15-Lipoxygenase Worsens Renal Fibrosis, Inflammation, and Metabolism in a Murine Model of Ureteral Obstruction

2021 ◽  
Author(s):  
John Ross Montford ◽  
Colin Bauer ◽  
Jeremy Rahkola ◽  
Julie A Reisz ◽  
Deana Floyd ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Murine Model ◽  
Ureteral Obstruction ◽  
Dynamic Change ◽  
Heme Iron ◽  
Macrophage Phenotype ◽  
Post Injury ◽  
Inflammatory Microenvironment ◽  
Anti Inflammatory ◽  
Knockout Animals

INTRODUCTION: 15-Lipoxygenase (15-LO) is a non-heme iron-containing dioxygenase that has both pro- and anti-inflammatory roles in many tissues and disease states. 15-LO is thought to influence macrophage phenotype; and silencing 15-LO reduces fibrosis after acute inflammatory triggers. The goal of this study was to determine if altering 15-LO expression influences inflammation and fibrogenesis in a murine model of unilateral ureteral obstruction (UUO). METHODS: C57BL/6J mice, 15-lipoxygenase knockout (Alox15-/-) mice, and 15-lipoxygenase transgenic overexpressing mice (15LOTG) were subjected UUO and kidneys were analyzed at 3, 10, and 14-days post injury. Histology for fibrosis, cytokine quantification, flow cytometry, and metabolomics were performed on injured tissues and controls. PD146176, a specific 15-LO inhibitor, was used to complement studies involving knockout animals. RESULTS: Compared to WT animals undergoing UUO, Alox15-/- mouse kidneys had less pro-inflammatory, pro-fibrotic message along with less fibrosis. PD146176 inhibited 15-LO, and resulted in reduced fibrosis similar to Alox15-/- mice. Flow cytometry revealed that Alox15-/- UUO-injured kidneys had a dynamic change in macrophage phenotype, with an early blunting of CD11bHiLy6CHi "M1" macrophages and increase in anti-inflammatory CD11bHiLy6CInt "M2c" macrophages and reduced expression of the fractalkine receptor, CX3CR1. Many of these findings were reversed when UUO was performed on 15LOTG mice. Metabolomics analysis revealed that WT kidneys developed a glycolytic shift post-injury, while Alox15-/- kidneys exhibited increased oxidative phosphorylation. CONCLUSIONS: 15-LO manipulation by genetic or pharmacologic means induces dynamic changes in the inflammatory microenvironment in the UUO-model and appears to be critical in the progression of UUO-induced fibrosis.

scholarly journals Time related effects of non-steroidal anti-inflammatory drugs on Achilles tendinopathy in a murine model

2017 ◽  
Vol 2 (2) ◽  
pp. 2473011416S0000 ◽  
Author(s):  
Adam Bitterman ◽  
Shuguang Gao ◽  
Katie Trella ◽  
Jun Li ◽  
Jorge Galante ◽  
...  
Keyword(s):  
Wound Healing ◽  
Murine Model ◽  
Achilles Tendinopathy ◽  
Early Postoperative Period ◽  
Post Injury ◽  
Thin Sections ◽  
Delayed Treatment ◽  
Systemic Mechanism ◽  
Inflammatory Drugs ◽  
Anti Inflammatory

