Tissue Inhibitors Of Matrix Metalloproteinases Research Articles

Ameliorative effects of high intensity interval training and Lactobacillus rhamnosus GG Protect against tetracycline-induced fatty liver in rats: a gene expression profiling comparative study.

Exercise training and probiotics have been suggested as a treatment for the prevention of chronic liver damage such as non-alcoholic fatty liver disease (NAFLD). Lactobacillus rhamnosus Gorbach‐Goldin (LGG) is one of the most widely used probiotic strains that decreases liver damage. Thus, this study aims to consider the ameliorative effects of high intensity interval training (HIIT) and LGG against tetracycline-induced fatty liver in rats. Eighty male Wistar rats were randomly divided into 8 groups of (n=10 each group): control, LGG, HIIT, LGG+HIIT, NAFLD, NAFLD+LGG, NAFLD+HIIT, and NAFLD+LGG+HIIT. The rats are treated by intraperitoneal injection with 140 mg/kg−1 tetracycline, an antibiotic previously known to induce steatosis. The exercise training groups performed HIIT 5 days/week for 5 weeks. 107 colony-forming units (cfu) of LGG were gavaged for LGG groups 5 days/week for 5 weeks. Probiotic supplementation in combination with interval training significantly decreased tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) mRNA and matrix metalloproteinase-2 (MMP-2) mRNA in the liver (p<0.05), while the levels of lysosomal acid lipase (LIPA) mRNA was significantly increased compared to NAFLD group. Also, compared with NAFLD group, NAFLD+LGG, NAFLD+HIIT and NAFLD+LGG+HIIT groups showed a significant decrease in hepatic monocyte chemoattractant protein-1 (MCP-1). Compared to LGG and LGG+HIIT groups, all NAFLD groups showed a significant decrease in apolipoprotein C3 (apoc3) in liver tissue (p<0.05). The results suggested that interval exercise with LGG supplementation minimizes cell destruction and inflammation in liver tissue due to NAFLD by improving gene expression profiles.

Sp1 induced gene TIMP1 is related to immune cell infiltration in glioblastoma

Tumor immune microenvironment exerts a profound effect on the population of infiltrating immune cells. Tissue inhibitor of matrix metalloproteinase 1 (TIMP1) is frequently overexpressed in a variety of cells, particularly during inflammation and tissue injury. However, its function in cancer and immunity remains enigmatic. In this study, we find that TIMP1 is substantially up-regulated during tumorigenesis through analyzing cancer bioinformatics databases, which is further confirmed by IHC tissue microarrays of clinical samples. The TIMP1 level is significantly increased in lymphocytes infiltrating the tumors and correlated with cancer progression, particularly in GBM. Notably, we find that the transcriptional factor Sp1 binds to the promoter of TIMP1 and triggers its expression in GBM. Together, our findings suggest that the Sp1-TIMP1 axis can be a potent biomarker for evaluating immune cell infiltration at the tumor sites and therefore, the malignant progression of GBM.

Poster 217: Local and Systemic Comparison of Senescent Profiles in Acute Knee Injury

Poster 217: Local and Systemic Comparison of Senescent Profiles in Acute Knee Injury

Evaluation of Serum Levels of Tissue Inhibitor Metalloproteinase-1and Matrix Metalloproteinase-1 in Egyptian Patients with Diabetic Nephropathy

We aimed to evaluate the levels of serum tissue inhibitor metalloproteinase-1 (TIMP-1) and matrix metalloproteinase-1 (MMP-1) in patients with type 2 diabetes (T2D)alone and in diabetic nephropathy(DN) and assess their relation with other clinical features. This study was conducted on 30 patients withT2D, 30 with diabetic nephropathy (DN) and 50 age- corresponded healthy individuals. Serum MMP-1 and TIMP-1 were measured with ELISA in patients and controls. Elevated serum concetrations of TIMP-1 and MMP-1 were recognized in DN group compared to patients with T2D alone as well as controls. Positive correlation was also observed between MMP-1, TIMP-1, FBG and HbA1c in DN group. In multiple linear regression analysis HBLA1c, TG and low HDL were selected as components significantly related to MMP-1 and TIMP-1 in DN patients. in conclusion, serum MMP-1 and TIMP-1were significantly increased in DN. Further, in DM type 2 both markers were not significantly increased than controls. These blood biomarkers are associated with DN and might be salutary in clinical discrimination of DN in patients with type 2 diabetes.

In vitro effects of Eucommia ulmoides and its active components on the growth, lipid metabolism and collagen metabolism of grass carp (Ctenopharyngodon idellus) hepatocyte and intramuscular fibroblast.

