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Nox2 and Nox4 mediate tumour necrosis factor-α-induced ventricular remodelling in mice.

Moe KT, Yin NO, Naylynn TM, Khairunnisa K, Wutyi MA, Gu Y, Atan MS, Wong MC, Koh TH, Wong P - J. Cell. Mol. Med. (2011)

Bottom Line: ROS was significantly decreased by inhibitors of NADPH oxidase, but not by inhibitors of other ROS production systems.Nox2 and Nox4 siRNA significantly attenuated TNF-α-induced ROS and upregulation of IL-1β and IL-6 in cardiomyocytes.Our study highlights a novel TNF-α-induced chronic ventricular remodelling mechanism mediated by sequential regulation of Nox2 and Nox4 subunits.

View Article: PubMed Central - PubMed

Affiliation: Research and Development Unit, National Heart Centre Singapore, Singapore. moe.kyaw.thu@nhc.com.sg

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Related in: MedlinePlus

Nox2 and Nox4 mediate TNF-α-induced upregulation of IL-1β and IL-6 genes and ROS production. Human adult cardiomyocytes were transfected with Nox2 and Nox4 siRNA using siPORT Amine transfection reagents. Total RNA was extracted from human adult cardiomyocytes 24 hrs after transfection. Human Nox2 and Nox4 mRNA levels were examined using real-time PCR. Nox2 (A) and Nox4 (B) mRNA levels after siRNA treatment were shown. In the other experiments, cells were treated with TNF-α (20 ng/ml) for 6 hrs at 37°C after treatment with Nox2 and Nox4 siRNA. IL-1β (C) and IL-6 mRNA (D) levels were examined with real-time PCR. TNF-α-induced ROS production in human adult cardiomyocytes was determined with CM-H2DCFDA following Nox2 and Nox4 siRNA treatment (E). Data were expressed as relative light unit (RLU) ± S.D. *P < 0.05 versus control values. #P < 0.05 versus TNF-α treatment values.
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fig06: Nox2 and Nox4 mediate TNF-α-induced upregulation of IL-1β and IL-6 genes and ROS production. Human adult cardiomyocytes were transfected with Nox2 and Nox4 siRNA using siPORT Amine transfection reagents. Total RNA was extracted from human adult cardiomyocytes 24 hrs after transfection. Human Nox2 and Nox4 mRNA levels were examined using real-time PCR. Nox2 (A) and Nox4 (B) mRNA levels after siRNA treatment were shown. In the other experiments, cells were treated with TNF-α (20 ng/ml) for 6 hrs at 37°C after treatment with Nox2 and Nox4 siRNA. IL-1β (C) and IL-6 mRNA (D) levels were examined with real-time PCR. TNF-α-induced ROS production in human adult cardiomyocytes was determined with CM-H2DCFDA following Nox2 and Nox4 siRNA treatment (E). Data were expressed as relative light unit (RLU) ± S.D. *P < 0.05 versus control values. #P < 0.05 versus TNF-α treatment values.

Mentions: Nox2 siRNA and Nox4 siRNA significantly decreased Nox2 and Nox4 mRNA levels by 45% and 80%, respectively, in adult human cardiomyocytes (Fig. 6A and B). However, Nox2 and Nox4 siRNA showed no changes in Nox4 and Nox2 mRNA, respectively (data not shown). The role of Nox2 and Nox4 in TNF-α-induced pro-inflammatory cytokine IL-1β and IL-6 expression, and ROS production was determined after knocking down Nox2 and Nox4. Nox2 siRNA significantly decreased TNF-α-induced upregulation of IL-1β by 50% and IL-6 by 25% (Fig. 6C and D). Nox4 siRNA also significantly decreased TNF-α-induced upregulation of IL-1β by 45% and IL-6 by 30%. Nox2 and Nox4 siRNA had no effect on IL-1β and IL-6 expression without TNF-α treatment. Control siRNA had no effect on TNF-α-induced upregulation of IL-1β and IL-6 (data not shown). Finally, Nox2 and Nox4 siRNA significantly decreased TNF-α-induced ROS production by 40% and 50%, respectively (Fig. 6E).


