Introduction

To investigate the effect of a novel soluble carbon monoxide-releasing molecule (CORM) on cisplatin (CP) induced cytotoxicity and ischemia reperfusion injury (IRI) in vitro.

Methods

The effects of CORM-3 (200 µM) were compared in normal kidney epithelial cells (HK-2, LLC-PK1) and renal cancer cells (Caki1, Caki2), which were treated with CP (50~200 μM) and induced IRI. To induce IRI condition, cell plates were placed anaerobic chamber (37°C, 95% N2, 5% CO2) for 48hrs, and then cell medium was changed complete medium and incubation in 37°C humidified CO2 incubator for 6hr. The effect of CORM-3 on stimulated IRI and CP treated normal cells/RCC was the determined by measuring the cell viability (CCK assay), TNF-α mRNA induction (Q-RT PCR), protein expression of cleaved caspase 3 and oxidative stress markers including Erk1/2, JNK, P38 (western blot).

Results

Viability after IRI were approximately 40% compared with control. Protective effect of CORM-3 on IRI in vitro model was dose-dependent. Cell viability was 40% recovered in 200 μM CORM-3 pretreated cells compared with control cells. Confluent normal cells and cancer cells were exposed for 24h to CP (50~200 µM) alone or in combined with CORM-3 (12.5 ~200 µM). Protective effects of CORM-3 on CP-treated cells were weaker than those of IRI model. TNF-α mRNA induction occurs following stimulate IRI or CP exposed cells and expression of TNF-α mRNA levels decreased in CORM-3 pretreated cells. Also, IRI or CP-induced activated oxidative stress markers decreased in CORM-3 pretreated cells. CORM-3 reduced expression of c-caspase-3 which is an apoptotic marker.

Conclusions

Our data demonstrate that protective effect of CORM-3 on CP-treated and IRI model in vitro. We suggest that CO attenuates IRI and CP induced cytotoxicity by amelioration of inflammatory and oxidative stress pathways. These effects were observed in not only normal kidney cells but also renal cancer cells.

Funding

none