Higher Levels Of MMP-9 Research Articles

Tear levels of apoptotic, matrix-degrading and antioxidant biomarkers in patients with and without keratoconus: A cross sectional study

PurposeTo assess the tear levels of a set of apoptotic, matrix-degrading and antioxidant biomarkers, including Metalloproteinase 9 (MMP9), High Mobility Group Box 1 (HMGB1) and Superoxide Dismutase 3-Extracellular (SOD3). MethodsSandwich-ELISA commercial kits were used to test the expression of the three tear biomarkers in the lacrimal fluid of eligible participants. Linear logistic regression analysis was performed todetermine whether the set of tear biomarkers could be associated with clinically manifest keratoconus. ROC curve analysis using 10-fold cross-validation was performedto evaluate the prediction accuracy of the model. ResultsEighty-one participants aged 30–48 years old were enrolled in this study; 48 were patients with keratoconus and 33 were age-matched healthy subjects. The linear combination of the three tear biomarkers levels (AUC = 0.811; CI 95 %: 0.712–0.911) accurately indicated the existence of keratoconus; higher levels of MMP9 (Odd Ratio: 1.069; CI 95 %: 1.029–1.130) and HMGB1 (OR: 1.011; CI 95 %: 1.003–1.022) and lower levels of SOD3 (OR: 0.994; CI 95 %: 0.989–0.997) were significantly associated with a higher probability of keratoconus. ConclusionMultivariable analysis of the set of tear levels of MMP9, HMGB1 and SOD3 biomarkers confirmed a chronic state of inflammation in the ocular surface of patients with keratoconus.

Differences in metalloproteinases and their tissue inhibitors in the cerebrospinal fluid are associated with delirium

BackgroundThe aetiology of delirium is not known, but pre-existing cognitive impairment is a predisposing factor. Here we explore the associations between delirium and cerebrospinal fluid (CSF) levels of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), proteins with important roles in both acute injury and chronic neurodegeneration.MethodsUsing a 13-plex Discovery Assay®, we quantified CSF levels of 9 MMPs and 4 TIMPs in 280 hip fracture patients (140 with delirium), 107 cognitively unimpaired individuals, and 111 patients with Alzheimer’s disease dementia. The two delirium-free control groups without acute trauma were included to unravel the effects of acute trauma (hip fracture), dementia, and delirium.ResultsHere we show that delirium is associated with higher levels of MMP-2, MMP-3, MMP-10, TIMP-1, and TIMP-2; a trend suggests lower levels of TIMP-4 are also associated with delirium. Most delirium patients had pre-existing dementia and low TIMP-4 is the only marker associated with delirium in adjusted analyses. MMP-2, MMP-12, and TIMP-1 levels are clearly higher in the hip fracture patients than in both control groups and several other MMP/TIMPs are impacted by acute trauma or dementia status.ConclusionsSeveral CSF MMP/TIMPs are significantly associated with delirium in hip fracture patients, but alterations in most of these MMP/TIMPs could likely be explained by acute trauma and/or pre-fracture dementia. Low levels of TIMP-4 appear to be directly associated with delirium, and the role of this marker in delirium pathophysiology should be further explored.

Secondary Prevention of Multifocal Atherosclerosis Combined with Chronic Obstructive Pulmonary Disease

The problem of secondary prevention of multifocal atherosclerosis (MAS) combined with chronic obstructive pulmonary disease (COPD), is a leading cause of death in the world. Therefore, the search for drugs capable of affecting the destructive mechanisms underlying both MAS and COPD is an extremely urgent problem today. Objective — to reduce the risks of destabilization of atherosclerotic plaques by reducing the levels of metallomatrix metalloproteases (MMP)-2 and MMP-9 in patients on MAS combined with COPD. Materials and methods. The study included 62 men (68.1 ± 4.2) years old with MAS. All patients with MAS had clinical and functional signs of injury in coronary, cerebral and femoral vascular territories, 30 of these patients (group MAS-2) additionally had clinical and functional signs of COPD (GOLD-2). The control group (CG) consisted of 18 practically healthy men, aged (65.4 ± 3.7) years. Examination of the patients included echocardiography, dopplerography of the vessels of the neck and arteries of the lower extremities, determination of walking distance and ankle-brachial index, Holter ECG monitoring, spirography and determination of MMP-2 and MMP-9 levels in blood. Patients of both groups were prescribed cilostazol (50 mg twice a day) and GABA aminalon (250 mg twice a day) on the background of basic treatment. The course of treatment lasted 16 weeks. Results and discussion. During the initial examination of patients, MAS-2 group showed significantly (p < 0.001) lower volumetric blood flow in the studied arteries, and significantly (p < 0.01) higher levels of MMP-2 and MMP-9, as compared to CG, as well as with patients of the MAS-1 group. After 16 weeks of treatment, with the addition of cilostazol and aminalon, the level of MMP in the blood significantly (p < 0.05) decreased in both groups, in particular, in the MAS-2 group, in patients with the combined pathology of MAS and COPD, a significant decrease was found MMP-2 by 23.6 % (p < 0.01), and MMP-9 by 12.1 % (p < 0.05). Volumetric blood flow indicators increased significantly (p < 0.05) in all studied vascular territories, which led to improvement in clinical manifestations of the disease — a decrease in the number of painful and painless episodes of myocardial ischemia, and an increase in walking distance. Conclusions. The use of a complex therapy of statins, cilostazol and GABA in patients with MAS combined with COPD allows to reduce significantly the levels of MMP-2 and MMP-9, which ensures the stability of atheromatous plaques and significantly improves blood supply in vascular territories with atherosclerotic lesions.

