TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, China; College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, China; College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
J Ethnopharmacol. 2025 Jan 10;336:118726. doi: 10.1016/j.jep.2024.118726. Epub 2024 Aug 23.
Sea buckthorn (Hippophae rhamnoides), a traditional Tibetan medicinal herb, exhibits protective effects against cardiovascular and respiratory diseases. Although Sea buckthorn extract (SBE) has been confirmed to alleviate airway inflammation in mice, its therapeutic effect and underlying mechanism on chronic obstructive pulmonary disease (COPD) requires further clarification.
To elucidate the alleviative effect and molecular mechanism of SBE on lipopolysaccharides (LPS)/porcine pancreatic elastase (PPE)-induced COPD by blocking ferroptosis.
The anti-ferroptotic effects of SBE were evaluated in human BEAS-2B bronchial epithelial cells using CCK8, RT-qPCR, western blotting, and transmission electron microscopy. Transwell was employed to detect chemotaxis of neutrophils. COPD model was induced by intranasally administration of LPS/PPE in mice and measured by alterations of histopathology, inflammation, and ferroptosis. RNA-sequencing, western blotting, antioxidant examination, flow cytometry, DARTS, CETSA, and molecular docking were then used to investigate its anti-ferroptotic mechanisms.
In vitro, SBE not only suppressed erastin- or RSL3-induced ferroptosis by suppressing lipid peroxides (LPOs) production and glutathione (GSH) depletion, but also suppressed ferroptosis-induced chemotactic migration of neutrophils via reducing mRNA expression of chemokines. In vivo, SBE ameliorated LPS/PPE-induced COPD phenotypes, and inhibited the generation of LPOs, cytokines, and chemokines. RNA-sequencing showed that p53 pathway and mitogen-activated protein kinases (MAPK) pathway were implicated in SBE-mediated anti-ferroptotic action. SBE repressed erastin- or LPS/PPE-induced overactivation of p53 and MAPK pathway, thereby decreasing expression of diamine acetyltransferase 1 (SAT1) and arachidonate 15-lipoxygenase (ALOX15), and increasing expression of glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11). Mechanistically, erastin-induced elevation of reactive oxygen species (ROS) was reduced by SBE through directly scavenging free radicals, thereby contributing to its inhibition of p53 and MAPK pathways. CETSA, DARTS, and molecular docking further showed that ROS-generating enzyme nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4) may be the target of SBE. Overexpression of NOX4 partially impaired the anti-ferroptotic activity of SBE.
Our results demonstrated that SBE mitigated COPD by suppressing p53 and MAPK pro-ferroptosis pathways via directly scavenging ROS and blocking NOX4. These findings also supported the clinical application of Sea buckthorn in COPD therapy.
沙棘（Hippophae rhamnoides），一种传统的藏药草药，具有预防心血管和呼吸系统疾病的作用。尽管沙棘提取物（SBE）已被证实可减轻小鼠气道炎症，但它在慢性阻塞性肺疾病（COPD）中的治疗效果和潜在机制仍需进一步阐明。
通过阻断铁死亡来阐明 SBE 对脂多糖（LPS）/猪胰弹性蛋白酶（PPE）诱导的 COPD 的缓解作用及其分子机制。
采用 CCK8、RT-qPCR、western blot 和透射电镜评估 SBE 对人 BEAS-2B 支气管上皮细胞的抗铁死亡作用。Transwell 用于检测嗜中性粒细胞的趋化性。通过组织病理学、炎症和铁死亡的改变，在小鼠中通过鼻内给予 LPS/PPE 来诱导 COPD 模型，并进行测量。然后使用 RNA 测序、western blot、抗氧化检测、流式细胞术、DARTS、CETSA 和分子对接来研究其抗铁死亡机制。
体外，SBE 通过抑制脂质过氧化物（LPOs）的产生和谷胱甘肽（GSH）的耗竭，不仅抑制了 erastin 或 RSL3 诱导的铁死亡，还通过降低趋化因子的 mRNA 表达抑制了铁死亡诱导的嗜中性粒细胞趋化性迁移。体内，SBE 改善了 LPS/PPE 诱导的 COPD 表型，并抑制了 LPOs、细胞因子和趋化因子的产生。RNA 测序显示，p53 途径和丝裂原活化蛋白激酶（MAPK）途径参与了 SBE 介导的抗铁死亡作用。SBE 抑制了 erastin 或 LPS/PPE 诱导的 p53 和 MAPK 途径的过度激活，从而降低了二胺乙酰转移酶 1（SAT1）和花生四烯酸 15-脂氧合酶（ALOX15）的表达，增加了谷胱甘肽过氧化物酶 4（GPX4）和溶质载体家族 7 成员 11（SLC7A11）的表达。机制上，SBE 通过直接清除自由基减少了 erastin 诱导的活性氧（ROS）的升高，从而抑制了 p53 和 MAPK 途径。CETSA、DARTS 和分子对接进一步表明，烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶 4（NOX4）可能是 SBE 的靶点。NOX4 的过表达部分削弱了 SBE 的抗铁死亡活性。
我们的研究结果表明，SBE 通过直接清除 ROS 和阻断 NOX4 抑制 p53 和 MAPK 促铁死亡途径来减轻 COPD。这些发现也支持沙棘在 COPD 治疗中的临床应用。
