Tissue Inhibitor Of Metalloproteinase Research Articles

Relevant MicroRNAs of MMPs and TIMPs with Certain Gut Microbiota Could Be Involved in the Invasiveness and Metastasis of Malignant Tumors

Malignant tumors are heterogeneous diseases characterized by uncontrolled cell proliferation, invasion, and/or metastasis. In particular, cancer stem cells within these tumors might be responsible for the invasiveness and the property of recurrence of malignancies. It has been reported that matrix metalloproteases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) could contribute to the development of tumor invasiveness. Remarkably, specific MMPs can drive the malignant phenotype of tumors counting the acquisition of stem cell feature, in which STATs signaling might play an important role. Furthermore, MMPs might be crucial for extra-cellular matrix remodeling in the pathological condition of cancer stem cells. Some immune responses against cancer stem cells in the connection to MMPs and/or TIMPs have been revealed, which might be an important issue in the treatment of aggressive cancers. MicroRNAs (miRNAs) have emerged as favorable biomarkers owing to their important roles in gene regulation and/or stability. Differential expression patterns of specific miRNA may be associated with different types of tumor. In addition, miRNAs can be detected in various body fluids, proposing easily available specimens for diagnostic purposes in tumor microenvironment. Remarkably, it has been shown that miRNA expression might be also related with the gut microbiota composition. Early detection and accurate monitoring of tumors are critical for effective treatment and/or improved patient outcomes. A better understanding of this mechanism of epigenetics could be of use to bring about innovative tactics for the treatment of malignant tumors.

Available Therapeutic Options for Corneal Neovascularization: A Review

Corneal neovascularization can impair vision and result in a poor quality of life. The pathogenesis involves a complex interplay of angiogenic factors, notably vascular endothelial growth factor (VEGF). This review provides a comprehensive overview of potential therapies for corneal neovascularization, covering tissue inhibitors of metalloproteinases (TIMPs), transforming growth factor beta (TGF-β) inhibitors, interleukin-1L receptor antagonist (IL-1 Ra), nitric oxide synthase (NOS) isoforms, galectin-3 inhibitors, retinal pigment epithelium-derived factor (PEDF), platelet-derived growth factor (PDGF) receptor inhibitors, and surgical treatments. Conventional treatments include anti-VEGF therapy and laser interventions, while emerging therapies such as immunosuppressive drugs (cyclosporine and rapamycin) have been explored. Losartan and decorin are potential antifibrotic agents that mitigate TGF-β-induced fibrosis. Ocular nanosystems are innovative drug-delivery platforms that facilitate the targeted release of therapeutic agents. Gene therapies, such as small interfering RNA and antisense oligonucleotides, are promising approaches for selectively inhibiting angiogenesis-related gene expression. Aganirsen is efficacious in reducing the corneal neovascularization area without significant adverse effects. These multifaceted approaches underscore the corneal neovascularization management complexity and highlight ideas for enhancing therapeutic outcomes. Furthermore, the importance of combination therapies and the need for further research to develop specific inhibitors while considering their therapeutic efficacy and potential adverse effects are discussed.

Kidney Fibrosis and Matrix Metalloproteinases (MMPs).

Chronic kidney disease (CKD) is a disorder that causes changes in both the structure and function of the kidneys, causing complications such as hypertension, edema, and oliguria. Renal fibrosis is also a common pathological feature of CKD. Matrix metalloproteinases (MMPs) are endopeptidases that degrade extracellular matrix (ECM) proteins. The proteinase domain consists of a zinc ion in the active site, which contributes to its stabilization with another zinc and three calcium structural ions. Many cellular processes are controlled by MMPs, such as cell-cell interactions and various signaling pathways, while they are also involved in degrading substrates on cell surfaces. Tissue inhibitors of metalloproteinases (TIMPs) are key regulators of metalloproteinases, and both are involved in regulating cell turnover, the regulation, and the progression of fibrosis and apoptosis in the tissue. MMPs play a role in renal fibrosis, such as the tubular cell epithelial-mesenchymal transition (TEM), activation of resident fibroblasts, endothelial-mesenchymal transition (EndoMT), and pericyte-myofibroblast transdifferentiation. This review aims to show the mechanisms through which MMPs contribute to renal fibrosis, paying particular attention to MMP-9 and the epithelial-mesenchymal transition.

