MMP-9 Gene Expression Research Articles

The effects of nanomicelle of curcumin on the matrix metalloproteinase (MMP-2, 9) activity and expression in patients with coronary artery disease (CAD): A randomized controlled clinical trial.

BACKGROUNDCoronary artery disease (CAD) is the most common type of cardiovascular disease. Increasing the expression and activity of matrix metalloproteinases (MMPs) facilitates vascular remodeling and cardiovascular complications. Curcumin (the active ingredient of turmeric) is a potent natural anti-inflammatory agent, with cardiovascular protective effects. The present study was a clinical trial for investigating the effects of curcumin on activity and gene expression of MMP-2 and MMP-9 in patients with CAD.METHODSIn this study, 70 patients with CAD (with 40%-50% stenosis) were randomly divided into two groups of curcumin (80 mg nanomicelle per day) and placebo. The intervention lasted 3 months. The activity levels of MMP-2 and MMP-9 in serum samples of patients were measured using gelatin zymography assay before and after the intervention. MMP-2 and MMP-9 gene expression in peripheral blood mononuclear cells (PBMCs) was also analyzed using real-time polymerase chain reaction (PCR). Statistical significance was set at P < 0.0500.RESULTSAfter 3 months of medication, the expression of MMP-9 produced by PBMCs significantly decreased in the curcumin group (0.811 ± 0.25) in comparison with the placebo group (2.23 ± 0.94) (P < 0.0001). Furthermore, the zymographic analysis showed that the administration of curcumin significantly inhibited the activity levels of MMP-2 (12469.7 ± 5308.64 pixels) and MMP-9 (14007.2 ± 5371.67 pixels) in comparison with that in patients receiving placebo (MMP-2: 17613.8 ± 5250.68 pixels; MMP-9: 20010.1 ± 3259.37 pixels) (P < 0.0500).CONCLUSIONOur results show that curcumin can significantly reduce the expression and activity of MMP-2 and MMP-9. Because of the anti-inflammatory effects of curcumin, this compound can be considered as a new strategy for the prevention of cardiovascular events.

Human Wharton's jelly stem cells inhibit endometriosis through apoptosis induction.

Endometriosis is a relatively benign disease characterized by endometrial tumors and uterus stroma. Apoptosis suppression is one of the most important pathological processes of endometriosis. Recently, several studies reported that human Wharton's jelly stem cells (hWJSCs) can inhibit growth and proliferation of various cancer cells through induction of apoptosis. Therefore, the aim of the present study was to investigate the effects of hWJSCs conditioned medium (hWJSC-CM) and cell-free lysate (hWJSC-CL) on endometriosis cells in vitro. In the present study, effects of different concentrations of hWJSC-CM and hWJSC-CL on viability and proliferation, morphological alterations, colony formation, migration, invasion, and apoptosis of endometriosis cells were evaluated. Our results showed that hWJSC-CM and hWJSC-CL decrease viability and proliferation, colony formation, migration, and invasion, as well as increase morphological alterations and apoptosis of endometriosis cells, in a concentration- and time-dependent manner. Decreased migration and invasion of treated endometriosis cells with hWJSC-CM and hWJSC-CL may be due to decrease of MMP-2 and MMP-9 gene expression. Moreover, induction of apoptosis in treated endometriosis cells can be due to regulation of apoptosis-related genes expression, including BAX, BCL-2, SMAC, and SURVIVIN. The results of the present study suggest that hWJSC-CM and hWJSC-CL can inhibit endometriosis cells at a mild-to-moderate level through various physiological mechanisms. However, further studies on animal models are necessary to achieve more accurate results.

Collagen/GAG scaffolds activated by RALA-siMMP-9 complexes with potential for improved diabetic foot ulcer healing.