Two experiments were conducted to investigate the in vitro effects of Eucommia ulmoides (E. ulmoides) and its active components on the growth, lipid metabolism and collagen metabolism of grass carp's (Ctenopharyngodon idellus) hepatocytes and intramuscular fibroblasts. In experiments 1 and 2 (Expt. 1, 2), hepatocytes and intramuscular fibroblasts were treated with 2.5, 5, 10, 20, 40 and 80 μg ml-1 of Eucommia bark extract (EBE), Eucommia leaf extract (ELE), pinoresinol diglucoside (PDG), chlorogenic acid (CGA), quercetin (QC) and aucubin (AU) for 24 h, respectively, then the cell growth, lipid and collagen metabolism-related gene expressions were evaluated. The results showed that the cell proliferation rate of hepatocytes and intramuscular fibroblasts was significantly improved by the supplementation of EBE, ELE, CGA, QC and AU. Moreover, triglyceride concentration of hepatocytes was significantly decreased by the EBE, ELE, CGA and QC supplementations compared to the control. Meanwhile, EBE, ELE, CGA, QC and AU supplementations significantly upregulated the relative gene expressions of insulin-like growth factor-1 (igf1), protein kinase B (akt), target of rapamycin (tor) and eukaryotic initiation factor 4E binding protein 1 (4ebp1) in hepatocytes, and ribosomal protein S6 kinase 1 (s6k1) transcription was significantly activated by ELE, CGA and QC supplementations. Nonetheless, phosphatidylinositol 3-kinase (pi3k) was unaffected by any of the supplements. In addition, the mRNA expressions of genes associated with lipid metabolism (peroxisome proliferator activated receptor α pparα, carnitine palmitoyltransferase 1 cpt1, adipose triglyceride lipase atgl, hormone-sensitive lipase hsl, peroxisome proliferator activated receptor γ pparγ) were significantly upregulated by EBE, ELE, CGA and QC. In intramuscular fibroblasts, the EBE, ELE, CGA, QC and AU supplementations significantly increased in vitro hydroxyproline concentrations, promoted the relative expressions of transforming growth factor-β1 (tgfβ1), connective tissue growth factor (ctgf), collagen type I alpha 1/2 chain (col1a1, col1a2), lysine oxidase (lox) and tissue inhibitor of matrix metalloproteinase-2 (timp2), and decreased matrix metalloproteinase-2 (mmp2) gene expression. Also, the gene expressions of drosophila mothers against decapentaplegic protein 2/4 (smad2, smad4) and proline hydroxylase (phd) were significantly upregulated by ELE, CGA, QC and AU supplementations. Based on the present in vitro results of grass carp, EBE, ELE, CGA, QC and AU improved the growth and lipid metabolism (except AU) in hepatocytes, and promoted the collagen deposition in intramuscular fibroblast, which is partly attributed to the signalling pathways of AKT/TOR, PPARα and TGF-β/Smads/CTGF.

Protective role of Achyranthes bidentata polysaccharides against chondrocyte extracellular matrix degeneration through lncRNA GAS5 in osteoarthritis.

Achyranthes bidentata polysaccharides (ABPS) is an active ingredient of the flowering plant Achyranthes bidentata that has been previously reported to be effective for the treatment of osteoarthritis (OA). However, the underlying molecular mechanism remain to be fully clarified. Emerging studies have shown that the long non-coding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) is involved in the pathogenesis of OA. Therefore, the present study aimed to investigate the potential mechanism of ABPS by focusing on its effects on the regulation of chondrocyte extracellular matrix (ECM) homeostasis, with particular emphasis on lncRNA GAS5. In the present study, the modified Hulth method was used to construct OA rats, which were gavaged with 400 mg/kg ABPS for 8 weeks. Histopathological changes in cartilage and subchondral bone were evaluated by hematoxylin-eosin staining and Safranin O-fast green staining. In in vitro experiments, IL-1β-treated chondrocytes were infected with Lenti-lncRNA GAS5. Fluorescence in situ hybridization assay was performed to measure the expression of the lncRNA GAS5 in chondrocytes. Moreover, the relative expression level of lncRNA GAS5 in cartilage tissue and chondrocytes was detected using reverse transcription-quantitative PCR. Western blot analysis was used to detect protein expression levels of MMP-9, MMP-13, TIMP-1, TIMP-3 and type II collagen in cartilage tissue and chondrocytes. The results indicated that ABPS delayed the degradation of the ECM by chondrocytes in addition to reducing lncRNA GAS5 expression both in vivo and in vitro. Furthermore, silencing of lncRNA GAS5 expression in IL-1β-treated chondrocytes downregulated the protein expression of MMP-9 and MMP-13 whilst upregulating the expression of tissue inhibitor matrix metalloproteinase (TIMP)-1, TIMP-3 and type II collagen. To conclude, the present study provides evidence that ABPS can inhibit the expression of lncRNA GAS5 in chondrocytes to regulate the homeostasis of ECM, which in turn may delay the occurrence of cartilage degeneration during OA.