Fibroblasts’ secretome from calcified and non-calcified dermis in Pseudoxanthoma elasticum differently contributes to elastin calcification

Pseudoxanthoma elasticum (PXE) is a rare disease characterized by ectopic calcification, however, despite the widely spread effect of pro/anti-calcifying systemic factors associated with this genetic metabolic condition, it is not known why elastic fibers in the same patient are mainly fragmented or highly mineralized in clinically unaffected (CUS) and affected (CAS) skin, respectively. Cellular morphology and secretome are investigated in vitro in CUS and CAS fibroblasts. Here we show that, compared to CUS, CAS fibroblasts exhibit: a) differently distributed and organized focal adhesions and stress fibers; b) modified cell-matrix interactions (i.e., collagen gel retraction); c) imbalance between matrix metalloproteinases and tissue inhibitor of metalloproteinases; d) differentially expressed pro- and anti-calcifying proteoglycans and elastic-fibers associated glycoproteins. These data emphasize that in the development of pathologic mineral deposition fibroblasts play an active role altering the stability of elastic fibers and of the extracellular matrix milieu creating a local microenvironment guiding the level of matrix remodeling at an extent that may lead to degradation (in CUS) or to degradation and calcification (in CAS) of the elastic component. In conclusion, this study contributes to a better understanding of the mechanisms of the mineral deposition that can be also associated with several inherited or age-related diseases (e.g., diabetes, atherosclerosis, chronic kidney diseases).

Effect of human epididymis protein 4 on hyperoxia-induced bronchial dysplasia in newborn rats

Objective The study aimed to elucidate the role and the underlying mechanism of human epididymis protein 4 (HE4) in the pathogenesis of hyperoxia-induced bronchial dysplasia in newborn rats. Methods Forty neonatal Sprague–Dawley (SD) rats were separated into two groups: a normal control group (20.8% oxygen concentration) and a hyperoxia-induced group (85% oxygen concentration). Three time intervals of 24 h, 3 days and 7 days were chosen for each group. Haematoxylin–eosin staining was used to identify the pathological alterations in the lung tissue of the SD rats. Enzyme-linked immunosorbent assay was used to evaluate plasma protein levels. Real-time reverse transcription polymerase chain reaction was used to determine messenger RNA (mRNA) expression. Results In newborn SD rats, hyperoxia intervention within 7 days may result in acute lung damage. In the plasma and tissue of newborn SD rats, hyperoxia induction may raise levels of HE4, matrix metalloproteinases (MMP) 9 and tissue inhibitors of metalloproteinases (TIMP) 1. We discovered that the HE4 protein activates the phosphorylation of extracellular regulated protein kinases (ERK) and p65, activates the downstream MMP9 signalling pathway, inhibits MMP9 mRNA expression, inhibits protein activity, reduces type I collagen degradation, increases collagen secretion and promotes matrix remodelling and fibrosis in neonatal rat primary alveolar type II epithelial cells by overexpressing and silencing the HE4 gene. Conclusion Through the ERK, MMP9 and TIMP1 signalling pathways, HE4 mediates the pathophysiological process of hyperoxia-induced lung damage in rats. Lung damage and lung basal remodelling are mediated by HE4 overexpression.

Non-TGFβ profibrotic signaling in ulcerative colitis after in vivo experimental intestinal injury in humans.

Although impaired regeneration is important in many gastrointestinal diseases including ulcerative colitis (UC), the dynamics of mucosal regeneration in humans are poorly investigated. We have developed a model to study these processes in vivo in humans. Epithelial restitution (ER) and extracellular matrix (ECM) regulation after an experimental injury of the sigmoid colonic mucosa was assessed by repeated high-resolution endoscopic imaging, histological assessment, RNA sequencing, deconvolution analysis, and 16S rDNA sequencing of the injury niche microbiome of 19 patients with UC in remission and 20 control subjects. Human ER had a 48-h lag before induction of regenerative epithelial cells [wound-associated epithelial (WAE) and transit amplifying (TA) cells] along with the increase of fibroblast-derived stem cell growth factor gremlin 1 mRNA (GREM1). However, UC deconvolution data showed rapid induction of inflammatory fibroblasts and upregulation of major structural ECM collagen mRNAs along with tissue inhibitor of metalloproteinase 1 (TIMP1), suggesting increased profibrotic ECM deposition. No change was seen in transforming growth factor β (TGFβ) mRNA, whereas the profibrotic cytokines interleukin 13 (IL13) and IL11 were upregulated in UC, suggesting that human postinjury responses could be TGFβ-independent. In conclusion, we found distinct regulatory layers of regeneration in the normal human colon and a potential targetable profibrotic dysregulation in UC that could lead to long-term end-organ failure, i.e., intestinal damage.NEW & NOTEWORTHY The study reveals the regulatory dynamics of epithelial regeneration and extracellular matrix remodeling after experimental injury of the human colon in vivo and shows that human intestinal regeneration is different from data obtained from animals. A lag phase in epithelial restitution is associated with induction of stromal cell-derived epithelial growth factors. Postinjury regeneration is transforming growth factor β-independent, and we find a profibrotic response in patients with ulcerative colitis despite being in remission.

Alginate-Encapsulated Mesenchymal Stromal Cells Improve Hind Limb Ischemia in a Translational Swine Model.

