-
Redox Biology Aug 2023Irisin is a newly discovered myokine which links exercise to inflammation and inflammation-related diseases through macrophage regulation. However, the effect of irisin...
INTRODUCTION
Irisin is a newly discovered myokine which links exercise to inflammation and inflammation-related diseases through macrophage regulation. However, the effect of irisin on the activity of inflammation related immune cells (such as neutrophils) has not been clearly described.
OBJECTIVES
The objective of our study was to explore the effect of irisin on the neutrophil extracellular traps (NETs) formation.
METHODS
Phorbol-12-myristate-13-acetate (PMA) was used to construct a classic neutrophil inflammation model that was used to observe the formation of NETs in vitro. We studied the effect of irisin on NETs formation and its regulation mechanism. Subsequently, acute pancreatitis (AP) was used to verify the protective effect of irisin in vivo, which was an acute aseptic inflammatory response disease model closely related to NETs.
RESULTS
Our study found that addition of irisin significantly reduced the formation of NETs via regulation of the P38/MAPK pathway through integrin αVβ5, which might be the one of key pathways in NETs formation, and which could theoretically offset the immunoregulatory effect of irisin. Systemic treatment with irisin reduced the severity of tissue damage common in the disease and inhibited the formation of NETs in pancreatic necrotic tissue of two classical AP mouse models.
CONCLUSION
The findings confirmed for the first time that irisin could inhibit NETs formation and protect mice from pancreatic injury, which further elucidated the protective effect of exercise on acute inflammatory injury.
Topics: Mice; Animals; Extracellular Traps; Pancreatitis; Fibronectins; Acute Disease; Neutrophils; Inflammation; Tetradecanoylphorbol Acetate
PubMed: 37392517
DOI: 10.1016/j.redox.2023.102787 -
Molecules and Cells Mar 2016The mechanism by which 12-O-tetradecanoylphorbol-13-acetate (TPA) bypasses cellular senescence was investigated using human diploid fibroblast (HDF) cell replicative...
The mechanism by which 12-O-tetradecanoylphorbol-13-acetate (TPA) bypasses cellular senescence was investigated using human diploid fibroblast (HDF) cell replicative senescence as a model. Upon TPA treatment, protein kinase C (PKC) α and PKCβ1 exerted differential effects on the nuclear translocation of cytoplasmic pErk1/2, a protein which maintains senescence. PKCα accompanied pErk1/2 to the nucleus after freeing it from PEA-15pS(104) via PKCβ1 and then was rapidly ubiquitinated and degraded within the nucleus. Mitogen-activated protein kinase docking motif and kinase activity of PKCα were both required for pErk1/2 transport to the nucleus. Repetitive exposure of mouse skin to TPA downregulated PKCα expression and increased epidermal and hair follicle cell proliferation. Thus, PKCα downregulation is accompanied by in vivo cell proliferation, as evidenced in 7, 12-dimethylbenz(a)anthracene (DMBA)-TPA-mediated carcinogenesis. The ability of TPA to reverse senescence was further demonstrated in old HDF cells using RNA-sequencing analyses in which TPA-induced nuclear PKCα degradation freed nuclear pErk1/2 to induce cell proliferation and facilitated the recovery of mitochondrial energy metabolism. Our data indicate that TPA-induced senescence reversal and carcinogenesis promotion share the same molecular pathway. Loss of PKCα expression following TPA treatment reduces pErk1/2-activated SP1 biding to the p21(WAF1) gene promoter, thus preventing senescence onset and overcoming G1/S cell cycle arrest in senescent cells.
Topics: Animals; Cell Proliferation; Cellular Senescence; Fibroblasts; Gene Expression Regulation; Humans; MAP Kinase Signaling System; Mice; Phosphorylation; Protein Kinase C beta; Protein Kinase C-alpha; Protein Transport; Tetradecanoylphorbol Acetate
PubMed: 26912086
DOI: 10.14348/molcells.2016.2362 -
Frontiers in Immunology 2023The infusion of -generated regulatory B cells may represent a promising novel therapeutic approach for a variety of autoimmune and hyperinflammatory conditions including...
INTRODUCTION
The infusion of -generated regulatory B cells may represent a promising novel therapeutic approach for a variety of autoimmune and hyperinflammatory conditions including graft-versus-host disease.
METHODS
Previously, we developed a protocol for the generation of a novel population of regulatory B cells, which are characterized by secretion of enzymatically active granzyme B (). This protocol uses recombinant interleukin 21 (IL-21) and goat-derived F(ab)'2 fragments against the human B cell receptor (anti-BCR). Generally, the use of xenogeneic material for the manufacturing of advanced therapy medicinal products should be avoided to prevent adverse immune reactions as well as potential transmission of so far unknown diseases.
