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Inhibitor of DNA Binding 1 Is Induced during Kidney Ischemia-Reperfusion and Is Critical for the Induction of Hypoxia-Inducible Factor-1α.

Wen D, Zou YF, Gao YH, Zhao Q, Xie YY, Shen PY, Xu YW, Xu J, Chen YX, Feng XB, Shi H, Zhang W - Biomed Res Int (2016)

Bottom Line: Moreover, the suppression of ID1 mRNA expression could lead to decreased expression and transcription of HIF-1α during hypoxia and reoxygenation.In addition, it was demonstrated that both ID1 and HIF-1α can regulate the transcription of twist.This study demonstrated that ID1 is induced in renal TECs during I/R and can regulate the transcription and expression of HIF-1α.

View Article: PubMed Central - PubMed

Affiliation: Department of Nephrology, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025, China.

ABSTRACT
In this study, rat models of acute kidney injury (AKI) induced by renal ischemia-reperfusion (I/R) and HK-2 cell models of hypoxia-reoxygenation (H/R) were established to investigate the expression of inhibitor of DNA binding 1 (ID1) in AKI, and the regulation relationship between ID1 and hypoxia-inducible factor 1 alpha (HIF-1α). Through western blot, quantitative real-time PCR, immunohistochemistry, and other experiment methods, the induction of ID1 after renal I/R in vivo was observed, which was expressed mainly in renal tubular epithelial cells (TECs). ID1 expression was upregulated in in vitro H/R models at both the protein and mRNA levels. Via RNAi, it was found that ID1 induction was inhibited with silencing of HIF-1α. Moreover, the suppression of ID1 mRNA expression could lead to decreased expression and transcription of HIF-1α during hypoxia and reoxygenation. In addition, it was demonstrated that both ID1 and HIF-1α can regulate the transcription of twist. This study demonstrated that ID1 is induced in renal TECs during I/R and can regulate the transcription and expression of HIF-1α.

No MeSH data available.


Related in: MedlinePlus

ID1 is required for HIF-1α induction in the H/R model. HK-2 cells were transiently transfected with specific siRNA for ID1 or siRNA for negative control and then subjected to H/R. (a) In cells transfected with ID1 siRNA, the expression of ID1 was markedly inhibited. HIF-1α protein expression was remarkably suppressed after exposure to hypoxia for 24 hours or to reoxygenation for 12 or 24 hours. β-actin expression was examined as the loading control. A representative blot from 3 independent experiments is shown. (b, c) In cells transfected with ID1 siRNA, relative mRNA levels of ID1 were efficiently reduced. Relative mRNA levels of HIF-1α were decreased significantly after 24 hours of hypoxia and 12 hours of reoxygenation. β-actin mRNA served as an internal control (n = 3). ∗P < .05 compared with the negative control (scramble). ∗∗P < .05 compared with the prehypoxia controls. ID1, inhibitor of DNA binding 1; HIF-1α, hypoxia-inducible factor-1 alpha; H/R, hypoxia-reoxygenation.
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fig6: ID1 is required for HIF-1α induction in the H/R model. HK-2 cells were transiently transfected with specific siRNA for ID1 or siRNA for negative control and then subjected to H/R. (a) In cells transfected with ID1 siRNA, the expression of ID1 was markedly inhibited. HIF-1α protein expression was remarkably suppressed after exposure to hypoxia for 24 hours or to reoxygenation for 12 or 24 hours. β-actin expression was examined as the loading control. A representative blot from 3 independent experiments is shown. (b, c) In cells transfected with ID1 siRNA, relative mRNA levels of ID1 were efficiently reduced. Relative mRNA levels of HIF-1α were decreased significantly after 24 hours of hypoxia and 12 hours of reoxygenation. β-actin mRNA served as an internal control (n = 3). ∗P < .05 compared with the negative control (scramble). ∗∗P < .05 compared with the prehypoxia controls. ID1, inhibitor of DNA binding 1; HIF-1α, hypoxia-inducible factor-1 alpha; H/R, hypoxia-reoxygenation.

Mentions: To study the function of ID1, siRNA was used to inhibit ID1 expression. The mRNA levels of ID1 were lowered significantly through the whole H/R process, and the protein levels were also reduced markedly (Figures 6(a) and 6(b)).