Cellular therapies have been investigated to improve blood flow and prevent amputation in peripheral artery disease with limited efficacy in clinical trials. Alginate-encapsulated mesenchymal stromal cells (eMSCs) demonstrated improved retention and survival and promoted vascular generation in murine hind limb ischemia through their secretome, but large animal evaluation is necessary for human applicability. We sought to determine the efficacy of eMSCs for peripheral artery disease-induced limb ischemia through assessment in our durable swine hind limb ischemia model. Autologous bone marrow eMSCs or empty alginate capsules were intramuscularly injected 2 weeks post-hind limb ischemia establishment (N=4/group). Improvements were quantified for 4 weeks through walkway gait analysis, contrast angiography, blood pressures, fluorescent microsphere perfusion, and muscle morphology and histology. Capsules remained intact with mesenchymal stromal cells retained for 4 weeks. Adenosine-induced perfusion deficits and muscle atrophy in ischemic limbs were significantly improved by eMSCs versus empty capsules (mean±SD, 1.07±0.19 versus 0.41±0.16, P=0.002 for perfusion ratios and 2.79±0.12 versus 1.90±0.62 g/kg, P=0.029 for ischemic muscle mass). Force- and temporal-associated walkway parameters normalized (ratio, 0.63±0.35 at week 3 versus 1.02±0.19 preligation; P=0.17), and compensatory footfall patterning was diminished in eMSC-administered swine (12.58±8.46% versus 34.85±15.26%; P=0.043). Delivery of eMSCs was associated with trending benefits in collateralization, local neovascularization, and muscle fibrosis. Hypoxia-cultured porcine mesenchymal stromal cells secreted vascular endothelial growth factor and tissue inhibitor of metalloproteinase 2. This study demonstrates the promise of the mesenchymal stromal cell secretome at improving peripheral artery disease outcomes and the potential for this novel swine model to serve as a component of the preclinical pipeline for advanced therapies.

Exploring the genetic causal association of TIMP3 on CKD and kidney function: a two-sample mendelian randomization.

Background: Numerous studies have demonstrated a positive association between the level of tissue inhibitor of metalloproteinase 3 (TIMP3) and chronic kidney disease (CKD). Nevertheless, whether those associations reflect causal links still to be determined. This study intended to research the causal relationship of TIMP3 with CKD and markers of kidney function, such as creatinine-based estimated glomerular filtration rate (eGFRcrea), cystatin C-based estimated glomerular filtration rate (eGFRcys), eGFRcrea in diabetics (eGFRcrea (DM)) and eGFRcrea in non diabetics (eGFRcrea (No DM)). Methods: In this study, we investigated the causal relationships between TIMP3 and CKD and kidney function markers using a two-sample Mendelian randomization (MR) technique. We used summary level datasets for TIMP3 and CKD from genome-wide association studies that we were able to access through the study by Suhre K and Pattaro C. Results: We found that TIMP3 had a significant positive causal effect on the risk of CKD (Inverse variance weighted (IVW):odds ratio (OR):0.962, 95% confidence interval (CI): (0.936-0.988),P:0.005). However TIMP3 levels had no significant effect on risk of eGFRcys (PIVW: 0.114),eGFRcrea (PIVW:0.333). After grouping patients based on their diabetes status, we found that genetically higher levels of TIMP3 had a significant impact on eGFRcrea in participants without diabetes (OR:1.003,95%CI (1.001-1.006),P IVW:0.007), but not in participants with diabetes (PIVW = 0.057). Heterogeneity and pleiotropy analyses were carried out to verify the accuracy of the MR findings. Their findings were all not statistically significant. Conclusion: Our study suggests that TIMP3 may be causally associated with CKD and eGFRcrea (No DM)in people of European ancestry. Strategies aimed to increase TIMP3 levels may provide new ways to delay the deterioration of renal function.