Matrix Metalloproteinase-Targeted SPECT/CT Imaging for Evaluation of Therapeutic Hydrogels for the Early Modulation of Post-Infarct Myocardial Remodeling.

Following myocardial infarction (MI), maladaptive upregulation of matrix metalloproteinase (MMP) alters extracellular matrix leading to cardiac remodeling. Intramyocardial hydrogel delivery provides a vehicle for local delivery of MMP tissue inhibitors (rTIMP-3) for MMP activity modulation. We evaluated swine 10-14days following MI randomized to intramyocardial delivery of saline, degradable hyaluronic acid (HA) hydrogel, or rTIMP-3 releasing hydrogel with an MMP-targeted radiotracer (99mTc-RP805), 201Tl, and CT. Significant left ventricle (LV) wall thinning, increased wall stress, reduced circumferential wall strain occurred in the MI region of MI-Saline group along with left atrial (LA) dilation, while these changes were modulated in both hydrogel groups. 99mTc-RP805 activity increased twofold in MI-Saline group and attenuated in hydrogel animals. Infarct size significantly reduced only in rTIMP-3 hydrogel group. Hybrid SPECT/CT imaging demonstrated a therapeutic benefit of intramyocardial delivery of hydrogels post-MI and reduced remodeling of LA and LV in association with a reduction in MMP activation.

AMMP-8 Oral Fluid PoC Test in Relation to Oral and Systemic Diseases.

The manuscript uses the previously published literature and highlights the benefits of active-matrix metalloproteinase (aMMP)-8 chairside/point-of-care (PoC) diagnostic tools as adjunctive measures in oral and systemic diseases. Previous studies suggest that as a biomarker, aMMP-8 is more precise than total MMP-8, MMP-9, MMP-2, MMP-3, MMP-13, MMP-7, MMP-1, calprotectin, myeloperoxidase (MPO), human neutrophil elastase (HNE), tissue inhibitor of matrix metalloproteinase (TIMP)-1, and bleeding of probing (BOP). Therefore, aMMP-8 could be implemented as the needed key biomarker for the new disease classification for both periodontitis and peri-implantitis. With a sensitivity to the tune of 75–85% and specificity in the range of 80–90%, lateral flow aMMP-8 PoC testing is comparable to catalytic protease activity assays for aMMP-8. The test can be further applied to estimate the glycemic status of an individual, to ascertain whether a person is at risk for COVID-19, in managing the oral side effects of radiotherapy carried in head and neck cancers, and in selected cases pertaining to reproductive health. In the future, aMMP-8 could find application as a potential systemic biomarker in diseases affecting the cardiovascular system, cancers, bacteremia, sepsis, diabetes, obesity, meningitis, as well as pancreatitis. The aMMP-8 PoCT is the first practical test in the emerging new dental clinical field, that is, oral clinical chemistry representing oral medicine, clinical chemistry, peri-implantology, and periodontology.

Improved anti-fibrotic effects by combined treatments of simvastatin and NS-398 in experimental liver fibrosis models.

Background/AimsEfficient anti-fibrotic therapies are required for the treatment of liver cirrhosis. Hydroxymethylglutaryl-coenzyme A reductase inhibitors (statins) and cyclooxygenase-2 (COX-2) inhibitors have been reported to have anti-fibrotic effects. Here, we investigated whether combined treatment with a statin and a COX-2 inhibitor has synergistic anti-fibrotic effects.MethodsThe effects of treatment strategies incorporating both simvastatin and a COX-2 inhibitor, NS-398, were investigated using an immortalized human hepatic stellate cell line (LX-2) and a hepatic fibrosis mouse model developed using thioacetamide (TAA) in drinking water. Cellular proliferation was investigated via 5-bromo-2-deoxyuridine uptake. Pro- and anti-apoptotic factors were investigated through Western blotting and real-time polymerase chain reaction analysis.ResultsThe evaluation of the anti-proliferative effects on LX-2 cells showed that the observed effects were more pronounced with combination therapy than with single-drug therapy. Moreover, hepatic fibrosis and collagen deposition decreased significantly in TAA-treated mice in response to the combined treatment strategy. The mechanisms underlying the anti-fibrotic effects of the combination therapy were investigated. The effects of the combination therapy were correlated with increased expression levels of extracellular signal-regulated kinase 1/2 signaling molecules, upregulation of the Bax/Bcl-2 signaling pathway, inhibition of the transforming growth factor-β signaling pathway, and inhibition of tissue inhibitor of matrix metalloproteinases 1 and 2.ConclusionsThe combination of simvastatin and NS-398 resulted in a synergistic anti-fibrotic effect through multiple pathways. These findings offer a theoretical insight into the possible clinical application of this strategy for the treatment of advanced liver diseases with hepatic fibrosis.

