Thrombospondin-like Motifs Research Articles

Association of ADAMTS proteoglycanases downregulation with IVF-ET outcomes in patients with polycystic ovary syndrome: a systematic review and meta-analysis

BackgroundA disintegrin and metalloproteinase with thrombospondin-like motifs (ADAMTS) is involved in inflammation and fertility in women with polycystic ovary syndrome (PCOS). This study aims to assess the role of ADAMTS level in the outcomes of in vitro fertilization and embryo transfer (IVF-ET) in women with PCOS, using a meta-analytic approach.MethodsWe systematically searched Web of Science, PubMed, EmBase, and the Cochrane library to identify potentially eligible studies from inception until December 2021. Study assess the role of ADAMTS levels in patients with PCOS was eligible in this study. The pooled effect estimates for the association between ADAMTS level and IVF-ET outcomes were calculated using the random-effects model.ResultsFive studies involving a total of 181 patients, were selected for final analysis. We noted that ADAMTS-1 levels were positively correlated to oocyte maturity (r = 0.67; P = 0.004), oocyte recovery (r = 0.74; P = 0.006), and fertilization (r = 0.46; P = 0.041) rates. Moreover, ADAMTS-4 levels were positively correlated to oocyte recovery (r = 0.91; P = 0.001), and fertilization (r = 0.85; P = 0.017) rates. Furthermore, downregulation of ADAMTS-1, ADAMTS-4, ADAMTS-5, and ADAMTS-9 was associated with elevated follicle puncture (ADAMTS-1: weighted mean difference [WMD], 7.24, P < 0.001; ADAMTS-4: WMD, 7.20, P < 0.001; ADAMTS-5: WMD, 7.20, P < 0.001; ADAMTS-9: WMD, 6.38, P < 0.001), oocytes retrieval (ADAMTS-1: WMD, 1.61, P < 0.001; ADAMTS-4: WMD, 3.63, P = 0.004; ADAMTS-5: WMD, 3.63, P = 0.004; ADAMTS-9: WMD, 3.20, P = 0.006), and Germinal vesicle oocytes levels (ADAMTS-1: WMD, 2.89, P < 0.001; ADAMTS-4: WMD, 2.19, P < 0.001; ADAMTS-5: WMD, 2.19, P < 0.001; ADAMTS-9: WMD, 2.89, P < 0.001). Finally, the oocytes recovery rate, oocyte maturity rate, fertilization rate, cleavage rate, good-quality embryos rate, blastocyst formation rate, and clinical pregnancy rate were not affected by the downregulation of ADAMTS-1, ADAMTS-4, ADAMTS-5, and ADAMTS-9 (P > 0.05).ConclusionsThis study found that the outcomes of IVF-EF in patients with PCOS could be affected by ADAMTS-1 and ADAMTS-4; further large-scale prospective studies should be performed to verify these results.

ADAMTS12 promotes migration and epithelial-mesenchymal transition and predicts poor prognosis for pancreatic cancer

BackgroundADAMTS (a disintegrin and metalloproteinase with thrombospondin-like motifs) family, a group of extracellular multifunctional enzymes, has been proven to play a pivotal role in the tumor. In pancreatic cancer, the role and mechanism of this family remain unclear. The present study aimed to figure out the hub gene of ADAMTSs and explore the exact roles in the prognosis and biological functions in pancreatic ductal adenocarcinoma (PDAC). MethodsWe used several databases to analyze the ADAMTS family and then screen out the hub genes. The expression of ADAMTS12 in 106 pairs of PDAC tumors and adjacent normal tissues was examined by immunohistochemistry, and its correlations with clinical parameters were further analyzed. The impacts of ADAMTS12 on the migration of PDAC cells were predicted by gene set enrichment analysis and confirmed by transwell assays. The potential impacts of ADAMTS12 on the epithelial-mesenchymal transition (EMT) were identified by database analysis and experimental proof of real-time quantitative polymerase chain reaction (qPCR) and Western blotting. ResultsOur study found that ADAMTS12 was a crucial gene in PDAC, and it was highly expressed in tumor tissues when compared to that in the adjacent tissues. ADATMS12 had predictive value of a poor prognosis for PDAC. The elevation of ADAMTS12 was parallel to the progression of PDAC. Inhibition of ADAMTS12 suppressed the migration of PDAC cells and interfered with the process of EMT. ConclusionsADAMTS12 is a crucial member of ADAMTSs in PDAC and a predictor of poor prognosis. Additionally, based on its impacts on migration and metastasis in PDAC and the relationship with EMT, ADAMTS12 plays a role of an oncogene in PDAC and may be a promising target for treatment.

