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Updated: Sep 26, 2026
Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
Published on: January 7, 2013
Cytochrome P450 2E1 null mice provide novel protection against cisplatin-induced nephrotoxicity and apoptosis
1Department of Pediatrics, University of Mississippi Medical Center, Jackson, Mississippi 39216, USA.
Background:
Reactive oxygen metabolites (ROM) are important mediators of cisplatin-induced nephrotoxicity and apoptosis. The site and source of generation of these metabolites are not well defined. Cytochrome P450 (CYP) are heme-containing enzymes that can generate ROM during the oxidative metabolism of exogenous and endogenous compounds. CYP2E1 was identified and localized to the kidney proximal tubule. There is evidence to suggest that CYP2E1 is involved in the generation of ROM.
Methods:
The current study was performed utilizing CYP2e1 null mice (CYP2e1-/-). Cisplatin nephrotoxicity was induced in mice by single intraperitoneal injection of cisplatin and animals were sacrificed 72 hours later. Renal function was assessed and various biochemical tests were performed, including histologic studies.
Results:
CYP2e1-/- demonstrated marked functional and histologic protection against cisplatin-induced renal injury. Incubation of CYP2e1-/- kidney slices with cisplatin resulted in significant decrease in the generation of ROM and attenuation of cytotoxicity as compared to that of wild-type mice (CYP2e1+/+). Cisplatin-induced apoptosis was also markedly reduced in the CYP2e1-/- mice. Direct incubation of cisplatin with the microsomes isolated from CYP2e1-/- kidney cortex produced significant decrease in the generation of hydrogen peroxide, catalytic iron content, and hydroxyl radical formation compared to CYP2e1+/+ microsomes.
Conclusion:
Our results thus demonstrate a pivotal role of CYP2E1 in cisplatin-induced nephrotoxicity and apoptosis. We postulate that the interaction of cisplatin with CYP2E1 results in the generation of ROM that causes renal injury and initiates apoptosis.
Insights
Cytochrome P450 2E1 (CYP2E1) plays a key role in cisplatin-induced kidney damage. Eliminating CYP2E1 protected mice from kidney injury and apoptosis, suggesting it generates harmful reactive oxygen metabolites.
Area of Science:
- Nephrology
- Toxicology
- Biochemistry
Background:
- Reactive oxygen metabolites (ROM) mediate cisplatin-induced nephrotoxicity and apoptosis.
- Cytochrome P450 (CYP) enzymes, including CYP2E1, can generate ROM during metabolism.
- CYP2E1 is present in kidney proximal tubules and implicated in ROM generation.
Purpose of the Study:
- To investigate the role of CYP2E1 in cisplatin-induced nephrotoxicity and apoptosis.
- To determine if CYP2E1 is a source of ROM contributing to kidney injury.
Main Methods:
- Utilized CYP2E1-null mice (CYP2E1-/-) and wild-type littermates (CYP2E1+/+).
- Induced nephrotoxicity via cisplatin injection and assessed renal function and histology 72 hours post-injection.
- Measured ROM generation, cytotoxicity, apoptosis, hydrogen peroxide, catalytic iron, and hydroxyl radical formation in kidney tissues and microsomes.
Main Results:
- CYP2E1-/- mice showed significant protection against cisplatin-induced renal injury, reduced ROM generation, and attenuated cytotoxicity.
- Cisplatin-induced apoptosis was markedly reduced in CYP2E1-/- mice.
- Kidney microsomes from CYP2E1-/- mice produced less hydrogen peroxide, catalytic iron, and hydroxyl radical upon cisplatin incubation compared to CYP2E1+/+.
Conclusions:
- CYP2E1 plays a pivotal role in cisplatin-induced nephrotoxicity and apoptosis.
- The interaction of cisplatin with CYP2E1 generates ROM, leading to renal injury and apoptosis initiation.
