目的 基于核因子 E2 相关因子 2（Nrf2）/谷胱甘肽过氧化物酶 4（GPX4）铁死亡途径探讨加味桃仁承气汤 对机械通气相关性肺损伤（ventilator-induced lung injury，VILI）大鼠的保护作用。方法 将大鼠随机分为对照 组、模型组、加味桃仁承气汤组、ML385（Nrf2 抑制剂）组、加味桃仁承气汤+ML385 组，每组 12 只。对照组大 鼠进行气管插管，但自主呼吸；其他组大鼠均需进行机械通气 4 h。机械通气前 7 d 进行给药处理，每日 1 次， 连续 7 d。机械通气结束后，ELISA 法检测肺泡灌洗液（BALF）中肿瘤坏死因子 α（TNF-α）、白细胞介素 6（IL-6） 水平；检测大鼠肺组织湿质量/干质量比值变化；检测肺组织病理；试剂盒检测大鼠肺组织中谷胱甘肽（GSH）、 丙二醛（MDA）、Fe2+ 含量；免疫荧光染色法检测肺组织中活性氧（ROS）、4-羟基壬烯醛（4-HNE）相对荧光强 度；检测肺组织中质载体家族 7 成员 11（SLC7A11）、Nrf2、GPX4 mRNA 及蛋白表达。结果 与对照组比较， 模型组大鼠肺组织破坏明显，BALF 中 TNF-α、IL-6 水平升高，肺组织湿质量/干质量比值、MDA、Fe2+ 含量及 ROS、4-HNE 相对荧光强度升高，GSH 含量、Nrf2、SLC7A11、GPX4 mRNA 及蛋白表达降低（P＜0.01）。与模 型组比较，加味桃仁承气汤组大鼠肺组织病理损伤有所改善，BALF 中 TNF-α、IL-6 水平降低，肺组织湿质 量/干质量比值、MDA、Fe2+含量及 ROS、4-HNE 相对荧光强度降低，GSH 含量、Nrf2、SLC7A11、GPX4 mRNA 及蛋白表达升高（P＜0.01）；ML385 组对应指标变化趋势与上述相反（P＜0.01）；加味桃仁承气汤+ML385 组肺组织病理损伤减轻，BALF 中 TNF-α、IL-6 水平降低，肺组织湿质量/干质量比值、MDA、Fe2+ 含量及 ROS、4-HNE 相对荧光强度降低，GSH 含量、Nrf2、SLC7A11、GPX4 mRNA 及 Nrf2、GPX4 蛋白表达升 高（P＜0.01，P＜0.05）。与 ML385 组比较，加味桃仁承气汤+ML385 组肺组织病理损伤有所缓解，BALF 中 TNF-α、IL-6 水平降低，肺组织湿质量/干质量比值、MDA、Fe2+ 含量及 ROS、4-HNE 相对荧光强度降低， GSH 含量、Nrf2、SLC7A11、GPX4 mRNA 及蛋白表达升高（P＜0.01）。与加味桃仁承气汤组比较，加味桃仁承 气汤+ML385 组大鼠肺组织病理损伤加剧，BALF 中 TNF-α、IL-6 水平升高，肺组织湿质量/干质量比值、 MDA、Fe2+ 含量及 ROS、4-HNE 相对荧光强度升高，GSH 含量、Nrf2、SLC7A11、GPX4 mRNA 及蛋白表达降 低（P＜0.01）。结论 加味桃仁承气汤可能通过激活 Nrf2/GPX4 通路抑制铁死亡进而改善大鼠 VILI。

Objective To explore the protective effect of modified Taoren Chengqi Decoction on ventilator-induced lung injury （VILI） in rats based on the nuclear factor-erythroid 2-related factor 2 （Nrf2） /glutathione peroxidase 4 （GPX4） -ferroptosis pathway. Methods Rats were randomly separated into the control group，the model group， the modified Taoren Chengqi Decoction group， the ML385 （Nrf2 inhibitor） group， and the modified Taoren Chengqi Decoction+ML385 group， with 12 rats in each group. The rats in control group underwent tracheal intubation and kept spontaneous breathing. The rats of other groups were subjected to mechanical ventilation for 4 hours. Seven days before mechanical ventilation，medication treatment was carried out once a day for seven days. After mechanical ventilation，ELISA was applied to detect the levels of tumor necrosis factor- α （TNF- α） and interleukin 6 （IL-6） in bronchoalveolar lavage fluid （BALF）. Lung wet/dry weight ratio and lung tissue pathology of rat were detected. The reagent kit was applied to detect the content of glutathione （GSH），malonaldehyde （MDA）， and Fe2+ in rat lung tissue. The relative fluorescence intensity of reactive oxygen species （ROS） and 4- hydroxynonenal（4-HNE） in lung tissue was detected by immunofluorescence staining. The mRNA and protein expressions of solute carrier family 7 member 11 （SLC7A11），Nrf2，GPX4 were detected. Results Compared with the control group，the lung tissue of rats in model group was severely damaged，the levels of TNF-α and IL-6 in BALF increased，lung wet/ dry weight ratio，content of MDA and Fe2+，and the relative fluorescence intensity of ROS and 4-HNE increased，but the content of GSH，the mRNA and protein expressions of Nrf2，SLC7A11，and GPX4 decreased （P＜0.01） . Compared with the model group， the pathological damage of lung tissue in the modified Taoren Chengqi Decoction group was improved，the levels of TNF-α and IL-6 in BALF decreased，lung wet/ dry weight ratio，content of MDA and Fe2+ ，the relative fluorescence intensity of ROS and 4-HNE decreased， the content of GSH， the mRNA and protein expressions of Nrf2， SLC7A11， and GPX4 increased （P＜0.01） . However，an opposite trend for corresponding indicators in the ML385 group was found （P＜0.01）. The pathological injury of lung tissue was alleviated，the levels of TNF-α and IL-6 in BALF decreased，lung wet/ dry weight ratio， content of MDA and Fe2+ ，and the relative fluorescence intensity of ROS and 4-HNE decreased，GSH content，the mRNA expression of Nrf2，SLC7A11 and GPX4，as well as the protein expression of Nrf2 and GPX4 increased in modified Taoren Chengqi Decoction+ML385 group （P＜0.01， P＜0.05）. Compared with ML385 group， the pathological injury of lung tissue was alleviated，the levels of TNF-α and IL-6 in BALF decreased，lung wet/ dry weight ratio，the content of MDA and Fe2+，and the relative fluorescence intensity of ROS and 4-HNE decreased， GSH content， the mRNA and protein expression of Nrf2， SLC7A11 and GPX4 increased in modified Taoren Chengqi Decoction +ML385 group （P＜0.01）. Compared with the modified Taoren Chengqi Decoction group，the pathological damage in lung tissue of rats was intensified，the levels of TNF-α and IL-6 in BALF increased，lung wet/ dry weight ratio， the content of MDA and Fe2+ ， the relative fluorescence intensity of ROS and 4-HNE increased，the content of GSH，the mRNA and protein expressions of Nrf2，SLC7A11，and GPX4 decreased （P＜ 0.01） in modified Taoren Chengqi Decoction+ML385 group. Conclusion Modified Taoren Chengqi Decoction may improve rat VILI by activating the Nrf2/GPX4 pathway to inhibit ferroptosis.

R285.5

河南省卫生健康委国家中医临床研究基地科研专项（2021JDZX2033）