Overexpression Of Matrix Metalloproteinases Research Articles

Bacterial Expression and Purification of Human Matrix Metalloproteinase-3 using Affinity Chromatography.

Matrix metalloproteinases (MMPs) belong to the family of metzincin proteases with central roles in extracellular matrix (ECM) degradation and remodeling, as well as interactions with several growth factors and cytokines. Overexpression of specific MMPs is responsible in several diseases such as cancer, neurodegenerative diseases, and cardiovascular disease. MMPs have been the center of attention recently as targets to develop therapeutics that can treat diseases correlated to MMP overexpression. To study the MMP mechanism in solution, more facile and robust recombinant protein expression and purification methods are needed for the production of active, soluble MMPs. However, the catalytic domain of most MMPs cannot be expressed in Escherichia coli (E. coli) in soluble form due to lack of posttranslational machinery, whereas mammalian expression systems are usually costly and have lower yields. MMP inclusion bodies must undergo the tedious and laborious process of extensive purification and refolding, significantly reducing the yield of MMPs in native conformation. This paper presents a protocol using Rosetta2(DE3)pLysS (hereafter referred to as R2DP) cells to produce matrix metalloproteinase-3 catalytic domain (MMP-3cd), which contains an N-terminal His-tag followed by pro-domain (Hisx6-pro-MMP-3cd) for use in affinity purification. R2DP cells enhance the expression of eukaryotic proteins through a chloramphenicol-resistant plasmid containing codons normally rare in bacterial expression systems. Compared to the traditional cell line of choice for recombinant protein expression, BL21(DE3), purification using this new strain improved the yield of purified Hisx6-pro-MMP-3cd. Upon activation and desalting, the pro domain is cleaved along with the N-terminal His-tag, providing active MMP-3cd for immediate use in countless in vitro applications. This method does not require expensive equipment or complex fusion proteins and describes rapid production of recombinant human MMPs in bacteria.

Novel Cinnamic Acid Derivatives as Potential Anticancer Agents: Synthesis, In Vitro Cytotoxicity and Molecular Docking Studies

Inhibition of matrix metalloproteinase-9 (MMP-9) can be an emerging strategy for a cancer cure as overexpression of MMP-9 is associated with several types of malignancies, including cancers related to lungs, breasts, and prostate. The present work was proposed to design and synthesize some novel cinnamic acid derivatives as potential anticancer agents. Novel cinnamic acid derivatives were designed using a structure-based drug design approach, and a series of 16 newer cinnamic acid analogs was prepared and evaluated for in vitro cytotoxicity (lung cancer cell line, A-549), followed by docking studies to explore binding interactions of designed compounds in the binding site of MMP-9. In the in vitro cytotoxicity assay, compound 5 was found to be most potent, with an IC50 value of 10.36 µM amongst the synthesized analogs. The designed molecules showed appreciable docking interactions and binding patterns with MMP-9 protein supporting the in vitro cytotoxicity outcomes. Compound 5 can be further explored as a possible lead molecule to develop potent and selective MMP-9 inhibitors as potential antineoplastic agents.

AGEs-induced MMP-9 activation mediated by Notch1 signaling is involved in impaired wound healing in diabetic rats

AimsTo elucidate the relationship between advanced glycation end products (AGEs), Notch1 signaling, nuclear factor-kappa B (NF-κB), and matrix metalloproteinase-9 (MMP-9) in diabetic wound healing in vitro and in vivo. MethodsWe incubated primary keratinocytes with AGEs alone or AGEs along with γ-secretase inhibitor DAPT, and established diabetic rat wound model by intraperitoneal streptozotocin treatment. The Notch1 signaling components and MMP-9 expression were detected by qPCR, western blotting and gelatin zymography. ResultsThe exposure of primary keratinocytes to AGEs led to a significant increase in Notch intracellular domain (NICD), Delta-like 4 (Dll4), and Hes1; however, Notch1 expression was inhibited by the RAGE siRNA. Furthermore, MMP-9 activation was up-regulated, secondary to AGEs treatment. In contrast, increased MMP-9 expression by AGEs-stimulation was eliminated after treatment with DAPT. NF-κB activation participated in the Notch1-modulated MMP-9 expression. Notably, in the diabetic animal model, inhibition of the Notch signaling pathway with DAPT attenuated NICD and MMP-9 overexpression, improved collagen accumulation, and ultimately accelerated diabetic wound healing. ConclusionsThese findings identified that activation of the Notch1/NF-κB/MMP-9 pathway, in part, mediates the repressive effects of AGEs on diabetic wound healing and that targeting this pathway may be a potential strategy to improve impaired diabetic wound healing.