Message in the bottle: regulation of the tumor microenvironment via exosome-driven proteolysis.

Exosomes comprise a subtype of extracellular vesicles involved in cell-to-cell communication, specifically by transporting biological molecules, such as proteins and nucleic acids, to either local or more distant recipient cells, thus triggering distinct biological behaviors. Included in the exosome cargo is frequently a wide range of proteolytic enzymes, such as the matrix metalloproteinases (MMPs), the disintegrin and metalloproteinases (ADAMs), and the ADAM with thrombospondin-like motifs (ADAMTSs), whose functions contribute to the development and progression of cancer. In recent years, extensive research on the potential use of exosomes in diagnostic and therapeutic applications for personalized medicine has emerged, but the targeting of the proteolytic cargo of exosomes has not been fully exploited in this direction. In this review, we aim to explore both the mechanistic and the translational importance of proteolytic enzymes carried by the tumor cell-derived exosomes, as well as their role in the acquisition and support of certain hallmarks of cancer.

Association of polymorphisms in ADAMTS-5 gene with the susceptibility to knee osteoarthritis: A protocol for systematic review and meta-analysis.

Background:To systematically review literature evidence to discover the association of ADAMTS5 (A Disintegrin And Metalloproteinase with Thrombospondin-like motifs 5) gene polymorphisms and the risk of developing KOA (knee osteoarthritis).Methods:We systematically searched the related randomized controlled trials in 4 databases from inception to August 2021, including the Embase, Web of Science, PubMed, and CNKI (Chinese National Knowledge Infrastructure) databases. No language and publication status restrictions. Two reviewers will independently screen all included studies, and the meta-analysis will be conducted using the Review Manager (RevMan 5.3, Cochrane Collaboration, Nordic Cochrane Center, Copenhagen, Denmark).Results:The gathered evidence suggests that there may be a close relationship between the SNP in the ADAMTS5 gene and KOA development. This study will provide a high-quality and convincing evaluation of the treatment of KOA from the consideration of ADAMTS5 gene and will be published in a peer-reviewed journal.Conclusion:ADAMTS5 polymorphism is likely an important risk factor for the development of KOA. Our study will provide a more accurate treatment method for the treatment of KOA.Trial registration number:CRD42021276317

Involvement of increased expression of chemokine C–C motif chemokine 22 (CCL22)/CC chemokine receptor 4 (CCR4) in the inflammatory injury and cartilage degradation of chondrocytes

CCL22, which could induce chondrocyte apoptosis, was identified to be overexpressed in damaged cartilage. This study was conducted with the aim of investigating the effects of CCL22 interference on chondrocyte injury. The osteoarthritis model was established by stimulating chondrocytes with LPS. The expressions of CCL22, CCR4, matrix metallopeptidase (MMP) 3, MMP9, MMP13, (a disintegrin and metalloproteinase with thrombospondin-like motifs) ADAMTS-4, collagen II and inflammatory cytokines were measured using quantitative reverse transcription PCR (RT-qPCR) and western blot. Besides, immunoprecipitation (IP) was employed to verify the binding of CCL22 and CCR4. After CCR4 was overexpressed, cell viability was observed using Cell Counting Kit-8 (CCK-8). Additionally, cell apoptosis as well as its related proteins was detected by TUNEL and western blot, respectively. ng What's more, glycosaminoglycan (GAG) level was detected using GAG kits. CCL22 and CCR4 expression increased noticeably in LPS-stimulated ATDC5 chondrocytes. CCL22 inhibition could suppress the expression of CCR4 in LPS-induced ATDC5 cells. Likewise, CCL22 inhibition could revive the activation of LPS-induced ATDC5 cells by regulating CCR4. In addition, CCL22 knockdown alleviated inflammatory response and cell apoptosis through CCR4. Furthermore, the cartilage degradation of ADTC5 cells could be relieved by CCL22 silence via regulating CCR4. CCL22/CCR4 expression was increased in osteoarthritic cartilage injury and participated in the inflammation and cartilage degradation of chondrocytes.