Impaired wound healing of diabetic foot ulcers has been linked to high MMP-9 levels at the wound site. Strategies aimed at the simultaneous downregulation of the MMP-9 level in situ and the regeneration of impaired tissue are critical for improved diabetic foot ulcer (DFU) healing. To fulfil this aim, collagen/GAG (Col/GAG) scaffolds activated by MMP-9-targeting siRNA (siMMP-9) were developed in this study. The siMMP-9 complexes were successfully formed by mixing the RALA cell penetrating peptide with siMMP-9. The complexes formulated at N:P ratios of 6 to 15 had a diameter around 100nm and a positive zeta potential about 40mV, making them ideal for cellular uptake. In 2 dimensional (2D) culture of human fibroblasts, the cellular uptake of the complexes surpassed 60% and corresponded to a 60% reduction in MMP-9 gene expression in low glucose culture. In high glucose culture, which induces over-expression of MMP-9 and therefore serves as an in vitro model mimicking conditions in DFU, the MMP-9 gene could be downregulated by around 90%. In the 3D culture of fibroblasts, the siMMP-9 activated Col/GAG scaffolds displayed excellent cytocompatibility and ~60% and 40% MMP-9 gene downregulation in low and high glucose culture, respectively. When the siMMP-9 complexes were applied to THP-1 macrophages, the primary cell type producing MMP-9 in DFU, MMP-9 gene expression was significantly reduced by 70% and 50% for M0 and M1 subsets, in 2D culture. In the scaffolds, the MMP-9 gene and protein level of M1 macrophages decreased by around 50% and 30% respectively. Taken together, this study demonstrates that the RALA-siMMP-9 activated Col/GAG scaffolds possess high potential as a promising regenerative platform for improved DFU healing.

In vivo antitumoral effect of 4-nerolidylcatechol (4-NC) in NRAS-mutant human melanoma

NRAS-mutations arise in 15–20% of all melanomas and are associated with aggressive disease and poor prognosis. Besides, the treatment for NRAS-mutant melanoma are not very efficient and is currently limited to immune checkpoints inhibitors or aggressive chemotherapy. 4-nerolidylcathecol (4-NC), a natural product extracted from Pothomorphe umbellata, induces apoptosis in melanoma cells by ROS production, DNA damage and increased p53 expression, in addition to inhibiting invasion in reconstructed skin. Moreover, 4-NC showed cytotoxicity in BRAF/MEKi-resistant and naive melanoma cells by Endoplasmic Reticulum (ER) stress induction in vitro. We evaluated the in vivo efficacy and the systemic toxicity of 4-NC in a NRAS-mutant melanoma model. 4-NC was able to significantly suppress tumor growth 4-fold compared to controls. Cleaved PARP and p53 expression were increased indicating cell death. As a proof of concept, MMP-2 and MMP-14 gene expression were decreased, demonstrating a possible role of 4-NC in melanoma invasion inhibition. Toxicological analysis indicated minor changes in the liver and bone marrow, but this toxicity was very mild when compared to other proteasome inhibitors and ER stress inductors already described. Our data indicate that 4-NC can counteract melanoma growth in vivo with minor adverse effects, suggesting further investigation as a potential NRAS-mutant melanoma treatment.

Expression of Matrix Metalloproteinases and Their Inhibitors in Endometrium: High Levels in Endometriotic Lesions.

Endometriosis is a condition defined as presence of endometrium outside of the uterine cavity. These endometrial cells are able to attach and invade the peritoneum or ovary, thus forming respectively the deep infiltrating endometriosis (DIE) and the ovarian endometrioma (OMA), the ectopic lesions feature of this pathology. Endometriotic cells display high invasiveness and share some features of malignancy with cancer cells. Indeed, the tissue remodeling underlining lesion formation is achieved by matrix metalloproteinases (MMPs) and their inhibitors. Therefore, these molecules are believed to play a key role in development and pathogenesis of endometriosis. This study investigated the molecular profile of metalloproteinases and their inhibitors in healthy (n = 15) and eutopic endometrium (n = 19) in OMA (n = 10) and DIE (n = 9); moreover, we firstly validated the most reliable housekeeping genes allowing accurate gene expression analysis in these tissues. Gene expression, Western blot, and immunofluorescence analysis of MMP2, MMP3, and MMP10 and their tissue inhibitors TIMP1 and TIMP2 demonstrated that these enzymes are finely tuned in these tissues. In OMA lesions, all the investigated MMPs and their inhibitors were significantly increased, while DIE expressed high levels of MMP3. Finally, in vitro TNFα treatment induced a significant upregulation of MMP3, MMP10, and TIMP2 in both healthy and eutopic endometrial stromal cells. This study, shedding light on MMP and TIMP expression in endometriosis, confirms that these molecules are altered both in eutopic endometrium and endometriotic lesions. Although further studies are needed, these data may help in understanding the molecular mechanisms involved in the extracellular matrix remodeling, a crucial process for the endometrial physiology.