Serum inflammatory factors such as MMP-9 are associated with post-percutaneous transluminal angioplastyacute myocardial infarction in coronary heart disease patients complicated by lower extremity arteriosclerosis obliterans.

This clinical study mainly analyzed the correlation of changes in serum inflammatory factors (IFs), such as matrix metalloproteinase (MMP)-9, hypersensitive C-reactive protein (hs-CRP), tumor necrosis factor (TNF)-α, and interleukin (IL)-6 with post-percutaneous transluminal angioplasty (PTA)acute myocardial infarction (AMI) in coronary heart disease (CHD)patients complicated by lower extremity arteriosclerosis obliterans (ASO). This retrospective study selected sixty ASO+CHD patients (ASO group) who underwent lower limb angioplasty between January 2014 and June 2016, as well as 50 concurrent healthy controls (HCs, HC group). According to the occurrence of AMI after PTA, cases were further subdivided into AMI (n = 18) and non-AMI (n = 42) groups. For all participants, IFs (MMP-9, hs-CRP, TNF-α, and IL-6) were detected on an empty stomach. The correlations of these IFs with the post-PTAAMI risk of ASO + CHD patients were analyzed using Pearson correlation coefficients, and their predictive value for AMI was visualized by receiver operating characteristic (ROC)curves. Finally, the prognostic factors of perioperative AMI in ASO+CHD patients were identified by multivariate analysis using the Cox model. MMP-9, hs-CRP, TNF-α and IL-6 presented statistically higher levels in the AMI group than in non-AMI and HC groups and were positively correlated with AMI. ROC analysis data showed that MMP-9, hs-CRP, TNF-α and IL-6 had better diagnostic performance, sensitivity and specificity for post-PTAAMI in patients with ASO+CHD. According to Cox multivariate analysis, high levels of MMP-9, hs-CRP and IL-6 increased the risk of perioperative AMI in ASO+CHD patients after PTA. This study shows a significant correlation between the changes of serum IFs (MMP-9, hs-CRP, IL-6, and TNF-α) and post-PTA AMI in ASO patients complicated by CHD. Patients with upregulated post-PTA levels of the above Ifs in serum are at an elevated risk of developing AMI, and active and effective control will help to prevent AMI.

MMP-9 EXPRESSION IN ORAL SQUAMOUS CELL CARCINOMA USING IMMUNOHISTOCHEMISTRY, WESTERN BLOTTING, AND REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION TECHNIQUES

Research into matrix-metalloproteinases (MMP) and their inhibition in certain cancers has moved significantly, studies on MMP expression in oral squamous cell carcinoma (OSCC) remains scarce. MMP-9, a gelatinase-degrading enzyme, destroys matrix proteins in gingivitis and adult periodontitis. MMP-9 genetic (mRNA) and protein profiles were examined to determine MMP-9 expression in oral cancer in this study. Our gender findings indicate that the majority of patients were males and consumers of various tobacco forms.The buccal mucosa was involved in 52% of OSCCs, followed by the tongue (33%), and the gingivobuccal sulcus (9%).The levels of MMP-9 detected in OSCC grades were higher than controls in the order of well-, followed by moderately- and lastly poorly-differentiated grades.Immunohistochemistryseemed to be a reliable approach for measuring MMP-9 expression in OSCC samples when compared to RT-qPCR and western blotting. The association of high MMP-9 levels with well-differentiated OSCC grade implies its potential as a biomarker for early detection of OSCC which could lead to the development of early OSCC treatments. More studies with larger samples might aid in the evaluation of the techniques used.

Expression of Matrix Metalloproteinases in the Circulating Immune Cells in Children with Helicobacter pylori Infection-Correlation with Clinical Factors.

