Protective Effects ofagainst Hydrogen Peroxide-Induced Oxidative Stress and Cell Death in Skin Keratinocytes.
Life (Basel). 2020 Oct 27 ;10(11). Epub 2020 Oct 27. PMID: 33120909
Seung Young Lee
BACKGROUND: (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(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 HO. 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 HO-induced comet tail formation, and inhibited HO-induced phosphorylation of the histoneγH2AX, 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 HO. 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 HO, 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 HO-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.