Excessive mechanical stretch‑mediated osteoblasts promote the catabolism and apoptosis of chondrocytes via the Wnt/β‑catenin signaling pathway.

Excessive biomechanical loading is considered an important cause of osteoarthritis. Although the mechanical responses of chondrocytes and osteoblasts have been investigated, their communication during mechanical loading and the underlying molecular mechanisms are not yet fully known. The present study investigated the effects of excessive mechanically stretched osteoblasts on the metabolism and apoptosis of chondrocytes, and also assessed the involvement of the Wnt/β-catenin signaling pathway. In the present study, rat chondrocytes and osteoblasts were subjected to mechanical tensile strain, and an indirect chondrocyte-osteoblast co-culture model was established. Reverse transcription-quantitative PCR and western blotting were performed to determine the expression levels of genes and proteins of interest. An ELISA was performed to investigate the levels of cytokines, including matrix metalloproteinase (MMP) 13, MMP 3, interleukin-6 (IL-6) and prostaglandin E2 (PG E2), released from osteoblasts. Flow cytometry was performed to detect the apoptosis of chondrocytes exposed to stretched osteoblast conditioned culture medium. The levels of MMP 13, IL-6 and PG E2 increased significantly in the supernatants of stretched osteoblasts compared with the un-stretched group. By contrast, the mRNA expression levels of Collagen 1a and alkaline phosphatase were significantly decreased in osteoblasts subjected to mechanical stretch compared with the un-stretched group. The mRNA expression level of Collagen 2a was significantly decreased, whereas the expression levels of MMP 13 and a disintegrin and metalloproteinase with thrombospondin-like motifs 5 were significantly increased in chondrocytes subjected to mechanical stretch compared with the un-stretched group. In the co-culture model, the results indicated that excessive mechanically stretched osteoblasts induced the catabolism and apoptosis of chondrocytes, which was partly inhibited by Wnt inhibitor XAV-939. The results of the present study demonstrated that excessive mechanical stretch led to chondrocyte degradation and inhibited osteoblast osteogenic differentiation; furthermore, excessive mechanically stretched osteoblasts induced the catabolism and apoptosis of chondrocytes via the Wnt/β-catenin signaling pathway.

Association of polymorphisms in ADAMTS-7 gene with the susceptibility to coronary artery disease - a systematic review and meta-analysis.

Objective: To systematically review literature evidence to discover the association of ADAMTS7 (A Disintegrin And Metalloproteinase with Thrombospondin-like motifs 7) polymorphisms and the risk of developing CAD (coronary artery disease).Data sources: A related literature search in online databases, including EMBASE, PubMed, and Web of Science was undertaken. The period covered was from 2007 to September 10, 2019.Results: Of 256 citations retrieved, nine relevant studies were selected for detailed evaluation. Five SNPs (rs3825807, rs1994016, rs4380028, rs79265682, and rs28455815) in ADAMTS7 gene were identified among included studies. There were 51,851 cases and 89,998 controls included in four studies for SNP rs3825807, 13,403 cases and 11,381 controls included in two studies for SNP rs1994016, 37,838 cases and 38,245 controls included in two studies for SNP rs4380028, 3,133 cases and 5,423 controls included in one study for SNP rs79265682, 103,494 cases and 198,684 controls included in one study for SNP rs28455815. We found most consistent evidence for an association with CAD on coronary angiogram with ADAMTS7 SNP rs3825807 risk allele A in contrast to control G allele, followed by rs4380028 (C vs. T allele), and rs1994016 (C vs. T allele).Conclusions: ADAMTS7 polymorphism is likely an important risk factor for development of CAD. Our data also suggest that the ADAMTS7 polymorphism may be a risk factor for CAD progression in patients who already have pathology in their coronary arteries.Review methods: We included all studies in English language that reported correlation between the ADAMTS7 polymorphism and CAD in human cases.