Aetiology of Primary Spontaneous Pneumothorax.

Air in the pleural cavity is termed pneumothorax. When this occurs in the absence of trauma or medical intervention, it is called spontaneous pneumothorax. Primary spontaneous pneumothorax typically occurs in young patients without known lung disease. However, the idea that these patients have “normal” lungs is outdated. This article will review evidence of inflammation and respiratory bronchiolitis on surgical specimens, discuss the identification of emphysema-like change (i.e., blebs and bullae), the concept of pleural porosity and review recent data on the overexpression of matrix metalloproteinases in the lungs of patients who have had pneumothorax.

GSK-3α/β Activity Negatively Regulates MMP-1/9 Expression to Suppress Mycobacterium tuberculosis Infection.

Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) infection is the deadliest infectious disease and a global health problem. Macrophages (Mφs) and neutrophils that can phagocytose Mtb represent the first line of immune response to infection. Glycogen synthase kinase-3α/β (GSK-3α/β) represents a regulatory switch in host immune responses. However, the efficacy and molecular mechanisms of how GSK-3α/β interacts with Mtb infection in Mφs remain undefined. Here, we demonstrated that Mtb infection downregulated GSK-3α/β activity and promoted matrix metalloproteinase-1 (MMP-1) and MMP-9 expressions in Mφs derived from acute monocytic human leukemia THP-1 cells (THP-1-Mφs). We confirmed the upregulation of MMP-9 expression in tissues of TB patients compared with patients of chronic inflammation (CI). In THP-1-Mφs and C57BL/6 mice, GSK-3α/β inhibitor SB216763 significantly increased MMP-1/9 production and facilitated Mtb load, while MMP inhibitors blocked MMP-1/9 expression and Mtb infection. Consistently, GSK-3α/β silencing significantly increased MMP-1/9 expression and Mtb infection, while overexpression of GSK-3α/β and constitutive activated GSK-3α/β mutants significantly reduced MMP-1/9 expression and Mtb infection in THP-1-Mφs. MMP-1/9 silencing reduced Mtb infection, while overexpression of MMP-1/9 promoted Mtb infection in THP-1-Mφs. We further found that GSK-3α/β inhibition increased Mtb infection and MMP-1/9 expression was blocked by ERK1/2 inhibitor. Additionally, we showed that protein kinase C-δ (PKC-δ) and mammalian target of rapamycin (mTOR) reduced GSK-3α/β activity and promoted MMP-1/9 production in Mtb-infected THP-1-Mφs. In conclusion, this study suggests that PKC-δ-mTOR axis suppresses GSK-3α/β activation with acceleration of MMP-1/9 expression through phospho-ERK1/2. These results reveal a novel immune escape mechanism of Mtb and a novel crosstalk between these critical signaling pathways in anti-TB immunity.

Prognostic value of matrix metalloproteinase 2 protein expression in ovarian cancer is age- and stage-dependent

