Modulatory effect of curcumin on ketamine-induced toxicity in rat thymocytes: Involvement of reactive oxygen species (ROS) and the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway

Svetlana Pavlovic; Zorica Jovic; Radmila Karan; Dane Krtinic; Gorana Rankovic; Mladjan Golubovic; Jelena Lilic; Voja Pavlovic

doi:10.17305/bjbms.2018.2607

Authors

Svetlana Pavlovic Department of Anesthesiology, Medical Faculty University of Nis, Nis, Serbia
Zorica Jovic Department of Pharmacology, Medical Faculty University of Nis, Nis, Serbia
Radmila Karan Department of Anesthesiology, Clinical Centre of Serbia, Belgrade, Serbia
Dane Krtinic Department of Pharmacology, Medical Faculty University of Nis, Nis, Serbia
Gorana Rankovic Department of Pharmacology, Medical Faculty University of Nis, Nis, Serbia
Mladjan Golubovic Department of Anesthesiology, Medical Faculty University of Nis, Nis, Serbia
Jelena Lilic Department of Anesthesiology, Medical Faculty University of Nis, Nis, Serbia
Voja Pavlovic Institute of Physiology, Medical Faculty University of Nis, Nis, Serbia

DOI:

https://doi.org/10.17305/bjbms.2018.2607

Keywords:

Ketamine, curcumin, toxicity, thymocytes, PI3K/Akt signaling pathway, anti-apoptotic effect, protective effect, apoptosis, reactive oxygen species

Abstract

Ketamine is a widely used anesthetic in pediatric clinical practice. Previous studies have demonstrated that ketamine induces neurotoxicity and has a modulatory effect on the cells of the immune system. Here, we evaluated the potential protective effect and underlying mechanisms of natural phenolic compound curcumin against ketamine-induced toxicity in rat thymocytes. Rat thymocytes were exposed to 100 µM ketamine alone or combined with increasing concentrations of curcumin (0.3, 1, and 3 μM) for 24 hours. Cell viability was analyzed with CCK-8 assay kit. Apoptosis was analyzed using flow cytometry and propidium iodide as well as Z-VAD-FMK and Z-LEHD-FMK inhibitors. Reactive oxygen species (ROS) production and mitochondrial membrane potential [MMP] were measured by flow cytometry. Colorimetric assay with DEVD-pNA substrate was used for assessing caspase-3 activity. Involvement of phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway was tested with Wortmannin inhibitor. Ketamine induced toxicity in cells, increased the number of hypodiploid cells, caspase-3 activity and ROS production, and inhibited the MMP. Co-incubation of higher concentrations of curcumin (1 and 3 μM) with ketamine markedly decreased cytotoxicity, apoptosis rate, caspase-3 activity, and ROS production in rat thymocytes, and increased the MMP. Application of Z-VAD-FMK (a pan caspase inhibitor) or Z-LEHD-FMK (caspase-9 inhibitor) with ketamine effectively attenuated the ketamine-induced apoptosis in rat thymocytes. Administration of Wortmannin (a PI3K inhibitor) with curcumin and ketamine significantly decreased the protective effect of curcumin on rat thymocytes. Our results indicate that ketamine-induced toxicity in rat thymocytes mainly occurs through the mitochondria-mediated apoptotic pathway and that the PI3K/Akt signaling pathway is involved in the anti-apoptotic effect of curcumin.

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Author Biographies

Svetlana Pavlovic, Department of Anesthesiology, Medical Faculty University of Nis, Nis, Serbia

Department of Anesthesiology
Zorica Jovic, Department of Pharmacology, Medical Faculty University of Nis, Nis, Serbia

Department of Pharmacology
Radmila Karan, Department of Anesthesiology, Clinical Centre of Serbia, Belgrade, Serbia

Department of Anesthesiology
Dane Krtinic, Department of Pharmacology, Medical Faculty University of Nis, Nis, Serbia

Department of Pharmacology
Gorana Rankovic, Department of Pharmacology, Medical Faculty University of Nis, Nis, Serbia

Department of Pharmacology
Mladjan Golubovic, Department of Anesthesiology, Medical Faculty University of Nis, Nis, Serbia

Department of Anesthesiology
Jelena Lilic, Department of Anesthesiology, Medical Faculty University of Nis, Nis, Serbia

Department of Anesthesiology

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