Effect of SPARC Suppression in Mice, Perfused Human Anterior Segments, and Trabecular Meshwork Cells

PurposeSecreted protein, acidic and rich in cysteine (SPARC) elevates intraocular pressure (IOP), increases certain structural extracellular matrix (ECM) proteins in the juxtacanalicular trabecular meshwork (JCT), and decreases matrix metalloproteinase (MMP) protein levels in trabecular meshwork (TM) endothelial cells. We investigated SPARC as a potential target for lowering IOP. We hypothesized that suppressing SPARC will decrease IOP, decrease structural JCT ECM proteins, and alter the levels of MMPs and/or their inhibitors.MethodsA lentivirus containing short hairpin RNA of human SPARC suppressed SPARC in mouse eyes and perfused cadaveric human anterior segments with subsequent IOP measurements. Immunohistochemistry determined structural correlates. Human TM cell cultures were treated with SPARC suppressing lentivirus. Quantitative reverse transcriptase polymerase chain reaction (PCR), immunoblotting, and zymography determined total RNA, relative protein levels, and MMP enzymatic activity, respectively.ResultsSuppressing SPARC decreased IOP in mouse eyes and perfused human anterior segments by approximately 20%. Histologically, this correlated to a decrease in collagen I, IV, and VI in both the mouse TM and human JCT regions; in the mouse, fibronectin was also decreased but not in the human. In TM cells, collagen I and IV, fibronectin, MMP-2, and tissue inhibitor of MMP-1 were decreased. Messenger RNA of the aforementioned genes was not changed. Plasminogen activator inhibitor 1 (PAI-1) was upregulated in vitro by quantitative PCR and immunoblotting. MMP-1 activity was reduced in vitro by zymography.ConclusionsSuppressing SPARC decreased IOP in mice and perfused cadaveric human anterior segments corresponding to qualitative structural changes in the JCT ECM, which do not appear to be the result of transcription regulation.

Preparation and evaluation of photoprotective kenaf seed oil nanocarriers-based cream of tocotrienol-rich fraction

Applications of nanotechnology and vegetable oil in topical formulation have emerged to provide benefits to develop effective and safe advanced biocosmetic prototypes. The study aimed to optimize the ratio of UV filters in kenaf (Hibiscus cannabinus L.) seed oil nanocarrier to form a sunscreen formulation. The nanocarriers were characterized based on particle size, polydispersity index, encapsulation efficiency, UV absorption, Raman microscopy, and high-resolution transmission electron microscopy analysis. The selected nanocarrier was added to a tocotrienol-rich fraction (TRF) cream and subjected to photoprotection and functional analysis to determine its skin beneficial properties based on cellular antioxidant capacity and relative protein expression on fibroblasts. The results revealed that nanocarriers with particle sizes ranging from 180 nm to 240 nm showed > 90% of encapsulation efficiency for UV filters. Sample F5 contained 1.25% of diethylamino hydroxybenzoyl hexyl benzoate (DHHB) and 0.75% of ethylhexyl triazone (EHT), having a spherical shape at particle size 204.80 nm with > 95% of UV filter encapsulation and high intensity in both UVA and UVB spectra. The cream produced from F5 and TRF (NLC-TRF cream) indicated no cytotoxicity and was able to provide sun protection factor (SPF) value of 53 and cellular antioxidant capacity of 45.13%. Besides, a significant protein upregulation (1.29-fold for fibroblast growth factor, 7.77-fold for vascular endothelial growth factor, 11.21-fold for transforming growth factor-β1, and 1.69-fold for tissue inhibitor matrix metalloproteinase-1) and downregulation for matrix metalloproteinase-1 and − 2 resulted after 24 hr of treatment. The application of nanocarriers in TRF cream can enhance the photoprotection, skin-protective properties in cellular antioxidant capacity, and protein expression which could be a promising delivery system in sunscreen formulations.

TARGETING THE HISTONE DEMETHYLASE JMJD2B/KDM4B AS A NOVEL THERAPEUTIC APPROACH IN IL-13-MEDIATED FIBROSIS IN AIRWAY FIBROBLASTS OF SEVERE ASTHMATICS

