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Quercetin inhibits IL-1β-induced inflammation, hyaluronan production and adipogenesis in orbital fibroblasts from Graves' orbitopathy.

Yoon JS, Lee HJ, Choi SH, Chang EJ, Lee SY, Lee EJ - PLoS ONE (2011)

Bottom Line: Management of Graves' orbitopathy (GO) is challenging, as no reliable, specific, and safe medical therapeutic agents have yet been developed.Treatment with noncytotoxic doses of quercetin inhibited accumulation of intracytoplasmic lipid droplets and resulted in a dose-dependent decrease in expression of peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein (C/EBP) α, and C/EBPβ proteins.In conclusion, inhibition of inflammation, hyaluronan production, and adipogenesis by the natural plant product quercetin in vitro provides the basis for further study of its potential use in the treatment of GO.

View Article: PubMed Central - PubMed

Affiliation: Institute of Vision Research, Department of Ophthalmology, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT
Management of Graves' orbitopathy (GO) is challenging, as no reliable, specific, and safe medical therapeutic agents have yet been developed. We investigated the effect of quercetin in primary cultured orbital fibroblasts from GO, targeting pathways of inflammation, aberrant accumulation of extracellular matrix macromolecules, and adipose tissue expansion. Quercetin significantly attenuated intercellular adhesion molecule-1 (ICAM-1), interleukin (IL) -6, IL-8, and cyclooxygenase (COX) -2 mRNA expression, and inhibited IL-1β-induced increases in ICAM-1, IL-6, and IL-8 mRNA. Increased hyaluronan production induced by IL-1β or tumor necrosis factor-α was suppressed by quercetin in a dose- and time-dependent manner. Treatment with noncytotoxic doses of quercetin inhibited accumulation of intracytoplasmic lipid droplets and resulted in a dose-dependent decrease in expression of peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein (C/EBP) α, and C/EBPβ proteins. In conclusion, inhibition of inflammation, hyaluronan production, and adipogenesis by the natural plant product quercetin in vitro provides the basis for further study of its potential use in the treatment of GO.

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Effect of quercetin on cell viability and apoptosis in preadipocyte orbital fibroblasts and differentiating orbital fibroblasts.(A) Orbital fibroblasts (1×105) of normal and Graves' orbitopathy (GO) patients were seeded into 24-well culture plates and treated with different concentrations of quercetin (10, 30, 50, or 100 µM) for 24 h. After treatment, assays with 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) were performed to test for viability. (B) An annexin V/FITC kit was used to detect phosphatidylserine externalization, as an index of apoptosis. Percentage of stained cells with annexin V was analyzed by flow cytometry. (C) Orbital fibroblasts (1×105) of GO patients were seeded into 24-well culture plates and treated with different concentrations of quercetin (10, 50, 100, or 200 µM) for 3 days in adipogenic medium containing adipogenesis inducers and rosiglitazone (10 µM). After treatment, MTT assays were performed. Results are expressed as percentage of untreated control values presented as mean ± standard deviation (SD). Assays were performed at least three times in triplicate; data from a representative experiment are shown, expressed as the differences between treated and untreated cells.
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pone-0026261-g001: Effect of quercetin on cell viability and apoptosis in preadipocyte orbital fibroblasts and differentiating orbital fibroblasts.(A) Orbital fibroblasts (1×105) of normal and Graves' orbitopathy (GO) patients were seeded into 24-well culture plates and treated with different concentrations of quercetin (10, 30, 50, or 100 µM) for 24 h. After treatment, assays with 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) were performed to test for viability. (B) An annexin V/FITC kit was used to detect phosphatidylserine externalization, as an index of apoptosis. Percentage of stained cells with annexin V was analyzed by flow cytometry. (C) Orbital fibroblasts (1×105) of GO patients were seeded into 24-well culture plates and treated with different concentrations of quercetin (10, 50, 100, or 200 µM) for 3 days in adipogenic medium containing adipogenesis inducers and rosiglitazone (10 µM). After treatment, MTT assays were performed. Results are expressed as percentage of untreated control values presented as mean ± standard deviation (SD). Assays were performed at least three times in triplicate; data from a representative experiment are shown, expressed as the differences between treated and untreated cells.

Mentions: To determine nontoxic, concentrations of quercetin in orbital fibroblasts, 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and annexin V-fluorescence isothiocyanate (FITC) apoptosis assay were performed. Exposure of cells to quercetin at ≤100 µM for 24 h neither decreased cell viability below 95% in both normal and GO orbital fibroblasts (Fig. 1A) nor induced a significant level of apoptosis (less than 7%; Fig. 1B). Therefore, treatment of cells with 100 µM quercetin for 24 h was used to determine effects on inflammation and hyaluronan production. For experiments testing suppression of adipogenesis, cells in adipogenic medium were treated with quercetin (10–200 µM) from days 1–3 of differentiation. Cell viability at day three in the presence of 100 µM quercetin was not significantly reduced compared to untreated cells, whereas 200 µM attenuated cell viability to 82.3% (Fig. 1C). Therefore noncytotoxic concentrations (50, 100 µM) of quercetin were used for 3 days in differentiating cells cultures to find the effect of quercetin on adipogenesis.


