Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Jul 24:5:12480.
doi: 10.1038/srep12480.

Inhibition of Mitochondrial Complex-1 Prevents the Downregulation of NKCC2 and ENaCα in Obstructive Kidney Disease

Yue Zhang  1 , Ying Sun  1 , Guixia Ding  1 , Songming Huang  1 , Aihua Zhang  1 , Zhanjun Jia  1
Affiliations

Inhibition of Mitochondrial Complex-1 Prevents the Downregulation of NKCC2 and ENaCα in Obstructive Kidney Disease

Yue Zhang et al. Sci Rep. .

Abstract

Ureteral obstruction with subsequent hydronephrosis is a common clinical complication. Downregulation of renal sodium transporters in obstructed kidneys could contribute to impaired urinary concentrating capability and salt waste following the release of a ureteral obstruction. The current study was undertaken to investigate the role of mitochondrial complex-1 inhibition in modulating sodium transporters in obstructive kidney disease. Following unilateral ureteral obstruction (UUO) for 7 days, a global reduction of sodium transporters, including NHE3, α-Na-K-ATPase, NCC, NKCC2, p-NKCC2, ENaCα, and ENaCγ, was observed, as determined via qRT-PCR and/or Western blotting. Interestingly, inhibition of mitochondrial complex-1 by rotenone markedly reversed the downregulation of NKCC2, p-NKCC2, and ENaCα. In contrast, other sodium transporters were not affected by rotenone. To study the potential mechanisms involved in mediating the effects of rotenone on sodium transporters, we examined a number of known sodium modulators, including PGE2, ET1, Ang II, natriuretic peptides (ANP, BNP, and CNP), and nitric oxide synthases (iNOS, nNOS, and eNOS). Importantly, among these modulators, only BNP and iNOS were significantly reduced by rotenone treatment. Collectively, these findings demonstrated a substantial role of mitochondrial dysfunction in mediating the downregulation of NKCC2 and ENaCα in obstructive kidney disease, possibly via iNOS-derived nitric oxide and BNP.

PubMed Disclaimer

Figures

Figure 1
Figure 1. mRNA expression of sodium transporters in obstructed kidneys following rotenone treatment.
(A) qRT-PCR analysis of NHE3. (B) qRT-PCR analysis of NCC. (C) qRT-PCR analysis of α-Na-K-ATPase. (D) qRT-PCR analysis of NKCC2. (E) qRT-PCR analysis of ENaCα. (F) qRT-PCR analysis of ENaCβ. (G) qRT-PCR analysis of ENaCγ. The presented data are means ± SE. N = 5 in each group.
Figure 2
Figure 2. Protein expression of total NKCC2 and p-NKCC2 in obstructed kidneys following rotenone treatment.
(A) Immunohistochemistry of NKCC2. (B) Western blot analysis of NKCC2 and p-NKCC2. (C) Densitometric analysis of NKCC2. (D) Densitometric analysis of p-NKCC2 normalized by β-actin. (E) Densitometric analysis of p-NKCC2 normalized by total NKCC2. The presented data are means ± SE. N = 4–5 in each group.
Figure 3
Figure 3. Protein expression of ENaCα in obstructed kidneys following rotenone treatment.
(A) Immunohistochemistry of ENaCα. (B) Western blot analysis of ENaCα. (C) Densitometric analysis of ENaCα. The presented data are means ± SE. N = 4–5 in each group.
Figure 4
Figure 4. Protein expression of ENaCβ and ENaCγ in obstructed kidneys following rotenone treatment.
(A) Western blot analysis of ENaCβ and ENaCγ. (B) Densitometric analysis of ENaCβ. (C) Densitometric analysis of ENaCγ (85 kDa). (D) Densitometric analysis of ENaCγ (70 kDa). The presented data are means ± SE. N = 4–5 in each group.
Figure 5
Figure 5. Protein expression of NHE3 and α-Na-K-ATPase in obstructed kidneys following rotenone treatment.
(A) Western blot analysis of NHE3 and α-Na-K-ATPase. (B) Densitometric analysis of NHE3. (C) Densitometric analysis of α-Na-K-ATPase. The presented data are means ± SE. N = 4–5 in each group.
Figure 6
Figure 6. Kidney contents of PGE2 and Ang II and mRNA expression of AGT and ET1 in obstructed kidneys following rotenone treatment.
(A) Kidney content of PGE2 determined via EIA. (B) Kidney content of Ang II determined via ELISA. (C) mRNA expression of AGT determined via qRT-PCR. (D) mRNA expression of ET1 determined via qRT-PCR. The presented data are means ± SE. N = 5 in each group.
Figure 7
Figure 7. mRNA expression of natriuretic peptides in obstructed kidneys following rotenone treatment.
(A) qRT-PCR analysis of ANP. (B) qRT-PCR analysis of BNP. (C) qRT-PCR analysis of CNP. The presented data are means ± SE. N = 5 in each group.
Figure 8
Figure 8. mRNA expression of nitric oxide synthases in obstructed kidneys following rotenone treatment.
(A) qRT-PCR analysis of iNOS. (B) qRT-PCR analysis of eNOS. (C) qRT-PCR analysis of nNOS. The presented data are means ± SE. N = 5 in each group.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Ahmadi F. et al. Contribution of stone size to chronic kidney disease in kidney stone formers. Int J Urol 22, 104–108 (2015). - PubMed
    1. Costalonga E. C. et al. Prostatic surgery associated acute kidney injury. World journal of nephrology 3, 198–209 (2014). - PMC - PubMed
    1. Decramer S. et al. Identification of urinary biomarkers by proteomics in newborns: use in obstructive nephropathy. Contrib Nephrol 160, 127–141 (2008). - PubMed
    1. Rule A. D., Krambeck A. E. & Lieske J. C. Chronic kidney disease in kidney stone formers. Clin J Am Soc Nephrol 6, 2069–2075 (2011). - PMC - PubMed
    1. Bianchi D., Vespasiani G. & Bove P. Acute kidney injury due to bilateral ureteral obstruction in children. World journal of nephrology 3, 182–192 (2014). - PMC - PubMed

Publication types

MeSH terms