Objective: Matrix metalloproteinase 2 (MMP2) has been associated with tumor development and invasion; however, the information available regarding its prognostic value in ovarian cancer (OC), especially in the Arabian Peninsula, is limited. The aim of this retrospective study was to analyze MMP2 protein expression and assess its prognostic value. Methods: In total, 245 formalin-fixed and paraffin-embedded (FFPE) primary OC tissue samples were randomly collected from patients with available clinicopathological data, including disease of all stages and all histological subtypes. MMP2 protein expression was measured using automated tissue microarray and immunohistochemistry techniques. Statistical analyses were performed using SPSS, with p << 0.05 considered statistically significant. Results: Cytoplasmic MMP2 protein expression patterns were higher in 53% of all tumor samples. The MMP2 expression profile was not significantly correlated with most clinicopathological features including age, tumor site, size, grade, and lymph node status (p >> 0.05). However, when adjusted according to the disease stage or patient age, MMP2 overexpression showed a significant indication of a poor outcome and recurrence as evaluated using univariate Kaplan–Meier analysis for disease-free survival (DFS) (p = 0.04 and p = 0.03, respectively, log-rank test), but not for disease-specific survival (DSS) (p >> 0.05, log-rank test). Conclusion: This study showed that MMP2 protein overexpression was a negative prognosticator in Saudi OC patients with advanced stage and/or young age. These results could pave the way towards more effective and personalized detection, prognosis, and management of OC.

Circ_0000620 acts as an oncogenic factor in gastric cancer through regulating MMP2 expression via sponging miR-671-5p

BackgroundGastric cancer (GC) is one of the most common cancers in the digestive system. Circular RNAs (circRNAs) have been found to function as important regulators in the pathogenesis of GC. This study focused on the biological role and molecular mechanism of circ_0000620 in GC progression.MethodsThe expression levels of circ_0000620, microRNA-671-5p (miR-671-5p) and Matrix MetalloProteinase 2 (MMP2) were measured by quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC) assay or western blot. The stability of circ_0000620 was confirmed by Ribonuclease R (RNase R) assay. The protein levels were determined by western blot assay. Cell viability, colony formation, cell migratory ability, cell invasive ability and tube formation capacity were respectively examined by CCK-8 assay, colony formation assay, wound healing assay, transwell invasion assay and tube formation assay. The interaction between miR-671-5p and circ_0000620 or MMP2 was validated by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay. The role of circ_0000620 in GC undefined was explored by xenograft tumor assay.ResultsCirc_0000620 was conspicuously upregulated in GC tissues and cells. Circ_0000620 knockdown reduced cell viability, colony formation, migration, invasion and tube formation capacity of GC cells in vitro. Furthermore, MMP2 was upregulated in GC and MMP2 overexpression reversed the anti-tumor response of circ_0000620 knockdown in GC progression. Moreover, circ_0000620 directly interacted with miR-671-5p and circ_0000620 downregulation regulated malignant behaviors of GC cells by upregulating miR-671-5p. In addition, silencing of circ_0000620 inhibited tumor growth in vivo.ConclusionsCirc_0000620 knockdown inhibited the malignant development of GC partly through modulating the miR-671-5p/MMP2 axis.

Matrix Metalloproteinase-7 Aggravated the Oxidized Low Density Lipoprotein-Induced Damage of Human Vascular Endothelial Cells

Introduction: Vascular endothelial injury could induce many cardiovascular diseases. Recently, some studies have indicated that matrix metalloproteinase-7 (MMP-7) was associated with the occurrence and development of cardiovascular diseases. However, whether higher levels of MMP-7 were associated with the occurrence of the vascular endothelial injury is unclear. Material and methods: In this study, ox-LDL was used for the simulation of vascular endothelial injury in HUVECs. Next, we detected the expression of MMP-7 in these cells. After that, we established the cell models with MMP-7 overexpression and knockdown, respectively. At last, the apoptosis and inflammation of HUVECs were detected with corresponding assays. Results: After the stimulation of ox-LDL, the expression of MMP-7 was enhanced compared to the control groups. After the stimulation of ox-LDL and the overexpression of MMP-7, the apoptosis rates of HUVECs were enhanced, while MMP-7 knockdown led to the decreased apoptosis rates of these cells. Furthermore, after the stimulation of ox-LDL and overexpression of MMP-7, the expression of inflammatory factors (IL-6, IL-1β and TNF-α) was promoted. Additionally, the expression of these proteins was repressed after knockdown of MMP-7. Conclusion: MMP-7 aggravated the ox-LDL-induced damage of HUVECs by promoting the apoptosis and inflammation of these cells.

Osteoblasts Regulate the Expression of ADAMTS and MMPs in Chondrocytes through ERK Signaling Pathway.

