J Korean Dent Soc Anesthesiol. 2014 Sep;14(3):157-165. English.
Published online May 27, 2016.
Copyright © 2014 Journal of the Korean Dental Society of Anesthesiology
Original Article

The Effect of Remifentanil Preconditioning on Injured Keratinocyte

Hun Pyo Hong, Cheul Hong Kim, Ji Young Yoon, Yong Deok Kim,* Bong Soo Park, Yong Ho Kim, and Ji Uk Yoon
    • Department of Dental Anesthesia and Pain Medicine, School of Dentistry, Pusan National University, Gyeongnam, Korea.
    • *Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University, Gyeongnam, Korea.
    • Department of Oral Anatomy, School of Dentistry, Pusan National University, Gyeongnam, Korea.
    • Department of Anesthesia and Pain Medicine, School of Medicine, Pusan National University, Gyeongnam, Korea.
Received September 19, 2014; Revised September 30, 2014; Accepted October 02, 2014.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Incisional site of surgical operation become transient ischemic state and then occur reoxygenation due to vasodilatation by inflammatory reaction, the productive reactive oxygen species (ROS) give rise to many physiologic results. Apoptosis have major role on elimination of inflammatory cell and formation of granulation tissue in normal wound healing process. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. After cardiopulmonary bypass for coronary artery surgery, remifentanil can also inhibit the release of biomarkers of myocardial damage. Here we investigated whether remifentanil pretreatment has cellular protective effect against hypoxia-reoxygenation in HaCaT human keratinocytes, if so, the role of apoptosis and autophagy on this phenomenon.

Methods

The HaCaT human keratinocytes were exposed to various concentrations of remifentanil (0.01, 0.05, 0.1, 0.5 and 1 ng/ml) for 2 h before hypoxia (RPC/HR group). These cells were cultured under 1% oxygen tension for 24h at 37°C. After hypoxia, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at 37°C. 3-MA/RPC/HR group was treated 3-methyladenine (3-MA), autophagy inhibitor for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with thiazolyl blue tetrazoliumbromide (MTT, amresco), showing the mitochondrial activity of living cells. To investigate whether the occurrence of autophagy and apoptosis, we used fluorescence microscopy and Western blot analysis.

Results

The viability against hypoxia-reoxygenation injury in remifentanil preconditioning keratinocytes were increased, and these cells were showed stimulated expression of autophagy 3-MA suppressed the induction of autophagy effectively and the protective effects on apoptosis. Atg5, Beclin-1, LC3-II and p62 were elevated in RPC/HR group. But they were decreased when autophagy was suppressed by 3-MA.

Conclusions

Remifentanil preconditioning showed the protective effect in human keratinocytes, and we concluded that autophagy may take the major role in the recovery of wound from hypoxia-reoxygenation injury. We suggest that further research is needed about the cell protective effects of autophagy.

Keywords
Remifentanil; Keratinocyte


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