Category: Basic Sciences/Biologics Introduction/Purpose: Tendinopathy has been identified to result from work and sports-related activities, in many patients. Common treatment for such degenerative tendon pathology includes non-steroidal anti-inflammatory drugs (NSAIDs). Although they provide pain relief, their mechanistic effects, on modulating inflammation associated with tendinopathy is poorly understood. Recent reports suggest impaired functional healing of rat rotator cuff tear repairs when treated in the early postoperative period with ibuprofen, while delayed treatment did not affect healing. We have now identified, using a murine model, a cascade of genes associated with innate inflammation, which influence premature fibrotic scarring without repair. The objective of this study was to evaluate the effects of orally administered ibuprofen on inflammation and wound-healing responses after initiation and during progression of Achilles tendinopathy in a murine model. Methods: All animal experimentation was carried out under IACUC approval. C57BL/6 wild-type male mice (12 wks) received two injections, two days apart, of 100ng rhTGF-β1 into the midportion of the Achilles tendon. Ibuprofen (IBU) was administered orally as described by Ezell et al5, for 7 days either 1 day (Early) or 8 days (Late) after the initiating injury. Experimental groups were 1) Tendinopathy + NO drug, 2) Tendinopathy + IBU Early, 3) Tendinopathy + IBU Late, 4) Naïve mice + IBU Early. Following sacrifice of the mice at 28 days post injury, Achilles tendons were harvested, and RNA was prepared as described by Trella et al6, for transcriptomic analyses using wound healing and NfKb target gene platforms (PAMM-121A, PAMM-225ZA Qiagen, Valencia, CA). Tendon histopathology was assessed on paraffin embedded thin sections using standard methods for HE and Safranin O staining. Results: Both early and late dosing times with ibuprofen prolonged the innate inflammation response genes (e.g., PGE Synthase, Interleukin1b, Interferon gamma, Cxcl3 chemokine), relative to untreated tendinopathy. Furthermore, early dosing also elevated collagen genes (Collagens 1, 3 and 5), and such a response has been associated with fibrotic scarring. Drug administration to naïve mice showed no significant effects on gene expression in the tendons. Conclusion: This study represents the first report of the time related effect of ibuprofen on mechanisms involved in healing of tendinopathy. If the drug is administered early after the initiating injury, a prolonged increase in inflammation and potential for scarring in the tendon was observed, whereas later administration resulted in persistent inflammation only. The data suggest that the anti-inflammatory action of such NSAID may occur outside of the tendon (such as the bone marrow) resulting in a dysregulated systemic mechanism that controls the innate (tendon) inflammation and healing pathways.

scholarly journals AMPKα1 is essential for Glucocorticoid Receptor triggered anti-inflammatory macrophage activation

2020 ◽  
Author(s):  
Giorgio Caratti ◽  
Thibaut Desgeorges ◽  
Gaëtan Juban ◽  
Mascha Koenen ◽  
Bozhena Kozak ◽  
...  
Keyword(s):  
Glucocorticoid Receptor ◽  
Transcriptional Control ◽  
Inflammatory Cells ◽  
Macrophage Phenotype ◽  
Post Injury ◽  
Inflammatory Phase ◽  
Anti Inflammatory ◽  
Amp Activated Protein Kinase ◽  
Inflammatory Macrophage

SummaryMacrophages are key immune cells which mediate both the acute inflammatory phase and the repair phase after tissue damage. Macrophages switch from pro-inflammatory to anti-inflammatory cells that sustain repair and return to tissue homeostasis. We show that the metabolic sensor, AMP-activated protein kinase (AMPK) is essential for glucocorticoid induction of an anti-inflammatory macrophage phenotype. While canonical gene regulation by glucocorticoids was not affected by loss of AMPK, we identified AMPK-dependent glucocorticoid-regulated genes in macrophages, related to efferocytosis. AMPK-deficient macrophages do not acquire phenotypic and functional anti-inflammatory features upon glucocorticoid exposure. We identified FOXO3 as an AMPK-dependent regulator of glucocorticoid activity in macrophages. Loss of AMPK in macrophages in vivo abrogates glucocorticoid anti-inflammatory actions during post-injury muscle regeneration and endotoxin induced acute lung injury. These data highlight that the glucocorticoid receptor is dependent on AMPK for its immunomodulatory actions in macrophages, linking their metabolic status to transcriptional control in resolving inflammation.

The antifibrotic and anti-inflammatory effects of icariin on the kidney in a unilateral ureteral obstruction mouse model

Phytomedicine ◽  
2019 ◽  
Vol 59 ◽  
pp. 152917 ◽  
Author(s):  
Hsin‐An Chen ◽  
Chang-Mu Chen ◽  
Siao-Syun Guan ◽  
Chih-Kang Chiang ◽  
Cheng-Tien Wu ◽  
...  
Keyword(s):  
Mouse Model ◽  
Ureteral Obstruction ◽  
Unilateral Ureteral Obstruction ◽  
Anti Inflammatory

scholarly journals SAMP1/Sku as a Murine Model for Tubulointerstitial Nephritis: a Study Using Unilateral Ureteral Obstruction