220 Litter performance, serum collagen, and hormone concentrations in prolapsed and non-prolapsed sows

Abstract Pelvic organ prolapse is one of the top leading causes of sow mortality today and widely considered a multifactorial issue. Previous reports observed prolapsed sows having decreased serum concentrations of various trace minerals and increased tumor necrosis factor-alpha compared with non-prolapsed sows. Additionally, collagen, hormones, and previous litter performance are other factors proposed to be associated with POP. A study was completed to determine differences in serum collagen type 1 (COL1), matrix metalloproteinase 1 (MMP-1), tissue inhibitor of metalloproteinase 1 (TIMP-1), as well as relaxin and estradiol differences in the serum concentration between prolapsed and non-prolapsed sows. Furthermore, previous litter performance, lactation days, pigs born, stillborns, mummified, and pigs weaned on prolapse rate were evaluated. The study utilized 44 prolapsed sows and 44 non-prolapsed sows of similar parity, location, management, and stage of production. Blood was collected from the prolapsed and non-prolapsed sows upon discovery. Serum samples were analyzed using radioimmunoassay for estradiol content. Additionally, COL1, MMP-1, TIMP-1, and relaxin concentrations were determined from the serum using ELISA testing. Data were analyzed using the PROC GLIMMIX procedure in SAS. Prolapsed sows had decreased (P < 0.05) COL1 and TIMP-1 concentrations in the serum compared with the non-prolapsed sows. Prolapsed sows had marginally greater (P < 0.10) MMP-1 concentration compared with non-prolapsed sows. There were no differences (P > 0.10) between prolapsed and non-prolapsed serum concentraions of sows for estradiol and relaxin. Prolapsed sows had longer lactation days (P < 0.05) in the litter before prolapsing compared with non-prolapsed sows. Additionally, stillborns in the previous litter were marginally greater (P = 0.06) in prolapsed sows when compared with the non-prolapsed sows. There were no differences (P > 0.10) in pigs born, mummified, or pigs weaned between prolapsed and non-prolapsed sows. This study suggests that there is a reduced concentration of serum COL1 and TIMP-1 found in prolapsed sows compared with non-prolapsed sows and a marginally increased MMP-1 concentration in prolapsed sows. This could influence the structural integrity of the connective tissue that supports the pelvic organs. We found no differences in serum estradiol or relaxin levels. Additionally, lactation days and stillborn pigs in the litter prior to prolapsing could be contributing factors to prolapse. This work further supports the idea that prolapse is a multifactorial issue.

Anti-Metastatic Effects of Standardized Polysaccharide Fraction from Diospyros kaki Leaves via GSK3β/β-Catenin and JNK Inactivation in Human Colon Cancer Cells.

A polysaccharide fraction from Diospyros kaki (PLE0) leaves was previously reported to possess immunostimulatory, anti-osteoporotic, and TGF-β1-induced epithelial-mesenchymal transition inhibitory activities. Although a few beneficial effects against colon cancer metastasis have been reported, we aimed to investigate the anti-metastatic activity of PLE0 and its underlying molecular mechanisms in HT-29 and HCT-116 human colon cancer cells. We conducted a wound-healing assay, invasion assay, qRT-PCR analysis, western blot analysis, gelatin zymography, luciferase assay, and small interfering RNA gene silencing in colon cancer cells. PLE0 concentration-dependently inhibited metastasis by suppressing cell migration and invasion. The suppression of N-cadherin and vimentin expression as well as upregulation of E-cadherin through the reduction of p-GSK3β and β-catenin levels resulted in the outcome of this effect. PLE0 also suppressed the expression and enzymatic activity of matrix metalloproteinases (MMP)-2 and MMP-9, while simultaneously increasing the protein and mRNA levels of the tissue inhibitor of metalloproteinases (TIMP-1). Furthermore, signaling data disclosed that PLE0 suppressed the transcriptional activity and phosphorylation of p65 (a subunit of NF-κB), as well as the phosphorylation of c-Jun and c-Fos (subunits of AP-1) pathway. PLE0 markedly suppressed JNK phosphorylation, and JNK knockdown significantly restored PLE0-regulated MMP-2/-9 and TIMP-1 expression. Collectively, our data indicate that PLE0 exerts an anti-metastatic effect in human colon cancer cells by inhibiting epithelial-mesenchymal transition and MMP-2/9 via downregulation of GSK3β/β-catenin and JNK signaling.

Epigenetic and biological age acceleration in children with atopic dermatitis

BackgroundAtopic dermatitis is a chronic inflammatory skin disease resulting from the complex interplay of genetic and environmental factors, meriting exploration using temporally-dynamic biomarkers. DNA methylation-based algorithms have been trained to accurately estimate biological age, and deviation of predicted age from true age (epigenetic age acceleration) has been implicated in several inflammatory diseases, including asthma. ObjectiveTo determine the role of epigenetic and biological aging, telomere length, and epigenetically-inferred abundance of seven inflammatory biomarkers in atopic dermatitis. MethodsWe performed DNA methylation-based analyses in a pediatric atopic dermatitis cohort (n=24, mean age 2.56±0.28y) and age-matched healthy subjects (n=24, mean age 2.09±0.15y) derived from blood, using five validated algorithms that assess epigenetic (Horvath, Skin&Blood) and biological age (PhenoAge, GrimAge), telomere length (TelomereLength), and inflammatory biomarker levels. ResultsEpigenetic and biological age, but not telomere length, were accelerated in atopic dermatitis patients for four algorithms: Horvath (+0.88 years; 95%CI 0.33-1.4; p=2.3x10-3), Skin&Blood (+0.95 years; 95%CI 0.67-1.2; p=1.8x10-8), PhenoAge (+8.2 years; 95%CI 3.4-13.0; p=1.3x10-3), and GrimAge (+1.8 years 95%CI 0.22-3.3; p=0.026). Moreover, patients had increased levels of beta-2-microglobulin (+47,584.4 ng/ml; p=0.029), plasminogen activation inhibitor 1 (+3,432.9 ng/ml; p=1.1x10-5) and cystatin C (+31,691 ng/ml; p=4.0x10-5), while levels of tissue inhibitor metalloproteinase 1 (-370.7 ng/ml; p=7.5x10-4) were decreased versus healthy subjects. ConclusionDNA methylation changes associated with epigenetic and biological aging, and inflammatory proteins appear early in life in pediatric atopic dermatitis and may be relevant clinical biomarkers of pathophysiology.