Degradative enzymes such as matrix metalloproteinase (MMP) and disintegrin metalloproteinase with platelet thrombin-sensitive protein-like motifs (ADAMTS) play a key role in the development of osteoarthritis (OA). We aimed to investigate the effects of OA subchondral osteoblasts on the expression of ADAMTS4, ADAMTS5, MMP-3, MMP-9, and MMP-13 in chondrocytes and the regulation of mitogen-activated protein kinase (MAPK) signaling pathway. A rat knee OA model was constructed by cutting the anterior cruciate ligament of the knee joints, and normal rat articular cartilage chondrocytes (N-ACC), OA rat articular cartilage chondrocytes (O-ACC), normal subchondral bone osteoblasts (N-SBO), and OA subchondral bone osteoblasts (O-SBO) were isolated and extracted. The expressions of O-ACC and O-SBO COL1 and COL2 were detected respectively. Chondrocytes were identified by immunofluorescence of COL2 and toluidine blue staining, and osteoblasts were identified by COL1 immunofluorescence, alkaline phosphatase (ALP), and Alizarin Red staining. Gene expression of COL1, COL2, and aggrecan in normal chondrocytes and OA chondrocytes, and gene expression of osteoblast ALP and osteocalcin (OCN) were detected by RT-PCR to identify the two chondrocytes and the two osteoblast phenotypes. The constructing N-ACC group, O-ACC group, N-ACC + N-SBO group, N-ACC + O-SBO group, O-ACC + N-SBO group, O-ACC + O-SBO group, I+ N-ACC + O-SBO group, and I + O-ACC + O-SBO group cell cultures, and the expression of ERK, ADAMTS4, ADAMTS5, MMP-3, MMP-9, and MMP-13 genes in chondrocytes cultured for 0, 24, 48, and 72 h were detected by RT-PCR. The protein expressions of pERK, ADAMTS4, ADAMTS5, MMP-3, MMP-9, and MMP-13 were detected by Western blot. · The X-ray showed that the knee joint space of the affected limb became narrow.. · The results of RT-PCR of COL2 and aggrecan gene in OA and normal chondrocytes suggest that the relative expression of COL2 in OA articular chondrocytes (0.24 ± 0.07) issignificantly lower than that in normal cartilage (0.61 ± 0.07) (p < 0.05). The relative expression of AGG (0.37 ± 0.16) in OA chondrocytes was significantly lower than that of normal chondrocytes AGG (1.30 ± 0.25) (p < 0.05). The expression of COL1 was very low, and was not statistically significant.. · The results of RT-PCR of the osteoblast ALP and OCN gene indicated that gene expression of ALP (12.30 ± 1.17) and OCN (20.47 ± 4.19)was upregulated when compared with the relative expression of ALP (4.66 ± 0.71) (p < 0.05) and OCN (12.17 ± 2.76) (p < 0.05) in normal osteoblasts, indicating that osteoblasts of OA have greater osteogenic potential than normal osteoblasts.. · The expressions of ADAMTS4, ADAMTS5, MMP-3, MMP-9, and MMP-13 genes and proteins in OA chondrocytes or normal chondrocytes were basically unchanged when they were cocultured with normal osteoblasts. Indirect coculture of OA osteoblasts and chondrocytes could promote the expression of ADAMTS4, ADAMTS5, MMP-3, MMP-9, and MMP-13 genes and proteins in chondrocytes. Overexpression of ADAMTS and MMP in coculture systems can be reversed by MAPK-ERK inhibitors.. · OA subchondral bone osteoblasts can promote the overexpression of ADAMTS and MMPs in chondrocytes.. · The ERK signaling pathway may be involved in the regulation of the effect of subchondral bone osteoblasts on chondrocytes..

Matrix metalloproteinase contribution in management of cancer proliferation, metastasis and drug targeting.

