Korean J Physiol Pharmacol.  2013 Oct;17(5):435-440. 10.4196/kjpp.2013.17.5.435.

Antiapoptotic Effect of Paricalcitol in Gentamicin-induced Kidney Injury

• Suh SH
• Lee KE
• Park JW
• Kim IJ
• Kim O
• Kim CS
• Choi JS
• Bae EH
• Ma SK
• Lee JU
• Kim SW

• KMID: 1500239
• DOI: http://doi.org/10.4196/kjpp.2013.17.5.435

Abstract

While the anti-apoptotic effect of paricalcitol has been demonstrated in various animal models, it is not yet clear whether paricalcitol attenuates the apoptosis in gentamicin (GM)-induced kidney injury. We investigated the effect of paricalcitol on apoptotic pathways in rat kidneys damaged by GM. Rats were randomly divided into three groups: 1) Control group (n=8), where only vehicle was delivered, 2) GM group (n=10), where rats were treated with GM (150 mg/kg/day) for 7 days, 3) PARI group (n=10), where rats were co-treated with paricalcitol (0.2 microg/kg/day) and GM for 7 days. Paricalcitol attenuated renal dysfunction by GM administration in biochemical profiles. In terminal deoxynucleotidyl transferase dUTP nick end labeling staining, increased apoptosis was observed in GM group, which was reversed by paricalcitol co-treatment. Immunoblotting using protein samples from rat cortex/outer stripe of outer medulla showed increased Bax/Bcl-2 ratio and cleaved form of caspase-3 in GM group, both of which were reversed by paricalcitol. The phosphorylated Jun-N-terminal kinase (JNK) expression was increase in GM, which was counteracted by paricalcitol. The protein expression of p-Akt and nitro-tyrosine was also enhanced in GM-treated rats compared with control rats, which was reversed by paricalcitol co-treatment. Paricalcitol protects GM-induced renal injury by antiapoptotic mechanisms, including inhibition of intrinsic apoptosis pathway and JNK.

Keyword

Apoptosis; Gentamicin; Kidney; Paricalcitol

MeSH Terms

Acute Kidney Injury*
Animals
Apoptosis
Caspase 3
DNA Nucleotidylexotransferase
Ergocalciferols
Gentamicins
Immunoblotting
Kidney*
Models, Animal
Phosphotransferases
Rats
Caspase 3
DNA Nucleotidylexotransferase
Ergocalciferols
Gentamicins
Phosphotransferases

Figure

• 

  Fig. 1 Immunoblotting of Bax and Bcl-2 in renal cortex/OSOM. GM treatment, compared with vehicle treatment, significantly decreased Bcl-2 expression, while co-administration of paricalcitol restored Bcl-2 expression level. Either of GM or paricalcitol did not affect the expression of Bax protein. The ratio of Bax to Bcl-2, accordingly, was increased in GM group, compared to control or PARI groups. The level of cytochrome C expression was not statically significant difference among the three groups. *p<0.05 vs. control. †p<0.05 vs. GM.

• 

  Fig. 2 Immunoblotting of caspase-3 in renal cortex/OSOM. Cleaved caspase-3 was upregulated by GM treatment, compared by vehicle treatment, which was reversed by paricalcitol co-administration. The ratio of cleavage form to total form caspase-3 was also increased in GM group, compared with the other groups. *p<0.05 vs. control. †p<0.05 vs. GM.

• 

  Fig. 3 TUNEL staining in renal cortex (×200 magnifications). Markedly increased numbers of TUNEL-positive cells in the kidney of GM group, compared with in those of control and PARI groups, were observed. *p<0.05 vs. control. †p<0.05 vs. GM.

• 

  Fig. 4 Immnublotting of JNK in renal cortex/OSOM. The expression of p-JNK was remarkably increased by GM treatment, compared with vehicle injection, which was reversed by paricalcitol co-treatment, while JNK expression was not affected by either agent. *p<0.05 vs. control. †p<0.05 vs. GM.

• 

  Fig. 5 Immnublotting of p-Akt in renal cortex/OSOM. The level of p-Akt was significantly increased in GM-treated rats compared with control rats, which was reversed by paricalcitol co-treatment. *p<0.05 vs. control. †p<0.05 vs. GM.

• 

  Fig. 6 the Immunoblotting of nitro-tyrosine in renal cortex/OSOM. GM treatment increased the level of nitro-tyrosine in rat kidneys, which alteration was attenuated by paricalcitol co-treatment. *p<0.05 vs. control. †p<0.05 vs. GM.

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