TYPE: Abstract TOPIC: Allergy and Airway PURPOSE: Asthma is a heterogeneous disorder characterized by chronic inflammation and remodeling of the airways. Asthma responses are driven mainly by type 2 immune responses, where IL-13 plays a key role in asthma pathogenesis. We previously identified JMJD2B as an IL-13-regulated epigenetic modifier in asthmatic airway fibroblasts. Therefore, this study aimed to target JMJD2B to potentially alleviate IL-13-mediated fibrosis in asthma. METHODS: Bronchial fibroblasts isolated from asthmatic and healthy individuals were stimulated with IL-13 and treated with JIB-04, a pan-selective inhibitor of histone demethylase(s). Gene and protein expression of extracellular matrix markers were assessed using qRT-PCR, western blotting and matrix metalloproteinases (MMPs) activity assay. Chromatin immunoprecipitation (ChIP) assay was used to determine the bindig of JMJD2B to promoter region of Tissue inhibitors of matrix metalloproteinases (TIMPs). RESULTS: Here we show that JIB-04 inhibited JMJD2B activity by reducing the demethylation of its downstream target, H3K36me3, in asthmatic fibroblasts. Inhibition of JMJD2B significantly affected the viability of the bronchial fibroblasts at 48 hours. JMJD2B inhibition was further associated with the downregulation of extracellular matrix (ECM) proteins such as MMP-2, MMP-9, collagen-1, and fibronectin, and upregulation of TIMP-2, at both the gene and protein levels. This was accompanied by inhibition of IL-13 mediated MMPs activity. JIB-04 further prevented the association of JMJD2B on TIMP-2 promoter region. CONCLUSIONS: Our results demonstrate the potential of JMJD2B inhibition in attenuating IL-13-mediated fibrosis in asthma. CLINICAL IMPLICATIONS: Therapeutic targeting of JMJD2B using JIB-04 is a promising candidate to alleviate IL-13-mediated responses in chronic disorders such as asthma. DISCLOSURE: Nothing to declare. KEYWORD: Asthma

FEATURES OF CHANGES IN THE LEVELS OF MATRIX METALLOPROTEINASES AND THEIR INHIBITORS IN DIABETIC PATIENTS WITH RESISTANT HYPERTENSION AFTER RENAL DENERVATION

Objective: The family of matrix metalloproteinases (MMPs) involved in fibrogenesis, but the effect of renal denervation (RDN) on their levels is currently insufficiently studied and highly controversial. The aim of study was to evaluate the change in MMPs and their inhibitors during 2 years after RDN in patients with resistant hypertension (RHTN) and type 2 diabetes mellitus (DM). Design and method: Forty three patients with RHTN and type 2 DM were included in single-arm prospective interventional studies (protocol numbers NCT01499810 and NCT02667912 on ClinicalTrials.gov) (mean age 60.6 ± 8.8 years, mean office (systolic/diastolic) blood pressure (BP) 169.7/89.1 ± 19.2/15.2mmHg, HbA1c 6.7 ± 1.4%, 17(40%) men). All patients were undergone to ambulatory 24-hour BP and assessment of lab testes (plasma concentrations of MMP-9, MMP-2, tissue inhibitor of MMP type 1 (TIMP1), TIMP-1/MMP-2 and TIMP-1/MMP-9 ratios) at baseline and at 6–12–24 months follow-up. The patients were advised not to change their medication regimen during the study. Results: At baseline 70%/16%/17% of patients had an elevated level of MMP-9/MMP-2/TIMP-1, respectively. At 6 months after RDN there was a significant decrease in mean 24-hour systolic/diastolic BP (SBP/DBP) (-8.8 [95%CI -1.3;-18.4]/-8.6 [95%CI -2.5;-4.7] mmHg, P &lt; 0.05). The number of responders (patients with decrease in 24-hour SBP 10 mmHg and more) was 26 (61%). There were no significant changes in the mean levels of MMPs, TIMT1 and their ratios at 6-12-24 months follow up. At the same time, at 2 years after RDN there was a significant decrease in level of MMP-9 in responders compared with non-responders (-319.6[-2.3;-636.8] ng/mL, p = 0.04 vs 66.6 [221.4;-354.5] ng/mL, p = 0.63). Conclusions: Our data indicate the decrease in the MMP-9 level at 2 years after RDN only in the responders, that may be due to a decrease in the hemodynamic load and peripheral vasodilation that no longer requires the involving of proteolytic systems for an improvement in tissue blood flow.

A quality of life, clinical and biochemical improvements after catheter ablation of persistent arrhythmia in patients with structural heart disease and arrhythmia-mediated cardiomyopathy.