H. pylori gastritis is strongly associated with the upregulation of the expression of several matrix metalloproteinases (MMPs) in the gastric mucosa. However, the role of MMP-2 and MMP-9, and their inhibitors (tissue inhibitors of metalloproteinases -TIMPs) produced by immune cells in infected children have not been clearly defined. Moreover, the effects of H. pylori eradication therapy on MMPs and TIMPs production has not been evaluated. A total of 84 children were studied: 24-with newly diagnosed H. pylori gastritis, 25-after H. pylori eradication therapy (17 of them after successful therapy), 24-with H. pylori-negative gastritis, and 11-controls. Plasma levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 by ELISA; MMPs and TIMPs expression in lymphocytes; neutrophils and monocytes in peripheral blood by multiparameter flow cytometry; and mucosal mRNA expression levels of MMPs and TIMP-1 in gastric biopsies by RT-PCR were evaluated. Children with H. pylori-related gastritis showed the following: (1) increased MMP-2 and TIMP-2 plasma levels, (2) increased intracellular expression of MMP-2 in the circulating lymphocytes and neutrophils, (3) low frequencies of circulating TIMP-1+ and TIMP-2+ leukocytes, and (4) high expression of mRNA for MMP-9 along with low expression of mRNA for MMP-2 in the gastric mucosa. Unsuccessful H. pylori eradication was associated with the following: (1) high plasma levels of MMP-9 and TIMP-1, (2) increased pool of TIMP-1+ lymphocytes as well as high expression of MMP-9 in circulating lymphocytes, and (3) high expression of mRNA for MMP-9 in the gastric mucosa. Our data suggest that MMPs are important contributors to stomach remodelling in children with H. pylori-related gastritis. Unsuccessful H. pylori eradication is associated with increased MMP-9 in plasma, circulating lymphocytes, and gastric mucosa.

Biomarkers of Collagen Metabolism Are Associated with Left Ventricular Function and Prognosis in Dilated Cardiomyopathy: A Multi-Modal Study.

Collagen cross-linking is a fundamental process in dilated cardiomyopathy (DCM) and occurs when collagen deposition exceeds degradation, leading to impaired prognosis. This study investigated the associations of collagen-metabolism biomarkers with left ventricular function and prognosis in DCM. DCM patients who underwent endomyocardial biopsy, blood sampling, and cardiac MRI were included. The primary endpoint included death, heart failure hospitalization, or life-threatening arrhythmias, with a follow-up of 6 years (5-8). In total, 209 DCM patients were included (aged 54 ± 13 years, 65% male). No associations were observed between collagen volume fraction, circulating carboxy-terminal propeptide of procollagen type-I (PICP), or collagen type I carboxy-terminal telopeptide [CITP] and matrix metalloproteinase [MMP]-1 ratio and cardiac function parameters. However, CITP:MMP-1 was significantly correlated with global longitudinal strain (GLS) in the total study sample (R = -0.40, p < 0.0001; lower CITP:MMP-1 ratio was associated with impaired GLS), with even stronger correlations in patients with LVEF > 40% (R = -0.70, p < 0.0001). Forty-seven (22%) patients reached the primary endpoint. Higher MMP-1 levels were associated with a worse outcome, even after adjustment for clinical and imaging predictors (1.026, 95% CI 1.002-1.051, p = 0.037), but CITP and CITP:MMP-1 were not. Combining MMP-1 and PICP improved the goodness-of-fit (LHR36.67, p = 0.004). The degree of myocardial cross-linking (CITP:MMP-1) is associated with myocardial longitudinal contraction, and MMP-1 is an independent predictor of outcome in DCM patients.

Diterpenoid tanshinone attenuates the metastasis of non-small-cell lung cancer (NSCLC) cells by inhibiting the Cavin-1-mediated ERK/Smad2 signaling pathway.

The objective of this study is to explore the potential therapeutic role of diterpenoid tanshinone (DT) against non-small cell lung cancer (NSCLC) in vitro and elucidate the molecular mechanism involved in tumor metastasis. Human NSCLC lines (A549 and NCI-H1299) were transfected with pcDNA3.1-Cavin-1 plasmids and corresponding controls. We tested the effects of DT on migration and invasion of lung cancer cells using transwell filters coated with fibronectin and Matrigel. Next, Quantitative Real-Time PCR and western blot were used to determine the transcriptional and protein levels of epithelial-mesenchymal transition (EMT) markers, transcription factors (Snail, Slug), and matrix metalloproteinases. As expected, Cavin-1 related to the enhanced ability of cell migration and invasion. DT not only inhibited the migratory and invasive capacity of Cavin-1-transfected NSCLC cells but also significantly increased the expression of ZEB1 and E-cadherin and decreased the level of N-cadherin, Vimentin, Snail, and Slug. Moreover, DT treatment obviously alleviated Cavin-1 overexpression-induced high levels of MMP2, MMP7, and MMP9 at both the protein and transcriptional levels. Furthermore, overexpressed Cavin-1 upregulated ERK and Smad2 signaling pathways in NSCLC cells, which were also strongly weakened by DT administration. Our results suggested that DT effectively attenuates Cavin-1-mediated NSCLC metastasis via the ERK/Smad2 signaling pathway.