Matrix metalloproteinases (MMPs) or matrixins, are members of a zinc-dependent endopeptidase family. They cause remodeling of the extracellular matrix (ECM) leading to numerous diseases. MMPs subfamilies possess: collagenases, gelatinases, stromelysins and membrane-type MMPs (MT-MMP). They consist of several domains; pro-peptide, catalytic, linker peptide and the hemopexin (Hpx) domains. MMPs are involved in initiation, proliferation and metastasis of cancer through the breakdown of ECM physical barriers. Overexpression of MMPs is associated with poor prognosis of cancer. This review will discuss both types of MMPs and current inhibitors, which target them in different aspects, including, biosynthesis, activation, secretion and catalytic activity. Several synthetic and natural inhibitors of MMPs (MMPIs) that can bind the catalytic domain of MMPs have been designed including; peptidomimetic, non-peptidomimetic, tetracycline derivatives, off-target MMPI, natural products, microRNAs and monoclonal antibodies.

The elevated expression of TLR4 and MMP9 in human abdominal aortic aneurysm tissues and its implication

BackgroundToll-like receptor 4 (TLR4) and matrix metalloproteinase 9 (MMP9) have been investigated to play significant roles in the formation of abdominal aortic aneurysm (AAA). But the reports on the expression pattern of TLR4 and MMP9 in human AAA specimens were relatively scant. The aim of this study was to make a detailed analysis of TLR4 and MMP9 expression in situ and their association with clinical parameters involved in human AAA.Methods40 AAA specimens were obtained from full-thickness aneurysmal tissues at the maximal dilation area during the open surgical repair, and 8 non-aneurysmal abdominal aortas from transplant donors served as controls. Expression of TLR4 and MMP9 protein was determined by immunohistochemistry.ResultsThere were increased levels of TLR4 and MMP9 expression in human AAA tissues. Compared with macrophages or SMCs, lymphocytes showed a higher positive rate of TLR4 and MMP9 staining, and an elevated ratio of high MMP9 expression (all P < 0.05). There existed a significant association between TLR4 and MMP9 expression (r = 0.767, P < 0.001), and both TLR4 and MMP9 levels were statistically related to circulating CRP. Moreover, TLR4 expression in situ indicated a positive correlation with its serum level (r = 0.654, P = 0.006). Multiple analysis revealed that high TLR4 expression in situ was associated with the risk of large AAA (OR = 6.211, 95%CI = 1.226–31.480, P = 0.027), while high MMP9 expression was correlated to the presence of thrombus within AAA (OR = 5.494, 95%CI = 1.181–25.562, P = 0.030), separately compared with their low expression.ConclusionsThis study confirmed the overexpression of TLR4 and MMP9 in human AAA tissues, and their close relationship implying in the pathogenesis of AAA. We further provided evidence that TLR4 had a potential effect on AAA size and MMP9 could influence the occurrence of thrombus within AAA.

Identification of Broad-Spectrum MMP Inhibitors by Virtual Screening.

Matrix metalloproteinases (MMPs) are the family of proteases that are mainly responsible for degrading extracellular matrix (ECM) components. In the skin, the overexpression of MMPs as a result of ultraviolet radiation triggers an imbalance in the ECM turnover in a process called photoaging, which ultimately results in skin wrinkling and premature skin ageing. Therefore, the inhibition of different enzymes of the MMP family at a topical level could have positive implications for photoaging. Considering that the MMP catalytic region is mostly conserved across different enzymes of the MMP family, in this study we aimed to design a virtual screening (VS) workflow to identify broad-spectrum MMP inhibitors that can be used to delay the development of photoaging. Our in silico approach was validated in vitro with 20 VS hits from the Specs library that were not only structurally different from one another but also from known MMP inhibitors. In this bioactivity assay, 18 of the 20 compounds inhibit at least one of the assayed MMPs at 100 μM (with 5 of them showing around 50% inhibition in all the tested MMPs at this concentration). Finally, this VS was used to identify natural products that have the potential to act as broad-spectrum MMP inhibitors and be used as a treatment for photoaging.

Overexpression of HOXA10 promotes the growth and metastasis of nasopharyngeal carcinoma.

