Ethanol induces IL-6 and TNF-α cytokine and iNOS and COX-2 enzyme gene expression in 3T3-L1 preadipocytes

Kiselova-Kaneva, Yoana; Tasinov, Oskan; Vankova, Deyana; Ivanova, Diana

doi:10.14748/ssm.v44i2.354

Cited by 7 publications

(4 citation statements)

References 19 publications

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“…Viability of treated cells was evaluated using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay 8 with some modifications. 9 10 Cells were seeded in 2 × 10 5 cells/well in 6-well plates with DMEM (with 4.5 g/L glucose, with phenol red and L-glutamine) (Lonza, Belgium) supplemented with 10% FBS, 100 U/mL penicillin/100 μg/mL streptomycin mixture. After 24 hours, culture medium was replaced and cells were treated with DMEM (with 4.5 g/L glucose, without phenol red and L-glutamine) (Lonza, Belgium) containing FP D12 or PT (0.00625 mg/mL, 0.0125 mg/mL, 0.025 mg/mL, 0.25 mg/mL, 2.5 mg/mL, 5 mg/mL).…”

Section: Methodsmentioning

confidence: 99%

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Ferrum phosphoricum D12 Treatment Affects J774A.1 Cell Proliferation, Transcription Levels of Iron Metabolism, Antioxidant Defense, and Inflammation-related Genes

et al. 2021

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Background Ferrum phosphoricum (FP) is prescribed as a homeopathic remedy to treat the early stages of fever and inflammation in cases of colds or flu, muscle fatigue and anemia. We aimed to analyze the molecular mechanisms of action of FP D12 on cell proliferation and mRNA expression of iron metabolism, antioxidant defense and inflammation-related genes in mouse J774A.1 macrophages. Methods Cell proliferation was examined using the MTT test. RT-qPCR analyses were performed to estimate gene expression changes. Relative gene expression levels were calculated using the 2–ΔΔCt method. The effect of treatment using FP D12 tablets was compared with that using placebo tablets (PT). Results FP D12 in low concentrations (0.0125 mg/mL to 0.025 mg/mL) significantly stimulated proliferation of J774A.1 cells by up to 11% (p < 0.01) versus control untreated cells and by up to 40% (p < 0.01) versus PT-treated cells in the respective concentration. FP D12 versus PT induced a significant increase in mRNA expression of ferritin light chain (Ftl1) (by 8-fold, p < 0.01), β-2-microglobulin (B2m) (by 2.5-fold, p < 0.05) and iron-responsive element binding protein 2 (Ireb2) (by 4-fold, p < 0.05), and induced a slight decrease in myosin IE (Myo1e) mRNA expression levels (by 0.4-fold, p < 0.01) in macrophages. A highly significant (r2 = 0.99, p < 0.05) correlation was observed between Ireb2 and B2m transcription levels. Significant stimulation of antioxidant enzyme Gpx-1 (by 1.27-fold, p < 0.01) in cells by 0.025 mg/mL FP D12, but a slight decrease (by 0.12-fold, p < 0.05) in 0.0125 mg/mL-treated cells, was observed. A significant increase in the gene expression of IL-1β (by 3.5-fold, р < 0.05) in macrophages was also detected. Conclusion Ferrum phosphoricum in D12 dilution potentially exhibits iron retention, antioxidant and immunomodulation activities, possibly by modulating transcription levels of related genes in non-stimulated mouse macrophages.

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Section: Methodsmentioning

confidence: 99%

“…Gene expression was analyzed using two step RT-qPCR as previously described. 10 Relative gene expression levels were calculated using the 2 -DDCt method. 11 The primer sequences (Sigma-Aldrich, Germany) used for each analyzed gene are presented in ►Table 1.…”

Section: Gene Expression Analysismentioning

confidence: 99%

Ferrum phosphoricum D12 Treatment Affects J774A.1 Cell Proliferation, Transcription Levels of Iron Metabolism, Antioxidant Defense, and Inflammation-related Genes

et al. 2021

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“…The concentration of EtOH used was determined based on an extensive literature survey (Kiselova‐Kaneva et al., ; Liu et al., ).…”

Section: Methodsmentioning

confidence: 99%

Protective Effects of Diallyl Sulfide Against Ethanol-Induced Injury in Rat Adipose Tissue and Primary Human Adipocytes

Kema

Khan

Jamal

et al. 2017

Alcohol Clin Exp Res

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Our study results prove that DAS is effective in ameliorating EtOH-induced damage to adipose tissue as evidenced by the reduction brought about by DAS in oxidative stress, ER stress, and proinflammatory gene expression levels. DAS treatment also regulated lipogenic gene expression levels, thereby reducing free fatty acid release. In conclusion, this study has clinical implications with respect to alcohol-induced adipose tissue injury among alcohol users.

