Matrix Metalloproteinase-9 Expression Research Articles

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.

Protective Effect of Rosmarinic Acid on Inflammatory Changes of Hepatic Cells in Non-alcoholic Steatohepatitis in Mice: Cyclooxygenase 2/ Prostaglandin E2 Pathway

Background and Objectives: Nonalcoholic Steatohepatitis (NASH) is one of the common liver diseases, characterized by excessive lipid accumulation (steatosis) and inflammation in the liver. Inflammation is one of main reasons for this disease which takes place from different pathways. Rosmarinic Acid (RA) is an antioxidant and anti-inflammatory product with positive effects on the improvement of liver injuries. This study aims to evaluate protective effect of RA against liver inflammation caused by NASH in male mice through affecting cyclooxygenase-2/ Prostaglandin E2 (COX-2/PEG2) pathway and Matrix Metalloproteinase-9 (MMP-9). Methods: In this study, 24 C57/BL6 male mice were used. They were divided into four groups of 6 including; Control, RA, NASH, and NASH+RA. The NASH was induced by a methionine/choline-deficient diet for 8 weeks. The RA was administrated intraperitoneally and daily for 8 weeks. After 8 weeks, the animals were sacrificed and their liver was removed. hematoxylin and eosin staining was preformed for histological evaluation and real-time PCR and ELISA assays were used for molecular studies. Results: The RA significantly reduced hepatic steatosis and inflammation as well as the expression of MMP-9 and COX-2 which led to the significant reduction of PEG2 level. Conclusion: It seems that RA can reduce liver inflammation and thereby attenuate NASH in male mice by reducing the expression of COX-2 enzyme and MMP-9 and the concentration of PEG2.

Experimental approaches for altering the expression of Abeta-degrading enzymes.

Cerebral clearance of amyloid β-protein (Aβ) is decreased in early-onset and late-onset Alzheimer's disease (AD). Aβ is cleared from the brain by enzymatic degradation and by transport out of the brain. More than 20 Aβ-degrading enzymes have been described. Increasing the degradation of Aβ offers an opportunity to decrease brain Aβ levels in AD patients. This review discusses the direct and indirect approaches which have been used in experimental systems to alter the expression and/or activity of Aβ-degrading enzymes. Also discussed are the enzymes' regulatory mechanisms, the conformations of Aβ they degrade, where in the scheme of Aβ production, extracellular release, cellular uptake, and intracellular degradation they exert their activities, and changes in their expression and/or activity in AD and its animal models. Most of the experimental approaches require further confirmation. Based upon each enzyme's effects on Aβ (some of the enzymes also possess β-secretase activity and may therefore promote Aβ production), its direction of change in AD and/or its animal models, and the Aβ conformation(s) it degrades, investigating the effects of increasing the expression of neprilysin in AD patients would be of particular interest. Increasing the expression of insulin-degrading enzyme, endothelin-converting enzyme-1, endothelin-converting enzyme-2, tissue plasminogen activator, angiotensin-converting enzyme, and presequence peptidase would also be of interest. Increasing matrix metalloproteinase-2, matrix metalloproteinase-9, cathepsin-B, and cathepsin-D expression would be problematic because of possible damage by the metalloproteinases to the blood brain barrier and the cathepsins' β-secretase activity. Many interventions which increase the enzymatic degradation of Aβ have been shown to decrease AD-type pathology in experimental models. If a safe approach can be found to increase the expression or activity of selected Aβ-degrading enzymes in human subjects, then the possibility that this approach could slow the AD progression should be examined in clinical trials.

Omental cancer-associated fibroblast-derived exosomes with low microRNA-29c-3p promote ovarian cancer peritoneal metastasis.