Nox2 and Nox4 mediate tumour necrosis factor-α-induced ventricular remodelling in mice.

Moe KT, Yin NO, Naylynn TM, Khairunnisa K, Wutyi MA, Gu Y, Atan MS, Wong MC, Koh TH, Wong P - J. Cell. Mol. Med. (2011)

Nox2 and Nox4 mediate TNF-α-induced upregulation of IL-1β and IL-6 genes and ROS production. Human adult cardiomyocytes were transfected with Nox2 and Nox4 siRNA using siPORT Amine transfection reagents. Total RNA was extracted from human adult cardiomyocytes 24 hrs after transfection. Human Nox2 and Nox4 mRNA levels were examined using real-time PCR. Nox2 (A) and Nox4 (B) mRNA levels after siRNA treatment were shown. In the other experiments, cells were treated with TNF-α (20 ng/ml) for 6 hrs at 37°C after treatment with Nox2 and Nox4 siRNA. IL-1β (C) and IL-6 mRNA (D) levels were examined with real-time PCR. TNF-α-induced ROS production in human adult cardiomyocytes was determined with CM-H2DCFDA following Nox2 and Nox4 siRNA treatment (E). Data were expressed as relative light unit (RLU) ± S.D. *P < 0.05 versus control values. #P < 0.05 versus TNF-α treatment values.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4373429&req=5

fig06: Nox2 and Nox4 mediate TNF-α-induced upregulation of IL-1β and IL-6 genes and ROS production. Human adult cardiomyocytes were transfected with Nox2 and Nox4 siRNA using siPORT Amine transfection reagents. Total RNA was extracted from human adult cardiomyocytes 24 hrs after transfection. Human Nox2 and Nox4 mRNA levels were examined using real-time PCR. Nox2 (A) and Nox4 (B) mRNA levels after siRNA treatment were shown. In the other experiments, cells were treated with TNF-α (20 ng/ml) for 6 hrs at 37°C after treatment with Nox2 and Nox4 siRNA. IL-1β (C) and IL-6 mRNA (D) levels were examined with real-time PCR. TNF-α-induced ROS production in human adult cardiomyocytes was determined with CM-H2DCFDA following Nox2 and Nox4 siRNA treatment (E). Data were expressed as relative light unit (RLU) ± S.D. *P < 0.05 versus control values. #P < 0.05 versus TNF-α treatment values.
Mentions: Nox2 siRNA and Nox4 siRNA significantly decreased Nox2 and Nox4 mRNA levels by 45% and 80%, respectively, in adult human cardiomyocytes (Fig. 6A and B). However, Nox2 and Nox4 siRNA showed no changes in Nox4 and Nox2 mRNA, respectively (data not shown). The role of Nox2 and Nox4 in TNF-α-induced pro-inflammatory cytokine IL-1β and IL-6 expression, and ROS production was determined after knocking down Nox2 and Nox4. Nox2 siRNA significantly decreased TNF-α-induced upregulation of IL-1β by 50% and IL-6 by 25% (Fig. 6C and D). Nox4 siRNA also significantly decreased TNF-α-induced upregulation of IL-1β by 45% and IL-6 by 30%. Nox2 and Nox4 siRNA had no effect on IL-1β and IL-6 expression without TNF-α treatment. Control siRNA had no effect on TNF-α-induced upregulation of IL-1β and IL-6 (data not shown). Finally, Nox2 and Nox4 siRNA significantly decreased TNF-α-induced ROS production by 40% and 50%, respectively (Fig. 6E).

Bottom Line: ROS was significantly decreased by inhibitors of NADPH oxidase, but not by inhibitors of other ROS production systems.Nox2 and Nox4 siRNA significantly attenuated TNF-α-induced ROS and upregulation of IL-1β and IL-6 in cardiomyocytes.Our study highlights a novel TNF-α-induced chronic ventricular remodelling mechanism mediated by sequential regulation of Nox2 and Nox4 subunits.

View Article: PubMed Central - PubMed

Affiliation: Research and Development Unit, National Heart Centre Singapore, Singapore. moe.kyaw.thu@nhc.com.sg

Show MeSH
Related in: MedlinePlus