Joint injury-induced angiogenesis and thrombospondin 2

Purpose: It is controversial whether inhibition of angiogenesis slows the progression of post-traumatic osteoarthritis (PTOA). The matricellular protein thrombospondin-2 (TSP2) is a candidate for inhibiting intraarticular angiogenesis after joint injury as it is a well-characterized endogenous anti-angiogenic protein. In addition, prior literature demonstrates the inhibition of matrix metalloproteinase (MMP) bioavailability by TSP2. However, little is known of the involvement of TSP2 in joint trauma, injury-induced angiogenesis and MMP activity, and PTOA progression. The purpose of this study was to characterize the endogenous TSP2 response following traumatic joint injury in mice. Methods: With IACUC approval, noninvasive ACLR was induced in adult TSP2-GFP reporter mice and C57Bl/6 mice via tibial compression. Limbs were processed for histology at 3d, 14d, and 28d post-ACLR (n=3-4/timepoint). The proportion of TSP2+ cells in synovium was quantified in injured and contralateral limbs of TSP2-GFP mice using k-means clustering of reporter intensity. ACLR-induced angiogenesis was assessed in C57Bl/6 mice via CD31 immunostaining and in vivo near-infrared imaging (NIR) utilizing the AngioSense-agent (Perkin Elmer), which enables in vivo assessment of vascular changes. Synovia from C57Bl/6 ACLR and contralateral knees were dissected, homogenized in Trizol, and analyzed for TSP2, MMP2, MMP3, MMP13 gene expression via rtPCR at 3d and 14d post-ACLR (n=5-8/timepoint), representing timepoints of early and late inflammation. To determine which cytokines induce TSP2 in fibroblast-like synoviocytes (FLS) and whether ACLR-associated in vivo inflammation affects the in vitro FLS cytokine response, primary mouse FLS were isolated from knee synovium of 7d post-ACLR and contralateral knees (n=3/group). FLS were induced with IL-1β and TNFα (10 ng/ml) for 48 hours and analyzed for TSP2 and MMP13 gene expression. Gene expression was normalized to ATP5b and paired t-tests performed. Results: ACLR induced intraarticular angiogenesis at 14d and 28d, assessed by influx of CD31+ cells. Abundant CD31+ staining was observed in synovium, menisci, and osteophytes of injured knees up to 28d post-ACLR (Figure. 1 A-C). An observed increase in in vivo AngioSense signal of the injured knee 7d post-ACLR (P=0.021) corroborates this finding and indicates acute vascular changes (Figure. 1D-E). ACLR also induced significant upregulation of MMP-2, -3, and -13 mRNA by synovium at 3d and 14d post-ACLR (Figure. 2K). In TSP2-GFP reporter mice, injured knees exhibited abundant synovial TSP2 expression, as early as 3d and up to 28d (at which point mice exhibit moderate PTOA severity; data not shown). At 3d and 14d, TSP2 expression is confined to the synovium, and by 28d, developing osteophytes are also TSP2(+). At all timepoints, synovial TSP2 expression is localized to the subintima. The intima of injured joints, although hyperplastic, remains TSP2(-). The cruciate ligaments, menisci, articular cartilage, and subchondral bone remain TSP2(-) (Figure. 2A-H). Quantitative image analysis indicates ACLR-induced TSP2-GFP expression in ∼27% of synovial cells at 14d (P=0.017) and ∼24% at 28d (P=0.018) post-ACLR, compared to near-zero expression in contralateral synovia (Figure. 2J). rtPCR confirms strong TSP2 upregulation in injured C57Bl/6 synovium at 3d (P=.006) and 14d (P=.007) (Figure. 2I). Surprisingly, both IL-1β and TNFα downregulated TSP2 in FLS from healthy (IL-1β - P=.009; TNFα - P=.001) and injured (IL-1β - P=.001; TNFα - P=.10) joints. TNFα-treated FLS from injured joints exhibited significantly less downregulation compared to healthy FLS (P=0.025) (Figure. 3B). Both IL-1β and TNFα induced a strong MMP13 response in healthy (P=0.002; P=0.004) and injured (P=0.001; P=0.006) FLS (Figure 3A), and there was no difference in the magnitude of MMP13 expression between healthy and injured FLS. Conclusions: TSP2 is strongly upregulated by the synovium following ACLR. This coincides with a synovial MMP-2, -3, and -13 response, increased intraarticular angiogenesis, and the time course of post-injury inflammation. Interestingly, TSP2 expression is confined to the subintimal region of the synovium. Our in vitro data shows that primary FLS do not upregulate TSP2 in response to IL-1β or TNFα, and interestingly, FLS from injured joints do not exhibit differentially-elevated TSP2 compared to FLS from contralateral joints, despite abundant in vivo synovial TSP2 expression following ACLR. Thus, ongoing work is identifying local TSP2+ cell populations and ligands that induce TSP2 expression in FLS. Given evidence on the anti-angiogenic capacity of TSP2, ongoing work is assessing angiogenesis, inflammation, catabolism, and PTOA severity in global TSP2-KO mice. Our ultimate objective is to mitigate injury-induced angiogenesis with exogenous TSP2 treatment.View Large Image Figure ViewerDownload Hi-res image Download (PPT)View Large Image Figure ViewerDownload Hi-res image Download (PPT)