Ovarian cancer (OC) is characterized by frequent widespread peritoneal metastasis. Cancer-associated fibroblasts (CAFs) represent a critical stromal component of metastatic niche and promote omentum metastasis in OC patients. However, the role of exosomes derived from omental CAFs in metastasis remains unclear. We isolated exosomes from primary omental normal fibroblasts (NFs) and CAFs from OC patients (NF-Exo and CAF-Exo, respectively) and assessed their effect on metastasis. In mice bearing orthotopic OC xenografts, CAF-Exo treatment led to more rapid intraperitoneal tumor dissemination and shorter animal survival. Similar results were observed in mice undergoing intraperitoneal injection of tumor cells. Among the miRNAs downregulated in CAF-Exo, miR-29c-3p in OC tissues was associated with metastasis and survival in patients. Moreover, increasing miR-29c-3p in CAF-Exo significantly weakened the metastasis-promoting effect of CAF-Exo. Based on RNA sequencing, expression assays, and luciferase assays, matrix metalloproteinase 2 (MMP2) was identified as a direct target of miR-29c-3p. These results verify the significant contribution of exosomes from omental CAFs to OC peritoneal metastasis, which could be partially due to the relief of MMP2 expression inhibition mediated by low exosomal miR-29c-3p.

Metformin Directly Binds to MMP-9 to Improve Plaque Stability

Vulnerable atherosclerotic plaque rupture is the principal mechanism that accounts for myocardial infarction and stroke. High matrix metalloproteinase-9 (MMP-9) expression and activity have been proven to lead to plaque instability. Metformin, a first-line treatment for type 2 diabetes, is beneficial to plaque vulnerability. However, the mechanism underlying its anti-atherogenic effect remains unclear. Molecular docking and surface plasmon resonance experiments showed that metformin directly interacts with MMP-9, and incubated MMP-9 overexpressing HEK293A cells with metformin (1 μmol·L-1) significantly attenuates MMP-9's activity using zymography and MMP activity assays. Moreover, metformin treatment drives MMP-9 degradation. Next, we constructed a carotid artery atherosclerotic plaque model and administered consecutive 14-day metformin (200 mg·kg-1·d-1) treatment by intragastric gavage. Immunofluorescence staining of the right carotid common artery and serum MMP activity assay results showed that metformin treatment decreased local plaque MMP-9 protein level and circulating MMP-9 activity, respectively. Histochemical staining revealed that after metformin treatment, the collagen content in plaque was significantly preserved, and the plaque vulnerability index decreased. These findings suggested that metformin improved atherosclerotic plaque stability by directly binding to MMP-9 and driving its degradation.

MMP11 is associated with the immune response and immune microenvironment in EGFR-mutant lung adenocarcinoma.

High expression of matrix metalloproteinase-11 (MMP11) is associated with various tumors and immune microenvironments. Conversely, poor response to immunotherapy in epidermal growth factor receptor (EGFR)-mutant lung adenocarcinoma (LUAD) patients is closely related to the characteristics of immune microenvironment. The Cancer Genome Atlas (TCGA)-LUAD database and our gathered clinical LUAD samples were used to examine the relationship between MMP11 expression and EGFR mutation. Then the correlation between MMP11 and immune response and the difference of immune cell infiltration in different groups were analyzed. Compared the differences in the immune microenvironment between the MMP11-positive and MMP11-negative expression groups using immunohistochemistry (IHC) and multiplex immunohistochemistry. The expression of MMP11 in samples with exon 19 deletions, exon 21 L858R or de novo exon 20 T790M mutations was higher than wild type, but there was no difference between the samples with uncommon mutation and the wild-type. The high MMP11 expression group had a higher Tumor Immune Dysfunction and Exclusion (TIDE) score. Pathways associated with enrichment in the extracellular matrix (ECM) were the main biological functions of differential genes between the high and low MMP11 groups. The IHC score of MMP11 in the EGFR-mutant group was higher than in the EGFR-wild group. In TCGA-LUAD, the high MMP11 group had a lower proportion of T cell CD8+ and NK cells activated. In the clinical samples, the infiltration levels of T cell CD8+ and NK cells in the tumor parenchyma of EGFR-mutant LUAD was lower in the MMP11-positive than in the MMP11-negative group. The expression levels of tumor cell PD-L1 were higher in the MMP11-positive expression group than in the MMP11-negative expression group, and the proportion of PD1+CD8+ T cells infiltrated was reduced in the MMP11-positive group compared to the MMP11-negative group. High expression of MMP11 was associated with EGFR mutations. Patients with EGFR-mutant LUAD with high expression of MMP11 responded poorly to immunotherapy, and the percentage of T cell CD8+ and NK cells in immune cell infiltration was lower in MMP11. Consequently, MMP11 is related to the immunological microenvironment of EGFR-mutant lung adenocarcinoma, which may be a predictor of possible immunotherapeutic response.