Clinical value of combined serum MMP-2, MMP-9 and TIMP-1 for the prognosis of perianal fistula patients who received minimally invasive surgery.

This study aimed to investigate the clinical value of combined serum matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) for the prognosis of perianal fistula patients. Patients diagnosed and treated for perianal fistula by minimally invasive surgery (MIS) were enrolled. The concentrations of serum MMP-2, MMP-9 and TIMP-1 were measured at 24 h after surgery. Different levels of wound secretion, growth of granulation tissue and wound pain were used as criteria to evaluate surgical incision healing. The receiver operating characteristic curve was used to analyze the predicted assessment value. The concentrations of serum MMP-2 and MMP-9 were significantly higher, while the concentrations of serum TIMP-1 at 24 h after surgery were significantly lower in the poor healing group than in the good healing group. It was further found that high levels of serum MMP-2 and MMP-9 were risk factors for poor healing, while high concentrations of serum TIMP-1 at 24 h after surgery were protective factors for poor healing. High concentrations of serum MMP-2 and MMP-9 and low concentrations of serum TIMP at 24 h after surgery are risk factors for poor healing in perianal fistula patients who receive MIS, and the combined test has a higher predictive value.

59 Predictors and Impact of Pneumonia on Adverse Outcomes in Inhalation Injury Patients

Abstract Introduction Next to age and burn size, the presence of inhalation injury (II) is the third mortality prognostic factor for burn injury. II can lead to pulmonary complications such as pneumonia, acute respiratory distress syndrome (ARDS), and acute lung injury (ALI), all of which have been hypothesized to increase mortality in II. Herein, we aimed to identify variables associated with the risk of developing pneumonia and to determine the impact of pneumonia on selected outcomes. Methods De-identified data from the Prospective Inhalation study (ISIS) were used. II was confirmed by fiberoptic bronchoscopy. Baseline demographics, injury data, burn size, laboratory data, radiographic results, treatment, and hospital course were recorded. Ventilator use, complications, including pneumonia, ARDS, ALI, and death, were tracked daily for seven days and then weekly until discharge. Univariate and multivariate analysis were performed. Variables included in multivariate analysis had a p value of 0.1 or less. A p value of &amp;lt; 0.05 was considered significant. Results Of the 108 ISIS study group, 106 had complete data and were included in the analysis. Median age was 48.5 y, 59.4% of the subjects were male, 76.4% White and 9.4% Black. Regarding complications, 46.2% of the subjects developed pneumonia, 11.3% ARDS, 9.4% ALI, and 17% died. On univariate analysis, subjects who developed pneumonia were more likely to be white (87.8% vs. 66.7%, p = 0.006), to present with higher total burn surface area (24.70 [8.00-49.50] vs. 2.85 [0.00-25.38], p = 0.002), and full thickness burn (55.1% vs. 31.6%, p = 0.018). On multivariate analysis, being white (p = 0.033) and receiving colloids in the first 24 h of admission (OR = 3.582 [1.157-11.087], p = 0.027) were associated with increased risk of pneumonia while higher level of MMP-9 in the plasma (OR = 0.999 [0.998-1], p = 0.041) lowered the risk. Admission radiographs, FB score, clinical examination findings, and other demographic factors were not related to developing pneumonia. Pneumonia was independently associated with increased risk of developing ARDS (OR = 8.496 [1.322-54.61], p = 0.024) but not with ALI or mortality. Pneumonia was also associated with a longer hospital length of stay (B = 15.2 [2.4-28], p = 0.020) and higher number of days on a ventilator (B = 15.6 [8.2-23], p &amp;lt; 0.001). Conclusions Pneumonia is associated with increased LOS and ventilator days but not mortality. Those who received colloids were 3.5 times more likely to develop pneumonia. The relationship between colloid and pneumonia requires more research, however, the search for other intervenable clinical variables to prevent pneumonia remains elusive. Applicability of Research to Practice As colloids may affect the glycocalyx and contribute to inflammation, their role in developing pneumonia needs to be elucidated.

