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

Zhang Y, Sun Y, Ding G, Huang S, Zhang A, Jia Z - Sci Rep (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Nephrology, Nanjing Children's Hospital, Affiliated with Nanjing Medical University, Nanjing 210008, China [2] Institute of Pediatrics, Nanjing Medical University, Nanjing, China [3] Nanjing Key Laboratory of Pediatrics, Nanjing Children Hospital, Affiliated with Nanjing Medical University, Nanjing 210008, China.

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.

No MeSH data available.


Related in: MedlinePlus

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.
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f2: 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.

Mentions: The immunohistochemistry results showed that the robust downregulation of NKCC2 protein expression in the obstructed kidney was entirely prohibited by rotenone treatment (Fig. 2A). Similarly, Western blotting revealed a striking reduction of NKCC2 levels and complete restoration upon rotenone administration (Fig. 2B,C). To investigate the status of NKCC2 phosphorylation, we further examined the levels of phosphorylated NKCC2 through Western blotting and observed a similar regulatory pattern to total NKCC2 (Fig. 2B,D). However, the ratio of p-NKCC2 to total NKCC2 was not affected by kidney obstruction or rotenone treatment (Fig. 2E), indicating that the change of p-NKCC2 was resulted from the alteration of total NKCC2. These results demonstrated a potent role of mitochondrial complex-I inhibition in inhibiting the downregulation of NKCC2 in the thick ascending limbs of obstructed kidneys.


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

Zhang Y, Sun Y, Ding G, Huang S, Zhang A, Jia Z - Sci Rep (2015)

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.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4513566&req=5

f2: 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.
Mentions: The immunohistochemistry results showed that the robust downregulation of NKCC2 protein expression in the obstructed kidney was entirely prohibited by rotenone treatment (Fig. 2A). Similarly, Western blotting revealed a striking reduction of NKCC2 levels and complete restoration upon rotenone administration (Fig. 2B,C). To investigate the status of NKCC2 phosphorylation, we further examined the levels of phosphorylated NKCC2 through Western blotting and observed a similar regulatory pattern to total NKCC2 (Fig. 2B,D). However, the ratio of p-NKCC2 to total NKCC2 was not affected by kidney obstruction or rotenone treatment (Fig. 2E), indicating that the change of p-NKCC2 was resulted from the alteration of total NKCC2. These results demonstrated a potent role of mitochondrial complex-I inhibition in inhibiting the downregulation of NKCC2 in the thick ascending limbs of obstructed kidneys.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Nephrology, Nanjing Children's Hospital, Affiliated with Nanjing Medical University, Nanjing 210008, China [2] Institute of Pediatrics, Nanjing Medical University, Nanjing, China [3] Nanjing Key Laboratory of Pediatrics, Nanjing Children Hospital, Affiliated with Nanjing Medical University, Nanjing 210008, China.

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.

No MeSH data available.


Related in: MedlinePlus