Pterostilbene attenuates hemin-induced dysregulation of macrophage M2 polarization via Nrf2 activation in experimental hyperglycemia.

Macrophages exhibit a high degree of plasticity that is physiologically relevant in wound healing, and disruption in normal macrophage response leads to delayed wound closure resulting in chronic wounds. Here, we attempt to discern macrophage responses to hemin via regulation of the nuclear factor-erythroid factor 2-related factor 2 (Nrf2) that could help us better understand the pathophysiology of diabetic foot ulcers (DFU). We demonstrate the alleviation of hemin-mediated Nrf2 suppression and M2 macrophage polarization by pterostilbene (PTS), a proven Nrf2 activator. IC-21 macrophages were treated with hemin under the normoglycemic or hyperglycemic environment with or without PTS and the expression levels of various markers, such as Nrf2 and its downstream target Heme Oxygenase-1 (HO-1), CD206, Ferroportin-1 among others were analyzed using qPCR and Western blot. Our results revealed that hemin under hyperglycemia reduced Nrf2 activation and its downstream targets, M2 polarization, and the induction of a proinflammatory cellular environment, and interestingly all of these were remedied by PTS treatment. Gelatin zymography of matrix metalloproteinase2 (MMP2) expression revealed that hemin under hyperglycemic condition significantly elevated MMP2 expression, which was reversed by PTS treatment. Further proteomic analysis using liquid chromatography with tandem mass spectrometry (LC-MS/MS) revealed a heightened cellular stress profile accompanying inflammation that was suppressed by PTS. This study has furthered our understanding on the role of Nrf2 in attenuating hemin-induced perturbations in macrophage responses and suggests a potential therapeutic target in the management of DFU.

Effects of riboflavin/ultraviolet-A scleral collagen cross-linking on regional scleral thickness and expression of MMP-2 and MT1-MMP in myopic guinea pigs.

To investigate the effects of scleral collagen cross-linking (SXL) using riboflavin and ultraviolet A (UVA) light on the scleral thickness of different regions and expression of matrix metalloproteinase 2 (MMP-2) and membrane-type MMP-1 (MT1-MMP) in guinea pigs with lens-induced myopia. Forty-eight 4-week-old guinea pigs were assigned to three groups (n = 16 per group): SXL group, lens-induced myopia (LIM) group, and control group. The sclera of the right eye of the guinea pig in the SXL group was surgically exposed, riboflavin was dropped on the treatment area for 10 minutes before the 30-minute UVA irradiation. The same surgical procedure was performed in the LIM group without UVA irradiation. The -10.00 D lenses were then placed on the right eyes of guinea pigs in the SXL and LIM groups for six weeks. The control group received no treatment. The left eyes were untreated in all groups. The ocular axial length (AXL) and refraction were measured at 4 weeks and 10 weeks of age. 10-week-old guinea pigs were sacrificed, and the right eyes were enucleated and evenly divided for preparation of hematoxylin and eosin (HE) stained sections, quantitative real-time polymerase chain reaction (qPCR) and western blotting. The scleral thickness of different regions was measured on HE stained sections. The temporal half of the sclera was harvested to measure the expression of MMP-2 and MT1-MMP by qPCR and western blotting. The AXL was significantly shorter, and the degree of myopic refraction was significantly lower in the SXL group than those in the LIM group at 10 weeks of age. The scleral thickness of the cross-linked area was significantly greater in the SXL group than that of the corresponding area in the LIM group, while the scleral thickness of the untreated nasal side was not significantly different between the SXL group and the LIM group. The expression of MMP-2 and MT1-MMP of the cross-linked sclera was significantly downregulated compared with that of the corresponding area in the LIM group. Riboflavin/UVA SXL could slow myopia progression and thicken the cross-linked sclera in guinea pigs, which might be related to the downregulation of MMP-2 and MT1-MMP expression during the scleral remodeling process.