Levels of Matrix Metalloproteinase 9 Are Elevated in Persons With Schizophrenia or Bipolar Disorder: The Role of Modifiable Factors

BackgroundMatrix metalloproteinases (MMPs) are a diverse set of enzymes associated with inflammation. MMP-9 is of particular interest because it has been associated with autoimmune and cardiopulmonary disorders, tobacco smoking, and obesity, prevalent in psychiatric populations. MethodsSensitive enzyme immunoassays measured MMP-9 in blood samples from 1121 individuals (mean age = 35.6 [SD = 13.0] years; 47.7% male; 440 with schizophrenia, 399 with bipolar disorder, and 282 without a psychiatric disorder). We estimated the odds of diagnosis associated with MMP-9, demographic variables, tobacco smoking, and obesity, and also the partial explained variance using regression methods. We also determined the association between psychiatric medications and MMP-9 levels. ResultsIndividuals with elevated MMP-9 levels had higher odds of schizophrenia or bipolar disorder compared with the nonpsychiatric group adjusted for demographic variables. Partial correlation analyses indicated the demographic-adjusted variance associated with MMP-9, smoking, obesity, and their interaction explained 59.6% for schizophrenia and 39.9% for bipolar disorder. Levels of MMP-9 were substantially lower in individuals receiving valproate, particularly relatively high doses. ConclusionsIndividuals with higher levels of MMP-9 have significantly higher odds of schizophrenia or bipolar disorder. Individuals receiving valproate had substantially lower levels of MMP-9, possibly related to its ability to inhibit histone deacetylation. A substantial portion of the variance in clinical disorders associated with MMP-9 can be attributed to smoking or obesity. Interventions to reduce smoking and obesity might reduce the morbidity and mortality associated with elevated MMP-9 levels and improve the health outcomes of individuals with these disorders.

Assessment of YME1L and mitofusin2 as a possible diagnostic and/or therapeutic target in hepatocellular carcinoma

Mitochondrial dynamics are a contraversal issue in hepatocellular carcinoma. The present study tries to illustrate the role of mitochondrial dynamics proteins (mitofusin-2 (Mfn2) and YME1L) in hepatocarcinogenesis. Five groups were used: the control group and three HCC groups (after 8, 16, and 24 weeks from DENA induction). The last group was treated with Sorafenib (SP) (10 mg/kg), via oral gavage for 4 weeks after cancer induction. This study revealed that Mfn-2 was downregulated and YME1l was overexpressed in different HCC groups. This dysregulation of mitochondrial dynamics proteins was associated with high hepatic levels of cyclin D1, MMP-9, and MDA and overexpression of ki67 as well as decreasing the hepatic expression of tissue inhibitor of matrix metalloproteinase-3 (Timp-3) and Bax. To confirm the possible role of Mfn2 and YME1L in HCC, we assessed the effect of sorafenib on these parameters and its related HCC characteristics. Sorafenib corrected the level of Mfn2 and YME1L and decreased tumor cell proliferation as well. We also elucidated that mitochondrial dynamics proteins (Mfn2 and YME1L) could be a good therapeutic target for HCC.

Markers of blood-brain barrier disruption increase early and persistently in COVID-19 patients with neurological manifestations.

Coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 infection is associated with disorders affecting the peripheral and the central nervous system. A high number of patients develop post-COVID-19 syndrome with the persistence of a large spectrum of symptoms, including neurological, beyond 4 weeks after infection. Several potential mechanisms in the acute phase have been hypothesized, including damage of the blood-brain-barrier (BBB). We tested weather markers of BBB damage in association with markers of brain injury and systemic inflammation may help in identifying a blood signature for disease severity and neurological complications. Blood biomarkers of BBB disruption (MMP-9, GFAP), neuronal damage (NFL) and systemic inflammation (PPIA, IL-10, TNFα) were measured in two COVID-19 patient cohorts with high disease severity (ICUCovid; n=79) and with neurological complications (NeuroCovid; n=78), and in two control groups free from COVID-19 history, healthy subjects (n=20) and patients with amyotrophic lateral sclerosis (ALS; n=51). Samples from COVID-19 patients were collected during the first and the second wave of COVID-19 pandemic in Lombardy, Italy. Evaluations were done at acute and chronic phases of the COVID-19 infection. Blood biomarkers of BBB disruption and neuronal damage are high in COVID-19 patients with levels similar to or higher than ALS. NeuroCovid patients display lower levels of the cytokine storm inducer PPIA but higher levels of MMP-9 than ICUCovid patients. There was evidence of different temporal dynamics in ICUCovid compared to NeuroCovid patients with PPIA and IL-10 showing the highest levels in ICUCovid patients at acute phase. On the contrary, MMP-9 was higher at acute phase in NeuroCovid patients, with a severity dependency in the long-term. We also found a clear severity dependency of NFL and GFAP levels, with deceased patients showing the highest levels. The overall picture points to an increased risk for neurological complications in association with high levels of biomarkers of BBB disruption. Our observations may provide hints for therapeutic approaches mitigating BBB disruption to reduce the neurological damage in the acute phase and potential dysfunction in the long-term.