THE EFFECTS OF SULINDAC ON CELL VIABILITY, CELL CYCLE AND ANJIOGENEZİS IN PHARYNGEAL CANCER CELL

Objective: The present study aimed to investigate the effects of Sulindac on cell viability, cell cycle and angiogenesis in pharyngeal cancer cell lines (FaDu). Methods: FaDu cells were incubated in a medium in a 5% CO2 incubator at 37°C, after which they were proliferated and passaged. IC50 concentration was used to determine the Sulindac dose. Cells were analyzed for cell viability, cell count and cell cycle after Sulindac administration. Immunohistochemistry (vascular endothelial growth factor receptor 2) and western blot (A disintegrin and metalloproteinase with thrombospondin-like motifs 1-ADAMTS1) analyses were used for angiogenesis assessment. Results: Cell viability decreased in pharyngeal cancer cells after Sulindac administration. In addition, FaDU cells were arrested in the G2/M phase. Sulindac was found to slightly increase vascular endothelial growth factor receptor 2 (VEGF-R2) and decrease ADAMTS1 levels in pharyngeal cancer cells. Conclusion: Sulindac showed positive results on cell proliferation in the treatment of pharyngeal cancer cells. However, it could cause a slight increase in angiogenesis.

MicroRNA-375 regulates oocyte in vitro maturation by targeting ADAMTS1 and PGR in bovine cumulus cells

MicroRNAs (miRNAs) have attracted increasing attention for their function in oocyte in vitro maturation (IVM). In this study, we aimed to explore the functional role and underlying mechanism of miR-375 in oocyte IVM. Cumulus-oocyte complexes (COCs) were cultured in standard cell culture conditions until they reach metaphase II (MII) stage. MiR-375 overexpression or knockdown was achieved by corresponding lentiviral transduction. Levels of miR-375, disintegrin and metalloproteinase with thrombospondin-like motifs 1 (ADAMTS1) mRNA and progesterone receptor (PGR) mRNA were detected by qRT-PCR. Western blotting was used to assess the expression of ADAMTS1 and PGR protein. The targeted interaction between miR-375 and ADAMTS1 or PGR was verified by dual-luciferase reporter assay and RNA immunoprecipitation assay. Our results demonstrate that miR-375 is downregulated, and ADAMTS1 and PGR are upregulated in cumulus cells during COC maturation. MiR-375 negatively regulates COC maturation. Moreover, ADAMTS1 and PGR are two targets of miR-375 in cumulus cells. ADAMTS1 or PGR knockdown represses COC maturation and miR-375 inhibits the expression levels of ADAMTS1 and PGR in cumulus cells. Additionally, miR-375 overexpression-mediated suppressive effect on COC maturation is abated by ADAMTS1 or PGR expression restoration. In conclusion, our study suggests that miR-375 represses oocyte IVM at least partially through targeting ADAMTS1 and PGR in cumulus cells, providing a novel insight for the involvement and underlying mechanism of miR-375 in oocyte IVM.

Association of ADAMTS4 and ADAMTS9 levels with cardiovascular risk in patients with primary hyperparathyroidism

ABSTRACT Purpose: Primary hyperparathyroidism (PHPT) has been associated with increased incidence of morbidity and mortality of the cardiovascular system. The pathogenetic mechanisms underlying this association are still not completely clear. A disintegrin and metalloproteinase with thrombospondin-like motifs (ADAMTS) play a critical role in atherosclerosis. This study aimed to investigate the levels of ADAMTS4 and ADAMTS9 and relationship between these proteoglycanases and cardiometabolic abnormalities in PHPT. Materials and Methods: A case-control study was performed in a training and research hospital. Fifty-six patients with PHPT and 61 healthy volunteers were recruited. The Framingham score was used to calculate cardiovascular risk (CVR). Serum ADAMTS levels were determined by a human enzyme-linked immunoassay in all subjects. Results: The ADAMTS9 concentration was significantly higher in patients with PHPT than in the control group (p < 0.05); however, there were no significant differences in ADAMTS4 levels between the groups (p > 0.05). In ROC curve analysis, PHPT can be predicted by the use of ADAMTS9 at a cut-off value of 30.7 pg/mL (69% sensitivity, 65% specificity). CVR score was significantly increased in the PHPT than controls (p < 0.05). ADAMTS4 and ADAMTS9 levels had positive correlations with CVR score (r = 0.322, p = 0.017; r = 0.275, p = 0.044 respectively). Conclusions: Based on the results of the present study, cardiovascular risk is increased in PHPT and associated with ADAMTS4 and ADAMTS9. Further efforts are needed to establish the function of ADAMTS proteases in both PHPT and atherosclerosis.