TEM1 up-regulates MMP-2 and promotes ECM remodeling for facilitating invasion and migration of uterine sarcoma

ObjectivesTo explore the correlation between tumor endothelial marker 1 (TEM1) and matrix metalloproteinase 2 (MMP-2) in uterine sarcoma and their roles in the progression of uterine sarcoma.MethodsUterine leiomyosarcoma (uLMS, n = 25) and uterine leiomyoma (n = 25) specimens were collected from a total of 50 patients. Immunohistochemistry assay was conducted to determine the expression of TEM1, MMP-2 and MMP-9. TEM1 over expression (hTEM1) and low expression (shRNA-TEM1) MES-SA cell lines were established as in vitro uterine sarcoma models. MMP-2 mRNA, protein expression and enzymatic activity were verified using qPCR, Western blot and gelatin zymography respectively. MMP-2 expression was downregulated using MMP-2 siRNA in hTEM1 MES-SA cells to better study the role of MMP-2. The invasive and migratory capacities of hTEM1, shRNA-TEM1, and hTEM1 treated with MMP-2 siRNA MES-SA cells were determined using transwell assays. Extracellular matrix (ECM) remodeling mediated by TEM1 was examined using cell-ECM adhesion and fluorescent gelatin-ECM degradation assays. The immunofluorescence of F-actin was examined to analyze the formation of invadopodia. Subcutaneous and intraperitoneal xenografts were established to validate the role of TEM1 in promoting uterine sarcoma metastasis.ResultsTEM1 and MMP-2 were expressed in 92% (n = 23) and 88% (n = 22) of uterine leiomyosarcoma specimens, respectively. Both TEM1 and MMP-2 were highly expressed in 100% (n = 17) of high stage (III-IV) uterine leiomyosarcoma specimens. In addition, TEM1 expression was positively correlated with MMP-2 expression in uterine leiomyosarcoma. The successful establishment of in vitro uterine sarcoma models was confirmed with qPCR and Western blotting tests. TEM1 promoted the invasion and metastasis of uterine sarcoma in vivo and in vitro. MMP-2 expression and activity were up-regulated in hTEM1 cells but down-regulated in shRNA-TEM1 cells. Importantly, MMP-2 knockdown impaired the invasive and migratory capacity of hTEM1 cells. TEM1 promoted ECM remodeling by increasing cell-ECM adhesion and ECM degradation. TEM1 overexpression also induced the formation of invadopodia.ConclusionTEM1 was co-expressed and positively correlated with MMP-2 in uterine leiomyosarcoma specimens. In addition, both TEM1 and MMP-2 were associated with tumor development. TEM1 promoted uterine sarcoma progression by regulating MMP-2 activity and ECM remodeling.

Development of In Vitro Dry Eye Models to Study Proliferative and Anti-Inflammatory Effects of Allogeneic Serum Eye Drops.

This study aimed to develop valid in vitro models for preclinical evaluation of proliferative and anti-inflammatory effects of human allogeneic serum eye drops for dry eye disease (DED) treatment. A DED wound healing model was developed by analyzing the influence of coating and serum concentrations on human corneal epithelial (HCE-T) wound closure. Further, intralaboratory variance, freeze-thaw cycle effects, donor variability and stability assays were conducted. Interleukin-1β (IL-1β) and tumor necrosis factor α (TNFα) were used to induce the gene expression of matrix metalloproteinase 9 (MMP9), cyclooxygenase 2 (COX2), transforming growth factor-β (TGFβ) and IL-1β. MMP9 induction was optimized using a design-of-experiments (DoE) approach and applied to examine serum under static and dynamic conditions. MMP9 protein expression was analyzed by ELISA. The DED wound healing model detected proliferative effects of serum down to 1% with a small intralaboratory variance. Serum stability was shown over six months, donor variance could be detected, and freeze-thaw cycle effects did not affect wound closure. Serum decreased MMP9 expression on the gene and protein levels. The induction method was successfully optimized using DoE modeling and transferred to a dynamic setting mimicking tear film fluidics. The DED wound healing and inflammatory DED model present useful in vitro models for the preclinical evaluation of allogeneic serum eye drops without the use of animal experiments.

