Liu Junya, Qin Song, Feng Banghai, Chen Miao, Mei Hong
Department of Critical Care Unit, Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou, China.
Department of Critical Care Medicine, Zunyi City Hospital of Traditional Chinese Medicine, Zunyi 563000, Guizhou, China. Corresponding author: Mei Hong, Email:
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2023 Nov;35(11):1177-1181. doi: 10.3760/cma.j.cn121430-20230324-00212.
To study whether wedelolactone can reduce hyperoxia-induced acute lung injury (HALI) by regulating ferroptosis, and provide a basic theoretical basis for the drug treatment of HALI.
A total of 24 C57BL/6J mice were randomly divided into normal oxygen control group, HALI model group and wedelolactone pretreatment group, with 8 mice in each group. Mice in wedelolactone pretreatment group were treated with wedelolactone 50 mg/kg intraperitoneally for 6 hours, while the other two groups were not given with wedelolactone. After that, the HALI model was established by maintaining the content of carbon dioxide < 0.5% and oxygen > 90% in the molding chamber for 48 hours, and the normal oxygen control group was placed in indoor air. After modeling, the mice were sacrificed and lung tissues were collected. The lung histopathological changes were observed under light microscope and pathological scores were performed to calculate the ratio of lung wet/dry mass (W/D). The levels of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-1β), superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) in lung tissues of mice in each group were determined. The protein expression of glutathione peroxidase 4 (GPX4) in lung tissue was detected by Western blotting.
Under light microscope, the alveolar structure of HALI model group was destroyed, and a large number of neutrophils infiltrated the alveolar and interstitial lung, and the interstitial lung was thickened. The pathological score of lung injury (score: 0.75±0.02 vs. 0.11±0.01) and the ratio of lung W/D (6.23±0.34 vs. 3.68±0.23) were significantly higher than those in the normal oxygen control group (both P < 0.05). Wedelolactone pretreated mice had clear alveolar cavity and lower neutrophil infiltration and interstitial thickness than HALI group. Pathological scores (score: 0.43±0.02 vs. 0.75±0.02) and W/D ratio (4.56±0.12 vs. 6.23±0.34) were significantly lower than HALI group (both P < 0.05). Compared with the normal oxygen control group, the levels of SOD (kU/g: 26.41±4.25 vs. 78.64±3.95) and GSH (mol/g: 4.51±0.33 vs. 12.53±1.25) in HALI group were significantly decreased, while the levels of MDA (mmol/g: 54.23±4.58 vs. 9.65±1.96), TNF-α (μg/L: 96.32±3.67 vs. 11.65±2.03), IL-6 (ng/L: 163.35±5.89 vs. 20.56±3.63) and IL-1β (μg/L: 72.34±4.64 vs. 15.64±2.47) were significantly increased, and the protein expression of GPX4 (GPX4/β-actin: 0.44±0.02 vs. 1.00±0.09) was significantly decreased (all P < 0.05). Compared with the HALI group, the levels of SOD (kU/g: 53.28±3.69 vs. 26.41±4.25) and GSH (mol/g: 6.73±0.97 vs. 12.53±1.25) were significantly higher in the wedelolactone pretreatment group, and the levels of MDA (mmol/g: 25.36±1.98 vs. 54.23±4.58), TNF-α (μg/L: 40.25±4.13 vs. 96.32±3.67), IL-6 (ng/L: 78.32±4.65 vs. 163.35±5.89), and IL-1β (μg/L: 30.65±3.65 vs. 72.34±4.64) were significantly lower (all P < 0.05), and protein expression of GPX4 was significantly higher (GPX4/β-actin: 0.68±0.04 vs. 0.44±0.02, P < 0.05).
Wedelolactone attenuates HALI injury by regulating ferroptosis.
研究水飞蓟宾葡甲胺是否能通过调节铁死亡来减轻高氧诱导的急性肺损伤（HALI），为HALI的药物治疗提供基础理论依据。
将24只C57BL/6J小鼠随机分为正常氧对照组、HALI模型组和水飞蓟宾葡甲胺预处理组，每组8只。水飞蓟宾葡甲胺预处理组小鼠腹腔注射50mg/kg水飞蓟宾葡甲胺6小时，其余两组不给予水飞蓟宾葡甲胺。之后，通过在成型箱中维持二氧化碳含量<0.5%、氧气含量>90% 48小时建立HALI模型，正常氧对照组置于室内空气中。建模后处死小鼠并收集肺组织。在光学显微镜下观察肺组织病理变化并进行病理评分，计算肺湿/干质量（W/D）比值。测定每组小鼠肺组织中肿瘤坏死因子-α（TNF-α）、白细胞介素（IL-6、IL-1β）、超氧化物歧化酶（SOD）、丙二醛（MDA）和谷胱甘肽（GSH）的水平。通过蛋白质印迹法检测肺组织中谷胱甘肽过氧化物酶4（GPX4）的蛋白表达。
光学显微镜下，HALI模型组肺泡结构破坏，大量中性粒细胞浸润肺泡和肺间质，肺间质增厚。肺损伤病理评分（评分：0.75±0.02 vs. 0.11±0.01）和肺W/D比值（6.23±0.34 vs. 3.68±0.23）显著高于正常氧对照组（均P<0.05）。水飞蓟宾葡甲胺预处理的小鼠肺泡腔清晰，中性粒细胞浸润和间质厚度低于HALI组。病理评分（评分：0.43±0.02 vs. 0.75±0.02）和W/D比值（4.56±0.12 vs. 6.23±0.34）显著低于HALI组（均P<0.05）。与正常氧对照组相比，HALI组SOD（kU/g：26.41±4.25 vs. 78.64±3.95）和GSH（mol/g：4.51±0.33 vs. 12.53±1.25）水平显著降低，而MDA（mmol/g：54.23±4.58 vs. 9.65±1.96）、TNF-α（μg/L：96.32±3.67 vs. 11.65±2.03）、IL-6（ng/L：163.35±5.89 vs. 20.56±3.63）和IL-1β（μg/L：72.34±4.64 vs. 15.64±2.47）水平显著升高，GPX4蛋白表达（GPX4/β-肌动蛋白：0.44±0.02 vs. 1.00±0.09）显著降低（均P<0.05）。与HALI组相比，水飞蓟宾葡甲胺预处理组SOD（kU/g：53.28±3.69 vs. 26.41±4.25）和GSH（mol/g：6.73±0.97 vs. 12.53±1.25）水平显著升高，MDA（mmol/g：25.36±1.98 vs. 54.23±4.58）、TNF-α（μg/L：40.25±4.13 vs. 96.32±3.67）、IL-6（ng/L：78.32±4.65 vs. 163.35±5.89）和IL-1β（μg/L：30.65±3.65 vs. 72.34±4.64）水平显著降低（均P<0.05），GPX4蛋白表达显著升高（GPX4/β-肌动蛋白：0.68±0.04 vs. 0.44±0.02，P<0.05）。
水飞蓟宾葡甲胺通过调节铁死亡减轻HALI损伤。