ПРИМЕНЕНИЕ ИНГИБИТОРОВ МЕТАЛЛОПРОТЕИНАЗ В КОМПЛЕКСНОМ ЛЕЧЕНИИ ВОСПАЛИТЕЛЬНЫХ ЗАБОЛЕВАНИЙ ПАРОДОНТА

Periodontitis is the most common inflammatory disease that leads to the destruction of the supporting tissues of the tooth. Therapeutic treatments aimed at reducing the effects of matrix metalloproteinases (MMPs) may be an effective adjunct to the treatment of periodontitis. The review examines the role of proteolytic enzyme inhibitors in the treatment of inflammatory periodontal diseases. General information about the ability of tetracyclines to inhibit MMPs is discussed. The purpose of the study was to examine the use of MMP inhibitors as a treatment for periodontal disease. Material and research methods. A scientific review of studies was carried out in Russian and English using information portals and platforms eLIBRARY.ru, Web of Science, PubMed and Scopus. The search was carried out using the following keywords: matrix metalloproteinases; oral diseases; tissue inhibitors of metalloproteinases. Of the 55 initially identified articles, after the initial analysis, 19 publications devoted to the study of MMP inhibitors associated with periodontal diseases were selected. Results of the study. Analysis of the literature allows us to conclude that the elimination of local irritants of periodontal tissues, the use of a therapeutic and prophylactic complex of measures using MMP inhibitors will have an advantage over traditional methods of treatment, will contribute to longer remission and prevent the progression of the inflammatory process in periodontal tissues, positively influence its course and reduce treatment time. The use of metalloproteinase inhibitors is an effective therapeutic strategy in the treatment of periodontal diseases. Conclusions. The therapeutic potential of metalloproteinase inhibitors may help prevent tissue damage in inflammatory periodontal diseases.

Colchicine modulates TIMP1, MMP1, and pMAP4 expression to enhance extracellular matrix degradation in 3T3 fibroblast-mediated myocardial infarction

Context: Cardiovascular disease is a leading cause of mortality worldwide. Following acute myocardial infarction (AMI), cardiac cells release many cytokines and proteolytic enzymes that regulate the extracellular matrix (ECM), including matrix metalloproteinases (MMPs), tissue inhibitor of metalloproteinases (TIMPs), and phosphorylated microtubule-associated protein 4 (Phospho/pMAP4). Aims: To analyze the role of colchicine in ECM regulation post-ischemic in cell line culture towards MMPs, TIMPs, and pMAP4. Methods: This in vitro experimental study was conducted on the 3T3 fibroblast cell line subjected to hypoxic conditions (CoCl2 treatment) and compared with 3T3 cells experiencing hypoxia/ischemic and colchicine treatment. MMP-1, TIMP-1, and pMAP4 expression were measured using flow cytometry. Data analysis was performed using One-Way ANOVA with a 95% confidence level. Results: Treatment with colchicine in the 3T3 fibroblast cell culture regulated the extracellular matrix by increasing TIMP-1 expression and reducing MMP-1 and pMAP4 expression (p<0.0001) following ischemic conditions induced by CoCl2. Conclusions: Colchicine effectively mitigates ECM degradation by enhancing TIMP-1 expression and inhibiting MMP-1 activity and pMAP4 expression post-ischemic conditions.

Matrix metalloproteinase-1 and matrix metalloproteinase-9 are highly expressed in the joint capsule of diabetic frozen shoulder

BackgroundThe pathophysiology of frozen shoulder (FS) involves abnormal expressions of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) that lead capsular fibrosis. However, there has been little concern for why diabetic FS has more protracted fibrotic condition. The objective of this study was to compare the expression levels of MMPs and TIMPs in the joint capsule of patients with diabetic and non-diabetic FS. Materials and methodsSamples of capsular tissue were collected from 20 patients with FS (10 diabetic patients; diabetic group, and 10 non-diabetic patients; non-diabetic group) and 10 patients (control group) with chronic anterior shoulder instability. Quantitative real-time RT-PCR and Western blot analysis were performed to determine the expression levels of mRNA and protein for MMP-1, 3, 9, 13, 14, and TIMP-1, 2. ResultsThe results of quantitative real-time RT-PCR showed significantly higher expression levels of all MMPs and TIMP-1 and significantly lower expression levels of TIMP-2 in the joint capsule of patients in the diabetic or non-diabetic groups compared with the control group. Significantly higher expression levels of MMP-1, 9, 14, and TIMP-1 were detected in the diabetic group compared with the non-diabetic group. The results of Western blot analysis showed significantly higher levels of MMP-3, 13, 14, and TIMP-1 in the joint capsule of patients in the diabetic or non-diabetic groups compared with the control group. However, no significant differences of protein levels of them were observed between diabetic and non-diabetic groups. ConclusionsThe findings of this study demonstrate the potential involvement of MMP-1 and 9 in the pathophysiology of diabetic FS. These findings may be helpful in identification of therapeutic targets for development of novel treatments for this protracted chronic fibrosing condition.

