to enhance the therapeutic effects of MSCs in a chronic disease setting; most likely a consequence of the ability of RLN to limit TGF-β1-induced matrix synthesis complemented by the MMP-promoting effects of MSCs.

Significance of circulating and crevicular matrix metalloproteinase-9 in rheumatoid arthritis-chronic periodontitis association.

In the recent years, statistically significant associations between rheumatoid arthritis (RA) and periodontal disease have been identified. Emerging as a chronic inflammatory joint disease, RA displays various features and pathogenetic events similar to chronic periodontitis (CP). The purpose of this study was to evaluate the utility of determining systemic and crevicular levels of metalloproteinase-9 (MMP-9) as potential biomarkers for association between RA and CP. A total of fifty-six patients were included in the study. The subjects were categorized into four groups as follows: healthy-control (n = 21), active RA (n = 16), CP (n = 14), and RA-CP association (n = 12). Assessment of serum and crevicular concentrations of total MMP-9 (active and pro-MMP-9) was based on ELISA technique. The results of this study showed statistically significant differences of serum MMP-9 between patients groups and control. Serum levels of MMP-9 were similar in RA and RA-CP associated patients. Gingival crevicular fluid (GCF) recorded increased MMP-9 levels in RA-CP association subjects as compared to CP. Considering that RA-CP association is characterized by a disregulation of the inflammatory response, MMP-9 may play a role in the pathogenesis of RA-CP association. MMP-9 is therefore a sensitive tool in the diagnosis and management of patients affected by this binomial association.

P. gingivalis lipopolysaccharide intensifies inflammation post-myocardial infarction through matrix metalloproteinase-9

Periodontal disease (PD) strongly correlates with increased mortality post-myocardial infarction (MI); however, the underlying mechanisms are unknown. Matrix metalloproteinase (MMP)-9 levels directly correlate with dysfunction and remodeling of the left ventricle (LV) post-MI. Post-MI, MMP-9 is produced by leukocytes and modulates inflammation. We have shown that exposure to Porphyromonas gingivalis lipopolysaccharide (PgLPS), an immunomodulatory molecule identified in PD patients, increases LV MMP-9 levels in mice and leads to cardiac inflammation and dysfunction. The aim of the study was to determine if circulating PgLPS exacerbates the LV inflammatory response post-MI through MMP-9 dependent mechanisms. We exposed wild type C57BL/6J and MMP-9−/− mice to PgLPS (ATCC 33277) for a period of 28days before performing MI, and continued to deliver PgLPS for up to 7days post-MI. We found systemic levels of PgLPS 1) increased MMP-9 levels in both plasma and infarcted LV resulting in reduced wall thickness and increased incidence of LV rupture post-MI and 2) increased systemic and local macrophage chemotaxis leading to accelerated M1 macrophage infiltration post-MI and decreased LV function. MMP-9 deletion played a protective role by attenuating the inflammation induced by systemic delivery of PgLPS. In conclusion, MMP-9 deletion has a cardioprotective role against PgLPS exposure, by attenuating macrophage mediated inflammation.

Increased CD147 (EMMPRIN) expression in the rat brain following traumatic brain injury

The extracellular matrix metalloproteinase inducer (EMMPRIN), or CD147, has been known to play a key regulatory role in vascular permeability and leukocyte activation by inducing the expression of matrix metalloproteinases (MMPs). The effects of traumatic brain injury on the expression of EMMPRIN remain poorly understood. In this study, we investigated changes in EMMPRIN expression in a rat model of fluid percussion injury (FPI) and examined the potential association between EMMPRIN and MMP-9 expression. Adult male rats were subjected to FPI. EMMPRIN expression was markedly up-regulated in the brain tissue surrounding the injured region 6–48h after TBI, as measured by immunoblot and immunohistochemistry. EMMPRIN expression was localized to inflammatory cells. The increase in EMMPRIN expression was temporally correlated with an increase in MMP-9 levels. These data demonstrate, for the first time, changes in CD147 and MMP-9 expression following TBI. These data also suggest that CD147 and MMP-9 may play a role in vascular injuries after TBI.

