Open AccessAspirin induces Nrf2‐mediated transcriptional activation of haem oxygenase‐1 in protection of human melanocytes from H 2 O 2 ‐induced oxidative stress
Author(s) -
Jian Zhe,
Tang Lingzhen,
Yi Xiuli,
Liu Bangmin,
Zhang Qian,
Zhu Guannan,
Wang Gang,
Gao Tianwen,
Li Chunying
Publication year - 2016
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/jcmm.12812
Subject(s) - oxidative stress , viability assay , reactive oxygen species , apoptosis , chemistry , intracellular , antioxidant , pharmacology , microbiology and biotechnology , biochemistry , biology
The removal of hydrogen peroxide (H 2 O 2 ) by antioxidants has been proven to be beneficial to patients with vitiligo. Aspirin (acetylsalicylic acid, ASA ) has antioxidant activity and has great preventive and therapeutical effect in many oxidative stress‐relevant diseases. Whether ASA can protect human melanocytes against oxidative stress needs to be further studied. Here, we investigated the potential protective effect and mechanisms of ASA against H 2 O 2 ‐induced oxidative injury in human melanocytes. Human melanocytes were pre‐treated with different concentrations of ASA , followed by exposure to 1.0 mM H 2 O 2 . Cell apoptosis, intracellular reactive oxygen species ( ROS ) levels were evaluated by flow cytometry, and cell viability was determined by an Cell Counting Kit‐8 assay. Total and phosphorylated NRF 2 expression, NRF 2 nuclear translocation and antioxidant response element ( ARE ) transcriptional activity were assayed with or without Nrf2‐si RNA transfection to investigate the possible molecular mechanisms. Concomitant with an increase in viability, pre‐treatment of 10‐90 μmol/l ASA resulted in decreased rate of apoptotic cells, lactate dehydrogenase release and intracellular ROS levels in primary human melanocytes. Furthermore, we found ASA dramatically induced NRF 2 nuclear translocation, enhanced ARE ‐luciferase activity, increased both p‐ NRF 2 and total NRF 2 levels, and induced the expression of haem oxygenase‐1 ( HO ‐1) in human melanocytes. In addition, knockdown of Nrf2 expression or pharmacological inhibition of HO ‐1 abrogated the protective action of ASA on melanocytes against H 2 O 2 ‐induced cytotoxicity and apoptosis. These results suggest that ASA protects human melanocytes against H 2 O 2 ‐induced oxidative stress via Nrf2‐driven transcriptional activation of HO ‐1.