Deregulation of HOX transcription factor family has frequently been observed in multiple human cancers; however, their role in nasopharyngeal carcinoma remains largely unclear. In the present study, we found that HOX gene family is consistently upregulated in nasopharyngeal carcinoma and identified HOXA10 as one of the mostly upregulated HOX genes. Importantly, we show that HOXA10 overexpression is associated with transcriptional activation of multiple oncogenes essential for nasopharyngeal carcinoma carcinogenesis, including S-phase kinase-associated protein 2 (SKP2), calcium/calmodulin-dependent protein kinase kinase 2 (CAMKK2), and matrix metalloproteinase 1 (MMP1). Mechanistically, the overexpression of SKP2 induces the degradation of cell cycle inhibitor p27, leading to rapid cell cycle progression and cell proliferation. The overexpression of CAMKK2 is associated with enhanced mTOR signaling activity to meet the increased demand for proteins synthesis in rapid growing nasopharyngeal carcinoma cells. Moreover, MMP1 overexpression facilitates nasopharyngeal carcinoma cell migration and invasion and contributes to cancer metastasis and progression. We thus concluded that HOXA10 overexpression promotes the growth and metastasis of nasopharyngeal carcinoma by transcriptionally activating various oncogenic pathways.

An in vitro investigation to understand the synergistic role of MMPs-1 and 9 on articular cartilage biomechanical properties

Matrix metalloproteinases (MMPs) play a crucial role in enzymatically digesting cartilage extracellular matrix (ECM) components, resulting in degraded cartilage with altered mechanical loading capacity. Overexpression of MMPs is often caused by trauma, physiologic conditions and by disease. To understand the synergistic impact MMPs have on cartilage biomechanical properties, MMPs from two subfamilies: collagenase (MMP-1) and gelatinase (MMP-9) were investigated in this study. Three different ratios of MMP-1 (c) and MMP-9 (g), c1:g1, c3:g1 and c1:g3 were considered to develop a degradation model. Thirty samples, harvested from bovine femoral condyles, were treated in groups of 10 with one concentration of enzyme mixture. Each sample was tested in a healthy state prior to introducing degradative enzymes to establish a baseline. Samples were subjected to indentation loading up to 20% bulk strain. Both control and treated samples were mechanically and histologically assessed to determine the impact of degradation. Young’s modulus and peak load of the tissue under indentation were compared between the control and degraded cartilage explants. Cartilage degraded with the c3:g1 enzyme concentration resulted in maximum 33% reduction in stiffness and peak load compared to the other two concentrations. The abundance of collagenase is more responsible for cartilage degradation and reduced mechanical integrity.

The Effect of B9 and B12 Vitamins Deficiency on Hypomethylation of MMP-9 gene Promoter Among Women With Preterm Parturition.

Preterm birth is one of the problems that pregnant women may encounter during pregnancy. Over-expression of Matrix metalloproteinases (MMPs) has the main role in the untimely remodeling of the extracellular matrix (ECM) and preterm delivery, but its exact mechanism is unknown. This study was undertaken to examine the methylation and expression patterns of this gene in women delivering preterm and also to analyze the correlation of serum B9 and B12 vitamins levels with expression and methylation of MMP-9 gene. The MMP-9 RNA and protein levels in 50 women delivering preterm and 50 women delivering at term were determined by Real-time PCR and enzyme-linked immunosorbent assay (ELISA) methods. Methylation pattern of this gene was evaluated by Epitect Methyl-II PCR assay and methylation-sensitive restriction enzymes combined with PCR amplification method; then, their correlation with serum B9 and B12 vitamins was studied. Placental MMP-9 RNA and protein levels were higher (P < 0.03, P < 0.01) in women delivering preterm as compared to those delivering at term. We also observed significant hypomethylation of promoter of this gene in preterm samples (P < 0.012). Moreover, significant negative correlation between B9 and B12 vitamins concentration and hypomethylation of MMP-9 gene was seen (r = - 0.68 P < 0.001). Our study suggests an impressive role of deficiency of B9 and B12 vitamins concentration on the hypomethylation and overexpression of MMP-9 gene which may lead to preterm parturition of pregnant women.

