High Expression Of Matrix Metalloproteinase-9 Research Articles

Gelatin Microspheres Based on H8-Loaded Macrophage Membrane Vesicles to Promote Wound Healing in Diabetic Mice.

Diabetic wound healing remains a worldwide challenge for both clinicians and researchers. The high expression of matrix metalloproteinase 9 (MMP9) and a high inflammatory response are indicative of poor diabetic wound healing. H8, a curcumin analogue, is able to treat diabetes and is anti-inflammatory, and our pretest showed that it has the potential to treat diabetic wound healing. However, H8 is highly expressed in organs such as the liver and kidney, resulting in its unfocused use in diabetic wound targeting. (These data were not published, see Table S1 in the Supporting Information.) Accordingly, it is important to pursue effective carrier vehicles to facilitate the therapeutic uses of H8. The use of H8 delivered by macrophage membrane-derived nanovesicles provides a potential strategy for repairing diabetic wounds with improved drug efficacy and fast healing. In this study, we fabricated an injectable gelatin microsphere (GM) with sustained MMP9-responsive H8 macrophage membrane-derived nanovesicles (H8NVs) with a targeted release to promote angiogenesis that also reduces oxidative stress damage and inflammation, promoting diabetic wound healing. Gelatin microspheres loaded with H8NV (GMH8NV) stimulated by MMP9 can significantly facilitate the migration of NIH-3T3 cells and facilitate the development of tubular structures by HUVEC in vitro. In addition, our results demonstrated that GMH8NV stimulated by MMP9 protected cells from oxidative damage and polarized macrophages to the M2 phenotype, leading to an inflammation inhibition. By stimulating angiogenesis and collagen deposition, inhibiting inflammation, and reducing MMP9 expression, GMH8NV accelerated wound healing. This study showed that GMH8NVs were targeted to release H8NV after MMP9 stimulation, suggesting promising potential in achieving satisfactory healing in diabetic treatment.

1-Kestose Blocks UVB-Induced Skin Inflammation and Promotes Type I Procollagen Synthesis via Regulating MAPK/AP-1, NF-κB and TGF-β/Smad Pathway.

Solar UVB irradiation cause skin photoaging by inducing the high expression of matrix metalloproteinase (MMPs) to inhibit the expression of Type1 procollagen synthesis. 1-Kestose, a natural trisaccharide, has been indicated to show a cytoprotective role in UVB radiation-induced-HaCaT cells. However, few studies have confirmed the anti-aging effects. In the present study, we evaluated the anti-photoaging and pathological mechanism of 1-kestose using Human keratinocytes (HaCaT) cells. The results found that 1-kestose pretreatment remarkably reduced UVB-generated reactive oxygen species (ROS) accumulation in HaCaT cells. 1-Kestose suppressed UVB radiation-induced MMPs expressions by blocking MAPK/AP-1 and NF-κB p65 translocation. 1-Kestose pretreatment increased Type 1 procollagen gene expression levels by activating TGF-β/Smad signaling pathway. Taken together, our results demonstrate that 1-kestose may serve as a potent natural trisaccharide for inflammation and photoaging prevention.

Early detection of cardiac fibrosis in diabetic mice by targeting myocardiopathy and matrix metalloproteinase 2

Early detection of myocardial fibrosis in diabetic cardiomyopathy (DCM) has significant clinical implications for diabetes management. In this study, we identified matrix metalloproteinase 2 (MMP2) as a potential biomarker for early fibrosis detection. Based on this finding, we designed a dual-targeting nanoparticle CHP-SPIO-ab MMP2 to specifically target myocardiopathy and MMP2, enabling sensitive fibrosis detection using magnetic resonance imaging (MRI). Our results demonstrate that collagen hyperplasia (early fiber formation) begins to develop in diabetic mice at 12 weeks old, with observable fibrosis occurring at 16 weeks old. Additionally, MMP2 expression significantly up-regulates around collagen starting from 12 weeks of age. T2 MRI analysis revealed significant T2% enhancement in the hearts of 12-week-old diabetic mice following administration of the CHP-SPIO-ab MMP2 probe, indicating noninvasive detection of fiber formation. Furthermore, after fibrosis treatment, a reduction in T2% signal was observed in the hearts of 16-week-old diabetic mice. These findings were supported by Sirius red and Prussian blue staining techniques. Overall, our study presents a promising strategy for early identification of myocardial fibrosis. Statement of significanceMyocardial damage typically exhibits irreversibility, underscoring the paramount importance of early fibrosis diagnosis. However, the clinical used T1 mapping for fibrosis detection still exhibits limitations in terms of sensitivity. Therefore, it is imperative to develop highly sensitive strategies for early cardiac fibrosis detection. Here, we investigated the development of myocardial fibrosis in diabetic mice, and designed a highly sensitive probe that specifically targets cardiomyopathy and high expression of MMP2 for the early diagnosis of fibrosis. The probe enables non-invasive detection of abnormalities through MRI imaging as soon as fiber deposition appear, which can be detected earlier than T1 mapping. This advancement holds great potential for clinical diagnosis of myocardial fibrosis using cardiac magnetic resonance.