Matrix metalloproteinases and their gene polymorphism in young ST-segment elevation myocardial infarction

BackgroundGenetic polymorphism in MMPs are associated with multiple adverse CV events. There is little evidence regarding role of MMPs and their genetic polymorphisms in young (<50 years) ST-segment elevation myocardial infarction (STEMI) patients. MethodsThis study included 100 young (18–50 years) STEMI patients and 100 healthy controls. Serum levels of MMP-3, MMP-9 and TIMP were estimated for both patients as well as controls. Additionally, genetic polymorphisms in the MMP-9 gene (−1562 C/T and R279Q) & MMP-3 gene (5A/6A-1612) was evaluated. All these patients were followed up for one year and major adverse cardiac events (MACE) were determined. ResultsSerum levels of MMP-3 (128.16 ± 115.81 vs 102.3 ± 57.28 ng/mL; P = 0.04), MMP-9 (469.63 ± 238.4 vs 188.88 ± 94.08 pg/mL; P < 0.0001) and TIMP (5.84 ± 1.93 vs 2.28 ± 1.42 ng/mL; P < 0.0001) were significantly higher in patients as compared to controls. Additionally, patients with genetic polymorphisms in the MMP genes (5A/5A, 6A/6A and the AG genotypes) had an increased risk of STEMI. Patients with MACE had significantly higher levels of MMP-9 (581.73 ± 260.93 vs 438.01 ± 223.38 pg/mL; P = 0.012). A cutoff value of 375.5 pg/mL of MMP-9 was best able to discriminate patients with STEMI and MACE with sensitivity of 77.3% and specificity of 57%. ConclusionNovel biomarkers such as MMP-3, MMP-9 and TIMP and their genetic polymorphism are associated with the susceptibility for STEMI in young individuals. Higher MMP-9 levels in STEMI patients with MACE suggests its potential role in predicting cardiac remodeling and left ventricular dysfunction.

Matrix metalloproteinase gene mutations and bioinformatics of telocytes in hepatocellular carcinoma.

Telocytes (TCs) have crucial functions to promote the metastasis of hepatocellular carcinoma (HCC) by over-expressing matrix metalloproteinases (MMPs), but the mechanism by which TCs secrete MMPs in the genome is still unknown. We first cultured and isolated primary TCs from distinct liver cancer tissues and hepatic hemangioma surrounding tissues (Control group). Their whole exon genes were tested by Illumina HiSeq family of platforms and by high-throughput sequencing as well as variant mutations. Moreover, immunohistochemistry, Western blot, and quantitative reverse transcription-polymerase chain reactionassays were utilized to assess the expression of MMPs. The perniciousness of signal-nucleotide polymorphism (SNP) mutations of proteins were predicted by the Polyphen-2 database. Divergent expression and overall survival (OS) of MMPs was screened by StarBase-Pan Cancer plate; and MMPs associated signaling pathways were found by Kyoto Encyclopedia Genes andGenomes. The "competing endogenous RNA(ceRNA)" network was constructed by Cytoscape software. We found that 12 specific types of SNP mutations related to 5 types of MMPs occurred in TCs of liver malignant tumors as a potential result of MMP1, MMP9, and MMP17 overexpression. High levels of MMP1, MMP7, and MMP9 represented poor OS in HCC, and an interactive network of MMPs is shown. Allele shifts of C/T (rs20544) and G/C (rs2250889) in MMP9 were risk factors for TCs in HCC by the prediction of the Polyphen-2 Database. (MMP9(-3 C/T)) mutation might be a genetic mechanism of upregulating MMP9 in TCs.

Upregulation of SGLT1 and 2 promotes oxidative stress in right atrial appendages of patients with low-grade inflammatory responses: potential role in atrial fibrillation