Fibroblast-Derived Extracellular Vesicle-Packaged Long Noncoding RNA Upregulated in Diabetic Skin Enhances Keratinocyte MMP-9 Expression and Delays Diabetic Wound Healing

Accurate communication between fibroblasts and keratinocytes is crucial for diabetic wound healing. Extracellular vesicles are being explored as essential mediators of intercellular communication in the skin. However, the mechanisms underlying wound healing mediated by fibroblast-derived extracellular vesicles (Fib-EVs) remain unclear. The present study evaluated the role of long noncoding RNA upregulated in diabetic skin (lnc-URIDS) packed in Fib-EVs in the wound healing of streptozotocin-induced diabetes and the potential mechanisms of the effects. We demonstrated that high glucose induced the enrichment of lnc-URIDS in Fib-EVs, facilitated the transfer of lnc-URIDS to primary rat epidermal keratinocytes, and increased the expression of matrix metalloproteinase-9. Mechanistically, the binding of lnc-URIDS to YTH domain family protein-2 enhanced the degradation of YTH domain family protein-2 in the lysosomes, which increased the translational activity of the messenger RNA of matrix metalloproteinase-9 and ultimately induced the degradation of collagen for wound healing. The results provided an insight into the crosstalk and cooperation between fibroblasts and keratinocytes in collagen homeostasis in diabetic wounds and clarified the mechanism by which lnc-URIDS degrades collagen for diabetic wound healing.

M2 macrophages enhance endometrial cell invasiveness by promoting collective cell migration in uterine adenomyosis

Research questionAre M2 macrophages implicated in endometrial invasiveness in adenomyosis? DesignSeventeen formalin-fixed paraffin-embedded uterine samples and 16 fresh endometrial biopsies were collected from women with or without adenomyosis. Double immunofluorescence was performed to determine the predominant macrophage population in adenomyosis between M1 and M2 phenotypes. The invasion capacity of endometrial cells was assessed by invasion assays and quantitative polymerase chain reaction for genes involved in cell motility and epithelial–mesenchymal transition (EMT). Specific mechanisms of invasion were investigated by immunohistochemistry for E-cadherin, N-cadherin and matrix metalloproteinase 9 (MMP9). ResultsOnly M2 macrophages were found to accumulate in adenomyosis, in higher numbers in both eutopic endometrium (P = 0.0109) and lesions (P = 0.0267) than healthy tissue. Co-culture with M2 macrophages significantly boosted invasion capacity in endometrial epithelial (P = 0.0002; P = 0.002) and stromal cells (P = 0.0469; P = 0.0047) from both adenomyosis patients and healthy controls. No gene expression differences indicating EMT were noted, either between co-cultured and control cells, or between healthy and adenomyotic cells. E- and N-cadherin protein expression did not differ significantly between endometrium from adenomyosis subjects and healthy tissue but MMP9 expression was increased in eutopic stroma from adenomyosis patients (P = 0.0492). In adenomyosis, both E-cadherin (P = 0.0379) and N-cadherin (P = 0.0196) were more extensively expressed in basal glands than functional glands. ConclusionsM2 macrophages accumulate in adenomyosis and enhance invasion capacity of adenomyotic and even healthy endometrial cells, implying that macrophage infiltration alone may be sufficient to promote the disease. This study failed to detect any changes pointing to EMT, suggesting an alternative mode of invasion. Strong E- and N-cadherin-positive intercellular junctions in basal (invasive) glands suggest the involvement of collective cell migration in the invasion process of endometrium.

Role of Matrix Metalloproteinase 9 in Predicting Lymph Node Metastases in Oral Squamous Cell Carcinoma.

Background Oral cancer is a common malignancy worldwide, with approximately 3,50,000 new cases diagnosed yearly. Out of many factors which affect the survival in patients with oral cancer, lymph node metastases are a major factor that reduces survival by 50%. Even though many biomarkers have been studied to predict lymph node metastasis, none have yet been accepted for routine use. Matrix metalloproteinases (MMPs) play a vital role in extracellular matrix (ECM) degradation, thus facilitating the invasive potential and the metastatic cascade of tumors. Of the different subtypes, multiple studies have demonstrated that matrix metalloproteinase 9 (MMP9) overexpression is often associated with the aggressive nature of the tumor. Therefore, this investigation is done to know the role of MMP9 in predicting lymph node metastasis in oral squamous cell carcinoma (OSCC). Aim To determine the immunohistochemical expression of MMP9 in OSCC and to find its association with lymph node metastasis. Settings and design It is a laboratory-based observational study. Materials and methods One hundred five histologically proven cases of OSCC were studied. Histopathological parameters like depth of invasion, presence of lymph node metastasis, grading, and TNM staging were done according to the 8th AJCC staging criteria. Both intensity and proportion of MMP9 expression were recorded. Statistical analysis For qualitative data, the Chi-square test was used as a test of significance. The p-value (probability that the result is true) of <0.05 was considered statistically significant after assuming all the rules of statistical tests. Results A higher expression of MMP9 was observed in 56.2% of cases and the higher expression correlated with the presence of lymph node metastases (p<0.001), an advanced stage of cancer (P <0.001), and grade of the tumor (p=0.003). Conclusion A positive association between MMP9 and lymph node metastasis and pathological TNM staging demonstrates MMP9 as a potential biomarker to predict the behavior of the tumor.