Network Pharmacological Analysis and Experimental Validation of the Effects of Silybin on Proliferation, Migration, and Immunotherapy of Papillary Thyroid Cancer.

The study aimed to use network pharmacology research and in vitro experiments to investigate the material basis and molecular mechanisms of silybin in the treatment of papillary thyroid carcinoma. Papillary thyroid cancer (PTC) has a decent prognosis; however, recurrence and metastasis are the leading causes of death in patients with PTC. A key research focus in thyroid cancer treatment is the inhibition of PTC proliferation, invasion, and migration. Silybin, the major active element in the traditional Chinese herb silymarin, has been used to treat a range of diseases, including cancer, but no study has been undertaken to determine whether it can help prevent PTC. In this study, we attempted to determine through network pharmacology and in vitro experiments if silybin inhibits the development of papillary thyroid cancer by inhibiting cell cycle and invasive migration. To predict the probable targets and underlying mechanisms of silybin against PTC, a network pharmacology research was performed. In vitro experiments were conducted to further evaluate silybin's anti-cancer properties and priority targets against PTC. The datasets revealed a total of 489 silybin targets acting on PTC, and functional enrichment analysis suggested that the target genes were enriched in functions and pathways related to PTC development, invasion, migration, and immunotherapy. By constructing these target PPI networks, the seven hub genes, fibronectin 1 (FN1), tissue inhibitor of metalloproteinases 1 (TIMP1), N-cadherin (CDH2), collagen type III alpha 1 chain (COL3A1), cyclin D1 (CCND1), AP-1 transcription factor subunit (JUN), and hepatocyte growth factor receptor (MET) were found. These hub genes were determined to be highly linked to a worse clinicopathological form, a higher risk of metastatic recurrence, and a worse prognosis of PTC. The common immunological checkpoint gene expression levels were positively correlated with the expression levels of the hub genes. Silybin decreased the proliferative and metastatic capacity of PTC cells, according to in vitro investigations. When PTC was treated with silybin, the FN1/AKT signaling pathway was blocked, CCND1 expression was reduced, and CDH2, Vimentin, Snail, Slug and PD-L1 expressions were dramatically reduced, while E-cadherin expression was significantly elevated. These findings provide preliminary evidence that silybin inhibits PTC cell proliferation, metastasis, and invasion by altering the FN1/AKT signaling pathway and inhibiting the EMT process. Silybin can reverse immunosuppression in papillary thyroid cancer by affecting immunological checkpoint gene expression levels. These studies provide a theoretical and experimental scientific basis for the potential anticancer effects of silybin on PTC.

Early prognostic markers to predict unsuccessful pregnancy in dairy cattle.

This study aimed to investigate maternal serum levels of some angiogenic factors and certain proteins in dairy cattle for (1) early prediction of unsuccessful fertilization and (2) early detection of possible pregnancy failures (early EM) after positive insemination Serum samples were collected from the same cattle at three distinct time points: 30 days before artificial insemination (B-AI), on the day of artificial insemination (AI), and 30 days after artificial insemination (A-AI). As a result of the pregnancy examination, the cows were divided into two main groups according to whether they were pregnant. The results showed that leucyl/cystinyl aminopeptidase (LNPEP) concentration was significantly decreased B-AI and Secreted frizzled-related proteins (SFRP-3), Vascular Endothelial Growth Factor (VEGF) and LNPEP levels were significantly decreased on day of AI, while PRL level was increased, and these data have prognostic significance as early indicator of the risk of potentially failed pregnancy. Additionally, a significant decrease in LNPEP, SFRP3, and VEGF levels, along with an increase in PRL levels was also observed in A-AI. These results suggest that these biomarkers can be used as a screening test to monitor the course of pregnancy. There were no significant differences in serum levels of Insulin-Like Growth Factor 2 (IGF-2), Tissue inhibitors of metalloproteinases (TIMP-1), angiopoietin (ANG), Endoglin (ENG), Fibroblast growth factor (FGF), Inhibine-A (INH-A) and Transforming growth factors-β1 (TGF-β1) between the evaluated periods neither unsuccessful nor the successful pregnancy groups. This is the first study reporting that the maternal serum levels of LNPEP, SFRP3, VEGF, and PRL have important roles in pregnancy success and may indicate whether insemination outcome will be successful B-AI and predict the risk of unsuccessful pregnancy after AI in dairy cattle. The increase in such studies will allow the development of more specific, practical, and applicable markers.