A trimethoxy flavonoid isolated from stem extract of Tabebuia chrysantha suppresses angiogenesis in angiosarcoma.

This research aimed to evaluate the antiangiogenic activity of isolated flavonoid 4a,5,8,8a-tetrahydro-5-hydroxy-3,7,8-trimethoxy-2-(3,4-dimethoxyphenyl) chromen-4-one (TMF) from Tabebuia chrysantha. STAT3-MMP9 signalling is a signal transduction mechanism that promotes angiogenesis in various cancers. The tumour xenografting chicken embryo chorioallantoic membrane (CAM) model-based ex vivo assay was used to evaluate the activity of TMF. The Western blot, densitometric analysis and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate the activity of the MMP9. Zebrafish embryos were used to evaluate embryotoxicity, and in vitro free radical scavenging activity of flavonoid was also elucidated. This research assessed the high level of STAT3, p-ERK, VEGF-R and MMP9 in the tissue extract of the control group, and also, the suppression of angiogenesis in the treatment groups was due to scavenged ROS and RNS, dephosphorylation of STAT3 and ERK, and suppression of MMP9 gene expression. The isolated flavonoid named TMF from T.chrysantha functions as specific regulators of target proteins of angiosarcoma. The STAT3-MMP9 signalling may be used as an effective prognostic marker of angiosarcoma.

TNF-related apoptosis-inducing ligand (TRAIL) promotes trophoblast cell invasion via miR-146a-EGFR/CXCR4 axis: A novel mechanism for preeclampsia?

IntroductionPreeclampsia (PE) is associated with failure of uterovascular transformation due to impaired trophoblast invasion. Previously, TNF-related apoptosis-inducing ligand (TRAIL) has been controversially reported to correlate with PE, but whether it regulates trophoblast invasion yet to be defined. MethodsWe treated HTR8/SVneo cells with sTRAIL at concentrations of 0.1 ng/mL, 1 ng/mL and 10 ng/mL for a 72-h time course and compared cell proliferation, apoptosis and invasion ability by CCK8 assay, flow cytometry analysis and Matrigel cell invasion assay, respectively. The expressions levels of miR-146a, CXCR4, EGFR and MMP2 were quantified by qRT-PCR and Western blot. Moreover, HTR8/SVneo cells were transfected with miR-146a mimics and miR-146a inhibitor, followed by analyzing invasion ability and gene expressions of CXCR4, EGFR and MMP2. Finally, both serum and placental samples from preeclamptic and gestational week-matched normotensive women were collected and assessed for the expression levels of TRAIL by ELISA assay and immunochemistry staining. ResultssTRAIL treatment of HTR8/SVneo cells resulted in no change in proliferation or apoptosis, but dose- and time-dependently enhanced invasion. TRAIL downregulated the expressions of miR-146a and upregulated CXCR4, EGFR and MMP2. Transfection of miR-146a resulted in the inhibition of invasion and downregulation of CXCR4, EGFR and MMP2. Lastly, the expression levels of TRAIL decreased in both serum and placenta of preeclamptic pregnancies and correlated with the disease severity. DiscussionTRAIL regulated miR-146a-CXCR/EGFR axis to promote the invasion of trophoblast like cells. Its deceased levels in preeclamptic sera and placenta, suggest that low levels of TRAIL might contribute to the pathogenesis of preeclampsia.

