College of Science, Department of Biological Sciences, King Faisal University, Al Ahsa, 31982., Saudi Arabia.
Department of Molecular Physiology, Zoology Department, Faculty of Science, Assiut University, Egypt.
Oxid Med Cell Longev. 2021 Apr 23;2021:5585303. doi: 10.1155/2021/5585303. eCollection 2021.
Atherosclerosis is a persistent inflammatory disorder specified by the dysfunction of the arteries, the world's leading cause of cardiovascular diseases. We sought to determine the effectiveness of KRL in B[a]P-induced oxidative stress and programmed cell death in endothelial cells. Western blotting, real-time PCR, DCFH2-DA, and TUNEL staining were performed to detect pPI3K, pAKT, Nrf2, HO-1, NQO-1, Bcl2, Bax, and caspase-3 on the HUVECs. Through the pretreatment of KRL, a drastic enhancement was observed in the cell viability of HUVECs, whereas DNA damage and generation of reactive oxygen species induced by B[a]P was suppressed. KRL's potential use as an antioxidant was observed to have a direct correlation with an antioxidant gene's augmented expression and the nuclear translocation activation of Nrf2, even during the event when B[a]P was found to be absent. In addition, this study proved that the signaling cascades of PI3K/AKT mediated Nrf2 translocation. Activation of suppressed nuclear Nrf2 and reduced antioxidant genes across cells interacting with an LY294002 confirmed this phenomenon. In addition, knockdown of Nrf2 by Nrf2-siRNA transfection abolished the protective effects of KRL on HUVECs cells against oxidative damage. Finally, the expression of apoptotic proteins also supported the hypothesis that KRL may inhibit endothelial dysfunction. This study showed that KRL potentially prevents B[a]P-induced redox imbalance in the vascular endothelium by inducing the Nrf2 signaling via the PI3K/AKT pathway.
动脉粥样硬化是一种以动脉功能障碍为特征的持续性炎症性疾病，是心血管疾病的主要全球病因。我们旨在确定 KRL 在 B[a]P 诱导的氧化应激和内皮细胞程序性细胞死亡中的作用。通过 Western blot、实时 PCR、DCFH2-DA 和 TUNEL 染色检测 HUVECs 中的 pPI3K、pAKT、Nrf2、HO-1、NQO-1、Bcl2、Bax 和 caspase-3。通过 KRL 的预处理，观察到 HUVECs 的细胞活力显著增强，而 B[a]P 诱导的 DNA 损伤和活性氧的产生受到抑制。观察到 KRL 作为抗氧化剂的潜在用途与抗氧化基因表达的增强以及 Nrf2 的核易位激活直接相关，即使在不存在 B[a]P 的情况下也是如此。此外，这项研究证明了 PI3K/AKT 介导的 Nrf2 易位信号级联。与 LY294002 相互作用的细胞中核 Nrf2 抑制和抗氧化基因减少的激活证实了这一现象。此外，通过 Nrf2-siRNA 转染敲低 Nrf2 消除了 KRL 对 HUVECs 细胞氧化损伤的保护作用。最后，凋亡蛋白的表达也支持了 KRL 可能抑制血管内皮功能障碍的假说。这项研究表明，KRL 通过 PI3K/AKT 通路诱导 Nrf2 信号，可能预防 B[a]P 诱导的血管内皮氧化还原失衡。
