Protective Effects of Oenothera biennis against Hydrogen Peroxide-Induced Oxidative Stress and Cell Death in Skin Keratinocytes

Seung Young Lee; Chul Hwan Kim; Buyng Su Hwang; Kyung-Min Choi; In-Jun Yang; Gi-Young Kim; Yung Hyun Choi; Cheol Park; Jin-Woo Jeong

doi:10.3390/life10110255

Protective Effects of Oenothera biennis against Hydrogen Peroxide-Induced Oxidative Stress and Cell Death in Skin Keratinocytes

Life (Basel). 2020 Oct 27;10(11):255. doi: 10.3390/life10110255.

Authors

Seung Young Lee¹, Chul Hwan Kim¹, Buyng Su Hwang¹, Kyung-Min Choi¹, In-Jun Yang², Gi-Young Kim³, Yung Hyun Choi⁴, Cheol Park⁵, Jin-Woo Jeong¹

Affiliations

¹ Nakdonggang National Institute of Biological Resources, 137, Donam 2-gil, Sangju-si, Gyeongsangbuk-do 37242, Korea.
² Department of Physiology, College of Oriental Medicine, Dongguk University, Gyeongju 780-714, Korea.
³ Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju 63243, Korea.
⁴ Department of Biochemistry, Dong-eui University College of Korean Medicine, Busan 47227, Korea.
⁵ Division of Basic Sciences, College of Liberal Studies, Dong-eui University, Busan 47340, Korea.

Abstract

Background: Oenothera biennis (evening primrose) produces bioactive substances with a diverse range of pharmacological functions. However, it is currently unknown whether extract prepared from the aerial parts of O. biennis (APOB) can protect the skin against oxidative stress.

Objective: The aim of this study is to investigate the protective effects of APOB against oxidative stress-induced damage in human skin keratinocytes (HaCaT) and elucidate the underlying mechanisms.

Methods: We pretreated HaCaT cells with various concentrations of APOB or the antioxidant N-acetyl-L-cysteine before applying H₂O₂. We then compared the cell viability, intracellular reactive oxygen species (ROS) production, and DNA and mitochondrial damage between pretreated and untreated control cells using a range of assays, flow cytometry, and Western blot analysis and also examined the reducing power and DPPH free radical scavenging activity of APOB.

Results: APOB pretreatment significantly increased cell viability, effectively attenuated H₂O₂-induced comet tail formation, and inhibited H₂O₂-induced phosphorylation of the histone γH2AX, as well as the number of apoptotic bodies and Annexin V-positive cells. APOB was found to have high reducing power and DPPH radical scavenging activity and also exhibited scavenging activity against intracellular ROS accumulation and restored the loss of mitochondrial membrane potential caused by H₂O₂. APOB pretreatment almost totally reversed the enhanced cleavage of caspase-3, the degradation of poly (ADP-ribose)-polymerase (PARP), DNA fragmentation that usually occurs in the presence of H₂O₂, and increased the levels of heme oxygenase-1 (HO-1), a potent antioxidant enzyme that is associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2).

Conclusions: APOB can protect HaCaT cells from H₂O₂-induced DNA damage and cell death by blocking cellular damage related to oxidative stress via a mechanism that affects ROS elimination and by activating the Nrf2/HO-1 signaling pathway.

Keywords: Nrf2/HO-1; Oenothera biennis; cell death; evening primrose; oxidative stress.

Grants and funding

NNIBR2020002106/This work was supported by a grant from the Nakdonggang National Institute of Biological Resources (NNIBR), funded by the Ministry of Environment (MOE) of the Republic of Korea