Antiproliferative and antimigratory effects of 3-(4-substituted benzyl)-5- isopropyl-5-phenylhydantoin derivatives in human breast cancer cells

In this study, a series of synthesized 3-(4-substituted benzyl)-5-isopropyl-5-phenylhydantoin derivatives as a potential antiproliferative and antimigratory agents were investigated. The possible antitumor mechanisms of investigated hydantoin derivatives were examined on human breast cancer cell line MDA-MB-231. The cells were treated with different concentrations of compounds (from 0.01 µM to 100 µM) during 24 h and 72 h. The proliferation index, nitric oxide production, apoptosis rate, and migration capacity were measured. The cell invasion potential was examined by measuring the level of MMP-9 and COX-2 gene expression. All tested compounds expressed antiproliferative activity and induced dose- and time-dependent increase in the level of nitrites. The investigated molecules significantly decreased cell survival rate, migration capacity and the expression levels of genes included in the process of tumor invasion. Obtained data suggest that the tested hydantoin derivatives express considerable antitumor activity by reducing cell division rate, elevating apoptosis level, and inhibiting the motility and invasiveness of breast cancer cells. The results obtained in this study indicate that investigated compounds express potential as a novel chemotherapeutic agents against breast cancer growth and progression.

Effect of a Peptide Construct on Differentiated Macrophage MMP-2 and MMP-9 Levels of Varicose Patients.

The Matrix Metalloproteinase (MMPs) secreted from macrophages can affect the extracellular matrix remodeling process and improve varicose veins. The aim of this study was to investigate the MMP-2 and MMP-9 gene expression and activity levels in the differentiated macrophages M2 of subjects with varicose veins, and to evaluate a peptide construct on their catalytic functions. The macrophages were differentiated from the monocytes using M-CSF. The MMP-2 and MMP-9 gene expression and activity levels were measured by RT-qPCR and Zymography techniques, respectively. A peptide construct (ESLCG) was predicted with bioinformatics tools, and was prepared for the study of enzyme functions as compared to Batimastat. Furthermore, the docking studies were obtained for the evaluation of interactions between peptide construct, Batimastat and enzyme 3D structures. The results showed significant increases in MMP2 and MMP9 gene expression levels (P<0.001 and P<0.004, respectively) and gelatinolytic activities (P<0.001 and P<0.0001, respectively) in the macrophages. In agreement with the inhibitory effects of Batimastat, the peptide construct inhibited the MMP-2 and MMP-9 gelatinolytic activities up to 6.8 and 6.5 folds in the concentration of 150 µM. The docking analyses showed that the Lys187, Arg98, Leu49, Gly189, Leu190, Met97, Tyr53 and Phe57 residues of MMP-2 and the Leu187, His190, Glu402, His401, His405 and His411 residues of MMP-9 are interacted with the atoms of Batimastat and ESLCG peptide. The ESLCG peptide may be applied as an inhibitor of MMP-2 and MMP-9 enzymes in the subjects with varicose veins.

Prediction of metastasis in oral squamous cell carcinoma through phenotypic evaluation and gene expression of E-cadherin, β-catenin, matrix metalloproteinase-2, and matrix metalloproteinase-9 biomarkers with clinical correlation.

CONTEXT:Controversies prevail regarding the true predictive role of epithelial–mesenchymal transition (EMT) biomarkers in metastasis of oral squamous cell carcinoma (OSCC). There is also limited research carried on till date wherein the protein and gene expression of EMT biomarkers have been investigated simultaneously in the Indian population.AIM:The aim of this study was to assess the gene expression and quantitative protein expression of EMT biomarkers using conventional method and MATLAB software and to determine if there is any difference in the expression between metastatic and nonmetastatic OSCCs with clinicopathologic correlation.SETTINGS AND DESIGN:Twenty metastatic and nonmetastatic OSCC tissue sections each were obtained from department archives. Gene expression and quantified protein expression of EMT markers were done and correlated with clinical parameters.SUBJECTS AND METHODS:Sections immunostained for EMT biomarkers were evaluated using semi-quantitative and quantitative (MATLAB) methods. Gene expression using semi-quantitative reverse transcriptase–polymerase chain reaction was done. These findings were correlated with clinical parameters.STATISTICAL ANALYSIS USED:Pearson's Chi-square test, Student's t-test, and univariate logistic regression analysis were performed using SPSS software.RESULTS:The low immunoexpression of E-cadherin and β-catenin and the high expression of matrix metalloproteinase (MMP)-2 and MMP-9 correlate with Stages III and IV showing high metastatic risk. Furthermore, the upregulated MMP-2 and MMP-9 gene expressions in advanced clinical stages of OSCC have high metastatic potential.CONCLUSIONS:This study is the first of its kind to employ texture and color segmentation in MATLAB to objectively assess the protein expression of EMT biomarkers. This research is instrumental in studying the protein and gene expressions of EMT markers with clinical correlation.