Abstract Introduction Atrial fibrillation (AF), the most common cardiac arrhythmia, is associated with increased morbidity and mortality. Inflammatory infiltrates have been observed in atrial tissues from AF patients suggesting that low-grade inflammation contributes to the pro-fibrotic, pro-remodeling and pro-senescence responses of the atrial tissue favoring AF. Although sodium glucose co-transporter2 inhibitors (SGLT2i) have shown beneficial effects in heart failure, the expression and role of SGLT2 in cardiac tissues remain unclear. Purpose This study examined the expression of SGLT1 and 2 in right atrial appendages (RAA) of cardiac patients, and, if so, to determine the underling mechanism and their functional role. Methods RAA were harvested from patients undergoing coronary artery bypass surgery or aortic valve replacement at the University Hospital of Strasbourg. Patients with comorbidities such as malignancies and chronic inflammatory diseases were excluded. The expression level of markers was assessed using RT-qPCR, Western blot analysis, and immunofluorescence staining, and the level of reactive oxygen species using dihydroethidium staining. Results Analysis of RAA samples evidenced up-to a 4-fold difference in the NOS3 mRNA level. Those with low levels of NOS3 mRNA showed high levels of ICAM1, F3, MMP9, TGF-β1, TP53, CDKN1A, CDKN2A, SLCA1, SLCA2, IL1B, IL6, TNF-a and CD68 whereas the contrary was observed in those with high levels of NOS3 mRNA. SGLT1/2 mRNA levels were negatively correlated with that of eNOS and positively with those of IL-1, IL-6, TNF- α, CD68, and the senescence marker p53. Western blot analysis of RAA indicated that those expressing low levels of eNOS protein showed high levels of ICAM-1, VCAM-1, TF, MMP-2, TGF-b, SGLT1, SGLT2, p-p65, p53, p21 and p16 proteins whereas the contrary was observed in those with high levels of eNOS protein. Immunofluorescence staining indicated that SGLT1/SGLT2 signals were colocalized with those of CD31, an endothelial cell marker, in RAA, and associated with those of CD68 and TNF-α. Samples with high levels of SGLT1 and 2 displayed higher levels of oxidative stress that were inhibited by the antioxidant NAC (N-acetylcysteine), VAS-2870 (NADPH oxidase inhibitor), perindoprilat (ACE inhibitor), losartan (AT1R antagonist), empagliflozin (selective SGLT2 inhibitor), sotagliflozin (dual SGLT1/2 inhibitor) and by infliximab (TNF-α receptor neutralizing antibody). Conclusion The findings indicate that high expression levels of SGLT1 and 2 are observed in RAA showing low eNOS levels associated with pro-inflammatory, pro-senescent, pro-remodeling and pro-fibrotic responses. They were further associated with increased levels of oxidative stress sensitive to inhibitors of either the local angiotensin system, TNF-alpha or SGLT2. Thus, inhibition of SGLT2 appears to be an interesting approach to reduce the stimulatory pro-oxidant signal in atrium related to low-grade inflammation. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Boehringer Ingelheim Pharma GmbH &amp; Co. KG

SGLT1/2 expression in the heart of transplanted patients is associated with inflammatory induction of oxidative stress: potential protective effect of SGLT2 inhibitors

Abstract Introduction Heart transplantation is a life-saving surgical procedure for patients with end-stage cardiac dysfunction, however, such procedures are usually at risk of rejection due to acute inflammatory activation. While such inflammatory induction leaves the newly transplanted heart at risk of functional and structural remodeling subsequent to oxidative damage, current anti-inflammatory treatment options expose the patient to an elevated risk for adverse reactions in addition to absence of cardio-protective effects. Therefore, novel therapies with anti-inflammatory and cardio-protective dual effects are needed. Recently, our group has reported that low-grade inflammation is associated with upregulation of SGLT1/2 in arteries of human with cardiovascular diseases. Yet, the role and function of SGLT2 in human cardiac tissue remains poorly understood. Aim This study focuses on the expression pattern of SGLT1/2 in cardiac biopsies of heart transplanted patients and aim to identifying their functional impact. Methods Routine endomyocardial Biopsies (23) were performed for the detection of acute rejection heart transplanted patients (less than 2 years) at our University Hospital. Gene expression levels were assessed using RT-qPCR, the in situ tissue localization of proteins by immunofluorescence staining, and the level of oxidative stress by dihydroethidium staining. Results Gene expression analysis revealed strong inflammatory reaction in 5 samples indicated by at least 20-fold higher levels of mRNA of IL1B, IL6, TNFA and CD68 compared to the other 18 samples and concomitant with high expression levels of SLC5A1, SLC5A2, AT1R, CYBA, NCF1, ICAM1, VCAM1, MMP2, MMP9 and TGFB1 in contrast to low levels of NOS3. In addition, increased levels of oxidative stress were observed in the same biopsies, which were diminished by the antioxidant N-acetylcysteine (NAC), NADPH oxidase inhibitor (VAS-2870), TNF-α receptor neutralizing antibody (infliximab), ACE inhibitor (perindoprilat), AT1R antagonist (losartan), dual SGLT1/2 inhibitor (sotagliflozin) and selective SGLT2 inhibitor (empagliflozin) with inhibitory effects reaching up to 80%. Immunofluorescence staining indicated signals for nitro-tyrosine, TNF-alpha, SGLT1 and 2 in several samples. Conclusion These findings indicate that both isoforms SGLT1 and SGLT2 are expressed in the transplanted human heart and suggest a pattern of expression associated with pro-inflammatory response. They further indicate a potential protective effect of SGLT2 inhibitors in transplanted hearts through mitigating oxidative stress and hence providing a possible novel therapy for heart transplantation recipients to preserve the heart function. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Boehringer Ingelheim Pharma GmbH &amp; Co. KG

Combined Silymarin and Cotrimoxazole Therapy Attenuates Pulmonary Fibrosis in Experimental Paracoccidioidomycosis.

