Remifentanil reduced the effects of hydrogen peroxide-induced oxidative stress in human keratinocytes via autophagy.
OBJECTIVE
Excess reactive oxygen species are detrimental to wound repair. Remifentanil decreases reactive oxygen species generation and inflammatory response; however, its effects on oxidative cell injury are not completely understood. Therefore, we investigated the effects of remifentanil on human keratinocytes under hydrogen peroxide-induced oxidative stress and the correlation of these effects with autophagy.


METHODS
Human keratinocytes (HaCaT cell line) were randomly assigned to four groups: control, hydrogen peroxide, remifentanil pretreatment + hydrogen peroxide, and 3-methyladenine + remifentanil pretreatment + hydrogen peroxide. The MTT assay was performed to analyze cell viability. Apoptotic cell death was measured by Hoechst staining. A scratch assay was used to measure cell migration. The role of autophagy was ascertained by autophagosome staining and western blot analysis of autophagy-related proteins.


RESULTS
Compared with the control group, the hydrogen peroxide group showed decreased cell viability, which was improved by remifentanil pretreatment. Hydrogen peroxide-induced apoptotic cell death and delayed cell migration were also improved by remifentanil pretreatment. Western blot analysis showed that the expression of autophagy-related proteins significantly increased in the remifentanil pretreatment + hydrogen peroxide group compared with that in the hydrogen peroxide group.


CONCLUSIONS
This study demonstrated that remifentanil pretreatment ameliorates hydrogen peroxide-induced oxidative injury in human keratinocytes. In addition, our results show that the antioxidative effect of remifentanil against hydrogen peroxide-induced oxidative injury was mediated by autophagy.
a Department of Dental Anesthesia and Pain Medicine, School of Dentistry , Pusan National University, Dental Research Institute , Yangsan , Republic of Korea.
Cheul Hong
CH
Kim
a Department of Dental Anesthesia and Pain Medicine, School of Dentistry , Pusan National University, Dental Research Institute , Yangsan , Republic of Korea.
Seong Soon
SS
Jeong
a Department of Dental Anesthesia and Pain Medicine, School of Dentistry , Pusan National University, Dental Research Institute , Yangsan , Republic of Korea.
Ji Young
JY
Yoon
b Department of Anesthesia and Pain Medicine, School of Medicine , Pusan National University , Yangsan , Republic of Korea.
Ji Uk
JU
Yoon
c Department of Oral Anatomy, School of Dentistry , Pusan National University , Yangsan , Republic of Korea.
Su Bin
SB
Yu
a Department of Dental Anesthesia and Pain Medicine, School of Dentistry , Pusan National University, Dental Research Institute , Yangsan , Republic of Korea.
Eun Jung
EJ
Kim
D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D015603:Keratinocytes / Q000187:drug effects*; D018384:Oxidative Stress / Q000187:drug effects*; D010880:Piperidines / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D000077208:Remifentanil
D010880:Piperidines; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D000077208:Remifentanil
Autophagy; keratinocytes; oxidative stress; remifentanil; wound healing