J Korean Dent Soc Anesthesiol.  2014 Sep;14(3):157-165. 10.17245/jkdsa.2014.14.3.157.

The Effect of Remifentanil Preconditioning on Injured Keratinocyte

• Hong HP
• Kim CH
• Yoon JY
• Kim YD
• Park BS
• Kim YH
• Yoon JU

• KMID: 2344989
• DOI: http://doi.org/10.17245/jkdsa.2014.14.3.157

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℃. After hypoxia, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at 37℃. 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.

Keyword

Remifentanil; Keratinocyte

MeSH Terms

Animals
Anoxia
Apoptosis
Autophagy
Biomarkers
Blotting, Western
Cardiopulmonary Bypass
Cell Survival
Coronary Vessels
Granulation Tissue
Humans
Keratinocytes*
Mice
Microscopy, Fluorescence
Nitric Oxide Synthase Type II
Oxygen
Reactive Oxygen Species
Vasodilation
Wound Healing
Wounds and Injuries
Biomarkers
Nitric Oxide Synthase Type II
Oxygen
Reactive Oxygen Species