The multifaceted roles of matrix metalloproteinases in lung cancer.

Though the matrix metalloproteinases (MMPs) are widely investigated in lung cancer (LC), however, almost no review systematically clarify their multi-faced roles in LC. We investigated the expression of MMPs and their effects on survival of patients with LC, the resistance mechanisms of MMPs in anti-tumor therapy, the regulatory networks of MMPs involved, the function of MMPs inducing CSCLs, MMPs-related tumor immunity, and effects of MMP polymorphisms on risk of LC. High expression of MMPs was mainly related to poor survival, high clinical stages and cancer metastasis. Role of MMPs in LC are multi-faced. MMPs are involved in drug resistance, induced CSCLs, participated in tumor immunity. Besides, MMPs polymorphisms may increase risk of LC. MMPs might be promising targets to restore the anti-tumor immune response and enhance the killing function of nature immune cells in LC.

Relationship between metalloproteinase-9 (MMP-9) expression and clinicopathology in colorectal cancer: a cross-sectional study.

According to the WHO's GLOBOCAN database, ~1,931,590 new colorectal cancer (CRC) cases and 915,607 CRC-related deaths occurred in 2020. The incidence of CRC in Indonesia is 8.6%, making it the fourth most common cancer. With CRC, matrix metalloproteinase-9 (MMP-9) has a role in tumour development and progression, such that patients with a higher MMP-9 expression had poorer survival. This study aimed to analyze the relationship between MMP-9 expression and clinicopathology in CRC patients. This was an analytic observational study with a cross-sectional research design. It was conducted from November 2021 to June 2022 with 52 patient tissue samples: these were subjected to MMP-9 immunohistochemistry stain, with the GeneTex (Irvine) MMP-9 monoclonal antibody. Patient data were collected with clinical variables based on medical records and histopathological examination by anatomy pathologists. Primary tumour location, cancer staging, and histopathology grading were associated with MMP-9 (P=0.016, P=0.001, P=0.049). The more proximal to the primary tumour, the higher the stage of cancer, and the higher the histopathological grade, thus the greater the expression of MMP-9. A significant relationship existed of primary tumour location, cancer staging, and histopathology grading with MMP-9 expression in CRC patients. MMP-9 expression could be a useful indicator for the clinical assessment of tumour biologic behaviour and prognosis in CRC patients.

Ectopic expression of matrix metalloproteinases and filopodia extension via JNK activation are involved in the invasion of blood tumor cells in Drosophila mxc mutant.

Drosophila mxcmbn1 mutant exhibits severe hyperplasia in larval hematopoietic tissue called the lymph glands (LGs). However, the malignant nature of these cells remains unknown. We aimed to identify if mxcmbn1 LG cells behave as malignant tumor cells and uncover the mechanism(s) underlying the malignancy of the mutant hemocytes. When mutant LG cells were allografted into normal adult abdomens, they continued to proliferate; however, normal LG cells did not proliferate. Mutant circulating hemocytes also attached to the larval central nervous system (CNS), where the basement membrane was disrupted. The mutant hemocytes displayed higher expression of matrix metalloproteinase (MMP) 1 and MMP2 and higher activation of the c-Jun N-terminal kinase (JNK) pathway than normal hemocytes. Depletion of MMPs or JNK mRNAs in LGs resulted in reduced numbers of hemocytes attached to the CNS, suggesting that the invasive phenotype involved elevated expression of MMPs via hyperactivation of the JNK pathway. Moreover, hemocytes with elongated filopodia and extra lamellipodia were frequently observed in the mutant hemolymph, which also depended on JNK signaling. Thus, the MMP upregulation and overextension of actin-based cell protrusions were also involved in hemocyte invasion in mxcmbn1 larvae. These findings contribute to the understanding of molecular mechanisms underlying mammalian leukemic invasion.

The Action of Angiocrine Molecules Sourced from Mechanotransduction-Related Endothelial Cell Partially Explain the Successful of Titanium in Osseointegration.

