Department of Biochemistry, College of Medicine, and Jeju Research Center for Natural Medicine, Jeju National University, Jeju 63243, Republic of Korea.
Mol Med Rep. 2024 Jul;30(1). doi: 10.3892/mmr.2024.13244. Epub 2024 May 17.
Physiological stress such as excessive reactive oxygen species (ROS) production may contribute normal fibroblasts activation into cancer‑associated fibroblasts, which serve a crucial role in certain types of cancer such as pancreatic, breast, liver and lung cancer. The present study aimed to examine the cytoprotective effects of luteolin (3',4',5,7‑tetrahydroxyflavone) against hydrogen peroxide (HO)‑generated oxidative stress in lung fibroblasts. To examine the effects of luteolin against HO‑induced damages, cell viability, sub‑G cell population, nuclear staining with Hoechst 33342, lipid peroxidation and comet assays were performed. To evaluate the effects of luteolin on the protein expression level of apoptosis, western blot assay was performed. To assess the antioxidant effects of luteolin, detection of ROS using HDCFDA staining, O‑ and ·OH using electron spin resonance spectrometer and antioxidant enzyme activity was performed. In a cell‑free chemical system, luteolin scavenges superoxide anion and hydroxyl radical generated by xanthine/xanthine oxidase and the Fenton reaction (FeSO/HO). Furthermore, Chinese hamster lung fibroblasts (V79‑4) treated with HO showed a significant increase in cellular ROS. Intracellular ROS levels and damage to cellular components such as lipids and DNA in HO‑treated cells were significantly decreased by luteolin pretreatment. Luteolin increased cell viability, which was impaired following HO treatment and prevented HO‑mediated apoptosis. Luteolin suppressed active caspase‑9 and caspase‑3 levels while increasing Bcl‑2 expression and decreasing Bax protein levels. Additionally, luteolin restored levels of glutathione that was reduced in response to HO. Moreover, luteolin enhanced the activity and protein expressions of superoxide dismutase, catalase, glutathione peroxidase, and heme oxygenase‑1. Overall, these results indicated that luteolin inhibits HO‑mediated cellular damage by upregulating antioxidant enzymes.
生理应激如过量活性氧（ROS）的产生可能导致正常成纤维细胞激活为癌相关成纤维细胞，其在某些类型的癌症如胰腺癌、乳腺癌、肝癌和肺癌中起着至关重要的作用。本研究旨在研究木樨草素（3'，4'，5，7-四羟基黄酮）对过氧化氢（HO）诱导的肺成纤维细胞氧化应激的细胞保护作用。为了研究木樨草素对 HO 诱导损伤的作用，进行了细胞活力、亚 G1 细胞群体、Hoechst 33342 核染色、脂质过氧化和彗星试验。为了评估木樨草素对凋亡蛋白表达水平的影响，进行了 Western blot 分析。为了评估木樨草素的抗氧化作用，通过 HDCFDA 染色检测 ROS、电子自旋共振光谱仪检测 O 和·OH 以及抗氧化酶活性检测。在细胞外化学系统中，木樨草素清除黄嘌呤/黄嘌呤氧化酶和 Fenton 反应（FeSO/HO）产生的超氧阴离子和羟基自由基。此外，HO 处理的中国仓鼠肺成纤维细胞（V79-4）显示细胞内 ROS 显著增加。HO 预处理显著降低了 HO 处理细胞中细胞内 ROS 水平和脂质和 DNA 等细胞成分的损伤。木樨草素增加了细胞活力，而 HO 处理后细胞活力受损，并阻止了 HO 介导的细胞凋亡。木樨草素抑制活性 caspase-9 和 caspase-3 水平，同时增加 Bcl-2 表达并降低 Bax 蛋白水平。此外，木樨草素恢复了对 HO 反应降低的谷胱甘肽水平。此外，木樨草素增强了超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶和血红素加氧酶-1 的活性和蛋白表达。总的来说，这些结果表明木樨草素通过上调抗氧化酶抑制 HO 介导的细胞损伤。