Effects of Tiarella polyphylla D. Don Callus Extract on Photoaging in Human Foreskin Fibroblasts Hs68 Cells

Ultraviolet (UV)-irradiation causes an overproduction of matrix metalloproteinases (MMPs) and collagen (COL) degradation causing skin sunburn, inflammation, or photoaging in human. In this study, we prepared callus from Tiarella polyphylla D. Don ( T. polyphylla) stem and its phytochemical profiles were analyzed using HPLC-MWD. The effects of T. polyphylla callus extract evaluated against on UVB-induced damage in human foreskin fibroblast (Hs68). Hs68 was exposed to UVB in the presence or absence of T. polyphylla callus extract at concentrations of 100 and 250 µg/mL. Cell damage caused by UVB was inhibited by T. polyphylla callus extract, which was tested by cell viability and caspase 3 activity in Hs68 cells. Further experiment revealed that T. polyphylla extract suppressed the level of MMP-1, but increased the level of type I procollagen. In addition, T. polyhylla callus extract inhibited UVB-mediated COL (-1 and -3) protein degradation and MMP (-1, 2, and -3) overexpression in Hs68. These results suggest that T. polyphylla callus extract has considerable potential as a cosmetic ingredient with anti-aging effects.

Cardiomyocyte‐Specific Transgenic MMP9 Overexpression Induces Cardiac Remodeling

Objective Activated matrix metalloproteinase-9 (MMP9), a collagenase, promotes cardiac remodeling leading to dysfunction. MMP9 is secreted by a wide variety of cells including cardiomyocytes. Although the role of MMP9 secreted from leukocytes and fibroblasts have been studied, very little is known about the role of cardiomyocyte-specific MMP9. Hypothesis Cardiomyocyte-specific MMP9 overexpression induces cardiac remodeling. Materials and Methods We have developed a novel cardiomyocytes-specific MMP9 transgenic (M9Tg) mouse strain. We used alpha-myosin heavy chain promoter and knocked in MMP9 gene using CRISPR-Cas9 method. After validation through genotyping, we performed gravimetric analyses and evaluated cardiac function by M-Mode echocardiography (short-axis), and hemodynamic changes by pressure-volume loop (open-chest, 4mm catheter, Transonic) studies. Results Eighteen-week old male M9Tg mice had a higher body weight and other organ's weight compared to the sibling WT. In four mice, the comparison of M9Tg/WT: body weight (g) = 26.8/24.8, heart weight (mg) = 119.0/107.0, Liver (mg) = 1309.5/1203.3, Kidney (mg) = 159.6/141.6, lung (mg) = 154.8/141.3). There was no significant change in the size of these organs relative to their body weight in M9Tg mice. The M-mode echocardiography analysis of three mice showed a decrease in cardiac output (WT= 15.8 ± 1.1, M9Tg =13.5 ± 0.8, P= 0.038) and a trend of decrease in fractional shortening (WT =27.4 ± 3.6, M9Tg = 24.9 ± 2.0, P = 0.231) and an increase in left ventricular mass (WT = 75.1 ± 4.1, M9Tg = 80.5 ± 3.8, P = 0.12). The pressure-volume loop study with two WT and three M9Tg mice showed an increase in dp/dt max (average, WT =5881.5, M9Tg = 7211.3 mmHg/s) and decrease in end systolic volume (WT = 22.1, M9Tg = 17.4 µL), suggesting a potential increase in cardiac contractility. Conclusion Cardiomyocyte-specific overexpression of MMP9 potentially increased left ventricular mass and cardiac contractility without altering fractional shortening. Further studies are ongoing to increase the sample size and examine the changes in the heart at the histological and molecular levels.

Role of Matrix Metalloproteinase in the Progression of Heart Failure: A Narrative Review

Heart failure (HF) is a cardiovascular disease with a complex pathological pathway and influenced by many factors. Such a complex pathological mechanism would impair cardiac function and structural stability. One that plays a role in maintaining the function and structure of the heart is the extracellular matrix (ECM), and disruption in the extracellular matrix has a role in causing cardiac dysfunction. ECM regulation is associated with matrix metalloproteinase (MMP). Overexpression of matrix metalloproteinases can lead to ECM degradation process which leads to cardiac remodelling. The role of MMP in heart failure is also related to the increased inflammatory response, which is one of the pathways for progression of heart failure. The close association of MMP with the development of heart failure makes MMP a potential biological marker. This article aims was to understand the role of MMP and its mechanisms in cardiac remodelling pathways leading to heart failure. This narrative review suggests that overexpression of MMP can lead to heart failure. Inflammation is one of the factors triggering the expression of MMP. Inflammation will increase the release of pro-inflammatory cytokines, thereby triggering MMP expression. MMP expression imbalance can damage collagen tissue through ECM degradation and damage the structure and function of the heart. MMP can also be used as a biological marker in heart failure cases. The application of MMP as a biological marker can be used to assess the degree of disease severity as well as a predictor of heart failure. In conclusion, MMP has an important role in the development process of heart failure and can be a biological marker in cases of heart failure.