Since Branemark's findings, titanium-based alloys have been widely used in implantology. However, their success in dental implants is not known when considering the heterogenicity of housing cells surrounding the peri-implant microenvironment. Additionally, they are expected to recapitulate the physiological coupling between endothelial cells and osteoblasts during appositional bone growth during osseointegration. To investigate whether this crosstalk was happening in this context, we considered the mechanotransduction-related endothelial cell signaling underlying laminar shear stress (up to 3 days), and this angiocrine factor-enriched medium was harvested further to use exposing pre-osteoblasts (pOb) for up to 7 days in vitro. Two titanium surfaces were considered, as follows: double acid etching treatment (w_DAE) and machined surfaces (wo_DAE). These surfaces were used to conditionate the cell culture medium as recommended by ISO10993-5:2016, and this titanium-enriched medium was later used to expose ECs. First, our data showed that there is a difference between the surfaces in releasing Ti molecules to the medium, providing very dynamic surfaces, where the w_DAE was around 25% higher (4 ng/mL) in comparison to the wo_DAE (3 ng/mL). Importantly, the ECs took up some of this titanium content for up to 3 days in culture. However, when this conditioned medium was used to expose pOb for up to 7 days, considering the angiocrine factors released from ECs, the concentration of Ti was lesser than previously reported, reaching around 1 ng/mL and 2 ng/mL, respectively. Thereafter, pOb exposed to this angiocrine factor-enriched medium presented a significant difference when considering the mechanosignaling subjected to the ECs. Shear-stressed ECs showed adequate crosstalk with osteoblasts, stimulating the higher expression of the Runx2 gene and driving higher expressions of Alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin. Mechanotransduction-related endothelial cell signaling as a source of angiocrine molecules also stimulated the higher expression of the Col3A1 gene in osteoblasts, which suggests it is a relevant protagonist during trabecular bone growth. In fact, we investigated ECM remodeling by first evaluating the expression of genes related to it, and our data showed a higher expression of matrix metalloproteinase (MMP) 2 and MMP9 in response to mechanosignaling-based angiocrine molecules, independent of considering w_DAE or the wo_DAE, and this profile reflected on the MMP2 and MMP9 activities evaluated via gelatin-based zymography. Complimentarily, the ECM remodeling seemed to be a very regulated mechanism in mature osteoblasts during the mineralization process once both TIMP metallopeptidase inhibitor 1 and 2 (TIMP1 and TIMP2, respectively) genes were significantly higher in response to mechanotransduction-related endothelial cell signaling as a source of angiocrine molecules. Altogether, our data show the relevance of mechanosignaling in favoring ECs' release of bioactive factors peri-implant, which is responsible for creating an osteogenic microenvironment able to drive osteoblast differentiation and modulate ECM remodeling. Taking this into account, it seems that mechanotransduction-based angiocrine molecules explain the successful use of titanium during osseointegration.

Comparison of MMP-9 Density Between Triple Negative and HER2 Enriched Breast Carcinoma Subtypes

Background
 Matrix metalloproteinase-9 (MMP-9) is a protease that plays an important role in various biological processes of breast cancer. This marker may be useful in determining the prognosis and become a promising therapy, especially in breast carcinoma with an aggressive subtype including HER2 enriched and triple negative. Both subtypes have high MMP-9 expression, but it is necessary to differentiate them because they have different biological and molecular aspects. This study aims to compare the density of MMP-9 between triple negative and HER2 enriched breast carcinoma subtypes and their relationship with clinicopathological characteristics.
  
 Methods
 This is a cross sectional study using 32 samples of paraffin blocks and slides of breast carcinoma patients with triple negative and HER2 Enriched subtypes at the Department of Anatomical Pathology Faculty of Medicine Sriwijaya University/Dr. Mohammad Hoesin Hospital Palembang for the period January 1, 2016 to December 31, 2019. Each sample was stained with MMP-9 antibodies immunohistochemistry (polyclonal, Abcam). Comparative analysis of MMP-9 density between the two subtypes was carried out using the median value based on the Mann-Whitney analysis. MMP-9 density was categorized as high and low based on the ROC curve and analyzed by using chi-square test to assess its association with clinicopathological characteristics.
  
 Results
 Mann-Whitney test showed that the MMP-9 density was higher in the HER2 enriched subtype (median 12.0) than triple negative (median 8.8) with a p value of 0.632. The relationship distribution of high MMP-9 density was found in the category of age group >40 years old (60.0%), positive lymphovascular invasion (86.7%) and histological grade III (80.0%) with p values of 0.798, 0.402 and 0.691.
  
 Conclusion
 MMP-9 density in HER2 enriched subtype was higher than triple negative subtype, but statistically not significant.