Paracoccidioidomycosis (PCM), which mainly affects rural workers, is a systemic mycosis caused by the Paracoccidioides genus that induces pulmonary sequelae in most adult patients, causing serious disability and impairing their quality of life. Silymarin is herbal medicine with an effective antifibrotic activity. Considering that in PCM, antifibrotic treatment is still not available in pulmonary fibrosis, we aimed to evaluate combined silymarin and cotrimoxazole (CMX) therapy via the intratracheal route in BALB/c mice infected with P. brasiliensis yeast. After 12 weeks of treatment, the lungs were collected for the determination of fungal burden, production of OH-proline, deposition of collagen fibers, pulmonary concentrations of cytokines, and expression of fibronectin, α-SMA, MMP-2, MMP-9, and TIMP-2. Spleen cell cultures were also performed. Our results showed that infected mice treated with combined silymarin/CMX showed lower deposition of collagen fibers in the lungs and lower pulmonary concentrations of hydroxyproline than the placebo groups. Decreased levels of TGF-β1 and fibronectin and high levels of MMP-2 and IFN-γ were also observed in this group of mice. Collectively, our findings indicate that the combination of antifungal treatment with silymarin has a potent antifibrotic effect associated with an immunomodulatory effect that potentializes the antifungal immune response.

FEATURES OF EXPRESSION OF GROWTH MEDIATORS AND MATRIX METALLOPROTEINASES AT THE LOCAL LEVEL IN PATIENTS WITH LOWER LIMB VARICOSE VEINS, CLINICAL CLASS C4 - C6

The purpose of the research was to study the features of the expression of growth mediators and matrix metalloproteinases in patients with lower limb varicose veins, clinical classes C4-C6. The study included 98 patients. The control group consisted of 102 healthy donors. The levels of VEGF, PDGF, matrix metalloproteinases (MMP-2, MMP-9), superoxide dismutase, and prolidase were studied. The average value of VEGF in pathologically altered vein (PAV) wall samples was 40.1 ± 6.2 ng/g of protein and was statistically significantly higher - 1.7 times compared with the same value in persons from CG 2 (23.0 ± 5.9 ng/g protein, p=0.001). Patients with lower limb varicose veins (LLVV) showed an inverse correlation between VEGF levels and SOD activity in blood plasma and PAV (r=-0.52; p=0.002 and r=-0.59; p=0.001, respectively). An increase in the level of PDGF correlated with an increase in the oxidative stress index (OSI) in the blood plasma and the PAV wall (r=0.59; p=0.022 and r=0.61; p&lt;0.001). In patients with LLVV, a high level of MMP-9 inversely correlated with the total oxidant capacity (TAC) in blood plasma and in the varicose vein wall (r=-0.48; p&lt;0.001 and r=-0.52; p=0.012). In patients with LLVV, there were statistically significant relationships between prolidase activity and OSI and TAC values ​​in the PAV wall (r=0.62; p=0.003 and r=-0.59; p&lt;0.001). It has been established that VEGF, PDGF, MMPs play a key role in the LLVV pathogenesis; the dynamics of these indicators can be used to predict the effectiveness of surgical treatment and the risk of recurrence.

Comparison of matrix metalloproteinase 9 and 14 levels in vitreous samples in diabetic and non-diabetic patients: a case control study

BackgroundMMP-9 plays a prominent role in inflammation and MMP-14 take part in angiogenesis. The objective of this study is to compare MMP-9 and MMP-14 levels between diabetic and non-diabetic patients.MethodsThe patients who scheduled for pars plana vitrectomy were included in our study. Patients are divided into 2 groups: the diabetic group and non-diabetic group. Age, gender, intraocular pressure(IOP), visual acuity (VA) were reported. Color fundus photography, fundus fluorescein angiography, optic coherence tomography (OCT) were performed before and after the operation. MMP-9 and MMP-14 levels in vitreous samples were analyzed with a reader device by ELISA method. Mann–Whitney U test and logistic regressions were used in statistical analysis, p < 0.05 accepted as statistically significant.Results70 eyes of 70 patients who received pars plana vitrectomy were enrolled in the study and divided into 2 groups: 34 patients in the diabetic group, 36 patients in the non-diabetic group. The average age of diabetic patients was 60.14 ± 10.20, and non-diabetic patients was 64.22 ± 11.16, respectively. The average MMP-9 (0.67 ± 0.66 ng/ml) and MMP-14 (0.16 ± 0.45 ng/ml) values in the diabetic group were significantly higher than the average MMP-9 (0.21 ± 0.05 ng/ml) and MMP-14 (and 0.07 ± 0.02 ng/ml) values in the non-diabetic group (P < 0.01). Also, it was observed that MMP-9 and MMP-14 levels increases as the diabetic disease duration increases. The risk of diabetes incidence increased with high levels of MMP-9 and MMP-14.ConclusionDue to the higher levels of MMP-9 and MMP-14 in the pathogenesis of diabetic retinopathy, these proteins may probably be among the therapeutic targets in the prevention and treatment of retinopathy.