Effect and mechanism of pirfenidone combined with 2-methoxy-estradiol perfusion through portal vein on hepatic artery hypoxia-induced hepatic fibrosis

PurposeThe aim of this study was to explore the effect and mechanism of pirfenidone (PFD) combined with 2-methoxyestradiol (2-ME2) perfusion through portal vein on hepatic artery hypoxia-induced hepatic fibrosis. Materials and methodsSprague-Dawley rats were divided into 5 groups (n ​= ​3/group): control group, hepatic artery ligation (HAL) group, HAL ​+ ​PFD (portal vein perfusion of PFD) group, HAL+2-ME2 (portal vein perfusion of 2-ME2) group and HAL ​+ ​PFD+2-ME2 group depending on whether they received HAL and/or portal vein perfusion (PFD and/or 2-ME2). Livers were harvested for pathology, western blotting (WB), and quantitative real-time PCR (qRT-PCR). ResultsSirius red staining showed that portal vein perfusion of drugs resulted in degradation of liver fibrosis. Immunohistochemistry showed decreased hypoxia-inducible factor-1 α (HIF-1α) and α-smooth muscle actin (α-SMA) after portal intravenous drugs infusion compared with HAL group (P ​< ​0.05). WB analysis showed increased Smad7 in HAL ​+ ​PFD group compared with HAL group (P ​< ​0.05). qRT-PCR analysis showed decreased matrix metallo-proteinase 2 (MMP2), transforming growth factor β1 (TGF-β1), monocyte chemoattractant protein-1 (MCP-1), and Collagen I mRNA in HAL ​+ ​PFD group except for tissue inhibitor of metalloproteinase-1 (TIMP-1) compared with HAL group (P ​< ​0.05). Compared with HAL ​+ ​PFD group, the addition of 2-ME2 did not lead to better results in qRT-PCR analysis. ConclusionsThe portal vein perfusion of PFD significantly reduced the hepatic artery hypoxia-induced fibrosis degree in treated rats by down-regulating the expression of HIF-1α, α-SMA, MMP2, TGF-β1, MCP-1, and Collagen I, as well as up-regulating the TIMP-1 expression and Smad7 protein level. Combined 2-ME2 infusion was not better than PFD alone.

The Potential Roles of CHI3L1 in Failed Autologous Arteriovenous Fistula in End-Stage Renal Disease.

Autologous arteriovenous fistula (AVF) is commonly placed for hemodialysis treatment. Recent studies show that increased baseline serum level of Chitinase-3-like protein 1 (CHI3L1) is independently associated with a higher risk of the early failure of forearm AVFs. However, the changes and mechanisms of CHI3LI in local vascular tissues of failed AVF have not be revealed. This study aims to conduct the expression and mechanism of CHI3L1 in vascular tissues from patients. Immunoreactivity of CHI3L1, matrix metalloproteinase 2 (MMP-2) and vascular endothelial growth factor-A (VEGF-A) were detected in vascular tissues collected from nine patients with AVF surgery. Due to the significant stenosis clinically, six of the nine patients received arteriovenous fistula reconstruction. The expression differences of CHI3L1 between the initial vascular tissues and failed AVF are significant (P < 0.05). Failed AVF due to stenosis shows intraluminal thrombus, collagen fiber rupture, fibrous connective tissue hyperplasia, tube wall thickening, neovascularization, scattered inflammatory cell infiltration in the tunica media as well as high CHI3L1 expression level, and the expression of MMP-2 (r = 0.9022, P = 0.0139) and VEGF-A (r = 0.8355, P = 0.0393) was positively correlated with CHI3L1. CHI3L1 expression in vascular tissues possibly plays an important role in AVF failure. MMP-2 and VEGF-A may participate in venous stenosis with CHI3L1.