Daurisoline attenuates H2O2-induced chondrocyte autophagy by activating the PI3K/Akt/mTOR signaling pathway

BackgroundOsteoarthritis (OA) is a chronic degenerative joint disease characterized by cartilage degeneration and intra-articular inflammation. Daurisoline (DAS) is an isoquinoline alkaloid isolated from Rhizoma Menispermi, whose antitumor and anti-inflammatory pharmacological effects have been demonstrated, but the effects of DAS on OA have rarely been researched. In this study, we aimed to explore the potential role of DAS in OA and its partial mechanism.Materials and methodsThe cytotoxicity of H2O2 and DAS toward chondrocytes was detected by the Cell Counting Kit-8 assay. Safranin O staining was used to detect chondrocyte phenotype changes. Cell apoptosis was measured by both flow cytometry and quantitative analysis of the protein levels of the apoptosis-related factors Bax, Bcl-2 and cleaved caspase-3 by western blot. Western blotting and immunofluorescence were used to assess the expression of the autophagy-related proteins LC3, Beclin-1 and p62. In addition, key signal pathway targets and matrix-degrading indicators were measured by western blot.ResultsOur results indicated that H2O2 induced human chondrocyte apoptosis and activated autophagy in a dose-dependent manner. DAS treatment dose-dependently reversed the expression of apoptosis-related proteins (Bax, Bcl-2 and cleaved caspase3) and the apoptosis rate induced by H2O2. Western blot and immunofluorescence analyses showed that DAS decreased the H2O2-induced upregulation of the autophagy marker Beclin-1 and the LC3 II/LC3 I ratio and upregulated the p62 protein level. Mechanistically, DAS inhibited autophagy through the activation of the classical PI3K/AKT/mTOR signaling pathway and protected chondrocytes from apoptosis. In addition, DAS alleviated the H2O2-induced degradation of type II collagen and the high expression of matrix metalloproteinase 3 (MMP3) and MMP13.ConclusionOur research demonstrated that DAS alleviated chondrocyte autophagy caused by H2O2 through activation of the PI3K/AKT/mTOR signaling pathway and protected chondrocytes from apoptosis and matrix degradation. In conclusion, these findings suggest that DAS may serve as a promising therapeutic strategy for OA.

OONO-/MMP2/MMP9 pathway-mediated apoptosis of porcine granulosa cells is associated with DNA damage.

The apoptosis of granulosa cells (GCs) is the main reason for porcine follicular atresia. This study provides a novel mechanism for peroxynitrite anion-mediated GC apoptosis and follicular atresia in porcine ovary. Granulosa cells play a crucial role in the development of follicles, and their cell apoptosis in the porcine ovary is a major contributor to follicular atresia. Here, we provide a new mechanism for follicular atresia by describing a crucial mechanism by which peroxynitrite anion (OONO-) may cause GC death. We discovered that nitric oxide, oxidative stress level, and OONO- were positively correlated with porcine follicular atresia, which was accompanied by high expression of matrix metalloproteinase 2 (MMP2) and MMP9. We created a model of OONO--induced apoptosis in GCs and discovered that OONO- could boost the expression of MMP2 and MMP9 and increase the expression of pro-apoptotic proteins and DNA damage. Furthermore, by inhibiting the activities of MMP2 and MMP9, we found that SB-3CT (a specific inhibitor for MMP2 and MMP9) alleviated the decrease in cell survival rates and DNA damage caused by OONO-, which may have been impacted by reducing the cleavage of PARP1 by MMP2 and MMP9. Therefore, our findings imply that OONO- can cause DNA damage to GCs, participating in mediating the expression of pro-apoptotic proteins and inhibiting DNA repair by preventing the activity of PARP1 through MMP2 and MMP9. These results help explain how OONO-/MMP2/MMP9 affects porcine follicular atresia and GC apoptosis.

Mechanism of Aspirin oxidative stress regulating interleukin-induced apoptosis in nucleus pulposus cells in a rat model of intervertebral disc degeneration.

Intervertebral disc degeneration (IDD) is an important cause of low back pain. Increase of reactive oxygen species (ROS), overexpression of inflammatory factors, and loss of extracellular matrix are important factors in the pathological changes of IDD. The present study aimed to investigate the mechanism of action of Aspirin regulating oxidative stress in IDD, so as to propose new treatment. Nucleus pulposus cells (NPCs) were isolated from the caudal intervertebral discs of Sprague Dawley (SD) rats under sterile conditions. The expression of ROS and inflammatory factors was detected sequentially, and the degree of degeneration of nucleus pulposus cells was observed by real-time fluorescence quantitative polymerase chain reaction (PCR) and cell immunofluorescence staining. In vivo, the caudal disc puncture model was used to induce degeneration, and a local injection of 10 or 100 µg/mL Aspirin was performed. The rats were sacrificed 1 week later, and the disc specimens of the tail vertebrae were collected for imaging, histomorphology, and immunohistochemical analysis. In vitro experiments showed that lipopolysaccharide (LPS) could significantly induce oxidative stress in NPCs and stimulate NPCs to secrete a large amount of ROS and inflammatory factors, which eventually leads to the reduction of collagen type II and polyglycoprotein gene expression in NPCs and the high expression of matrix metalloproteinase (MMP). Consequently, NPCs degeneration occurs. Our results clarified the important role of oxidative stress in IDD and proved that LPS can be used as a drug to alleviate oxidative stress and intervene in the IDD process.

