Protective effects ofNigella sativaextract against H₂O₂‐induced cell death through the inhibition of DNA damage and cell cycle arrest in human umbilical vein endothelial cells (HUVECs)

Abstract

Oxidative stress is known to induce cytotoxicity and apoptosis in endothelial cells and indorse development of atherosclerosis. The aim of this research was to assess the cytoprotective effects of ethanolic extract of Nigella sativa (NSE) against H2O2‐induced cell death in human umbilical vein endothelial cells (HUVECs) and also study the probable mechanisms through which NSE exhibited cyto‐protection. The cytotoxicity was measured by exposing the HUVECs with NSE (10–200 μg/ml) and H2O2 (25–1000 μM) for 24 h. Then, the HUVECs were pretreated with noncytotoxic doses (10–50 μg/ml) of NSE for 24 h before administration of 200 μM H2O2 for 24 h. The MTT, NRU, and morphological assays were performed to assess the cytotoxicity and cyto‐protection. Potential antioxidant activity of NSE on oxidative stress marker (glutathione [GSH] and lipid peroxidation [LPO]) was also evaluated. The fluorescence probe, DCF‐DA, and Rh123 were applied to measure the reactive oxygen species (ROS) level and mitochondrial membrane potential. Moreover, flow cytometric analysis and comet assay were used to study the cell cycle arrest and DNA damage, respectively. The concentrations (10, 30, and 50 μg/ml) of NSE were found to protect HUVECs against H2O2 (200 μM)‐induced cytotoxicity in HUVECs. Pretreatment of HUVECs with NSE significantly reduced the LPO and ROS levels and restored the GSH and loss of MMP induced by H2O2. Furthermore, NSE inhibited H2O2‐induced cell cycle arrest and cellular DNA damage in HUVECs. Altogether, these results suggest that NSE can prevent H2O2‐induced cell death, and NSE could be a potential candidate that can prevent HUVECs against toxicants.

Nigella sativa L. Phytochemistry and Pharmacological Activities: A Review (2019–2021)

“…In addition to the mentioned effects, anti-hemolytic activity has also been noticed [ 31 ] ( Table 1 ). These results are in accordance with those obtained in the study of Farshori et al, 2021 [ 32 ] where the pretreatment of human umbilical vein endothelial cells (HUVEC) with different doses of the ethanolic extract of the NS seeds protected the cells from the negative effect of the H 2 O 2 which confirms the antioxidant effect of the plant. The radiations have an enormous interactive capacity with human tissue and can cause free radical generation by radiolysis of water molecules which induce damage at the level of the DNA and cell death.…”

Arzanol, a natural phloroglucinol α‐pyrone, protects HaCaT keratinocytes against H2O2‐induced oxidative stress, counteracting cytotoxicity, reactive oxygen species generation, apoptosis, and mitochondrial depolarization

“…A dose‐dependent decrease in HaCaT cell viability was previously reported in HaCaT cells exposed to H 2 O 2 (100–500 μM) for 12 h, by MTT assay, with a 50% cell mortality at the concentration of 350 μM H 2 O 2 (Zhang et al, 2020), whereas a survival rate of about 60% was reported by Park et al (2020) for HaCaT cells exposed to 500 μM H 2 O 2 for 24 h. In our study, the H 2 O 2 cytotoxic effect was also confirmed by microscopic observation, which allowed to assess the presence of apoptotic features and decreased cell density. Similar morphological alterations were observed in other cell types exposed to H 2 O 2 ‐damage, such as human umbilical vein endothelial cells, neuroblastoma cell line SK‐N‐MC, and HepG2 cells (Bayati et al, 2011; Farshori et al, 2021; Liao et al, 2022). Pre‐treatment with arzanol preserved HaCaT cells from H 2 O 2 cytotoxicity, and significant greater cell viability was observed in arzanol‐treated cells in comparison with those no‐treated exposed to the highest concentrations of oxidants.…”

“…A dose-dependent decrease in HaCaT cell viability was previously reported in HaCaT cells exposed to H 2 O 2 (100-500 μM) for 12 h, by MTT assay, with a 50% cell mortality at the concentration of 350 μM H 2 O 2 (Zhang et al, 2020), whereas a survival rate of about 60% was reported by Park et al (2020) for HaCaT cells exposed to 500 μM H 2 O 2 for 24 h. In our study, the H 2 O 2 cytotoxic effect was also confirmed by microscopic observation, which allowed to assess the presence of apoptotic features and decreased cell density. Similar (Bayati et al, 2011;Farshori et al, 2021;Liao et al, 2022 (Park et al, 2020).…”

Protective effects of fruit extract of Rosa canina and quercetin on human umbilical vein endothelial cell injury induced by hydrogen peroxide

“…It is well established that oxidation of low‐density lipoprotein (LDL) particles contributes to the formation of oxidized LDL, which can be cytotoxic and cause the initial disruption of endothelial cells and subsequent injury to vascular walls (Siti et al, 2015 ; Tesfamariam & DeFelice, 2007 ). Previous studies showed several natural active substances had protective effect on vascular endothelial cells (Farshori et al, 2021 ; Guo et al, 2021 ; Liao et al, 2021 ). Numerous studies have shown that antioxidants originating from plants, such as vitamins E and C, tocopherols, β‐carotene, and polyphenols, may be protective against cardiovascular disorders (Siti et al, 2015 ; Wang et al, 2013 ).…”

“…It is well established that oxidation of low-density lipoprotein (LDL) particles contributes to the formation of oxidized LDL, which can be cytotoxic and cause the initial disruption of endothelial cells and subsequent injury to vascular walls (Siti et al, 2015;Tesfamariam & DeFelice, 2007). Previous studies showed several natural active substances had protective effect on vascular endothelial cells (Farshori et al, 2021;Guo et al, 2021;Liao et al, 2021).…”