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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

HIF-1α is required for ID1 induction in the H/R model. HK-2 cells were transiently transfected with specific siRNA for HIF-1α or siRNA for negative control and then subjected to H/R. (a) In cells transfected with HIF-1α siRNA, the expression of HIF-1α was markedly inhibited and ID1 was suppressed, although not remarkable. β-actin served as the loading control. A representative blot from 3 independent experiments is shown. (b, c) In cells transfected with HIF-1α siRNA, relative mRNA levels of HIF-1α were efficiently reduced. Relative mRNA levels of ID1 were decreased significantly at the prehypoxia, 12-hour and 24-hour time points for hypoxia, and after 12 hours of reoxygenation. β-actin mRNA was used as the internal control (n = 3). ∗P < .05 compared with the negative control (scramble). ID1, inhibitor of DNA binding 1; HIF-1α, hypoxia-inducible factor-1 alpha; H/R, hypoxia-reoxygenation.
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fig5: HIF-1α is required for ID1 induction in the H/R model. HK-2 cells were transiently transfected with specific siRNA for HIF-1α or siRNA for negative control and then subjected to H/R. (a) In cells transfected with HIF-1α siRNA, the expression of HIF-1α was markedly inhibited and ID1 was suppressed, although not remarkable. β-actin served as the loading control. A representative blot from 3 independent experiments is shown. (b, c) In cells transfected with HIF-1α siRNA, relative mRNA levels of HIF-1α were efficiently reduced. Relative mRNA levels of ID1 were decreased significantly at the prehypoxia, 12-hour and 24-hour time points for hypoxia, and after 12 hours of reoxygenation. β-actin mRNA was used as the internal control (n = 3). ∗P < .05 compared with the negative control (scramble). ID1, inhibitor of DNA binding 1; HIF-1α, hypoxia-inducible factor-1 alpha; H/R, hypoxia-reoxygenation.

Mentions: Subsequently, siRNAs for HIF-1α were used to investigate whether ID1 was mediated by HIF-1α. The expression of HIF-1α was inhibited and this outcome was confirmed by western blot and qRT-PCR (Figures 5(a) and 5(b)). Cells in which HIF-1α expression was suppressed showed reduced expression of ID1 through H/R, and mRNA levels were significantly downregulated (Figures 5(a) and 5(c)). The results indicate that ID1 is induced in the H/R model and is regulated by HIF-1α.


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)

HIF-1α is required for ID1 induction in the H/R model. HK-2 cells were transiently transfected with specific siRNA for HIF-1α or siRNA for negative control and then subjected to H/R. (a) In cells transfected with HIF-1α siRNA, the expression of HIF-1α was markedly inhibited and ID1 was suppressed, although not remarkable. β-actin served as the loading control. A representative blot from 3 independent experiments is shown. (b, c) In cells transfected with HIF-1α siRNA, relative mRNA levels of HIF-1α were efficiently reduced. Relative mRNA levels of ID1 were decreased significantly at the prehypoxia, 12-hour and 24-hour time points for hypoxia, and after 12 hours of reoxygenation. β-actin mRNA was used as the internal control (n = 3). ∗P < .05 compared with the negative control (scramble). 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

fig5: HIF-1α is required for ID1 induction in the H/R model. HK-2 cells were transiently transfected with specific siRNA for HIF-1α or siRNA for negative control and then subjected to H/R. (a) In cells transfected with HIF-1α siRNA, the expression of HIF-1α was markedly inhibited and ID1 was suppressed, although not remarkable. β-actin served as the loading control. A representative blot from 3 independent experiments is shown. (b, c) In cells transfected with HIF-1α siRNA, relative mRNA levels of HIF-1α were efficiently reduced. Relative mRNA levels of ID1 were decreased significantly at the prehypoxia, 12-hour and 24-hour time points for hypoxia, and after 12 hours of reoxygenation. β-actin mRNA was used as the internal control (n = 3). ∗P < .05 compared with the negative control (scramble). ID1, inhibitor of DNA binding 1; HIF-1α, hypoxia-inducible factor-1 alpha; H/R, hypoxia-reoxygenation.
Mentions: Subsequently, siRNAs for HIF-1α were used to investigate whether ID1 was mediated by HIF-1α. The expression of HIF-1α was inhibited and this outcome was confirmed by western blot and qRT-PCR (Figures 5(a) and 5(b)). Cells in which HIF-1α expression was suppressed showed reduced expression of ID1 through H/R, and mRNA levels were significantly downregulated (Figures 5(a) and 5(c)). The results indicate that ID1 is induced in the H/R model and is regulated by HIF-1α.

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