The Role of Plasmin/Plasminogen in Experimental Polymicrobial Sepsis

Introduction: Sepsis is characterized by systemic inflammation and increased coagulation. Although plasmin activity is increased at the onset of sepsis, increased levels of plasminogen activator inhibitor-1 can counter-regulate it triggering an imbalance of the procoagulant and fibrinolytic systems, which results in disseminated intravascular coagulation. Objective: To evaluate the role of the plasminogen/plasmin (Plg/Pla) system in experimental sepsis. Material and Methods: Plg-deficient (Plg-/-) mice, Plg+/+ littermate controls and wild-type (WT) C57Bl/6 mice were subjected to Cecal Ligation and Puncture (CLP) to induce sepsis. In this model, a double puncture is made through the cecum using either a 30 or 18-gauge needle to induce sub-lethal or lethal sepsis, respectively. Survival rates were assessed. Inflammatory parameters and bacterial counts were evaluated in peritoneal lavage fluid and blood 12h after CLP. In other experiments WT mice were treated with Pla or Plg, employing two different therapeutic protocols. Results: In WT mice, the survival rate in sub-lethal sepsis was more than 80% at 6 days after CLP, while lethal sepsis all mice died by 2 days. Increased inflammatory infiltrates were found in peritoneal cavities of mice subjected to lethal sepsis compared to sub-lethal sepsis. The plasma levels of Plg were reduced in lethal sepsis when compared to sub-lethal sepsis, while the levels of a marker for sepsis severity, IL-6, increased. In the sub-lethal model of sepsis, the survival rate of Plg-/- mice was 40% while all Plg+/+ mice survived. In this model, increased inflammatory infiltration, predominantly neutrophils and M1 macrophages, was present in peritoneal cavities of Plg-/- mice 12h after CLP, accompanied by increased IL-6 and ALT levels, compared with Plg+/+ littermates. No genotype-dependent difference in the levels of TNF-α and IL-10 were observed. The bacterial clearance in Plg-/- mice and Plg+/+ littermates were similar. In WT mice, exogenous Pla (10 µg/mice, i.p.) administered 3h after lethal sepsis did not modify the lethality rate, but decreased inflammatory infiltration (neutrophils and M1 macrophages) at the infectious site with reduced levels of CXCL1 and ALT. No differences in the levels of TNF-α, IL-1-6 and IL-10 were observed after Pla treatment. Pla administration reduced bacterial counts in blood with no differences in peritoneal fluid. In a separate protocol, administration of Pla or Plg (10 µg/mouse, i.p.) in two bolus injections, at both 6 and 12h after lethal CLP, resulted in a significant reduction in lethality rate (~30% survival) when compared to the vehicle group (no survival). Interestingly, co-injection of Plg in combination with a broad-spectrum antibiotic (imipenem 30mg/kg, i.p.) protected approximately 80% of mice, compared with 60% protection with antibiotic alone. Conclusion: Plg/Pla exhibit a protective effect in sepsis by reducing inflammatory parameters, neutrophil recruitment and tissue damage. Keywords: sepsis, plasmin, plasminogen system, CLP model. Disclosures No relevant conflicts of interest to declare.

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Stem Cell-Derived Exosomes Ameliorate Doxorubicin-Induced Muscle Toxicity through Counteracting Pyroptosis

Pharmaceuticals ◽

10.3390/ph13120450 ◽

2020 ◽

Vol 13 (12) ◽

pp. 450

Author(s):

Fatima Bianca A. Dessouki ◽

Rakesh C. Kukreja ◽

Dinender K. Singla

Keyword(s):