Arrhythmia-mediated cardiomyopathy (AMC) is an essential clinical situation that is commonly underdiagnosed. Successful arrhythmia control leads to improvement in health-related quality of life (HRQoL) and heart failure (HF) symptoms in patients with structural heart disease (SHD). The study aimed to evaluate the impact of catheter ablation (CA) of persistent arrhythmia on HRQoL, biochemical and clinical parameters HF in patients with SHD and AMC. Patients with SHD, on optimal medical treatment, with persistent arrhythmia and strong suspicion of AMC, scheduled for CA were prospectively enrolled. Study procedures included: HRQoL measurement (the Minnesota Living with Heart Failure Questionnaire [MLHFQ] and the EuroQol Research Foundation [EQ-5D-3L] questionnaire), biomarkers (N-terminal pro-B-type natriuretic peptide [NT-proBNP], troponin T [TnT], matrix metalloproteinase-9 [MMP-9], soluble suppression of tumorigenesis-2 [sST2], tissue inhibitor of matrix metalloproteinase-1 [TIMP-1]), transthoracic echocardiography and clinical assessment. At 6 months, 30/35 (86%) patients were free of persistent arrhythmia. Patients who underwent successful CA had a significant improvement in HRQoL: MLHFQ (median [interquartile range, IQR], -22 [-28; -11]; P <0.001), EQ5D-3L score (mean [standard deviation], 21.8 (16.8); P <0.001); EQ5D-3L index (median [IQR], 0.09 (0.05; 0.18); P <0.001). A significant decrease in injury bio markers was observed: NT-proBNP (median [IQR], -414 [-1397; -318] pg/ml; P <0.001), TnT (median [IQR], -2.27 (-8.52; 0.55) ng/l; P <0.01) but not in fibrosis biomarkers: (median [IQR], sST2: 2.20 [-5.4; 4.3] ng/ml; P = 0.741, MMP-9: 34 [-376; 283] ng/ml; P = 0.881, TIMP-1: 11.1 [-17.1; 31.9] ng/ml; P = 0.215). There was a significant increase of left ventricular ejection fraction (LVEF) (mean [SD], 9.8 [5.9] %; P <0.01). Successful CA significantly improved clinical status, LVEF, and HRQoL of patients with SHD and AMC.

Treatment of Abdominal Aortic Aneurysm Utilizing Adipose-Derived Mesenchymal Stem Cells in a Porcine Model

IntroductionThe current treatment paradigm of abdominal aortic aneurysms (AAA) focuses on observing patients until their disease reaches certain thresholds for intervention, with no preceding treatment available. There is an opportunity to develop novel therapies to prevent further aneurysmal growth and decrease the risk of a highly morbid rupture. We used a porcine model of aortic dilation to assess the ability of human adipose-derived mesenchymal stem cells (MSCs) to attenuate aortic dilation. Materials and methodsTwelve Yorkshire pigs received periadventitial injections (collagenase and elastase) into a 4-cm segment of infrarenal aorta. Animals were treated with either 1 × 106 MSCs placed onto Gelfoam or treated with media as a control. Aortic diameters were measured at the time of surgery and monitored at postoperative day (POD) 7 and 14 with ultrasound. Animals were sacrificed on POD 21. Aortic tissue was harvested for histopathological analyses and immunohistochemistry. Groups were compared with paired t-tests or Mann–Whitney U-tests. ResultsAll animals survived until POD 21. The mean aortic diameter was reduced in the aortic dilation + MSC treatment group compared to aortic dilation control animals (1.10 ± 0.126 versus 1.48 cm ± 0.151, P < 0.001). Aortic media thickness was reduced in the aortic dilation group compared to the aortic dilation + MSC group (609.14 IQR 445.21-692.93 μm versus 643.55 IQR 560.91-733.88 μm, P = 0.0048). There was a significant decrease in the content of collagen and alpha-smooth muscle actin and elastin perturbation in the aortic dilation group as compared to the aortic dilation + MSC group. Immunohistochemistry demonstrated an increased level of vascular endothelial growth factor, tissue inhibitor of matrix metalloproteinase 1, and tissue inhibitor of matrix metalloproteinase 3 expression in the aorta of aortic dilation + MSC animals. ConclusionsStem cell therapy suppressed the aortic dilation in a porcine model. Animals from the aortic dilation group showed more diseased gross features, histologic changes, and biochemical properties of the aorta compared to that of the aortic dilation + MSC treated animals. This novel finding should prompt further investigation into translatable drug and cell therapies for aneurysmal disease.

Correlation of Matrix Metalloproteinase-9 and Tissue Inhibitor Matrix Metalloproteinase-1 on Lactate Concentration in Sepsis patients Admitted to Intensive Care Unit