Altered Bone Mechanics, Architecture and Composition in the Skeleton of TIMP-3-Deficient Mice.

Tissue inhibitor of metalloproteinases-3 (TIMP-3) maintains a healthy extracellular matrix by regulating matrix metalloproteinases (MMP), disintegrin-metalloproteinases (ADAM), and disintegrin-metalloproteinases with ThromboSpondin-like motifs (ADAMTS) activity. Currently, there is a need for a comprehensive understanding of the effects of TIMP-3 on the bone quality and integrity. In this study, we examined the mechanical, morphological, and compositional properties of TIMP-3 knock out (Timp-3 -/-) mouse bone. We hypothesize that the lack of TIMP-3 plays an important role in maintaining the overall bone integrity. Mechanical properties of humeri, lumbar vertebrae, and femurs from Timp-3 -/- mice were determined using 3-point bending, compression, and notched 3-point bending, respectively. Morphological properties of the humeral cortical and trabecular bone and the caudal vertebrae cortical bone were evaluated using micro-computed tomography, while the composition of the femoral cortical and trabecular bone was examined using Fourier transform infrared spectroscopic imaging. Our results revealed that the integrity of the Timp-3 -/- bone is compromised due to changes in its composition, structure, and mechanics. Reductions in the yield and ultimate load and stress capacity, and loss in bone fracture toughness were attributed to reduced density and thickness, and increased porosity of cortical bone. Thin trabeculae were dense, highly connected, and closely packed in Timp-3 -/- bone. Furthermore, altered cortical and trabecular bone mineralization and increased compositional heterogeneity were found in Timp-3 -/- bone, all being indicative of high bone remodeling. In conclusion, this study suggests that the lack of TIMP-3 is detrimental to bone development and maintenance.

The impact of parathyroidectomy on serum ADAMTS1, ADAMTS4 levels, insulin resistance, and subclinical cardiovascular disease in primary hyperparathyroidism.

Primary hyperparathyroidism has been associated with increased incidence of morbidity and mortality of the cardiovascular system. The etiopathogenetic mechanisms underlying this association are still not completely clear. Accumulating evidence suggested that a disintegrin and metalloproteinase with thrombospondin-like motifs (ADAMTS) has a role in the development of inflammation and atherosclerosis. In this study, we aimed to determine whether there is a change in serum levels of ADAMTS1, ADAMTS4, carotid intima-media thickness, and cardiovascular risk score after the surgery and also whether there is a relationship between ADAMTS levels and cardiovascular risk score in hypercalcemic primary hyperparathyroidism patients. The study included the 48 consecutive newly diagnosed patients with primary hyperparathyroidism. The patients were evaluated before and six months after parathyroidectomy. The Framingham score is used to calculate cardiovascular risk. Serum ADAMTS levels were determined by a human enzyme-linked immunoassay in all subjects. The fasting glucose, fasting insulin levels and HOMA values were decreased significantly in all patients after surgery compared to the pretreatment values (p < 0.05). ADAMTS1, ADAMTS4, and carotid intima-media thickness levels were significantly lower after surgical correction of primary hyperparathyroidism compared to the preoperative values (p < 0.05). cardiovascular risk score was decreased after parathyroidectomy however, the difference were not statistical significant (p > 0.05). There were statistically significant relationship between cardiovascular risk score and waist/hip ratio, calcium, LDL-cholesterol, carotid intima-media thickness, ADAMTS4 values. Based on the results of the present study, fasting glucose, fasting insulin levels, ADAMTS1, ADAMTS4, and carotid intima-media thickness might be an additional parameters during the management of patients with primary hyperparathyroidism, since these factors might improve after surgery.