Muscle Function ◽

Muscular Atrophy ◽

Embryonic Stem ◽

Mouse Embryonic Fibroblast ◽

Negative Control ◽

M1 Macrophages ◽

Tnf Α ◽

Embryonic Fibroblast ◽

Muscle Toxicity

Doxorubicin (Dox)-induced muscle toxicity (DIMT) is a common occurrence in cancer patients; however, the cause of its development and progression is not established. We tested whether inflammation-triggered cell death, “pyroptosis” plays a role in DIMT. We also examined the potential role of exosomes derived from embryonic stem cells (ES-Exos) in attenuating DIMT. C57BL/6J mice (10 ± 2 wks age) underwent the following treatments: Control (saline), Dox, Dox+ES-Exos, and Dox+MEF-Exos (mouse-embryonic fibroblast-derived exosomes, negative control). Our results demonstrated that Dox significantly reduced muscle function in mice, which was associated with a significant increase in NLRP3 inflammasome and initiation marker TLR4 as compared with controls. Pyroptosis activator, ASC, was significantly increased compared to controls with an upregulation of specific markers (caspase-1, IL-1β, and IL-18). Treatment with ES-Exos but not MEF-Exos showed a significant reduction in inflammasome and pyroptosis along with improved muscle function. Additionally, we detected a significant increase in pro-inflammatory cytokines (TNF-α and IL-6) and inflammatory M1 macrophages in Dox-treated animals. Treatment with ES-Exos decreased M1 macrophages and upregulated anti-inflammatory M2 macrophages. Furthermore, ES-Exos showed a significant reduction in muscular atrophy and fibrosis. In conclusion, these results suggest that DIMT is mediated through inflammation and pyroptosis, which is attenuated following treatment with ES-Exos.

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Klotho protects chondrocyte viability via FOXO1/3 in osteoarthritis

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v20i9.24 ◽

2021 ◽

Vol 20 (9) ◽

pp. 1961-1968

Author(s):

Wei Wei ◽

Liefeng Ji ◽

Wanli Duan ◽

Jiang Zhu

Keyword(s):

Oxidative Stress ◽

Survival Rate ◽

Protective Effect ◽

Collagen I ◽

Inflammatory Factors ◽

Ros Production ◽

Chondrocyte Viability ◽

Tnf Α ◽

Collagen Ii

Purpose: To investigate the effect of Klotho and FOXO1/3 on the CH viability in OA.Methods: The survival rate of CHs, Klotho and FOXO1/3 protein expression, and ROS production were measured in the OA cartilages of different degenerative phases. H2O2 was also used to injure CHs, and the cell viability, Klotho and FOXO1/3 expressions, as well as ROS levels were investigated to clarify the effect of exogenic Klotho on the injured CHs. Additionally, in order to verify the role of FOXO1/3 in Klotho-treated CHs, SOD2, GPX1, inflammatory factors, collagen I/II, SOX9, and Runx-2 levels were analyzed by silencing FOXO1 and FOXO3 expression via siRNA transfection.Results: Klotho and FOXO1/3 expressions significantly decreased, and ROS production increased in severely human OA cartilage (p <0.05). Besides, H2O2 affected CHs viability with the suppression of Klotho and FOXO1/3 expression but ROS production was elevated. Exogenic Klotho application partly reversed the injury caused by H2O2. Furthermore, Klotho treatment of the injured CHs contributed to SOD2 and GPX1 expressions, and suppressed IL-1β, IL-6, TNF-α and MMP-13 production, resulting in the upregulation of collagen II and SOX9 as well as downregulation of collagen I and Runx-2. However, the protective effect of Klotho was weakened by FOXO1 and FOXO3 gene silencing.Conclusion: Klotho protects CHs viability by suppressing oxidative stress and inflammation, which is associated with the mediation of FOXO1 and FOXO3. These findings provide new insights into the treatment of OA.

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Genetic variants of interferon-gamma and its mRNA expression and inflammatory parameters in the pathogenesis of vitiligo

Biochemistry and Cell Biology ◽

10.1139/bcb-2016-0228 ◽

2017 ◽

Vol 95 (4) ◽

pp. 474-481 ◽

Cited By ~ 7

Author(s):

Rehab A. Karam ◽

Haidy E. Zidan ◽

Mohamed H. Khater

Keyword(s):