BACKGROUND: Lactate level has been used not only as a biomarker for diagnosis and guiding treatment of sepsis but also as predictor of poor clinical outcomes. Elevated lactate does not specifically reflect cellular damage and this condition can be seen in other metabolic disorders. Matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are two new promising biomarkers that have been reported to elevate significantly in sepsis. These two biomarkers can reflect physiological changes in tissue and cellular levels. AIM: This study aims to identify the correlation of MMP-9, TIMP-1, and MMP-9/TIMP-1 on lactate levels in sepsis patients. METHODS: This was a cross-sectional study conducted in two hospitals, Adam Malik General Hospital, Medan, and Grand Medistra Hospital, Deli Serdang, between April 2020 and May 2021. The inclusion criteria in this study were adult sepsis patients who were admitted to ICU, with Sequential Organ Failure Assessment (SOFA) or quick SOFA score ≥ 2. We recorded the characteristics, MMP-9, TIMP-1, and lactate concentration before given any intervention. The data were then analyzed to find the correlation. RESULTS: Sixty-four patients were included in this study which consisted of almost equal men and women. The mean age of the subjects was 52.16 ± 16.25 years old. There was no correlation between MMP-9 and TIMP-1 toward lactate concentration (p = 0.466 and p = 0.65, respectively). CONCLUSION: Our study showed no correlation between MMP-9 and TIMP-1 toward lactate concentration.

Genipin, an Inhibitor of UCP2 as a Promising New Anticancer Agent: A Review of the Literature.

Genipin is a protein cross-linking agent extracted from Gardenia (Gardenia jasminoides Ellis) fruits. This fruit has conventionally been used as a Chinese herbal medicine for the treatment of inflammation and jaundice and as an edible colorant in oriental countries. Uncoupling protein (UCP)-2 is a member of the family of uncoupling proteins, which are anion transporters positioned in the mitochondrial inner membrane. Genipin has been shown to have hepatoprotective activity, acting as an effective antioxidant and inhibitor of mitochondrial UCP2, and is also reported to exert significant anticancer effects. In this review, the author presents the latest progress of genipin as an anticancer agent and concisely describes its various mechanisms of action. In brief, genipin inhibits UCP2 to attenuate generation of reactive oxygen species (ROS), leading to ROS/c-Jun N-terminal kinase-dependent apoptosis of cancer cells. Genipin also increases the tissue inhibitors of matrix metalloproteases (MMP)-2, a kind of tumor promoter in a variety of cancers, as well as induces caspase-dependent apoptosis in in vitro and in vivo models. These findings suggest that genipin can serve as a promising novel antitumor agent that could be applicable for chemotherapy and/or chemoprevention for cancers.

Susceptibility to kidney fibrosis in mice is associated with early growth response-2 protein and tissue inhibitor of metalloproteinase-1 expression

Patients with chronic kidney disease and experimental animal models of kidney fibrosis manifest diverse progression rates. Genetic susceptibility may contribute to this diversity, but the causes remain largely unknown. We have previously described kidney fibrosis with a mild or severe phenotype in mice expressing transforming growth factor-beta1 (TGF-β1) under the control of a mouse albumin promoter (Alb/TGF-β1), on a mixed genetic background with CBAxC57Bl6 mice. Here, we aimed to examine how genetic background may influence kidney fibrosis in TGF-β1 transgenic mice, and in the unilateral ureteral obstruction (UUO) and subtotal nephrectomy (SNX) mouse models. Congenic C57Bl6(B6)-TGFβ and CBAxB6-TGFβ (F1) transgenic mice were generated and survival, proteinuria, kidney histology, transcriptome and protein expressions were analyzed. We investigated the kidneys of B6 and CBA mice subjected to UUO and SNX, and the effects of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) neutralization on the fibrotic process. CBAxB6-TGFβ mice developed severe kidney fibrosis and premature death, while B6-TGF-β mice had mild fibrosis and prolonged survival. Kidney early growth response factor-2 (EGR2) and TIMP-1 expression were induced only in CBAxB6-TGFβ mice. Similar strain-dependent early changes in EGR2 and TIMP-1 of mice subjected to UUO or SNX were observed. TIMP-1 neutralization invivo hindered fibrosis both in transgenic mice and the SNX model. EGR2 over-expression in cultured HEK293 cells induced TIMP-1 while EGR2 silencing hindered TGF-β induced TIMP-1 production in HK-2 cells and ureteral obstructed kidneys. Finally, EGR2 and TIMP1 was increased in human kidneys manifesting focal segmental glomerulosclerosis suggesting a correlation between animal studies and patient clinical settings. Thus, our observations demonstrate a strong relationship between genetic background and the progression of kidney fibrosis, which might involve early altered EGR2 and TIMP-1 response, but the relationship to patient genetics remains to be explored.

