Senolytic effects of quercetin in an in vitro model of pre-adipocytes and adipocytes induced senescence

AbstractThe dysfunction of adipose tissue with aging and the accumulation of senescent cells has been implicated in the pathophysiology of chronic diseases. Recently interventions capable of reducing the burden of senescent cells and in particular the identification of a new class of drugs termed senolytics have been object of extensive investigation. We used an in vitro model of induced senescence by treating both pre-adipocytes as well as mature adipocytes with hydrogen peroxide (H2O2) at a sub-lethal concentration for 3 h for three consecutive days, and hereafter with 20 uM quercetin at a dose that in preliminary experiments resulted to be senolytic without cytotoxicity. H2O2 treated pre-adipocytes and adipocytes showed typical senescence-associated features including increased beta-galactosidase activity (SA-ß-gal) and p21, activation of ROS and increased expression of pro-inflammatory cytokines. The treatment with quercetin in senescent pre-adipocytes and adipocytes was associated to a significant decrease in the number of the SA-β-gal positive cells along with the suppression of ROS and of inflammatory cytokines. Besides, quercetin treatment decreased miR-155-5p expression in both models, with down-regulation of p65 and a trend toward an up-regulation of SIRT-1 in complete cell extracts. The senolytic compound quercetin could affect AT ageing by reducing senescence, induced in our in vitro model by oxidative stress. The downregulation of miRNA-155-5p, possibly through the modulation of NF-κB and SIRT-1, could have a key role in the effects of quercetin on both pre-adipocytes and adipocytes.

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Effect of mesenchymal stem cells combined with chondroitin sulfate in an osteoarthritis in vitro model

10.21203/rs.3.rs-23610/v2 ◽

2020 ◽

Author(s):

Saúl Pérez-Castrillo ◽

María Luisa González-Fernández ◽

Jessica Álvarez-Suárez ◽

Jaime Sánchez-Lázaro ◽

Marta Esteban-Blanco ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Chondroitin Sulfate ◽

Inflammatory Cytokines ◽

In Vitro Model ◽

Promising Alternative ◽

Cell Based Therapy ◽

Vitro Model ◽

Pro Inflammatory Cytokines

Abstract Introduction: Osteoarthritis (OA) is a degenerative joint disease which affects the whole joint structure. Many authors have focused on the factors responsible for the development of inflammatory processes involved in OA. Adipose tissue-derived mesenchymal stem cells (ASCs) represent a promising alternative of cell-based therapy strategy in the treatment of OA which could be combined with any other drug. Chondroitin sulfate plays a protective role in the joint based on the decrease of pro-inflammatory cytokines, thus having an important role in activating and inhibiting the metabolic pathways in chondrocytes. Aims: The effectiveness of chondroitin sulfate and ASCs combined in an in vitro model of OA has been evaluated in this study. Materials: Cytokines and factors which are involved in OA as well as specific cartilage gene expression after adding ASCs and chondroitin sulfate have been discussed in detail. Results: Our results show a decrease in the expression of all genes related to the pro-inflammatory cytokines analysed. Although there was no increase in the expression of the specific genes of the cartilage matrix, such as collagen type II and aggrecan. Conclusions: This study shows the effectiveness of association of ASCs and chondroitin sulfate for the treatment of OA.

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Mesangial cells are key contributors to the fibrotic damage seen in the lupus nephritis glomerulus

Journal of Inflammation ◽

10.1186/s12950-019-0227-x ◽

2019 ◽

Vol 16 (1) ◽

Cited By ~ 2

Author(s):

Rachael D. Wright ◽

Paraskevi Dimou ◽

Sarah J. Northey ◽

Michael W. Beresford

Keyword(s):

Lupus Nephritis ◽

Inflammatory Cytokines ◽

Lupus Erythematosus ◽

In Vitro Model ◽

Mesangial Cells ◽

Vitro Model ◽

Tgf Β1 ◽

Onset Systemic Lupus Erythematosus ◽

Pro Inflammatory Cytokines

Abstract Background Lupus nephritis (LN) affects up to 80% of juvenile-onset systemic lupus erythematosus patients. Mesangial cells (MCs) comprise a third of the glomerular cells and are key contributors to fibrotic changes within the kidney. This project aims to identify the roles of MCs in an in vitro model of LN. Methods Conditionally immortalised MCs were treated with pro-inflammatory cytokines or with patient sera in an in vitro model of LN and assessed for their roles in inflammation and fibrosis. Results MCs were shown to produce pro-inflammatory cytokines in response to a model of the inflammatory environment in LN. Further the cells expressed increased levels of mRNA for extracellular matrix (ECM) proteins (COL1A1, COL1A2, COL4A1 and LAMB1), matrix metalloproteinase enzymes (MMP9) and tissue inhibitors of matrix metalloproteinases (TIMP1). Treatment of MCs with serum from patients with active LN was able to induce a similar, albeit milder phenotype. Treatment of MCs with cytokines or patient sera was able to induce secretion of TGF-β1, a known inducer of fibrotic changes. Inhibition of TGF-β1 actions through SB-431542 (an activin A receptor type II-like kinase (ALK5) inhibitor) was able to reduce these responses suggesting that the release of TGF-β1 plays a role in these changes. Conclusions MCs contribute to the inflammatory environment in LN by producing cytokines involved in leukocyte recruitment, activation and maturation. Further the cells remodel the ECM via protein deposition and enzymatic degradation. This occurs through the actions of TGF-β1 on its receptor, ALK5. This may represent a potential therapeutic target for treatment of LN-associated fibrosis.