Quercetin inhibits IL-1β-induced inflammation, hyaluronan production and adipogenesis in orbital fibroblasts from Graves' orbitopathy.

Yoon JS, Lee HJ, Choi SH, Chang EJ, Lee SY, Lee EJ - PLoS ONE (2011)

Effect of quercetin on cell viability and apoptosis in preadipocyte orbital fibroblasts and differentiating orbital fibroblasts.(A) Orbital fibroblasts (1×105) of normal and Graves' orbitopathy (GO) patients were seeded into 24-well culture plates and treated with different concentrations of quercetin (10, 30, 50, or 100 µM) for 24 h. After treatment, assays with 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) were performed to test for viability. (B) An annexin V/FITC kit was used to detect phosphatidylserine externalization, as an index of apoptosis. Percentage of stained cells with annexin V was analyzed by flow cytometry. (C) Orbital fibroblasts (1×105) of GO patients were seeded into 24-well culture plates and treated with different concentrations of quercetin (10, 50, 100, or 200 µM) for 3 days in adipogenic medium containing adipogenesis inducers and rosiglitazone (10 µM). After treatment, MTT assays were performed. Results are expressed as percentage of untreated control values presented as mean ± standard deviation (SD). Assays were performed at least three times in triplicate; data from a representative experiment are shown, expressed as the differences between treated and untreated cells.
© Copyright Policy
Related In: Results  -  Collection

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pone-0026261-g001: Effect of quercetin on cell viability and apoptosis in preadipocyte orbital fibroblasts and differentiating orbital fibroblasts.(A) Orbital fibroblasts (1×105) of normal and Graves' orbitopathy (GO) patients were seeded into 24-well culture plates and treated with different concentrations of quercetin (10, 30, 50, or 100 µM) for 24 h. After treatment, assays with 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) were performed to test for viability. (B) An annexin V/FITC kit was used to detect phosphatidylserine externalization, as an index of apoptosis. Percentage of stained cells with annexin V was analyzed by flow cytometry. (C) Orbital fibroblasts (1×105) of GO patients were seeded into 24-well culture plates and treated with different concentrations of quercetin (10, 50, 100, or 200 µM) for 3 days in adipogenic medium containing adipogenesis inducers and rosiglitazone (10 µM). After treatment, MTT assays were performed. Results are expressed as percentage of untreated control values presented as mean ± standard deviation (SD). Assays were performed at least three times in triplicate; data from a representative experiment are shown, expressed as the differences between treated and untreated cells.
Mentions: To determine nontoxic, concentrations of quercetin in orbital fibroblasts, 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and annexin V-fluorescence isothiocyanate (FITC) apoptosis assay were performed. Exposure of cells to quercetin at ≤100 µM for 24 h neither decreased cell viability below 95% in both normal and GO orbital fibroblasts (Fig. 1A) nor induced a significant level of apoptosis (less than 7%; Fig. 1B). Therefore, treatment of cells with 100 µM quercetin for 24 h was used to determine effects on inflammation and hyaluronan production. For experiments testing suppression of adipogenesis, cells in adipogenic medium were treated with quercetin (10–200 µM) from days 1–3 of differentiation. Cell viability at day three in the presence of 100 µM quercetin was not significantly reduced compared to untreated cells, whereas 200 µM attenuated cell viability to 82.3% (Fig. 1C). Therefore noncytotoxic concentrations (50, 100 µM) of quercetin were used for 3 days in differentiating cells cultures to find the effect of quercetin on adipogenesis.

Bottom Line: Management of Graves' orbitopathy (GO) is challenging, as no reliable, specific, and safe medical therapeutic agents have yet been developed.Treatment with noncytotoxic doses of quercetin inhibited accumulation of intracytoplasmic lipid droplets and resulted in a dose-dependent decrease in expression of peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein (C/EBP) α, and C/EBPβ proteins.In conclusion, inhibition of inflammation, hyaluronan production, and adipogenesis by the natural plant product quercetin in vitro provides the basis for further study of its potential use in the treatment of GO.

View Article: PubMed Central - PubMed

Affiliation: Institute of Vision Research, Department of Ophthalmology, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT
Management of Graves' orbitopathy (GO) is challenging, as no reliable, specific, and safe medical therapeutic agents have yet been developed. We investigated the effect of quercetin in primary cultured orbital fibroblasts from GO, targeting pathways of inflammation, aberrant accumulation of extracellular matrix macromolecules, and adipose tissue expansion. Quercetin significantly attenuated intercellular adhesion molecule-1 (ICAM-1), interleukin (IL) -6, IL-8, and cyclooxygenase (COX) -2 mRNA expression, and inhibited IL-1β-induced increases in ICAM-1, IL-6, and IL-8 mRNA. Increased hyaluronan production induced by IL-1β or tumor necrosis factor-α was suppressed by quercetin in a dose- and time-dependent manner. Treatment with noncytotoxic doses of quercetin inhibited accumulation of intracytoplasmic lipid droplets and resulted in a dose-dependent decrease in expression of peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein (C/EBP) α, and C/EBPβ proteins. In conclusion, inhibition of inflammation, hyaluronan production, and adipogenesis by the natural plant product quercetin in vitro provides the basis for further study of its potential use in the treatment of GO.

Show MeSH
Related in: MedlinePlus