Clinical significance of ADAMTS1, ADAMTS5, ADAMTS9 aggrecanases and IL-17A, IL-23, IL-33 cytokines in polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders among women of reproductive age. A disintegrin and metalloproteinase with thrombospondin-like motifs (ADAMTS) are involved in inflammation and fertility. The aim of this investigation was to evaluate the serum levels of ADAMTS1, ADAMTS5, ADAMTS9, IL-17, IL-23, IL-33 and to find out the relationship between these inflammatory cytokines and ADAMTSs in PCOS patients. A case-control study was performed in a training and research hospital. Eighty patients with PCOS and seventy-eight healthy female volunteers were recruited in the present study. Serum ADAMTS and IL levels were determined by a human enzyme-linked immunoassay (ELISA) in all subjects. The IL-17A, IL-23 and IL-33 levels were significantly higher in the PCOS patients compared to the controls (p<0.05). We could not find significant difference between the groups in terms of ADAMTS1, ADAMTS5 and ADAMTS9 levels. IL-17A had positive correlations with LDL cholesterol and IL-33 and negative correlations with ADAMTS1, ADAMTS5, and ADAMTS9. IL-33 had positive correlation with LDL cholesterol and IL-17A. In ROC curve analysis, PCOS can be predicted by the use of IL-17A, IL-23 and IL-33 which at a cut-off value of 8.37pg/mL (44% sensitivity, 83% specificity), 26.75pg/mL (36% sensitivity, 64% specificity) and 14.28pg/mL (83% sensitivity, 39% specificity), respectively. The results of the study might suggest that ADAMTS and IL molecules have a role in the pathogenesis of the PCOS. Further efforts are needed to establish causality for ADAMTS-IL axis.

Increased ADAMTS1 mediates SPARC-dependent collagen deposition in the aging myocardium.

Secreted protein acidic and rich in cysteine (SPARC) is a collagen-binding matricellular protein highly expressed during fibrosis. Fibrosis is a prominent component of cardiac aging that reduces myocardial elasticity. Previously, we reported that SPARC deletion attenuated myocardial stiffness and collagen deposition in aged mice. To investigate the mechanisms by which SPARC promotes age-related cardiac fibrosis, we evaluated six groups of mice (n = 5-6/group): young (3-5 mo old), middle-aged (10-12 mo old), and old (18-29 mo old) C57BL/6 wild type (WT) and SPARC-null (Null) mice. Collagen content, determined by picrosirius red staining, increased in an age-dependent manner in WT but not in Null mice. A disintegrin and metalloproteinase with thrombospondin-like motifs 1 (ADAMTS1) increased in middle-aged and old WT compared with young, whereas in Null mice only old animals showed increased ADAMTS1 expression. Versican, a substrate of ADAMTS1, decreased with age only in WT. To assess the mechanisms of SPARC-induced collagen deposition, we stimulated cardiac fibroblasts with SPARC. SPARC treatment increased secretion of collagen I and ADAMTS1 (both the 110-kDa latent and 87-kDa active forms) into the conditioned media as well as the cellular expression of transforming growth factor-β1-induced protein (Tgfbi) and phosphorylated Smad2. An ADAMTS1 blocking antibody suppressed the SPARC-induced collagen I secretion, indicating that SPARC promoted collagen production directly through ADAMTS1 interaction. In conclusion, ADAMTS1 is an important mediator of SPARC-regulated cardiac aging.

Induction of proteinases in the human preovulatory follicle of the menstrual cycle by human chorionic gonadotropin

To explore the temporal expression in granulosa and theca cells of key members of the MMP and ADAMTS families across the periovulatory period in women to gain insight into their possible roles during ovulation and early luteinization. Experimental prospective clinical study and laboratory-based investigation. University medical center and private IVF center. Thirty-eight premenopausal women undergoing surgery for tubal ligation and six premenopausal women undergoing assisted reproductive techniques. Administration of hCG and harvesting of follicles by laparoscopy and collection of granulosa-lutein cells at oocyte retrieval. Expression of mRNA for matrix metalloproteinase (MMPs) and the A disintegrin and metalloproteinase with thrombospondin-like motifs (ADAMTS) in human granulosa cells and theca cells collected across the periovulatory period of the menstrual cycle and in cultured granulosa-lutein cells after hCG. Localization of MMPs and ADAMTSs by immunohistochemistry. Expression of MMP1 and MMP19 mRNA increased in both granulosa and theca cells after hCG administration. ADAMTS1 and ADAMTS9 mRNA increased in granulosa cells after hCG treatment, however, thecal cell expression for ADAMTS1 was unchanged, while ADAMTS9 expression was decreased. Expression of MMP8 and MMP13 mRNA was unchanged. Immunohistochemistry confirmed the localization of MMP1, MMP19, ADAMTS1, and ADAMTS9 to the granulosa and thecal cell layers. The collection of the dominant follicle throughout the periovulatory period has allowed the identification of proteolytic remodeling enzymes in the granulosa and theca compartments that may be critically involved in human ovulation. These proteinases may work in concert to regulate breakdown of the follicular wall and release of the oocyte.

