Korean J Physiol Pharmacol.  2008 Aug;12(4):149-153. 10.4196/kjpp.2008.12.4.149.

Expression of Endothelin-1 and Its Receptors in Cisplatin-Induced Acute Renal Failure in Mice

• Lee S
• Ahn D

• KMID: 1838352
• DOI: http://doi.org/10.4196/kjpp.2008.12.4.149

Abstract

Endothelin-1 (ET-1) is unequivocally elevated in the kidney with ischemic acute renal failure (ARF), whereas ET receptors (ET(A)R and ET(B)R) are variably expressed. Although renal functional and structural changes are similar between ischemic and nephrotoxic ARF, there are few reports on the alteration in the ET system in nephrotoxic ARF. This study was, therefore, undertaken to investigate changes in renal expression of ET-1 and its receptors in nephrotoxic ARF induced by cisplatin. Mice were intraperitoneally injected with 16 mg of cisplatin/kg at a single dose, and the expression of mRNA and protein was then quantified by real-time RT-PCR and Western blot, respectively. Immunohistochemistry was conducted for localization. Three days after treatment, ET-1 transcript in cisplatin- treated mice was thirteen times higher than that in controls, whereas ET-1 peptide was increased by 1.5-fold. Cisplatin caused a 2-fold increase in the levels of ET(A)R mRNA and protein. Most of the increased immunoreactive ET-1 and ET(A)R were localized in damaged tubules. Neither the expression of ET(B)R mRNA nor the abundance and immunoreactive level of ET(B)R protein were changed. The findings suggest that the individual components of the renal ET system are differentially regulated in cisplatin-induced nephrotoxic ARF.

Keyword

Cisplatin; Acute renal failure; Endothelin; Endothelin receptor

MeSH Terms

Acute Kidney Injury
Animals
Blotting, Western
Cisplatin
Endothelin-1
Endothelins
Immunohistochemistry
Kidney
Mice
Receptors, Endothelin
RNA, Messenger
Cisplatin
Endothelin-1
Endothelins
RNA, Messenger
Receptors, Endothelin

Figure

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  Fig. 1. Relative expression of ET-1, ETAR, and ETBR genes in the kidney of the control and cisplatin-treated mice. Real-time RT-PCR was performed as described in Materials and Methods. Relative expression was calculated by a comparative CT method. Values are mean±SD of 3 separate experiments. ∗p<0.05 vs. matched control.

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  Fig. 2. ET-1 contents in kidney homogenates of the control and cisplatin-treated mice. Values are mean±SD of 6 animals. ∗p<0.05 vs. control.

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  Fig. 3. Representative immunoblots (upper panel) and densitometric analysis (lower panel) of ET-1, ETAR, and ETBR proteins. Each blot was loaded with 50 ug of protein. Band density was normalized to β-actin. Values are mean±SD of 3 separate experiments. ∗p<0.05 vs. respective control.

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  Fig. 4. Immunostaining for ET-1, ETAR, and ETBR in the renal cortex of control and cisplatin-treated mice. ET-1, ETAR, and ETBR are diffusely expressed in tubular segments. Note that intense immuno-staining for ET-1 and ETAR is found in the cisplatin- treated group. Magnification: ×100.

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