MMP-9 and CCR7 as Possible Predictors of Lymph Node Metastasis in Laryngeal Squamous Cell Carcinoma.

The expression of matrix metalloproteinase-9 (MMP-9) and chemokine receptor 7 (CCR7) is significantly associated with tumor invasion and metastasis. Little is known regarding the potential of these markers in predicting cancer metastasis in Laryngeal Squamous Cell Carcinoma (LSCC). Therefore, this study aimed to dissect the potential of these markers in predicting the lymph node metastasis in LSCC patients. Sixty tissue samples were obtained from the patients diagnosed pathologically with LSCC who underwent partial or total laryngectomy. The expression of MMP-9 and CCR7 was measured using the immunohistochemistry staining in the tissue samples of LSCC patients. The ROC (receiver operating characteristic) curve was used to determine the most significant cut-off points of expression according to the highest sensitivity and specificity of both the markers to predict the lymph node metastasis in LSCC. Then, the relationship between the clinicopathology features and the expression of MMP-9 and CCR7 was evaluated. The expression of both MMP-9 and CCR7 was significantly correlated with the lymph node metastasis in LSCC (P<0.001). Furthermore, CCR7 expression exhibited the highest prediction accuracy (AUC 95.7%) and sensitivity (100%) in predicting the lymph node metastasis in LSCC compared to that of MMP-9 (AUC 92.9%, sensitivity 90%). We also found that patients with larger tumor size (> 4 cm) had significantly higher expression of MMP-9 and CCR7 (P<0.002 and P<0.001, respectively). The Elevated expression level of CCR7 statistically correlated with higher MMP-9 expression (P<0.001). MMP-9 and CCR7 might be beneficial as predictors of lymph node metastasis in LSCC patients.

Supramolecular peptide hydrogel doped with nanoparticles for local siRNA delivery and diabetic wound healing

High expression of matrix metalloproteinase 9 (MMP-9) has been demonstrated to significantly impair the healing of diabetic wounds. Three-dimensional hydrogels have been widely used for local siRNA delivery and wound healing via silencing MMP-9 expression. However, the negatively charged nature of siRNA released from the hydrogel dramatically weakens the cellular uptake of siRNA, thereby inducing suboptimal therapeutic outcomes. To address this issue, we herein developed a supramolecular peptide hydrogel doped with nanoparticles (NPs) for local MMP-9 siRNA (siMMP-9) delivery and diabetic wound healing. For this siRNA delivery system, siMMP-9 was encapsulated into NPs made with an amphiphilic cationic lipid-like compound and then embedded into a supramolecular peptide hydrogel self-assembled from an amphiphilic peptide. After administration to the diabetic wounds as dressing, the peptide hydrogel could not only dramatically prolong the retention of siRNA-loaded NPs at the wound tissues, but also gradually release the siRNA-loaded NPs from its porous network to improve the siRNA uptake by keratinocytes, leading to efficient MMP-9 silencing and significant improvement of diabetic wound healing. The siRNA delivery system developed in this work could be used as an effective tool to regulate the expression of other diabetic wound-associated genes and improve the diabetic wound healing.

Diagnostic and prognostic values of MMP-9 expression in ovarian cancer: A study based on bioinformatics analysis and meta-analysis.