Evidence for Decreased Expression of ADAMTS-1 Associated With Impaired Oocyte Quality in PCOS Patients

Polycystic ovary syndrome (PCOS) is the most common cause of dysfunctional ovulation-affecting female fertility. A disintegrin and metalloproteinase with thrombospondin-like motifs (ADAMTS-1) is required for normal ovulation and subsequent fertilization, and the expression of ADAMTS-1 may be altered in PCOS granulosa cell (GC)-reflecting abnormalities in ovulatory signaling. The purpose of this paper is to analyze the differential expression of ADAMTS-1 in PCOS patients associated with impaired oocyte quality. A prospective comparative experimental study was performed at a clinical reproductive medicine center. Women with PCOS (n = 40) and normovulatory controls (n = 40) undergoing controlled ovarian hyperstimulation and in vitro fertilization were recruited in our study. Differential expression of ADAMTS-1 in GCs was analyzed with immunocytochemistry in PCOS patients and normal controls, and ADAMTS-1 mRNA expression was quantified by RT-PCR. Furthermore, the correlation between ADAMTS-1 mRNA and oocyte quality was analyzed. The expression of ADAMTS-1 was decreased in PCOS patients compared with normally ovulating women and was closely related to lower oocyte recovery, oocyte maturity, and fertilization rate. Our study provides evidence that the dysregulated expression of ADAMTS-1 in PCOS may influence oocyte quality-via GCs-oocyte paracrine and endocrine mechanism.

Luteinizing hormone stimulation of in vitro ovulation in brook trout (Salvelinus fontinalis) involves follicle contraction and activation of proteolytic genes

Luteinizing hormone (LH) is an essential hormone for the stimulation of the ovulatory process in vertebrates. However, little is known in fish regarding the different mechanisms induced by LH during ovulation that facilitate the rupture of the follicle wall and the subsequent expulsion of the mature oocyte. In this study, the effects of salmon LH (sLH) on in vitro ovulation were investigated in brook trout (Salvelinus fontinalis) isolated follicles. sLH significantly stimulated in vitro ovulation and contraction of brook trout preovulatory follicles. In order to investigate the possible involvement of proteolytic events in the ovulatory action of LH, the expression of genes known to have a crucial role in the degradation of follicle wall structure was examined. Our results show that sLH clearly stimulated the mRNA expression levels of matrix metalloproteinases (MMPs; including mmp2 and mmp19) and other enzymes with proteolytic action during ovulation, such as a disintegrin and metalloproteinase with thrombospondin-like motifs 1 (adamts1) and plasminogen (plg), in brook trout preovulatory follicles. In addition, the expression of mmp2, adamts1 and plg increased in brook trout follicles during the progression of LH-induced ovulation. Interestingly, the expression of tissue inhibitor of matrix metalloproteinase 2 (timp2), a known regulator of MMP2 activity, paralleled that of mmp2, suggesting the existence of a controlled mechanism of MMP2 action. Therefore, the known increase in proteolytic activity during ovulation in fish could be the result of the stimulation of the expression of proteolytic enzymes by LH in preovulatory follicles. We propose that LH may stimulate ovulation in brook trout follicles by stimulating proteolysis of the follicle wall and by stimulating follicle contraction.