Matrix Metallopeptidase 3 Polymorphisms: Emerging genetic Markers in Human Breast Cancer Metastasis.

Matrix metallopeptidase 3 or MMP3, is a zinc-dependent proteolytic enzyme that is involved in various physiological processes via modification of the extracellular matrix. In particular, its over-expression has been associated with cancer metastasis and tumor growth in various cancers including breast cancer. MMP3 gene expression is regulated by several factors such as DNA polymorphisms which also serve as risk factors for breast cancer. As such, DNA polymorphisms of MMP3 have the potential to be utilized as genetic biomarkers for prediction and prognosis of metastatic breast cancer. Presently, genome-wide association studies of MMP3 gene polymorphisms which are associated with breast cancer risk and patient survival in a variety of populations are reviewed. In order to understand the potential role of MMP3 polymorphisms as genetic markers for breast cancer metastasis, the domain structure of MMP3, the regulation of its expression and its role in breast cancer metastasis are also briefly discussed in this review. The emergence of MMP3 gene polymorphisms as prognostic biomarker candidates for breast cancer metastasis may contribute towards improving targeted therapies and categorization of breast cancer cases in order to provide a better and more accurate prognosis.

Type-I interferon regulates matrix metalloproteinases clearance of the bovine endometrial spheroid.

This study investigated the effect of type-I interferon (IFN) on the expression of matrix metalloproteinases (MMPs) of the bovine endometrial stromal cells (BES) and epithelial cells (BEE). The cells were separated and purified from the caruncles and cultured in DMEM/F-12 containing 10% fetal bovine serum. Spheroids were generated by using ascorbate. Zymograms of the supernatant showed that BEE predominantly expressed MMP-9, whereas MMP-2 was expressed in BES and homo-spheroids. While MMPs expression was not detected in hetero-spheroids. Real-time quantitative PCR revealed that type-I IFN and P4 suppressed the gene expression of MMP-2 and MMP-9 in hetero-spheroids, respectively. On the other hand, gelatin zymography analysis of the supernatant showed that type-I IFN strongly promote the clearance of MMPs. While zymograms of the MMPs stocked in the hetero-spheroids were significantly reduced by type-I IFN. Phenylmethanesulfonyl fluoride and leupeptin (both are serine proteinase inhibitors) significantly repressed the clearance of MMP-2 and MMP-9 induced by type-I IFN. Moreover, collagen fibers in hetero-spheroids significantly decreased after the treatment with type-I IFN. In conclusion, it was suggested that type-I IFN participate in the tissue remodeling by regulation the clearance of MMPs.

Photoaging Protective Effects of Ranunculus bulumei Methanol Extract.

Ultraviolet B (UVB) radiation is the main cause of photoaging processes including cellular senescence, skin drying, collagen degradation, melanogenesis, and inflammation. These responses occur because UVB induces a change in expression of aging-related genes through regulation of signal pathways such as that of mitogen-activated protein kinases- (MAPKs-) activator protein 1 (AP-1). Ranunculus bulumei, which is used as an herb in Indonesia, belongs to the Ranunculaceae family, which has been reported to perform various physiological effects including antioxidant and anti-inflammation. However, data on the pharmaceutical and cosmeceutical utility of Ranunculus bulumei have not been reported. Therefore, we evaluated the antiaging efficacy of RB-ME, a methanol extract of Ranunculus bulumei. Rb-ME attenuated MMP9 and COX-2 gene expression but enhanced SIRT1 and type-1 collagen in UVB-irradiated HaCaT cells. Rb-ME regulated these gene expressions through inhibition of p38 phosphorylation and inactivation of AP-1. In addition, mRNA expression of HAS-2 and -3, which are involved in skin hydration, was elevated in Rb-ME-treated HaCaT cells. Rb-ME also inhibited melanogenesis by suppression of tyrosinase, MITF, and TYRP-1 mRNA in B16F10 cells under α-MSH treatment. Taken together, these results indicate that Rb-ME has a protective effect on some UVB-induced skin photoaging events such as inflammation, collagen degradation, cellular senescence, skin drying, and melanin production through inhibition of the p38-AP-1 signal cascade, indicating that Rb-ME can be used as an active ingredient for antiaging cosmetics.

Overexpression of DDR1 Promotes Migration, Invasion, Though EMT-Related Molecule Expression and COL4A1/DDR1/MMP-2 Signaling Axis.