Elevated Serum ◽

Serum Levels ◽

Intercellular Adhesion Molecule ◽

Control Group ◽

Intercellular Adhesion Molecule 1 ◽

Inflammatory Parameters ◽

Increased Risk ◽

Tnf Α ◽

Ifn Γ

Although genetics plays an essential role in the pathogenesis of vitiligo, vitiligo pathogenesis is still unclear. Our aim was to investigate the role of IFN-γ expression and polymorphism in vitiligo susceptibility and whether intercellular adhesion molecule-1 (ICAM-1), tumor necrosis factor (TNF)-α, and TNF-β play a role in vitiligo pathogenesis as important inflammatory parameters. Eighty-five patients with vitiligo and 90 controls were investigated for IFN-γ gene expression by quantitative real-time PCR and genotyped for IFN-γ +874T/A (rs2430561) and IFN-γ +2109A/G (rs1861494) gene polymorphisms by sequence-specific primer (SSP)-PCR and PCR-restriction fragment length polymorphism (RFLP), respectively. Serum levels of inflammatory parameters were measured using ELISA. Frequencies of the +874 TT genotype and T allele were significantly higher in patients with active vitiligo than in stable patients (P = 0.01 and 0.03, respectively). Calculation of odds ratio suggested a 1.7-fold increased risk of vitiligo in individuals having the TA haplotype. We observed overexpression of IFN-γ mRNA with elevated serum levels of IFN-γ, ICAM-1, TNF-α, and TNF-β in patients with vitiligo when compared with the control group (P = 0.001, for all). In addition, these levels were elevated in patients with active vitiligo compared with stable patients with vitiligo (P = 0.008, 0.006, 0.01, 0.01, and 0.03, respectively), which suggests the involvement of these cytokines in disease activity. In conclusion, IFN-γ is a promising immunological marker in vitiligo pathogenesis.

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Renoprotective role of bariatric surgery in patients with established CKD

Clinical Kidney Journal ◽

10.1093/ckj/sfaa266 ◽

2020 ◽

Author(s):

Enrique Morales ◽

Esteban Porrini ◽

Marina Martin-Taboada ◽

Sergio Luis Lima ◽

Rocío Vila-Bedmar ◽

...

Keyword(s):

Bariatric Surgery ◽

Renal Function ◽

Glomerular Hyperfiltration ◽

Effective Measure ◽

Inflammatory Parameters ◽

Tnf Α ◽

Effective Option ◽

Obese Subjects

Abstract Background Bariatric surgery (BS) has been postulated as the most effective measure for weight reduction. Weight loss improves metabolic parameters and exerts changes in renal function that lead to the amelioration of absolute or relative glomerular hyperfiltration, a condition that may renoprotective in the long term. However, few studies have demonstrated the influence of bariatric surgery in patients with severe obesity and chronic kidney disease (CKD). Our objective was to analyze the evolution of renal function, adipose tissue-derived molecules and inflammatory parameters in patients with CKD after BS. Methods This is an observational and prospective study. Thirty patients were screened and 12 were included between January 2016 to January 2018 with a 24-month follow-up. Glomerular filtration rate (GFR) was determined by plasma iohexol clearance. Adipokines, cytokines, circulating hormones and fibrotic parameters were evaluated before and 12 months after bariatric surgery using the Bioplex system. Results The mean age was 50.6 years and 58.3% were males. Seven patients had a BMI> 40 kg/m2 and 66.7% were diabetic. Twenty-four months following bariatric surgery there was a significant decrease in body weight (36.4%). Proteinuria decreased by 63.7 ± 28.2%. Measured GFR significantly diminished from before surgery to month 24 after surgery (94 ± 44 to 79 ± 44 ml/min, p 0.03). There was a significant decrease in adipocyte-derived molecules (leptin, vifastin) as well as in pro-inflammatory cytokines (IL1-β, TNF-α, IL-6, MCP-1) and other circulating factors (VEGF and TGFß isoforms). Conclusions BS is an effective option to prevent kidney damage in obese subjects with CKD due to the improvement of glomerular hyperfiltration, adipocyte cytokines metabolic and inflammatory parameters.

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IL-10 deficiency protects hamsters from Leptospira infection

Infection and Immunity ◽

10.1128/iai.00584-21 ◽

2021 ◽

Author(s):

Xufeng Xie ◽

Tianbao Lv ◽

Dianjun Wu ◽

Haozhe Shi ◽

Shilei Zhang ◽

...

Keyword(s):

Gene Expression ◽

Survival Rate ◽

Zoonotic Disease ◽

Protective Effects ◽

Subclinical Infection ◽

Expression Levels ◽

Tnf Α ◽

Gene Expression Levels

Leptospirosis is a globally spread zoonotic disease with outcomes ranging from subclinical infection to fatal Weil syndrome. In addition to antibiotics, some immune activators have shown protective effects against leptospirosis. However, the unclear relationship between Leptospira and cytokines, has limited the development of antileptospiral immunomodulators. In this study, the particular role of IL-10 in leptospirosis was explored by using IL-10 defective (IL-10 -/- ) hamsters. After Leptospira infection, an improved survival rate, reduced leptospiral burden and alleviation of organ lesions were found in IL-10 -/- hamsters compared with WT hamsters. In addition, the gene expression levels of IL-1β, IL-6 and TNF-α and the NO level were higher in IL-10 -/- hamsters than in WT hamsters. Our results indicate that IL-10 deficiency protects hamsters from Leptospira infection.