This study aims to explore the expression of matrix metalloproteinase-9 (MMP-9) associated with both diagnostic and prognostic value in ovarian cancer by meta-analysis and bioinformatics analyses. We investigated the prognostic value of MMP-9 expression in ovarian cancer based on The Cancer Genome Atlas. Five databases were used to collect records about MMP-9 expression related to diagnostic and prognostic values in ovarian cancer from inception to June 2022. Using Stata 15.0 software, hazard ratio (HR) and odds ratio (OR) were calculated as the effect index of prognosis. We chose the pooled sensitivity, specificity, and area under the curve (AUC) to judge the diagnostic utility of MMP-9 for ovarian cancer. A total of 23 studies on prognosis, and five studies on diagnosis were entered into the meta-analysis. These suggest that high MMP-9 expression was detrimental to the overall survival of patients with ovarian cancer (HR = 1.34; 95% confidence interval (CI) 1.08∼1.66; P<0.01). High MMP-9 expression increased the risk of tumor stage (OR = 3.66; 95% CI 1.89∼7.07), but was not related to the tumor grade of ovarian cancer (P>0.05). The pooled analysis of serum MMP-9 diagnosing for ovarian cancer gave the pooled sensitivity, specificity, and AUC the values of 0.72 (95% CI 0.61∼0.81), 0.81 (95% CI 0.77∼0.85), and 0.84 (95% CI 0.81∼0.87), respectively. High MMP-9 expression can increase the tumor stage, and a correlation exists between high MMP-9 expression and poor prognosis in patients with ovarian cancer. Also, serum MMP-9 has a good diagnostic value for ovarian cancer.

Platelet–neutrophil hybrid membrane-coated gelatin nanoparticles for enhanced targeting ability and intelligent release in the treatment of non-alcoholic steatohepatitis

Non-alcoholic steatohepatitis (NASH) is the major form of chronic liver disease in adults; however, there are no approved drugs for NASH. In this study, we designed the PNM-G-PV method, in which gelatin nanoparticles (G) are loaded with pioglitazone and vitamin E (G-PV) and then encapsulated by the surfaces of platelet–neutrophil hybrid membranes (PNM). Inherited from the natural source cells, the PNM show immune evading ability due to the surface marker comprising a number of “do not eat me” signals and has dual inflammatory enrichment capabilities due to specific surface adhesion molecules. By functionalizing the gelatin nanoparticle biomimetic surfaces, PNM-G can enhance the targeting to inflammatory sites and enrich liver tissue. The high expression of matrix metalloproteinase-9 (MMP-9) at the NASH site enables the gelatin nanoparticles to intelligently respond to degradation and then release vitamin E and pioglitazone for drug treatment. We performed an in vivo analysis of these nanoparticles to monitor changes in triacylglycerol metabolism in liver tissues and assessed the therapeutic efficacy of PNM-G-PV in a NASH rat model. The results showed that PNM-G-PV exhibited better therapeutic efficacy than therapies using G-PV or PV alone. This work explores a new biomedical use of PNM-G-PV and a promising NASH treatment protocol based on a new drug delivery system.

Association and prognostic significance of alpha-L-fucosidase-1 and matrix metalloproteinase 9 expression in esophageal squamous cell carcinoma.

BACKGROUNDAlpha-L-fucosidase-1 (FUCA1) has been demonstrated to play opposing regulatory roles in adenocarcinoma and non-adenocarcinoma. Moreover, recent studies reported that FUCA1 could decrease the invasion capability by downregulating matrix metalloproteinase 9 (MMP-9) expression. However, the potential role and prognostic significance of FUCA1 in esophageal squamous cell carcinoma (ESCC) have not yet been explored.AIMTo evaluate the status, association, and prognostic value of FUCA1 and MMP-9 expression in ESCC. METHODSPatients who underwent esophagectomy for ESCC between January 1, 2014, and December 31, 2014 at Sun Yat-Sen University Cancer Center were enrolled. The expression status of FUCA1 and MMP-9 in cancerous tissues was detected using immunohistochemistry. In addition, the expression profiles of the FUCA1 and MMP-9 genes in non-metastatic ESCC were extracted from The Cancer Genome Atlas (TCGA) database. RESULTSHigh expression of FUCA1 and MMP-9 was found in 90 patients (75.6%) and 62 patients (52.1%), respectively. In the high FUCA1 expression group, the constituent ratios of patients with stage III disease (61.1% vs 37.9%, P = 0.029), lymphatic invasion (62.2% vs 31.0%, P = 0.003), and high MMP-9 expression (60.0% vs 27.6%, P = 0.002) were significantly higher than those in the low FUCA1 expression group. In Kaplan-Meier univariate analysis, advanced tumor-node-metastasis stage (III, P = 0.001), positive regional lymph node metastasis (N+, P = 0.002), high FUCA1 expression (P = 0.001), and high MMP-9 expression (P = 0.002) were potential predictors of shorter overall survival (OS), which was similar to the results analyzed based on the TCGA database. Further Cox multivariate regression analyses still demonstrated that FUCA1 and MMP-9 expression levels were independent prognostic factors of OS [hazard ratio (HR): 0.484, 95% confidence interval (CI): 0.239-0.979; P = 0.044; and HR: 0.591, 95%CI: 0.359-0.973, P = 0.039, respectively]. CONCLUSIONFUCA1 cooperation with MMP-9 may have a major role in affecting the ESCC invasion and metastatic capability, and serve as a valuable prognostic biomarker in ESCC.