RESULTS
In the present work we demonstrated that phorbol-12-myristate-13-acetate (PMA/TPA), a phorbol ester with a particular analogy to the second messenger diacylglycerol (DAG), is a potent enhancer of IL-21-induced differentiation of pre-activated B cells into . The percentage of after stimulation of pre-activated B cells with IL-21 and PMA/TPA was not significantly lower compared to stimulation with IL-21 and anti-BCR.
DISCUSSION
Given that PMA/TPA has already undergone encouraging clinical testing in patients with certain haematological diseases, our results suggest that PMA/TPA may be a safe and feasible alternative for manufacturing of .
Topics: Humans; B-Lymphocytes, Regulatory; Granzymes; Tetradecanoylphorbol Acetate
PubMed: 37588597
DOI: 10.3389/fimmu.2023.1194880 -
Methods in Molecular Biology (Clifton,... 1998
Topics: Acridines; Buffers; Cell Separation; Humans; In Vitro Techniques; Luminescent Measurements; Luminol; Phagocytes; Photometry; Respiratory Burst; Tetradecanoylphorbol Acetate
PubMed: 9680620
DOI: 10.1385/0-89603-520-4:179 -
Carcinogenesis Jun 1988Dehydroepiandrosterone, a naturally occurring adrenal steroid, is a highly effective tumor chemopreventive agent in laboratory mice and rats, inhibiting spontaneous...
Dehydroepiandrosterone, a naturally occurring adrenal steroid, is a highly effective tumor chemopreventive agent in laboratory mice and rats, inhibiting spontaneous breast cancer and chemically induced tumors of the lung, colon, skin, liver and thyroid. Dehydroepiandrosterone blocks three processes that have been implicated in experimental tumorigenesis: (i) carcinogen activation through the mixed-function oxidases, (ii) 12-O-tetradecanoylphorbol-13-acetate stimulation of superoxide anion production in neutrophils, and (iii) 12-O-tetradecanoylphorbol-13-acetate stimulation of [3H]thymidine incorporation in mouse epidermis. All of these effects of dehydroepiandrosterone very likely result from glucose-6-phosphate dehydrogenase inhibition and a lowering of the NADPH cellular pool. It is now reported that oral administration of dehydroepiandrosterone (0.2% in the diet) for two weeks inhibits the stimulation in prostaglandin E2 content in mouse epidermis produced by topical application of 12-O-tetradecanoylphorbol-13-acetate. Two synthetic steroids, 16 alpha-fluoro-5-androsten-17-one and 16 alpha-fluoro-5 alpha-androstan-17-one, which are more potent inhibitors of the above three processes in tumorigenesis and are also more effective than dehydroepiandrosterone in inhibiting skin papilloma development in the mouse, are more active in suppressing prostaglandin E2 induction by 12-O-tetradecanoyl-phorbol-13-acetate. These two structural analogs, which also lack specific side-effects associated with dehydroepiandrosterone treatment, may find application as cancer chemopreventive drugs in humans.
Topics: Animals; Dehydroepiandrosterone; Dinoprostone; Mice; Prostaglandins E; Skin; Structure-Activity Relationship; Tetradecanoylphorbol Acetate
PubMed: 2967126
DOI: 10.1093/carcin/9.6.1099 -
Nature Feb 1991
Topics: Animals; Cell Line; Epidermal Growth Factor; Mice; Protein Synthesis Inhibitors; Proto-Oncogenes; Signal Transduction; Tetradecanoylphorbol Acetate
PubMed: 2000146
DOI: 10.1038/349747c0 -
Folia Histochemica Et Cytobiologica 2013TPA (12-O-tetradecanoylphorbol-1, 3-acetate) can induce cell apoptosis and cause PKB (protein kinase B) degradation correlated with its phosphorylation in gastric cancer...
12-O-tetradecanoylphorbol-1,3-acetate-induced degradation of protein kinase B via ubiquitin-proteasomal pathway depends on its Ser473 phosphorylation in gastric cancer cells.
TPA (12-O-tetradecanoylphorbol-1, 3-acetate) can induce cell apoptosis and cause PKB (protein kinase B) degradation correlated with its phosphorylation in gastric cancer cells. We investigated whether the ubiquitin-proteasomal pathway is involved in TPA-induced PKB degradation. The results showed that TPA could induce PKB ubiquitination by inhibiting its phosphorylation at the serine 473 site. Moreover, MG132 (26S proteasome inhibitor) partially inhibited TPA-induced degradation of PKB. Taken together, TPA could degrade PKB via the ubiquitin-proteasomal pathway, and the suppression of PKB phosphorylation at the serine 473 site might be a prerequisite for the TPA-induced ubiquitination in gastric cancer cells.
