Effect of Carbon Monoxide Donor CORM-2 on Vitamin D&lt;sub&gt;3&lt;/sub&gt; Metabolism

Feger, Martina; Fajol, Abul; Lebedeva, Aleksandra; Meissner, Adrian; Michael, Diana; Voelkl, Jakob; Alesutan, Ioana; Schleicher, Erwin; Reichetzeder, Christoph; Hocher, Berthold; Qadri, Syed M.; Läng, Florian

doi:10.1159/000355730

Cited by 28 publications

(25 citation statements)

References 76 publications

(93 reference statements)

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“…Total RNA was isolated from mouse kidney tissues by using Trifast Reagent (Peqlab Biotechnologie GmbH, Germany) according to the manufacturer's instructions [41. ]Reverse transcription of 2 µg RNA was performed using oligo(dT) 12-18 primers (Invitrogen, Life Technologies GmbH, Germany) and SuperScriptIII Reverse Transcriptase (Invitrogen, Life Technologies GmbH, Germany).…”

Section: Methodsmentioning

confidence: 99%

Down-Regulation of Renal Klotho Expression by Shiga Toxin 2

Feger¹,

Mia²,

Pakladok³

et al. 2014

Kidney Blood Press Res

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Background/Aims: Shiga toxin 2 may trigger classical hemolytic uremic syndrome (HUS) eventually leading to renal failure. Klotho, a transmembrane protein, protease and hormone mainly expressed in kidney is involved in the regulation of renal phosphate excretion and also retains renal protective effects. Renal failure is associated with renal depletion of klotho. The present study explored the influence of Shiga toxin 2 on renal klotho expression. Methods: Mice were injected with either solvent or Shiga toxin 2 and urinary flow rate and phosphate excretion were determined in metabolic cages. Renal transcript levels were measured by quantitative RT-PCR and renal protein abundance by Western blotting. Plasma concentrations of 1,25(OH)₂D₃ and FGF23 were determined by ELISA and plasma phosphate and urea concentrations by photometry. Results: Shiga toxin 2 treatment was followed by increase of plasma urea concentration, urinary flow rate and renal phosphate excretion but not of plasma phosphate concentration. Shiga toxin 2 treatment strongly decreased klotho mRNA expression and klotho protein abundance in renal tissue. Shiga toxin 2 treatment further increased tumor necrosis factor (TnfE) mRNA levels, as well as protein abundance of phosphorylated p38 MAPK in renal tissue. The treatment significantly increased renal Cyp27b1 and decreased renal Cyp24a1 mRNA levels without significantly altering plasma 1,25(OH)₂D₃ levels. Shiga toxin 2 treatment was further followed by increase of plasma FGF23 concentrations. Conclusion: Shiga toxin 2 treatment stimulated TnfE transcription, down-regulated renal klotho expression and increased FGF23 formation, effects presumably contributing to renal tissue injury. i 2015 S. Karger AG, Basel

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Section: Methodsmentioning

confidence: 99%

Down-Regulation of Renal Klotho Expression by Shiga Toxin 2

Feger¹,

Mia²,

Pakladok³

et al. 2014

Kidney Blood Press Res

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“…All PCRs were performed in duplicate, and mRNA fold changes were calculated by the 2 -ΔΔCt method using GAPDH as internal reference [37,38]. …”

Section: Methodsmentioning

confidence: 99%

Up-Regulation of Hepatic Alpha-2-HS-Glycoprotein Transcription by TestosteroneviaAndrogen Receptor Activation

Voelkl

Pakladok

Lin

et al. 2014

Cell Physiol Biochem

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Background/Aims: Fetuin-A (alpha-2-HS-glycoprotein, AHSG), a liver borne plasma protein, contributes to the prevention of soft tissue calcification, modulates inflammation, reduces insulin sensitivity and fosters weight gain following high fat diet or ageing. In polycystic ovary syndrome, fetuin-A levels correlate with free androgen levels, an observation pointing to androgen sensitivity of fetuin-A expression. The present study thus explored whether the expression of hepatic fetuin-A is modified by testosterone. Methods: HepG2 cells were treated with testosterone and androgen receptor antagonist flutamide, and were silenced with androgen receptor siRNA. To test the in vivo relevance, male mice were subjected to androgen deprivation therapy (ADT) for 7 weeks. AHSG mRNA levels were determined by quantitative RT-PCR and fetuin-A protein abundance by Western blotting. Results: In HepG2 cells, AHSG mRNA expression and fetuin-A protein abundance were both up-regulated following testosterone treatment. The human alpha-2-HS-glycoprotein gene harbors putative androgen receptor response elements in the proximal 5 kb promoter sequence relative to TSS. The effect of testosterone on AHSG mRNA levels was abrogated by silencing of the androgen receptor in HepG2 cells. Moreover, treatment of HepG2 cells with the androgen receptor antagonist flutamide in presence of endogenous ligands in the medium significantly down-regulated AHSG mRNA expression and fetuin-A protein abundance. In addition, ADT of male mice was followed by a significant decrease of hepatic Ahsg mRNA expression and fetuin-A protein levels. Conclusions: Testosterone participates in the regulation of hepatic fetuin-A expression, an effect mediated, at least partially, by androgen receptor activation.

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“…The house-keeping gene Tbp (TATA binding protein) was amplified to standardize the amount of sample RNA. Relative quantification of gene expression was achieved using the ΔCT method as described earlier [29,30].…”

Section: Real-time Pcrmentioning

confidence: 99%