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“…Quantitative gene expression analysis was performed using two-step real-time qPCR as previously described (4). Primer sequences were as follows: mouse β-actin forward 5`-ACG GCC AGG TCA TCA CTA TTG-3`, reverse CAA GAA GGA AGG CTG GAA AAG; mouse COX-2 forward 5`-TGA GCA ACT ATT CCA AAC CAG C-3`, reverse 5`-GCA CGT AGT CTT CGA TCA CTA TC-3`; mouse iNOS forward 5`-GGC AGC CTG TGA GAC CTT TG-3`, reverse 5`-GCA TTG GAA GTG AAG CGT TTC-3`; mouse GCL catalytic subunit forward 5`-AAT GGA GGC GAT GTT CTT GAG-3`, reverse 5`-CAG AGG GTC GGA TGG TTG G-3`.…”

Section: Gene Expression Analysismentioning

confidence: 99%

Changes in COX-2, iNOS and GCL gene expression in 3T3-L1 preadipocytes incubated in macrophage conditioned medium

Kiselova-Kaneva

Tasinov

Vankova

et al. 2013

SSM

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PURPOSE: Obesity is recognized as a major risk factor for a number of diseases, including metabolic syndrome and type 2 diabetes mellitus. It is characterized with increased macrophage infiltration in adipose tissue. Macrophages secrete numerous inflammatory mediators and contribute to the increased inflammatory status of adipose tissue which is associated with increased reactive oxygen species generation and oxidative stress. In cell biology conditioned nutrient mediums (CMs) are used to assess the complex impact of the secretory products of a particular cell type on the behaviour of other cells. The aim of the present study was to examine the effect of CM obtained from J744A.1 macrophage cell culture on expression in 3T3-L1 preadipocytes of some enzymes related to antioxidant defense and the inflammatory response. MATERIAL AND METHODS: mRNA levels of inducible cyclooxygenase (COX-2), inducible NO synthase (iNOS) and glutamate-cysteine-ligase (GCL) were measured. 3T3-L1 cells were treated with increasing CM concentrations (10-30%) for 24 hours. Changes in gene expression levels were analyzed by real-time qPCR method. Calculations were performed using 2-ΔΔCt method. RESULTS: The higher CM concentration contributed to increased expression levels of the genes examined. Treatment with CM in concentrations of 10, 20 and 30% resulted in 2.3-, 2.6-and 2.7-fold increase in COX-2 expression (p<0,01), respectively. Both 10% and 20% CM up-regulated iNOS by 1,7 times (p<0,001). The highest CM concentration of 30% stimulated the enzyme by two times (p<0,01). GCL mRNA levels were by 2,5 times higher than these of the untreated controls when stimulated with 30% CM (p<0,05). CONCLUSION: CM from J744A.1 cell culture may induce inflammatory and oxidative stress-related response in 3T3-L1 preadipocytes thus demonstrating a possible link between macrophage infiltration and local inflammation in adipose tissue.

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Ethanol induces IL-6 and TNF-α cytokine and iNOS and COX-2 enzyme gene expression in 3T3-L1 preadipocytes

Cited by 7 publications

References 19 publications

Ferrum phosphoricum D12 Treatment Affects J774A.1 Cell Proliferation, Transcription Levels of Iron Metabolism, Antioxidant Defense, and Inflammation-related Genes

Ferrum phosphoricum D12 Treatment Affects J774A.1 Cell Proliferation, Transcription Levels of Iron Metabolism, Antioxidant Defense, and Inflammation-related Genes

Protective Effects of Diallyl Sulfide Against Ethanol-Induced Injury in Rat Adipose Tissue and Primary Human Adipocytes

Changes in COX-2, iNOS and GCL gene expression in 3T3-L1 preadipocytes incubated in macrophage conditioned medium

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