Purpose:Discoidin domain receptor 1 (DDR1) belongs to a novel class of receptor tyrosine kinases. Previous evidence indicates that DDR1 overexpression promotes the aggressive growth of bladder cancer (BC) cells. This study aimed to investigate the molecular mechanisms by which DDR1 influences BC.Methods:DDR1 was transfected into human BC RT4 cells. DDR1, COL4A1, and MMP-2 expression in 30 BC tissues and paired adjacent tissues were examined by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry. Transwell assays were conducted to determine cell migration and invasion. RT-PCR and western blot (WB) were also used to measure the DDR1, COL4A1, MMP-2, and EMT-related gene (ZEB1 and SLUG) expression in RT4 cells after DDR1 overexpression.Results:COL4A1 and MMP-2 interacted with DDR1 in the PPI network. RT-PCR and immunohistochemistry results showed that both mRNA and protein levels of DDR1 and COL4A1 were significantly increased in BC tissue, while the expression of MMP-2 was increased only at the mRNA level (P < 0.05). Overexpression of DDR1 in RT4 cells significantly promoted their migratory and invasive capabilities in vitro (P < 0.05). Moreover, overexpression of DDR1 in RT4 cells increased the mRNA and protein expression of ZEB1, SLUG, COL4A1, and MMP-2 (P < 0.01). DDR1-mediated migration and invasion of RT4 cells were reversed after COL4A1-siRNA treatment.Conclusion:DDR1 may be a potential therapeutic target in BC patients.

Apelin-APJ axis inhibits TNF-alpha-mediated expression of genes involved in the inflammatory response in periodontal ligament cells

Periodontitis is an inflammatory disease of the supportive tissues surrounding the teeth, and is characterized by irreversible destruction of the gingiva, periodontal ligament (PDL), and alveolar bone, which results in the loss of teeth. In the present study, we elucidated the correlation between periodontitis and apelin (APLN), an adipokine and a regulatory peptide, respectively, which are involved in inflammation and bone remodeling. The expression of APLN is negatively correlated with periodontitis progression in gingival tissue. In addition, treatment with TNF-α downregulated the expression of APLN in PDL cells and gingival fibroblasts, indicating the protective role played by APLN against periodontitis progression. The overexpression of APLN or treatment with exogenous APLN suppressed the TNF-α- mediated catabolic gene expression of MMP1, IL6 , and PTGS2 in PDL cells. Moreover, the inhibition of the APLNAPJ axis by ML221, an APJ inhibitor, induced catabolic gene expression in PDL cells. Thus, the results of this study provided evidence to support APLN as a regulatory factor of the inflammatory response during periodontitis.

The effect of opine on matrix metalloproteinase expression in mice with breast cancer

Regarding the anti-inflammatory and anti-tumour effects of arginine and its derivatives, this study evaluates matrix metalloproteinase (MMPs) expression in an animal model of breast cancer following administration of octopine. In this study, 40 animals of Balb/C mice were divided into 5 groups: the healthy control, the cancer control, the cancer group receiving 50 mg of octopine, the cancer group receiving 100 mg of octopine and the cancer group receiving 150 mg of octopine for 3 weeks. 4T1 cell line was used to induce cancer. Biopsy specimens were enrolled from mice and MMP-1, MMP-3 and MMP-9 gene expression evaluated using real-time PCR, while these protein amounts were measured using immunohistochemistry and ELISA methods. Data were analysed using one-way ANOVA, Kruskal-Wallis and Mann–Whitney U tests (p < .05). The results showed that 100 mg octopine consumption had significant decreasing effect on MMP-9 expression (p = .02) in the treatment group compared with cancerous non-treated mice. Furthermore, results from immunohistochemistry and ELISA confirmed this effect, the protein amount of MMP-9 was significantly decreased in group treating with 100 mg octopine (.005). The use of octopine has a beneficial effect on reducing MMP-9 in mice breast cancer.