Inhibitor of DNA Binding 1 Is Induced during Kidney Ischemia-Reperfusion and Is Critical for the Induction of Hypoxia-Inducible Factor-1α.

Wen D, Zou YF, Gao YH, Zhao Q, Xie YY, Shen PY, Xu YW, Xu J, Chen YX, Feng XB, Shi H, Zhang W - Biomed Res Int (2016)

ID1 is required for HIF-1α induction in the H/R model. HK-2 cells were transiently transfected with specific siRNA for ID1 or siRNA for negative control and then subjected to H/R. (a) In cells transfected with ID1 siRNA, the expression of ID1 was markedly inhibited. HIF-1α protein expression was remarkably suppressed after exposure to hypoxia for 24 hours or to reoxygenation for 12 or 24 hours. β-actin expression was examined as the loading control. A representative blot from 3 independent experiments is shown. (b, c) In cells transfected with ID1 siRNA, relative mRNA levels of ID1 were efficiently reduced. Relative mRNA levels of HIF-1α were decreased significantly after 24 hours of hypoxia and 12 hours of reoxygenation. β-actin mRNA served as an internal control (n = 3). ∗P < .05 compared with the negative control (scramble). ∗∗P < .05 compared with the prehypoxia controls. ID1, inhibitor of DNA binding 1; HIF-1α, hypoxia-inducible factor-1 alpha; H/R, hypoxia-reoxygenation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835634&req=5

fig6: ID1 is required for HIF-1α induction in the H/R model. HK-2 cells were transiently transfected with specific siRNA for ID1 or siRNA for negative control and then subjected to H/R. (a) In cells transfected with ID1 siRNA, the expression of ID1 was markedly inhibited. HIF-1α protein expression was remarkably suppressed after exposure to hypoxia for 24 hours or to reoxygenation for 12 or 24 hours. β-actin expression was examined as the loading control. A representative blot from 3 independent experiments is shown. (b, c) In cells transfected with ID1 siRNA, relative mRNA levels of ID1 were efficiently reduced. Relative mRNA levels of HIF-1α were decreased significantly after 24 hours of hypoxia and 12 hours of reoxygenation. β-actin mRNA served as an internal control (n = 3). ∗P < .05 compared with the negative control (scramble). ∗∗P < .05 compared with the prehypoxia controls. ID1, inhibitor of DNA binding 1; HIF-1α, hypoxia-inducible factor-1 alpha; H/R, hypoxia-reoxygenation.
Mentions: To study the function of ID1, siRNA was used to inhibit ID1 expression. The mRNA levels of ID1 were lowered significantly through the whole H/R process, and the protein levels were also reduced markedly (Figures 6(a) and 6(b)).

Bottom Line: Moreover, the suppression of ID1 mRNA expression could lead to decreased expression and transcription of HIF-1α during hypoxia and reoxygenation.In addition, it was demonstrated that both ID1 and HIF-1α can regulate the transcription of twist.This study demonstrated that ID1 is induced in renal TECs during I/R and can regulate the transcription and expression of HIF-1α.

View Article: PubMed Central - PubMed

Affiliation: Department of Nephrology, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025, China.

ABSTRACT
In this study, rat models of acute kidney injury (AKI) induced by renal ischemia-reperfusion (I/R) and HK-2 cell models of hypoxia-reoxygenation (H/R) were established to investigate the expression of inhibitor of DNA binding 1 (ID1) in AKI, and the regulation relationship between ID1 and hypoxia-inducible factor 1 alpha (HIF-1α). Through western blot, quantitative real-time PCR, immunohistochemistry, and other experiment methods, the induction of ID1 after renal I/R in vivo was observed, which was expressed mainly in renal tubular epithelial cells (TECs). ID1 expression was upregulated in in vitro H/R models at both the protein and mRNA levels. Via RNAi, it was found that ID1 induction was inhibited with silencing of HIF-1α. Moreover, the suppression of ID1 mRNA expression could lead to decreased expression and transcription of HIF-1α during hypoxia and reoxygenation. In addition, it was demonstrated that both ID1 and HIF-1α can regulate the transcription of twist. This study demonstrated that ID1 is induced in renal TECs during I/R and can regulate the transcription and expression of HIF-1α.

No MeSH data available.


Related in: MedlinePlus