MicroRNA-152 Regulates Endometrial Serous Carcinoma Cell Motility by Suppressing Matrix Metalloproteinase 10 Expression.

MicroRNA-152 (miR-152) expression has been reported to be associated with poor prognosis in patients with endometrial serous carcinoma (ESC). However, the function of miR-152 in ESCs is not fully understood. The present study aimed to investigate the involvement of miR-152 in ESC progression. The influence of miR-152 overexpression on cell proliferation and motility was assessed by transfecting two human ESC cell lines, USPC-1 and SPAC-1-L, with a miR-152 precursor. MiR-152 overexpression increased apoptosis and inhibited the proliferation of the two ESC cell lines. Cell motility was also suppressed in both cell lines following precursor transfection. Conversely, miR-152 inhibitor transfection led to an increase in cell migration ability, suggesting the involvement of miR-152 in ESC cell motility. Results of the analysis of publicly available messenger RNA dataset indicated that high expression of matrix metalloproteinase 10 (MMP10), one of the predicted targets of miR-152 by microRNA target prediction database, was a poor prognostic factor for ESC. In vitro examination results revealed that miR-152 overexpression reduced MMP10 expression, and knockdown of MMP10 significantly reduced cell motility. This study elucidates the function of miR-152 as a tumor suppressor in ESCs. We demonstrated that miR-152 plays an important role in ESC cell motility by regulating MMP10 expression.

Investigating the cerebral vascular integrity in a chronic cerebral hypoperfusion mouse model of vascular dementia: Pathophysiology and the intermittent fasting treatment.

Chronic cerebral hypoperfusion (CCH) is an important pathophysiological mechanism in Vascular Dementia (VaD). The heterogeneous nature of CCH complicates the establishment of a stand-alone pharmacological therapy against the disease. Intermittent Fasting (IF) is neuroprotective in physiological and disease conditions, but its effects against CCH-induced BBB and vascular damage remains unknown. We investigate the effects of IF in alleviating vascular damage in a CCH model. We hypothesize that CCH would decrease BBB and vascular integrity and these changes are prevented by IF. Male C57BL/6 mice were subjected to ad libitum feeding (AL) or IF (16h fasting per day) for 4 months before surgery. In both the fasted and fed groups, CCH was experimentally induced by the bilateral carotid artery stenosis (BCAS) surgery via microcoil insertion for 30days. As controls, SHAM groups were prepared. BBB permeability, tight junction, extracellular matrix components and microvascular changes were determined from the mouse brains. AL-BCAS mice had a significantly higher extent of Evans Blue extravasation compared to AL-SHAM mice, while the IF-BCAS had significant alleviation in the extent of BBB permeability compared to their corresponding AL-BCAS timepoints, indicating that IF preserves the physiological permeability of the BBB. AL-BCAS mice also had lower gene expression of tight junction proteins (ZO-1, Occludin, Claudin-5 and JAM-A), higher expression of matrix metalloproteinases (MMP-2 and MMP-9), lower density of microvascular structures in the brain as compared to as compared to AL-SHAM, and IF-BCAS mice showed opposing trends in these proteins compared to their corresponding AL-BCAS timepoints. This further suggests that IF has a protective effect in the brain against CCH. IF has neuroprotective effects in a mouse CCH model of VaD by preserving BBB permeability and microvascular density. IF is effective in human and rodent studies of several diseases which have been reported to be risk factors of cognitive decline. Other significant findings from animal studies of IF demonstrate positive effects against mechanisms of dementia. Although IF has yet to be assessed in VaD, we suggest that IF may be an effective means of preventing the onset or suppressing the development of VaD.

Rivaroxaban attenuates valvular inflammation and tissue remodeling in patients with severe aortic stenosis