Syzygium cumini reduces oxidative stress and matrix metalloproteinase-2 level in endothelial cells induced by plasma from preeclamptic patients

Objective: This study aimed to investigate whether an ethanolic extract of Syzygium cumini is able to decrease lipid peroxidation and matrix metalloproteinase (MMP)-2 expression in endothelial cells exposed to plasma from preeclamptic patients. Methods: Endothelial cells were obtained from human umbilical vascular endothelial cells. At confluency, endothelial cells were divided into five groups, which included control (untreated), endothelial cells exposed to 2% plasma from preeclamptic patients (PP), endothelial cells exposed to PP in the presence of ethanolic extract of S.cumini (PP+SC) at the doses of 100, 200 and 400 ppm. Analysis of malondialdehyde (MDA) level as a marker of lipid peroxidation was done colorimetrically. Analysis of MMP-2 level was done by enzyme linked immunosorbent assay (ELISA) technically. Results: This increase in MDA was significantly attenuated by both the 200 and 400 ppm treatments of S.cumini extract. Plasma from PP significantly increased MMP-2 levels compared to untreated cells. This increase in MMP-2 was significantly attenuated by the 200 and 400 ppm doses of the extract. Conclusion: In conclusion, Syzygium cumini extract prohibits the increase in oxidative stress and MMP-2 in endothelial cells induced by plasma from preeclamptic patients. Therefore this may provide a natural therapy for attenuating the maternal endothelial dysfunction found in this disease.

Abstract 393: Chronic Nebivolol Treatment Attenuates MMP Activity and Oxidative Stress and Improves Renovascular Hypertension-induced Cardiac Hypertrophy

Nebivolol (Nebi) is the most selective cardiac β1-adrenergic receptor blocker and exerts antioxidant effects. Reactive oxygen species (ROS) enhance cardiac matrix metalloproteinase activity (MMPs) and contribute to hypertension-induced left ventricular hypertrophy (LVH). However, it is uncertain whether Nebi decreases MMP-2 levels and cardiac remodeling associated with hypertension as a result of its antioxidant properties. Hypertension was induced by clipping the left renal artery (2K1C). Six weeks after surgery, hypertensive and sham rats were treated with Nebi, Metoprolol (Meto) or Vehicle (by gavage) for four weeks. Systolic blood pressure (SBP) was monitored weekly by tail-cuff plethysmography. LV structural changes were studied in hematoxylin/eosin sections. MMP levels and activity were determined by immunofluorescence and in situ zymography. ROS and nitrotyrosine levels were evaluated by dihydroethidium and immunohistochemistry, respectively. Similar reduction in SBP was found with both treatments (202±5 mmHg in 2K1C+Vehicle versus 161±19 and 162±18 mmHg in 2K1C Nebi or Meto groups, respectively; P&lt;0.05). Both treatments reduced LV remodeling and ROS formation (P&lt;0.05). Both drugs decreased nitrotyrosine levels (3.9±0.2, 2.3±0.3, and 2.4±0.3 arbitrary units, respectively in 2K1C+vehicle, 2K1C+Meto, and 2K1C+Nebi groups, respectively; P&lt;0.05). Increased MMP-2 levels (16.0±2.0 versus 7.0±0.9 arbitrary units; P&lt;0.05) and activity (8.0±0.4 versus 6.0±0.1 arbitrary units; P&lt;0.05) were observed in the 2K1C+vehicle group when compared to the sham group. These alterations were normalized by both treatments (similar levels to those found in the sham group). No significant changes were observed in the sham groups. These findings suggest that different β-blockers exert important antioxidant effects that may underlie the lower LV MMP activity and cardiac remodeling in 2K1C hypertension.

Treponema denticola associates with increased levels of MMP‐8 and MMP‐9 in gingival crevicular fluid

The aim was to assess the association between the presence of site-specific subgingival micro-organisms and the levels of matrix metalloproteinases-8 and matrix metalloproteinases-9 (MMP-8 and MMP-9) in gingival crevicular fluid (GCF). The patient group consisted of 56 subjects with periodontitis and the control group of 43 subjects without periodontitis. GCF samples from four test sites for each subject were collected. Polymerase chain reaction was used to detect the presence of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, and Treponema denticola. MMP-8 concentrations were analyzed by a time-resolved immunofluorometric assay, and MMP-9 levels were determined by enzyme-linked immunosorbent assay. Student's unpaired t-test, chi-square test, and Fisher's exact P-value were calculated. The presence of T.denticola in the test sites was significantly higher in the patient group than in the control group. The presence of T.forsythia and T.denticola was associated with increased levels of MMP-8 in the test sites. Respectively, site-specific presence of T.denticola was associated with an increase in MMP-9 levels in three of the four test sites. The presence of subgingival micro-organisms in GCF, particularly T.denticola, appeared to induce a host response with an increased release of MMP-8 and MMP-9 in the test sites.