PECULIARITIES OF DYNAMICS OF INDICATORS OF PROTEINS OXIDATIVE MODIFICATION AND MATRIX METALLOPROTEINASE-9 ACTIVITY IN PATIENTS WITH PARANOID SCHIZOPHRENIA DEPENDING ON THE DISEASE DURATION.

The aim: The objective of the research was to study the indicators of oxidative modification of proteins (OMP) and the activity of matrix metalloproteinase-9 (MMP-9) in patients with paranoid schizophrenia depending on the disease duration. Materials and methods: 320 patients were included in the examination. 20 patients were with "Primary psychotic episode" (Comparison Group) and 300 patients were diagnosed with "Paranoid schizophrenia" (Experimental Group): 60 of them have suffered from this disease for a duration from 3 to 5 years (Subgroup I ); 60 patients have suffered for a period from 6 to 10 years (Subgroup II); 60 individuals - from 11 to 15 years (Subgroup III); 60 patients have suffered for a duration from 16 to 20 years (Subgroup IV); 60 patients - from 21 years and longer (Subgroup V). Results: The presented data showed that the levels of OMP indicators in Subgroup I constituted 0.826±0.046 conventional units at a wavelength of 356 nm; 0.864±0.051 conventional units at a wavelength of 370 nm; 0.444±0.019 conventional units at a wavelength of 430 nm; 0.176±0.007 conventional units at a wavelength of 530 nm, which is 1.99; 1.6; 1.13 and 1.43 times higher than in the Comparison Group. The content of OMP products was higher by 2.24; 1.74; 1.17, and 1.43 times in Subgroup II, respectively, by 2.4; 1.80; 1.36 and 1.46 times in Subgroup III, respectively; by 2.5; 1.9; 1.4; 1.6 times in Subgroup IV, respectively; by 2.5; 2.02; 1.54; 1.7 times in Subgroup V, respectively. The conducted correlation analysis indicated a direct correlation between OMP indicators and the disease duration. The concentration of MMP-9 in the patients of the Comparison Group was equal to 892.84±87.80 pg/ml, which was 11.2% less compared to the Experimental Subgroup I, where this indicator was 992.84±67.50 pg/ml. MMP-9 constituted 1092.53±47.20 pg/ml on average in the patients of Subgroup II, which was 22.36% higher than in the Comparison Group. This indicator was 1702.84±37.60 pg/ml in Subgroup III, which was 90.7% higher than in the Comparison Group. It constituted 1492.84±47.29 pg/ml in Subgroup IV, which was 67.2% higher than in the Comparison Group; and 2037.21±57.80 pg/ ml in Subgroup V, which was more than two times higher than in the Comparison Group (p<0.05). The conducted correlation analysis showed a direct relation between MMP-9 expression and the increase in OMP indicators. This relation was more significant between MMP-9 and OMP products of a neutral nature. The correlation strength between MMP-9 and OMP products of a basic nature was somewhat less significant. Conclusions: According to the results of the conducted analysis, the examined patients had the signs of decompensation of reactive-adaptive biomolecular mechanisms which activated radical reactions with the subsequent accumulation of oxidation products.

ANNONA SQUAMOSA LEAF EXTRACT INHIBITS THE MIGRATION OF HUMAN CERVICAL CANCER CELLS THROUGH MMP-9 EXPRESSION

To determine the effect of giving a combination of Annona squamosa leaf extract and cisplatin on matrix metalloproteinase-9 expression and inhibition of cell migration in HeLa cells. The sample in this study was divided into 6 groups: group 1 as the control group, group 2 as the single cisplatin (Cis) group, group 3-5 as the different combinations with ASL 12.5, 25 mg/mL, and 50 mg/mL respectively, and group 6 as the single ASL 75 mg/mL group. This research included the cytotoxicity test with MTT assay and MMP-9 expression using immunofluorescence method cell migration with scratch wound healing assay. In this study, the expression of MMP-9 decreased significantly in Annona squamosa leaf extract 25 mg/mL + Cis. The cytotoxicity activity of Annona squamosa leaf extract increased significantly in the ASL 12.5+Cis group. The combination of ASL 25+Cis significantly inhibited cell migration. The combination extract was more effective than a single dose of Cis or a single dose of the extract alone (p &lt; 0.05). The novelty of this study can be used for combined cervical cancer therapy. The combination of cisplatin with an extract with the active ingredient acetogenin can induce cytotoxicity and reduce MMP-9 expression in HeLa cells. Acetogenins are also found in bark, seed, and fruit extracts; however, the extract material from the leaves is easier to obtain and process. Keywords: Annona Squamosa , Cervical Cancer, Cell Migration, Matrix Metalloproteinase-9 DOI： https://doi.org/10.35741/issn.0258-2724.58.3.37

Expression of laminin-1 and matrix metalloproteinase-9 in benign and malignant endometrium.