2005 ◽  
Vol 54 (1) ◽  
pp. 53-60 ◽  
Author(s):  
Akira YABUKI ◽  
Michie MAEDA ◽  
Mitsuharu MATSUMOTO ◽  
Ryozo KAMIMURA ◽  
Taku MASUYAMA ◽  
...  
Keyword(s):  
Murine Model ◽  
Ureteral Obstruction ◽  
Tubulointerstitial Nephritis ◽  
Unilateral Ureteral Obstruction

Abstract 1727: Apolipoprotein E (ApoE) Induces an Anti-inflammatory Phenotype in Macrophages

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Daniel Baitsch ◽  
Ralph Telgmann ◽  
Georg Varga ◽  
Carsten Muller-Tidow ◽  
Martine Bot ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Apolipoprotein E ◽  
Kinase Inhibitor ◽  
Plasma Cholesterol ◽  
Macrophage Phenotype ◽  
Anti Inflammatory ◽  
M2 Phenotype ◽  
Functional Phenotype ◽  
Expression And Activity ◽  
Phenotype Expression

Apolipoprotein E (apoE) exerts anti-atherogenic effects by promoting cholesterol efflux and hepatic lipoprotein clearance. However, apoE retains protective effects even under experimental settings, in which its influence on plasma cholesterol is negligible suggesting that this lipoprotein inhibits atherosclerosis independently from cholesterol transport. To gain further insight into mechanisms underlying apoE-mediated atheroprotection, we investigated its effect on the functional phenotype of RAW 264.7 macrophages overexpressing either of two apoE receptors: ApoER2/LRP8 or VLDL-R. Incubation of ApoER2/LRP8- or VLDL-R-expressing macrophages with apoE downregulated markers of pro-inflammatory M1 functional phenotype (expression and activity of iNOS, production of IL-12), whereas markers of anti-inflammatory M2 phenotype (expression and activity of arginase-I, production of IL-1RA) were upregulated. In addition, macrophage responses typical for M1 phenotype (migration, generation of reactive oxygen species, antibody-dependent cell cytotoxicity) were suppressed in ApoER2/LRP8- or VLDL-R-expressing cells in the presence of apoE. Finally, apoE prevented LPS- and IFN-γ-induced activation of ApoER2/LRP8- or VLDL-R-expressing macrophages as documented by reduced production of IL-12, TNF-α and MCP-1, reduced expression and activity of iNOS and COX2, and reduced activation and/or phosphorylation of NF-κB, IκB and STAT1. The modulatory effects of apoE on macrophage phenotype were inhibited by SB220025, a p38MAP kinase inhibitor, and PP1A, a tyrosine kinase inhibitor. Accordingly, apoE induced tyrosine kinase-dependent activation of p38MAP kinase in ApoER2/LRP8- or VLDL-R-expressing macrophages. Under in vivo conditions, apoE −/− mice transplanted with apoE-producing wild-type bone marrow presented with increased plasma IL-1RA levels. In addition, peritoneal macrophages from transplanted animals demonstrated enhanced M2 phenotype (increased IL-1RA production and CD206 expression). We conclude that apoE signalling over ApoER2/LRP8 or VLDL-R promotes macrophage conversion from pro-inflammatory M1 to anti-inflammatory M2 phenotype. This effect may represent a novel anti-atherogenic activity of apoE.

Fate alteration of bone marrow-derived macrophages ameliorates kidney fibrosis in murine model of unilateral ureteral obstruction

2018 ◽  
Vol 34 (10) ◽  
pp. 1657-1668 ◽  
Author(s):  
Ying Yang ◽  
Xiaojian Feng ◽  
Xinyan Liu ◽  
Ying Wang ◽  
Min Hu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Ureteral Obstruction ◽  
Renal Fibrosis ◽  
Kidney Diseases ◽  
Unilateral Ureteral Obstruction ◽  
Immunofluorescent Staining ◽  
Enzyme Linked Immunosorbent Assay ◽  
Pathological Feature ◽  
Kidney Fibrosis ◽  
Anti Inflammatory