Violation of Allowable Work Intensity does not Augment Acute Kidney Injury Risk During Simulated Occupational Heat Stress

We recently demonstrated that violation of the National Institute of Occupational Safety and Health (NIOSH) heat stress recommendations by exceeding the allowable wet bulb globe temperature (WBGT) for a given moderate work intensity and work:rest ratio augmented acute kidney injury (AKI) risk. However, the role of work intensity (at a fixed work:rest ratio) on AKI risk during NIOSH compliant and non-compliant scenarios remains largely unknown. This study tested the hypothesis that violating the NIOSH recommendations by exceeding the allowable work intensity at a given WBGT and fixed work:rest ratio would augment AKI risk. Twelve healthy adults completed two NIOSH recommendation compliant trials and one noncompliant trial consisting of a 4 h exposure to a fixed WBGT and work intensity. Work-rest ratio was fixed at 30 min of walking and 30 min of rest each hour. Work intensity (i.e., Hprod) was prescribed as a function of WBGT— 412±51 W (27.3±0.3°C; high intensity compliant [Chigh]), 290±75 W (31.6±0.2°C; low intensity compliant [Clow]), and 410±61 W (31.7±0.2°C; high intensity noncompliant [NChigh]). All subjects were provided 237 mL of sport drink every 15 min and drank ad libitum. TC is presented as peak across the 4 h exposure. Urine samples were collected preexposure, and immediately and 1 h postexposure. Urinary inflammatory (monocyte chemotactic protein-1 [MCP-1]), oxidative stress (thioredoxin 1 [TRX-1]), and AKI risk (the product of insulin-like growth factor-binding protein 7 and tissue inhibitor of metalloproteinase 2 [IGFBP7·TIMP-2]) biomarkers were measured and normalized to urine flow rate. Urinary biomarkers were log transformed before statistical analysis and data are presented as mean ± SD. Peak Tc was higher during the NChigh trial (38.3±0.4°C) compared to the compliant trials (Chigh: 38.0±0.3°C; Clow: 37.8±0.4°C; p≤0.0095). Body weight change from pre- to postexposure did not differ between trials (p=0.7817). MCP-1 increased from pre- to immediately postexposure in all trials (p≤0.0031) but was not different between trials (p=0.1354). TRX-1 increased from pre- to immediately postexposure in all trials (p≤0.0085) but was not different between trials (p=0.6372). [IGFBP7·TIMP-2] increased from pre- to immediately postexposure in all trials (p=0.0454) but the increase was not different between trials (Chigh: -1.7±2.6 LOG [ng/mL2]/1000; Clow: -1.6±2.4 LOG [ng/mL2]/1000; NChigh:-1.2±2.3 LOG [ng/mL2]/1000; p=0.2076). Simulated occupational heat stress elevates AKI risk, likely due to both inflammation and oxidative stress. Violating the NIOSH recommendations by exceeding either the allowable WBGT (i.e., NChigh versus Chigh) or work intensity (i.e., NChigh versus Clow) resulted in a modest elevation in peak Tc but did not modify AKI risk and/or the urinary inflammatory and oxidative stress responses. RCT: NCT04767347; Supported By: R01OH011528. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Ad Libitum Drinking Does Not Modify Renal Oxidative Stress or Inflammation During Simulated Occupational Heat Stress