Chronic Pressure Overload and Ovariectomy Alters the Regulation of Cardiac Fibrosis in a Chamber‐Specific Manner in Yucatan Mini‐Swine

Cardiac fibrosis, a hallmark feature of heart failure with preserved ejection fraction (HFpEF), is increased in postmenopausal women and may occur due to imbalances in the regulation of collagen metabolism. Our goal was to evaluate the effect of the loss of sex hormones via ovariectomy (OVX) on the development of aortic‐banding (AB) induced heart failure and cardiac fibrosis in 4 groups of female Yucatan mini‐swine: control (N=7); OVX (N=6); AB (N=7); and AB‐OVX (N=7). A 2x2 ANOVA (AB X OVX) was used to determine significance at the p &lt; 0.05 and 0.10 levels. Biochemical evaluation of total collagen demonstrated an increase due to OVX (main effect) in the left ventricle (LV) independent of AB. In contrast, total collagen was increased due to AB (main effect) in the right ventricle (RV) independent of OVX. These results led to the hypothesis that collagen is differentially regulated in a chamber‐specific manner due to imbalances in its metabolism. Collagen I and III mRNA levels were evaluated using qRT‐PCR. Collagen I mRNA was increased by AB (main effect) in both the LV and the RV. Collagen III mRNA levels were not altered in the LV, but were decreased due to OVX (main effect) in the RV. Collagen I and III protein levels, evaluated via western blot, were not significantly different in the LV, demonstrating a disconnect whereby changes to collagen I mRNA did not directly translate to alterations in protein. In the RV, collagen I protein levels were increased due to AB (main effect), while collagen III protein levels were unaltered. Thus, in contrast to the LV, a direct relationship between mRNA and protein was observed in the RV for collagen I. Given alterations to mRNA did not always match changes in protein level, collagen degradation was evaluated. Collagen degradation is regulated by matrix metalloproteinases (MMPs), which degrade collagen, and tissue inhibitors of matrix metalloproteinases (TIMPs), which inhibit MMPs. MMP‐2 and ‐9 activity/abundance was evaluated with gelatin zymography, MMP‐14 activity was evaluated via a fluorogenic substrate assay, and TIMP2 protein was evaluated through western blot. In the LV, there was a significant interaction for MMP‐2 and MMP‐9 whereby activity/abundance due to AB was dependent upon the presence of sex hormones. This interaction was not observed in the RV. MMP‐14 activity was decreased due to AB (main effect) in the LV, but not altered in the RV. TIMP2 protein levels were decreased due to OVX (main effect) in the LV, with no differences observed in the RV. In conclusion, these results demonstrate chamber‐specific differences in the regulation of cardiac fibrosis including: 1) An imbalance in collagen metabolism in the LV consisting of collagen mRNA levels not directly translating to protein, with dysregulation of MMP/TIMP protein and activity levels resulting from both OVX and AB; and 2) in the RV, an increase in collagen I mRNA and protein due to AB with no alterations to bioregulators of collagen degradation. These findings demonstrate important differences in the regulation of fibrosis due to the loss of sex hormones and chronic pressure overload between the LV and RV that may be critical to understanding the pathophysiology of HFpEF in postmenopausal women.

Folic Acid Levels During Pregnancy Regulate Trophoblast Invasive Behavior and the Possible Development of Preeclampsia.

BackgroundOne of the unique features of placentation is its similarity to tumorigenesis yet being very well regulated. It allows rapid proliferation, migration, and invasion of mononuclear trophoblast cells into the maternal uterus and remodeling the maternal vasculature. This pseudomalignant nature of trophoblastic cells is strictly regulated and its importance becomes evident in abnormal pregnancies that are characterized by aberrant trophoblast proliferation/invasion like preeclampsia. In addition to this, the importance of folic acid supplementation during pregnancy is well documented. We aimed to analyze the molecular and epigenetic regulation of the pseudomalignant nature of placentation via folic acid levels.MethodsPlacental tissue samples were collected from different pregnancies in three different gestational stages. We estimated the impact of folic acid levels on global methylation, LINE1 methylation, and expression of DNMTs in all three gestational stages in pregnant women and preeclampsia pregnancies. We also analyzed the effect of folic acid supplementation on trophoblastic invasion using placental derived cells viz, JEG-3 and HTR-8/SVneo cell line and verified the molecular and epigenetic mechanisms involved in this regulation.ResultsDevelopment of preeclampsia was observed to be associated with lower folate levels in placental tissue, higher global methylation level, and higher expression of DNMT1and DNMT3A. Folic acid supplementation was found to increase the invasive potential of placental trophoblasts by almost two folds which were associated with the decreased expression of tumor suppressor genes and tissue inhibitors of matrix metalloproteinases; and increased expression of oncogenes, telomerase gene, and matrix metalloproteinases. These folic acid-mediated changes were observed to be regulated by CpG methylation in the case of many genes. Folic acid supplementation was also observed to significantly decrease global methylation in placental trophoblasts related to decreasing expression of DNMT1 and DNMT3A.ConclusionLower folic acid levels are associated with preeclampsia development and folic acid supplementation regulates the invasive potential of placental trophoblasts as mediated by various epigenetic changes in the placenta suggesting the protective effect of folic acid against preeclampsia.