Topics: Cell Line, Tumor; HEK293 Cells; Humans; Phosphorylation; Phosphoserine; Proteasome Endopeptidase Complex; Proteolysis; Proto-Oncogene Proteins c-akt; Signal Transduction; Stomach Neoplasms; Tetradecanoylphorbol Acetate; Ubiquitin
PubMed: 23690212
DOI: 10.5603/FHC.2013.002 -
International Journal of Molecular... Nov 2022Neutrophil extracellular traps (NETs) are found in patients with various diseases, including cardiovascular diseases. We previously reported that copper-oxidized...
Neutrophil extracellular traps (NETs) are found in patients with various diseases, including cardiovascular diseases. We previously reported that copper-oxidized low-density lipoprotein (oxLDL) promotes NET formation of neutrophils, and that the resulting NETs increase the inflammatory responses of endothelial cells. In this study, we investigated the effects of high-density lipoproteins (HDL) on NET formation. HL-60-derived neutrophils were treated with phorbol 12-myristate 13-acetate (PMA) and further incubated with oxLDL and various concentrations of HDL for 2 h. NET formation was evaluated by quantifying extracellular DNA and myeloperoxidase. We found that the addition of native HDL partially decreased NET formation of neutrophils induced by oxLDL. This effect of HDL was lost when HDL was oxidized. We showed that oxidized phosphatidylcholines and lysophosphatidylcholine, which are generated in oxLDL, promoted NET formation of PMA-primed neutrophils, and NET formation by these products was completely blocked by native HDL. Furthermore, we found that an electronegative subfraction of LDL, LDL(-), which is separated from human plasma and is thought to be an in vivo oxLDL, was capable of promoting NET formation. These results suggest that plasma lipoproteins and their oxidative modifications play multiple roles in promoting NET formation, and that HDL acts as a suppressor of this response.
Topics: Humans; Lipoproteins, HDL; Extracellular Traps; Phospholipids; Endothelial Cells; Lipoproteins, LDL; Tetradecanoylphorbol Acetate
PubMed: 36430470
DOI: 10.3390/ijms232213992 -
Veterinary Pathology Sep 2013Assessment of the skin tumor-promoting potential of 12-O-tetradecanoylphorbol-13-acetate (TPA) after initiation with 7,12-dimethylbenz[a]anthracene (DMBA) was conducted... (Comparative Study)
Comparative Study
Assessment of the skin tumor-promoting potential of 12-O-tetradecanoylphorbol-13-acetate (TPA) after initiation with 7,12-dimethylbenz[a]anthracene (DMBA) was conducted using rasH2 transgenic (Tg) mice and their nontransgenic (non-Tg) littermates. Mice were treated with DMBA (50 μg/100 μL acetone) on clipped back skin at the commencement of the study, and 1 week thereafter, TPA was applied at 8 μg/200 μL or 4 μg/200 μL acetone, once or twice weekly, for 7 weeks. Skin nodules were observed in the rasH2 Tg mice from week 4, and the incidence reached 100% at weeks 5 and 6. The number of skin nodules (multiplicity) in the 8-μg twice-weekly, 8-μg once-weekly, 4-μg twice-weekly, and 4-μg once-weekly groups was 62.4, 46.2, 62.6, and 36.9, respectively. The non-Tg mice also developed skin nodules, but the sensitivity to induction in the rasH2 Tg mice was higher. No nodules were observed in the acetone groups, but single nodules were apparent in the no-treatment rasH2 Tg and non-Tg groups. In conclusion, skin promotion effects could be detected within only 8 weeks in the rasH2 mice, and the concentration of 4 μg TPA once weekly was sufficient as a positive control. This short-term skin carcinogenesis bioassay using rasH2 mice could represent a useful tool for the assessment of drug and chemical safety with cutaneous treatment.
Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Biological Assay; Carcinogenesis; Dose-Response Relationship, Drug; Female; Humans; Mice; Mice, Transgenic; Skin Neoplasms; Tetradecanoylphorbol Acetate
PubMed: 23610217
DOI: 10.1177/0300985813486811 -
IUBMB Life Nov 2008
Review
Topics: Canada; Catalysis; Enzyme Activation; History, 20th Century; History, 21st Century; Models, Biological; Phosphorylation; Protein Binding; Protein Conformation; Protein Kinase C; Tetradecanoylphorbol Acetate
PubMed: 18683875
DOI: 10.1002/iub.118