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Reactive oxygen species induce Cox-2 expression via TAK1 activation in synovial fibroblast cells.

Onodera Y, Teramura T, Takehara T, Shigi K, Fukuda K - FEBS Open Bio (2015)

Bottom Line: Furthermore, we elucidated a novel mechanism, in which oxidative stress induced phosphorylation of MAPKs and NF-κB through TAK1 activation and resulted in increased Cox-2 and prostaglandin E2 expression.Finally, we demonstrated that ROS-induced Cox-2 expression was inhibited by supplementation of an antioxidant such as N-acetyl cysteamine and hyaluronic acid in vitro and in vivo.From these results, we conclude that oxidative stress is an important factor for generation of Cox-2 in synovial fibroblasts and thus its neutralization may be an effective strategy in palliative therapy for chronic joint diseases.

View Article: PubMed Central - PubMed

Affiliation: Division of Cell Biology for Regenerative Medicine, Institute of Advanced Clinical Medicine, Kindai University Faculty of Medicine, Osaka, Japan.

ABSTRACT
Oxidative stress within the arthritis joint has been indicated to be involved in generating mediators for tissue degeneration and inflammation. COX-2 is a mediator in inflammatory action, pain and some catabolic reactions in inflamed tissues. Here, we demonstrated a direct relationship between oxidative stress and Cox-2 expression in the bovine synovial fibroblasts. Furthermore, we elucidated a novel mechanism, in which oxidative stress induced phosphorylation of MAPKs and NF-κB through TAK1 activation and resulted in increased Cox-2 and prostaglandin E2 expression. Finally, we demonstrated that ROS-induced Cox-2 expression was inhibited by supplementation of an antioxidant such as N-acetyl cysteamine and hyaluronic acid in vitro and in vivo. From these results, we conclude that oxidative stress is an important factor for generation of Cox-2 in synovial fibroblasts and thus its neutralization may be an effective strategy in palliative therapy for chronic joint diseases.

No MeSH data available.


Related in: MedlinePlus

H2O2 addition induced Cox-2 expression in SFs. (A) Estimation of intracellular ROS after H2O2 stimulation to the SFs by APF staining. (B) qPCR assay for Cox-2 mRNA expression after H2O2 treatment. (C) Cox-2 protein expression in the H2O2-treated SFs shown by WB. (D) Increasing of PGE2 expression by H2O2 stimulation in dose dependent manner was detected by ELISA assay.
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f0005: H2O2 addition induced Cox-2 expression in SFs. (A) Estimation of intracellular ROS after H2O2 stimulation to the SFs by APF staining. (B) qPCR assay for Cox-2 mRNA expression after H2O2 treatment. (C) Cox-2 protein expression in the H2O2-treated SFs shown by WB. (D) Increasing of PGE2 expression by H2O2 stimulation in dose dependent manner was detected by ELISA assay.

Mentions: We first determined if the ROS induced Cox-2 generation in the SFs. H2O2 clearly increased intracellular ROS levels in a dose dependent manner (Fig. 1A). Similarly, qRT-PCR revealed that the Cox-2 mRNA expression increased depending on the concentration of H2O2 (Fig. 1B). To confirm the expression of Cox-2 in protein levels, we performed WB assay and observed significant increase of Cox-2 proteins when the cells were stimulated by H2O2 at 50 μM and 100 μM (Fig. 1C). We then performed ELISA assay for PGE2 downstream molecule of Cox-2. The results were consistent with ELISA, in which significant up-regulation of PGE2 (Fig. 1D).


Reactive oxygen species induce Cox-2 expression via TAK1 activation in synovial fibroblast cells.

Onodera Y, Teramura T, Takehara T, Shigi K, Fukuda K - FEBS Open Bio (2015)

H2O2 addition induced Cox-2 expression in SFs. (A) Estimation of intracellular ROS after H2O2 stimulation to the SFs by APF staining. (B) qPCR assay for Cox-2 mRNA expression after H2O2 treatment. (C) Cox-2 protein expression in the H2O2-treated SFs shown by WB. (D) Increasing of PGE2 expression by H2O2 stimulation in dose dependent manner was detected by ELISA assay.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476901&req=5

f0005: H2O2 addition induced Cox-2 expression in SFs. (A) Estimation of intracellular ROS after H2O2 stimulation to the SFs by APF staining. (B) qPCR assay for Cox-2 mRNA expression after H2O2 treatment. (C) Cox-2 protein expression in the H2O2-treated SFs shown by WB. (D) Increasing of PGE2 expression by H2O2 stimulation in dose dependent manner was detected by ELISA assay.
Mentions: We first determined if the ROS induced Cox-2 generation in the SFs. H2O2 clearly increased intracellular ROS levels in a dose dependent manner (Fig. 1A). Similarly, qRT-PCR revealed that the Cox-2 mRNA expression increased depending on the concentration of H2O2 (Fig. 1B). To confirm the expression of Cox-2 in protein levels, we performed WB assay and observed significant increase of Cox-2 proteins when the cells were stimulated by H2O2 at 50 μM and 100 μM (Fig. 1C). We then performed ELISA assay for PGE2 downstream molecule of Cox-2. The results were consistent with ELISA, in which significant up-regulation of PGE2 (Fig. 1D).

Bottom Line: Furthermore, we elucidated a novel mechanism, in which oxidative stress induced phosphorylation of MAPKs and NF-κB through TAK1 activation and resulted in increased Cox-2 and prostaglandin E2 expression.Finally, we demonstrated that ROS-induced Cox-2 expression was inhibited by supplementation of an antioxidant such as N-acetyl cysteamine and hyaluronic acid in vitro and in vivo.From these results, we conclude that oxidative stress is an important factor for generation of Cox-2 in synovial fibroblasts and thus its neutralization may be an effective strategy in palliative therapy for chronic joint diseases.

View Article: PubMed Central - PubMed

Affiliation: Division of Cell Biology for Regenerative Medicine, Institute of Advanced Clinical Medicine, Kindai University Faculty of Medicine, Osaka, Japan.

ABSTRACT
Oxidative stress within the arthritis joint has been indicated to be involved in generating mediators for tissue degeneration and inflammation. COX-2 is a mediator in inflammatory action, pain and some catabolic reactions in inflamed tissues. Here, we demonstrated a direct relationship between oxidative stress and Cox-2 expression in the bovine synovial fibroblasts. Furthermore, we elucidated a novel mechanism, in which oxidative stress induced phosphorylation of MAPKs and NF-κB through TAK1 activation and resulted in increased Cox-2 and prostaglandin E2 expression. Finally, we demonstrated that ROS-induced Cox-2 expression was inhibited by supplementation of an antioxidant such as N-acetyl cysteamine and hyaluronic acid in vitro and in vivo. From these results, we conclude that oxidative stress is an important factor for generation of Cox-2 in synovial fibroblasts and thus its neutralization may be an effective strategy in palliative therapy for chronic joint diseases.

No MeSH data available.


Related in: MedlinePlus