PEPTIDE EXTRACTS FROM TEMPEH PROTECT AGAINST PHOTOAGING VIA DOWN-REGULATING MATRIX METALLOPROTEINASE-3 GENE EXPRESSION

Photoaging damages skin tissue via activation of matrix metalloproteinases (MMPs) and MMP-3 induces activation of MMP-1 and 9. Tempeh is made from fermented soybean in Indonesia and has been known for its high protein content. This study was aimed to investigate the effectiveness of peptide extracts from tempeh against photoaging at gene level in UVB-treated mice. Peptide was extracted from soybean and black soybean tempeh, and identified for its amino acid profiling. Photoaging model was made by gradient UVB exposure to seven groups of mice for 4 weeks. Mice were supplemented with 100 and 300 mg/kg body weight dosages of peptide extracts from soybean and black soybean tempeh. Dorsal skin tissue was collected for histology staining and quantitative polymerase chain reaction (qPCR). This present study found that both tempeh peptide extracts were rich in glutamic acid and branched-chain amino acids. Histology staining showed that tempeh protected mice skin from photoaging. At 100 and 300 mg/kg, both tempeh peptide extracts significantly attenuated MMP-3gene expression that lead to the decrease level of MMP-1 and MMP-9 gene expression. These results suggest that supplementation of both tempeh peptide extracts may possess anti-photoaging effect via altering MMP-3 gene expression in UVB-treated mice.

Fucoxanthin enhances 5-FU chemotherapeutic efficacy in colorectal cancer cells by affecting MMP-9 invasive proteins

Background : Drug resistance is a major inconvenient lowering the traditional chemotherapeutic efficacy and highly undesirable therapeutic problem which crucially editing faster in colorectal cancer. Fucoxanthin is a naturally orange-carotenoid, predominantly found in edible brown algae and justify considered as nutritional ingredient possess with powerfully new strategy to enhance concurrent drug chemotherapy. Obviously studied exhibited that fucoxanthin is very good well documented as potential anti-cancer activity, remarkable numerous biological activities. Accordingly, it has prominently gain research to enhancing the understand in molecular mechanism details associating cancer therapy. Objectives : This study was undertaken to assess the anti-cancer activity and explored the molecular mechanism of fucoxanthin on the inhibition of cell proliferation and cell invasion. Methods : SW-620 cells were cultivated with fucoxanthin with for 24, 48, 72, 96 hr and co-treatment with 5-FU to evaluate synergistic potential. The cell viability of cancerous cells were determined by MTT colorimetric assay. The inhibitory effect of cell invasion and adhesion was measured in the presence of fucoxanthin with 5-FU in various concentrations. To determine MMP-9 gene and protein expression after treated cells by RT-PCR and ELISA assay Results : The results illustrated that fucoxanthin profoundly inhibited cell proliferation of SW-620 cells accompanied by growth arrest and diminished invasive ability, which mediated at one least part by the down-regulation of MMP-9 mRNA and protein expression. Especially, fucoxanthin extremely attenuated the anti-proliferative effect of established 5-FU by modulating the habitually hallmark of cancerous cells. Conclusions : This results are raising up the capacity of fucoxanthin to eradicate cancer cells and the possibility that fucoxanthin could become a promising marine natural active constituents from seaweeds. Critical outcome of data in our studies will be serving as preliminary results for further studies marine drug in experimental model and well-controlled clinical trials.

Staphylococcus aureus on the effect of expression of MMPs/TIMPs and uPA system in bovine mammary fibroblasts.

Bovine mammary fibrosis is characteristic of chronic in injury in response to diverse pathogens. Staphylococcus aureus (S.aureus) is a frequent cause of mastitis in bovine and is prone to persistent infection. Diverse studies have shown MMPs/TIMPs and uPA system as a potent target for the treatment of fibrosis. However, pathogenesis of S. aureus-induced mammary fibrosis has not been completely defined. BMFBs treated with heat-inactivated S. aureus (105, 106, and 108CFU/mL) for 6, 12, 24, and 48h. Total RNA and protein were isolated from the treatments and controls of BMFBs samples. MMP-1, MMP-2, MMP-3, MMP-9, MMP-13, TIMP-1, TIMP-2, COL Ⅰ, uPA, uPAR and PAI-1 gene and protein expression were examined by RT-qPCR and Western blot analysis. Gelatin zymography assay was performed to assess the levels of MMP-2 and MMP-9 enzyme secreted. BMFBs treated with heat-inactivated S. aureus increased mRNA and protein expression levels of MMP-1, MMP-2, MMP-3, MMP-9 and MMP-13, and heat-inactivated S. aureus induced TIMP-1, TIMP-2 and COL Ⅰ expression. There was a clear activation of MMP-2 in the presence of heat-inactivated S. aureus in the conditioned medium from the BMFBs, whereas MMP-9 was no significantly altered. Moreover, uPA system was activated in BMFBs to S. aureus. Activation of the uPA system together with its impact on the MMPs levels could play a significant role in S. aureus-induced BMFBs with mechanism of ECM metabolism, MMPs/TIMPs and uPA system could participate in bovine mammary fibrosis.