AbstractBackgroundRenal fibrosis is a key pathological feature and final common pathway leading to end-stage kidney failure in many chronic kidney diseases. Myofibroblast is the master player in renal fibrosis. However, myofibroblasts are heterogeneous. Recent studies show that bone marrow-derived macrophages transform into myofibroblasts by transforming growth factor (TGF)-β-induced macrophage–myofibroblast transition (MMT) in renal fibrosis.MethodsTGF-β signaling was redirected by inhibition of β-catenin/T-cell factor (TCF) to increase β-catenin/Foxo in bone marrow-derived macrophages. A kidney fibrosis model of unilateral ureteral obstruction was performed in EGFP bone marrow chimera mouse. MMT was examined by flow cytometry analysis of GFP+F4/80+α-SMA+ cells from unilateral ureteral obstruction (UUO) kidney, and by immunofluorescent staining of bone marrow-derived macrophages in vitro. Inflammatory and anti-inflammatory cytokines were analysis by enzyme-linked immunosorbent assay.ResultsInhibition of β-catenin/TCF by ICG-001 combined with TGF-β1 treatment increased β-catenin/Foxo1, reduced the MMT and inflammatory cytokine production by bone marrow-derived macrophages, and thereby, reduced kidney fibrosis in the UUO model.ConclusionsOur results demonstrate that diversion of β-catenin from TCF to Foxo1-mediated transcription not only inhibits the β-catenin/TCF-mediated fibrotic effect of TGF-β, but also enhances its anti-inflammatory action, allowing therapeutic use of TGF-β to reduce both inflammation and fibrosis at least partially by changing the fate of bone marrow-derived macrophages.

scholarly journals Probiotic Conditioned Media Induces an Anti-Inflammatory Macrophage Phenotype In Vivo and In Vitro

2011 ◽  
Vol 140 (5) ◽  
pp. S-19
Author(s):  
Michelle Taylor ◽  
Vandana Gambhir ◽  
Curtis Noordhof ◽  
Oliver Jones ◽  
Shu-Mei He ◽  
...  
Keyword(s):  
Conditioned Media ◽  
Macrophage Phenotype ◽  
Anti Inflammatory ◽  
Inflammatory Macrophage

scholarly journals DOP53 PTPN2 and TiO2 in the pathogenesis of inflammatory bowel disease

2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S092-S092
Author(s):  
J Conde ◽  
M Schwarzfischer ◽  
E Katkeviciute ◽  
J Häfliger ◽  
A Niechcial ◽  
...  
Keyword(s):  
Inflammatory Bowel Disease ◽  
Flow Cytometry ◽  
Titanium Dioxide ◽  
Bowel Disease ◽  
Myeloid Cells ◽  
Food Additive ◽  
Inflammasome Activation ◽  
Inflammatory Bowel ◽  
Anti Inflammatory

Abstract Background Inflammatory bowel disease (IBD) is caused by a complex interaction among genetic, immunological, bacterial and environmental factors. In this scenario, protein tyrosine phosphatase non-receptor type-2 (PTPN2) has been recognised as a risk factor for the development of IBD and functional studies revealed a major role for this protein in the development of experimental colitis through the regulation of the inflammasome, among other processes. In the same way, a potential relationship between diet components and IBD was suggested. In fact, it was reported that the food additive titanium dioxide (TiO2) was able to induce inflammasome activation in vitro and increase colitis severity in vivo. These observations led us to hypothesise a putative relationship between PTPN2 and TiO2 that could explain the effects of this microparticle in the pathogenesis of bowel inflammation. Methods DSS colitis model was performed in mice lacking PTPN2 in myeloid cells and their wild-type littermates, treated or not with titanium dioxide. After that, histology studies, flow cytometry, expression analysis, ELISA and barrier function experiments were performed. Also, bone marrow-derived macrophages (BMDMs) were used for in vitro studies. Results Titanium dioxide was able to exacerbate DSS-induced colitis, especially in mice lacking PTPN2 in myeloid cells. Flow cytometry analysis of the lamina propria revealed significant changes in different immune cell populations such as macrophages. In vitro experiments using BMDMs revealed that TiO2 induced the activation of the inflammasome. Also, this microparticle down-regulated the expression of the anti-inflammatory cytokine IL-10 and these effects were mainly mediated by JNK and ERK kinases. Conclusions The food additive titanium dioxide seems to play a negative role in colitis development by affecting the production of pro- and anti-inflammatory mediators in macrophages. This study reveals a new mechanism by which a certain component of the diet modulates intestinal inflammation.

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