Prolonged heat exposure increases acute kidney injury (AKI) risk likely due to renal oxidative stress and inflammation. However, the effect of occupational heat stress (OHS) and the modifying role of hydration on renal oxidative stress and inflammation is unknown. This study tested the hypothesis that compared to fluid restriction ad libitum drinking consistent with OHS recommendations will attenuate renal oxidative stress and inflammation during simulated OHS. In a randomized, crossover design, 13 healthy adults (5 women) completed two 2 h OHS simulations consisting of of 8 circuits of treadmill walking (10 min) and rowing (5 min) at an oxygen uptake of 1.2 L/min in a wet bulb globe temperature of 33.1±0.2°C. In the drinking trial (Drink), subjects were provided 237 mL of a non-caloric sport drink (Gatorade Zero) every 15 min and drank ad libitum. In the fluid restriction trial (NoDrink), no fluid was provided. Rectal temperature and body weight were recorded. Urine and blood samples were collected pre-, post- and 1 h post- exposure. Inflammatory (monocyte chemotactic protein-1, MCP-1) and oxidative stress (thioredoxin 1, TRX-1) markers were measured in urine and serum, and fractional excretion (FE) was calculated. AKI risk was quantified as the product of urine insulin-like growth factor-binding protein 7 and urine tissue inhibitor of metalloproteinase 2 ([IGFBP7•TIMP-2]). Data are presented as mean ± SD. Peak rectal temperature was not different between trials (Drink: 38.5±0.4°C, NoDrink: 38.6±0.4°C, p=0.346). Percent changes in body weight were lower in Drink (1.3±0.8% vs. 2.8±0.9%, p<0.001). FETRX-1 was elevated at post- (0.50±0.00%) compared to pre- (0.36±0.00%, p=0.011) and 1 h post- (0.29±0.13%, p=0.011). FEMCP-1 was elevated at post- (1.25±0.18%) compared to pre- (1.02±0.17%, p=0.006) but not 1 h post- (1.27±0.03%, p>0.999). FETRX-1 and FEMCP-1 did not change significantly between trials (trial x time: p³0.622). [IGFBP7•TIMP-2] was higher at 1 h post- (17.5±1.0 (ng/mL)2/1000) compared to pre- (0.2±0.1 (ng/mL)2/1000, p<0.001) and post- (1.1±0.4 (ng/mL)2/1000, p<0.001) but did not differ between trials (trial x time: p=0.556). Peak changes in [IGFBP7•TIMP-2] were not significantly correlated with peak changes in FETRX-1 (r=0.126, p=0.557) or FEMCP-1 (r=0.292, p=0.166) and were correlated with peak changes in serum MCP-1 (r=0.714, p<0.001) but not serum TRX-1 (r=0.227, p=0.286). Two hours of simulated OHS increased renal inflammation (FEMCP-1), oxidative stress (FETRX-1), and AKI risk ([IGFBP7•TIMP-2]), while ad libitum drinking did not modify FEMCP-1, FETRX-1, or [IGFBP7•TIMP-2]. Lack of correlations between peak changes in [IGFBP7•TIMP-2] and FEMCP-1, FETRX-1, or serum TRX-1, but the strong positive correlation with peak serum MCP-1 suggests a role for systemic inflammation in OHS induced AKI risk. NCT05458843 Funding: R01OH011528. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Synthesis of amide derivatives containing the imidazole moiety and evaluation of their anti-cardiac fibrosis activity.

Three series of N-{[4-([1,2,4]triazolo[1,5-α]pyridin-6-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-yl]methyl}acetamides (14a-d, 15a-n, and 16a-f) were synthesized and evaluated for activin receptor-like kinase 5 (ALK5) inhibitory activities in an enzymatic assay. The target compounds showed high ALK5 inhibitory activity and selectivity. The half maximal inhibitory concentration (IC50) for phosphorylation of ALK5 of 16f (9.1 nM), the most potent compound, was 2.7 times that of the clinical candidate EW-7197 (vactosertib) and 14 times that of the clinical candidate LY-2157299. The selectivity index of 16f against p38α mitogen-activated protein kinase was >109, which was much higher than that of positive controls (EW-7197: >41, and LY-2157299: 4). Furthermore, a molecular docking study provided the interaction modes between the target compounds and ALK5. Compounds 14c, 14d, and 16f effectively inhibited the protein expression of α-smooth muscle actin (α-SMA), collagen I, and tissue inhibitor of metalloproteinase 1 (TIMP-1)/matrix metalloproteinase 13 (MMP-13) in transforming growth factor-β-induced human umbilical vein endothelial cells. Compounds 14c and 16f showed especially high activity at low concentrations, which suggests that these compounds could inhibit myocardial cell fibrosis. Compounds 14c, 14d, and 16f are potential preclinical candidates for the treatment of cardiac fibrosis.

Structure identification of selenium-biofortified soybean peptides and its effect on TGF-β mediated liver fibrosis pathway

Selenium-biofortified soybean peptides (SeSPs) has been verified to relieve CCl4-induced liver fibrosis in vivo previously. In the present study, the structures of SeSPs were characterized by HPLC-ESI-MS/MS and thirty-eight selenium-containing peptides sequences were identified. KLPISeM was screened out as the most promising peptide to inhibit TGF-β/Smads pathway with stable binding site to TGF-β in molecular docking. Besides, homo sapiens liver cell line (L-O2) and hepatic stellate cells from rat (HSC-T6) were treated with KLPISeM/KLPIM and TGF-β1 to explore the regression fibrosis mechanisms of peptides. Results showed that the apoptosis trend of L-O2 was inhibited while HSC-T6 apoptosis was promoted by KLPISeM. Smad2/3/4 gene levels were reduced by 60.36%, 59.08% and 71.82%, respectively and Smad7 gene expression was upregulated significantly (p < 0.05) after incubation with KLPISeM. More importantly, KLPISeM exhibited strong up-regulation effect on matrix metallo-proteinase (MMPs) and suppressive effect on tissue inhibitor of metalloproteinase gene/protein expressions in HSC-T6. These results suggested that KLPISeM could relieve liver fibrosis by blocking TGF-β/Smads and regulating MMPs/TIMPs pathway.