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Endotoxin But Not Hypoxia Increases The Expression Of Pro-inflammatory Cytokines From Alveolar Macrophages, ATII, And Endothelial Cells In An In-vitro Model Of The Alveolar Barrier

10.1164/ajrccm-conference.2010.181.1_meetingabstracts.a1294 ◽

2010 ◽

Author(s):

Emel Baloglu ◽

Nuray Yazihan ◽

Peter Bartsch ◽

Heimo Mairbaurl

Keyword(s):

Endothelial Cells ◽

Inflammatory Cytokines ◽

Alveolar Macrophages ◽

In Vitro Model ◽

Vitro Model ◽

Pro Inflammatory Cytokines

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Alliin diminished pro-inflammatory cytokines in supernatants of an in vitro model of adipose tissue.

Frontiers in Immunology ◽

10.3389/conf.fimmu.2013.02.01054 ◽

2013 ◽

Vol 4 ◽

Author(s):

Lopez-Roa Rocio ◽

Quintero-Fabian Saray ◽

Viveros-Paredes Juan ◽

Garcia-Iglesias Trinidad ◽

Ortu�o-Sahagun Daniel

Keyword(s):

Adipose Tissue ◽

Inflammatory Cytokines ◽

In Vitro Model ◽

Vitro Model ◽

Pro Inflammatory Cytokines

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Peptidergic properties expressed in vitro by embryonic neuroblasts after neural induction

Development ◽

10.1242/dev.105.3.529 ◽

1989 ◽

Vol 105 (3) ◽

pp. 529-540

Author(s):

F. Pituello ◽

P. Deruntz ◽

L. Pradayrol ◽

A.M. Duprat

Keyword(s):

Culture Medium ◽

In Vitro Model ◽

Neural Induction ◽

Defined Medium ◽

Neuronal Precursor Cells ◽

Cell Extracts ◽

Neuronal Precursors ◽

Immunocytochemical Techniques ◽

Vitro Model

As an immediate consequence of neural induction, some neuroectodermal cells acquire the ability to develop a number of characteristic neuronal features, without requiring any subsequent embryonic cues (Duprat et al. 1987). Thus, adrenergic, cholinergic and gabaergic traits are expressed in cultures of neural fold and neural plate isolated from amphibian embryos immediately after induction and grown in a defined medium. The aim of the present study was to determine, using the same in vitro model, their abilities to develop peptidergic phenotypes. Using immunocytochemical techniques, we show that substance P-, enkephalin- (leu-enkephalin, metenkephalin), and somatostatin- like immunoreactivities are expressed in subpopulations of neurones grown in vitro, whereas VIP (vasoactive intestinal polypeptide) is not detected under the same conditions. The appearance and development of the somatostatinergic phenotype has been quantified by RIA both in cell extracts and in the culture medium. Somatostatin-like immunoreactivity (SLI) undetectable at the late gastrula stage, can be measured in cells after 4 days of culture and continues to increase over the next 10 days. In culture medium, SLI is present at a constant level from day 4 up to day 14. These data reveal that some neuronal precursor cells acquire, during neural induction, the potentiality to biosynthesize, store and release neuropeptides. Furthermore, the expression of these peptidergic phenotypes in distinct subpopulations of neurones suggests that certain neuronal precursors become committed to different metabolic pathways at the earliest steps of neurogenesis.

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An in vitro model for investigation of vitamin A effects on wound healing

International Journal for Vitamin and Nutrition Research ◽

10.1024/0300-9831/a000692 ◽

2021 ◽

pp. 1-6

Author(s):

Hoda Keshmiri Neghab ◽

Mohammad Hasan Soheilifar ◽

Gholamreza Esmaeeli Djavid

Keyword(s):

Wound Healing ◽

Endothelial Cell ◽

Vitamin A ◽

In Vitro Model ◽

Cellular Proliferation ◽

Endothelial Cell Migration ◽

Normal Fibroblast ◽

Anti Inflammatory ◽

Vitro Model

Abstract. Wound healing consists of a series of highly orderly overlapping processes characterized by hemostasis, inflammation, proliferation, and remodeling. Prolongation or interruption in each phase can lead to delayed wound healing or a non-healing chronic wound. Vitamin A is a crucial nutrient that is most beneficial for the health of the skin. The present study was undertaken to determine the effect of vitamin A on regeneration, angiogenesis, and inflammation characteristics in an in vitro model system during wound healing. For this purpose, mouse skin normal fibroblast (L929), human umbilical vein endothelial cell (HUVEC), and monocyte/macrophage-like cell line (RAW 264.7) were considered to evaluate proliferation, angiogenesis, and anti-inﬂammatory responses, respectively. Vitamin A (0.1–5 μM) increased cellular proliferation of L929 and HUVEC (p < 0.05). Similarly, it stimulated angiogenesis by promoting endothelial cell migration up to approximately 4 fold and interestingly tube formation up to 8.5 fold (p < 0.01). Furthermore, vitamin A treatment was shown to decrease the level of nitric oxide production in a dose-dependent effect (p < 0.05), exhibiting the anti-inflammatory property of vitamin A in accelerating wound healing. These results may reveal the therapeutic potential of vitamin A in diabetic wound healing by stimulating regeneration, angiogenesis, and anti-inﬂammation responses.

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