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The Role of β-1.3/1.6-D-Glucan From Ganoderma lucidum Mycelium Extract in Ulcerative Colitis

The Indonesian Journal of Gastroenterology Hepatology and Digestive Endoscopy ◽

10.24871/2232021234-239 ◽

2022 ◽

Vol 22 (3) ◽

pp. 234-239

Author(s):

Marcellus Simadibrata ◽

Aditya Rachman ◽

Saskia Aziza Nursyirwan ◽

Murdani Abdullah ◽

Rabbinu Rangga Pribadi ◽

...

Keyword(s):

Ulcerative Colitis ◽

Side Effects ◽

Ganoderma Lucidum ◽

Animal Studies ◽

Alternative Therapy ◽

Drug Efficacy ◽

Mucosal Inflammation ◽

Inflammatory Parameters ◽

Tnf Α

Ulcerative colitis (UC) is an idiopathic inflammatory disease that affects the colon. Current pharmacological modalities to treat UC have various side effects; therefore, there is a demand to develop a new alternative medicine that can reduce side effects and increase drug efficacy. One candidate for alternative therapy is Polysaccharide Peptide which is extracted from Ganoderma lucidum mycelium. This Polysaccaharide has an active compound of Β-1,3/1,6-D-Glucan which has strong immunomodulatory and anti-inflammatory properties. Various studies have reported that Ganoderma lucidum polysaccharides can reduce inflammatory markers such as TNF-α, IFN-γ, and IL-17A, which is produced by colonic mucosal inflammation. In addition, β-1,3/1,6-D-Glucan has shown improvements in inflammatory parameters and intestinal immunological barrier function animal studies with artificial colitis and requires further research in humans before clinical applications.

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A methanolic extract of Trigonella foenum-graecum (fenugreek) seeds regulates markers of macrophage polarization.

Functional Foods in Health and Disease ◽

10.31989/ffhd.v5i12.216 ◽

2015 ◽

Vol 5 (12) ◽

pp. 417 ◽

Cited By ~ 2

Author(s):

Nurudeen Hassan ◽

Cathryn Withycombe ◽

Maninder Ahluwalia ◽

Andrew Thomas ◽

Keith Morris

Keyword(s):

Methanolic Extract ◽

Macrophage Polarization ◽

Diabetic Patients ◽

M1 Macrophages ◽

Fenugreek Seeds ◽

Tissue Microenvironment ◽

Trigonella Foenum Graecum ◽

Tnf Α ◽

Cellular Mediators

Background: Macrophages are key cellular mediators in diabetes-related inflammation. Molecular cues such as cytokines found in the tissue microenvironment regulates the polarization of macrophages into an M1 (pro-inflammatory) or M2 (immunoregulatory) phenotype. Recent evidence suggests that M1 macrophages in diabetic patients may contribute to the complications associated with the disease such as atherosclerosis. Trigonella foenum- graecum (Tfg: fenugreek) seeds have been used in traditional medicine in Asia, Africa and the Middle-East for their alleged anti-diabetic properties.Objective: To identify the molecular mechanism(s) through which Tfg seeds exert their effects, we investigated the role of a crude methanolic extract of Tfg (FME) seeds on macrophage polarization in vitro.Method: THP-1 macrophages (Mϕ) were treated with gBSA in the presence/absence of FME and the release and expression of M1 and M2 markers/cytokines were analysed. The role of FME on NF-κB activity was also explored using transfected HEK-293T cells.Results: This study found that the FME significantly (P<0.05) decreased gBSA-induced secretion of M1 cytokines (TNF-α, IL-1β, IL-6 and IL-8) in THP-1 Mϕ cells. In the presence of gBSA, FME also significantly increased the gene expression of the M2 marker Dectin-1, but had no effect on IL-10, IL-1Ra. FME also significantly decreased TNF-α induced NF-kB reporter activity.Conclusion: These results suggest that FME can regulate the expression of M1 and M2 markers in THP-1 Mϕ cells. This may be potentially through the modulation of NF-kB activity. Further work should be carried out to identify precise mechanism(s) involved in the effects of FME and Tfg seeds.Keywords: chronic inflammation, macrophage polarization, diabetes, glycated BSA, THP-1 cells, Trigonella foenum graecum, fenugreek seeds, NF-κB,

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