Laminin-1 and matrix metalloproteinase (MMP)-9 may play roles in the progression from benign to malignant endometrium, so we aimed to investigate their levels of expression in these tissues. This case-control study was conducted at a tertiary care center between January 2014 and December 2016. Paraffin blocks of 50 specimens of benign endometrium with proliferative (n = 20), secretory (n = 11), and atrophic (n = 5) endometrium; simple endometrial hyperplasia without atypia (n = 12); and endometrial polyp (n = 2) histology and 49 specimens of malignant endometrium with endometrioid (n = 40), serous (n = 7), clear cell (n = 1), and undifferentiated (n = 1) types were immunostained with laminin-1 and MMP-9 antibodies and assessed for basement membrane continuity for laminin-1 and the percentage and intensity of MMP-9 expression in epithelial cytoplasm. : Laminin-1 continuity in the basement membrane was higher in benign (92%) compared to malignant (16.3%) endometrium (p < 0.0001) without any difference between the subgroups within each group (p > 0.05). All atrophic endometria and endometrial polyps and 23.5% of low grade endometrioid and none of the other endometrial cancers showed uninterrupted basement membrane staining with laminin-1. All cases in malignant endometrium expressed MMP-9 with either low or high immunoreactivity while none of the cases in benign endometrium showed a high staining with MMP-9 (p < 0.01). Proliferative and hyperplastic endometrium together with grade 1 endometrioid cancer expressed MMP-9 better than the atrophic endometrium (p < 0.05). The immunoreactivity with MMP-9 increased gradually from secretory to hyperplastic endometrium and serous carcinoma (p < 0.05). MMP-9 expression in all types of cancers except grade 1 endometrioid and clear cell compared to proliferative endometrium was significantly higher (p < 0.05) and increased from proliferative to grade 2 endometrioid, grade 3 endometrioid, serous and undifferentiated endometrial carcinoma. Gradual increments in MMP-9 expression and basement membrane laminin-1 discontinuity may indicate progression from normal to hyperplastic and to low- and high-grade cancerous endometrium.

The effect of ascorbic acid on histopathological, biochemical, pharmacological, and immunological toxicity of chronic lead acetate exposure on the spleen in a rat model

Objective To evaluate the effect of vitamin C on histopathological, biochemical, and immunotoxicity of chronic lead exposure in the spleen of a rat model.Methods The rats were divided into five groups of 10 rats each: group I received normal saline orally as a control group; groups II and III received lead acetate for 4 and 8 weeks, respectively; and groups IV and V received lead acetate and vitamin C for 4 and 8 weeks, respectively. The spleen was excised and processed for light, electron microscopic, histopathological, and biochemical analyses. Quantitative assessments of matrix metalloproteinase-2 (MMP-2), MMP-9, interleukin-2 (IL-2), IL-6, and tumor necrosis factor-alpha gene expressions were performed by real-time PCR.Results The examination of control and vitamin C with lead acetate supplemented groups revealed normal splenic architecture. In contrast, the spleen of lead-intoxicated groups exhibited degenerative changes in the spleen, with a significantly decreased expression of IL-2, glutathione peroxidase, superoxide dismutase, and hemoglobin (P<0.05), with significantly increased proinflammatory cytokine (IL-6 and tumor necrosis factor-alpha) expressions, concomitantly with increased oxidative products (malondialdehyde) and protease enzymes (MMP-2 and MMP-9) in the spleen tissues. The coadministration of vitamin C with lead for 4 weeks markedly resolved these changes.Conclusion This study may specify the efficiency of vitamin C in lead toxicity prevention in the spleen, represented by the reduced splenic harmful changes produced by lead administration.

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.