MicroRNA-124 facilitates lens epithelial cellapoptosis by inhibiting SPRY2 and MMP-2.

Age-related cataract (ARC) is the primary cause of blindness worldwide. Abnormal expression of microRNAs (miRNAs/miRs) has been reported to be associated with multiple diseases, including ARC. However, the potential role of miR-124 in ARC remains unclear. The present study used the human lens epithelial cell line, SRA01/04, to investigate the potential role of miR-124 in ARC. Reverse transcription-quantitative PCR analysis was performed to detect the expression levels of miR-124, protein sprouty homolog 2 (SPRY2) and matrix metalloproteinase-2 (MMP-2) in ARC tissues, while western blotting was performed to detect the protein levels of SPRY2 and MMP-2. Cell viability and apoptosis of SRA01/04 cells were assessed via Cell Counting Kit-8 and TUNEL assays, respectively. The interaction between miR-124 and SPRY2 or MMP-2 was confirmed via the dual-luciferase reporter and RNA immunoprecipitation assays. The results of the present study demonstrated that miR-124 expression was significantly upregulated in ARC tissues, and knockdown of miR-124 increased SRA01/04 cell viability and suppressed apoptosis. In addition, SPRY2 and MMP-2 expression was decreased in ARC tissues, and were demonstrated to directly bind to miR-124. Overexpression of SPRY2 or MMP-2 increased SRA01/04 cell viability and repressed apoptosis, the effects of which were reversed following overexpression of miR-124. Taken together, these results suggested that miR-124 facilitates lens epithelial cell apoptosis by modulating SPRY2 or MMP-2 expression, providing a novel treatment approach for ARC.

Hypertension induces compensatory left ventricular hypertrophy by a mechanism involving gap junction lateralization and overexpression of CD36, PKC and MMP-2.

Hypertension-induced left ventricular hypertrophy evolves initially as an adaptive response meant to minimize ventricular wall stress. The mechanisms involved in the preservation of the cardiac function during the “compensatory” phase of the left ventricular hypertrophy are still unclear. Therefore, we aimed at uncovering fine changes that aid the heart to cope with the increased stress in hypertension. Male golden Syrian hamsters were given NG-nitro-L-arginine methyl ester (L-NAME) for 16 weeks, and they became hypertensive (HT), developing left ventricular hypertrophy with no impaired contractility or fibrosis. As compared to age-matched control hamsters, the hypertrophied left ventricles in L-NAME-induced HT hamsters exhibited the following structural and molecular changes: (i) accumulation of lipid droplets (LDs) within cardiomyocytes and relocation of gap junctions to the lateral membrane of cardiomyocytes or close to mitochondria (revealed by electron microscopy); (ii) overexpression of the cluster of differentiation 36 (CD36) fatty acid transporter, protein kinase C (PKC), and matrix metalloproteinase-2 (MMP-2), enhanced activation of the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway, and unchanged expression of the connexin 43 (Cx43) and N-cadherin junctional proteins (assessed by Western blot); (iii) increased protein carbonyl content, assessed with a 2,4-Dinitrophenylhydrazine (DNPH)-based spectrophotometric assay, indicative of an enhanced reactive oxygen species (ROS) production; and (iv) augmented MMP-2 activity (determined by gelatin zymography). These changes may participate in an orchestrated adaptive hypertrophic growth response that helps to maintain cardiac performance, in HT hamsters. Together, these findings could provide support for designing future strategies meant to prevent the transition from compensatory left ventricular hypertrophy to decompensated heart failure.