Intracellular calcium oscillations in articular chondrocytes induced by basic calcium phosphate crystals lead to cartilage degradation

Basic calcium phosphate (BCP) crystals, including octacalcium phosphate (OCP), carbonated-apatite (CA) and hydroxyapatite (HA) crystals are associated with destructive forms of osteoarthritis. Mechanisms of BCP-induced cartilage breakdown remain incompletely understood. We assessed the ability of BCP to induce changes in intracellular calcium (iCa(2+)) content and oscillations and the role of iCa(2+) in BCP-induced cartilage degradation. Bovine articular chondrocytes (BACs) and bovine cartilage explants (BCEs) were stimulated with BCP or monosodium urate (MSU) crystals. iCa(2+) levels were determined by spectrofluorimetry and oscillations by confocal microscopy. mRNA expression of matrix metalloproteinase 3 (MMP-3), a disintegrin and metalloprotease with thrombospondin-like motifs 4 (ADAMTS-4) and ADAMTS-5 was assessed by quantitative real-time PCR. Glycosaminoglycan (GAG) release was measured in the supernatants of BCE cultures. All three BCP crystals significantly increased iCa(2+) content. OCP also induced iCa(2+) oscillations. Rate of BACs displaying iCa(2+) oscillations increased over time, with a peak after 20min of stimulation. OCP-induced iCa(2+) oscillations involved both extracellular Ca(2+) (eCa(2+)) influx and iCa(2+) stores. Indeed, OCP-induced iCa(2+) oscillations decreased rapidly in Ca(2+)-free medium. Both voltage- and non-voltage-dependent Ca(2+) channels were involved in eCa(2+) influx. BCP crystal-induced variation in iCa(2+) content was associated with BCP crystal-induced cartilage matrix degradation. However, iCa²(+) was not associated with OCP crystal-induced mRNA expression of MMP-3, ADAMTS-4 or ADAMTS-5. BCP crystals can induce variation in iCa(2+) content and oscillations in articular chondrocytes. Furthermore, BCP crystal-induced changes in iCa(2+) content play a pivotal role in BCP catabolic effects on articular cartilage.

The roles of ADAMTS metalloproteinases in tumorigenesis and metastasis

The human ADAMTS (a disintegrin and metalloproteinase with thrombospondin-like motifs) family of 19 secreted, multidomain proteolytic enzymes is involved in a wide range of biological processes including ECM assembly and degradation, hemostasis, organogenesis and the regulation of angiogenesis. Defects in certain family members give rise to inherited human genetic diseases, while aberrant expression of other ADAMTSs has been linked to the pathogenesis of arthritis and cancer. Several ADAMTSs act as tumor or metastasis suppressors whose functions are lost either by mutation or epigenetic silencing during tumor progression. This review looks in depth at the involvement of ADAMTSs as positive and negative mediators in cancer growth and spread.

Anti-arthritic Action Mechanisms of Natural Chondroitin Sulfate in Human Articular Chondrocytes and Synovial Fibroblasts

To clarify the exact anti-arthritic action mechanisms of chondroitin sulfate (CS), we evaluated the effects of CS derived from shark cartilage (CS-SC) composed mainly of chondroitin-6-sulfate and porcine trachea cartilage (CS-PC) composed mostly of chondrotin-4-sulfate on the functions of human articular chondrocytes and synovial fibroblasts. Both CS-SC and CS-PC (from 1 to 100 mug/ml) effectively suppressed the interleukin (IL)-1beta (10 ng/ml)-enhanced gene expression of aggrecanase-1/a disintegrin and metalloproteinase with thrombospondin-like motifs (ADAMTS)-4 and aggrecanase-2/ADAMTS-5 in articular chondrocytes embedded in alginate beads and synovial fibroblasts. In addition, CS-SC and CS-PC overcame the IL-1beta-mediated suppression of the aggrecan core protein mRNA, and suppressed the IL-1beta-enhanced collagenase-3/matrix metalloproteinase (MMP)-13 gene expression in chondrocytes. CS-PC, but not CS-SC effectively recovered the IL-1beta-reduced gene expression of tissue inhibitor of metalloproteinases (TIMP)-3 in chondrocytes, and enhanced the production of TIMP-1 in synovial fibroblasts. It is noteworthy that CS is able to modulate the function of synovial fibroblasts as well as that of chondrocytes. Therefore, CS is very likely to be multifunctional chondroprotective material for degenerative arthritic diseases.