Abstract Background It has been demonstrated in a mouse model of atherosclerosis that non-vitamin K antagonist oral anticoagulants (NOACs) attenuate atherosclerotic plaque progression. However, it is not known whether NOACs can inhibit aortic stenosis (AS) progression. Aims To evaluate whether rivaroxaban at therapeutic concentrations can impair valvular inflammation and valve remodeling in patients with severe AS. Methods We recruited 38 patients with isolated severe AS aged 68±8 years (mean gradient 54 mmHg, max gradient 86 mmHg), including 18 individuals with AS and atrial fibrillation taking rivaroxaban (20 mg/daily for at least 3 years). Stenotic aortic valves were obtained during valve replacement surgery. Valvular expression of NFκB, matrix metalloproteinase 9 (MMP-9) and bone morphogenetic protein 2 (BMP-2) was evaluated by immunostaining. Primary cultures of valve interstitial cells (VICs) obtained from stenotic aortic valves were stimulated with TNF-α to induce inflammation. In parallel, VICs were cultured with TNF-α and rivaroxaban at therapeutic concentrations (1 and 10 μg/ml, corresponding to about 2.5 mg and 25 mg daily in vivo). The expression of NFκB, interleukin 1β (IL-1β), reactive oxygen species (ROS), MMP-9 and BMP-2 in VICs was assessed. The control VICs were cultured without any additives. All VICs were cultured for 72 hours and experiments were repeated 3 times. The fluorescence intensity was computed as the ratio (%) of positively and negatively stained areas. Results We observed decreased valvular expression of NFκB (−54%), MMP-9 (−32%) and BMP-2 (−23%) in patients taking rivaroxaban compared to those without such treatment. TNF-α stimulation up-regulated expression of NFκB, IL-1β and ROS (+58%; +34%; +38%; all p&amp;lt;0.01) accompanied by higher expression of MMP-9 and BMP-2 (+49% for MMP-9 and +32% for BMP-2; both p&amp;lt;0.01), when compared to control cells. Rivaroxaban at a concentration of 10 μg/ml decreased the expression of ROS (−31%, p&amp;lt;0.01), inflammatory markers (NFκB −47% and IL-1β −22%, both p&amp;lt;0.01) and MMP-9 (−39%, p&amp;lt;0.01) compared to VICs cultured without rivaroxaban. Similarly, BMP-2 expression was down regulated after 10 μg/ml of rivaroxaban (−26%, p&amp;lt;0.01). Even low dose of rivaroxaban decreased VICs expression of NFκB, IL-1β and ROS by 38%, 18% and 25%, respectively (all p&amp;lt;0.01) as compared to cells stimulated with TNF-α only. Comparable effect was observed for MMP-9 and BMP-2 expression (−33%; −22%; both p&amp;lt;0.01). Conclusions Rivaroxaban can decrease valvular inflammation and its remodelling and significantly inhibits inflammation, extracellular matrix activity and calcification of VICs. Our study suggests that NOACs, even at a low dose, could slow the rate of AS progression, at least in AS patients with mild-to-moderate AS and indications for anticoagulant therapy. Clinical relevance of these findings requires further studies. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Science Centre (NCN)

Study on the Mechanism of Capillary Leakage Caused by Hypoxia-Inducible Factor-1α through Inducing High Expression of Matrix Metalloproteinase-9.

Purposes This study mainly explored the mechanism of capillary leakage caused by hypoxia-inducible factor-1α through inducing high expression of matrix metalloproteinase-9. Method. We established a monolayer endothelial cell model by culturing human umbilical vein endothelial cells (HUVEC) in vitro, used tumor necrosis factor (TNFα) and HIF-1α inhibitor 2-methoxyestradiol (2ME2) to act on HUVEC, and at the same time constructed siRNA-transfected HUVEC to interfere with the expression of HIF-1α. The permeability of monolayer endothelial cells was measured by transwell chamber method, the concentration of MMP-9 in the supernatant was measured by ELISA method, the expression of key molecules related to permeability (HIF- 1α, MMP-9, claudin-5, and ZO-1) was measured by RT-PCR and Western blot method, and the localization and expression of claudin-5 and ZO-1 were measured by immunofluorescence method. We searched for 7 HIF-1α hypoxia response elements within 4000 bp before the transcription start site in the MMP-9 promoter region, constructed the MMP-9 promoter-luciferase reporter gene recombinant plasmid, transfected and stimulated HUVEC with TNFα, and detected the effect of 7 hypoxia response element plasmids on the transcription activity of MMP-9 promoter. Results Under the action of TNFα, the permeability of monolayer endothelial cells increased, and the concentration of MMP-9 in the cell supernatant increased. 2ME2 and HIF-1α-siRNA transfection can improve the above situation (P < 0.05). 2ME2 and HIF-1α-siRNA transfection can inhibit the high expression of HIF-1α and MMP-9 caused by TNFα, thereby increasing the expression of claudin-5 and ZO-1 (P < 0.05). 2ME2 and HIF-1α-siRNA transfection can reduce the inhibition of TNFα on the expression of cell membrane protein claudin-5 and tight junction protein ZO-1. Element 1, element 5, and element 7 are the sites where HIF-1α interacts with MMP-9 at the transcription level. Conclusion This study shows that HIF-1α can increase the permeability of monolayer epithelial cells by inducing the high expression of MMP-9, leading to capillary leakage. Its target is at the −3798 bp, −1878 bp, and −1489 bp points of the transcription initiation site in the MMP-9 promoter region.