摘要
目的 基于谷胱甘肽过氧化物酶4(GPX4)信号通路探讨银杏叶提取物(GBE)对慢性阻塞性肺疾病(COPD)大鼠肺组织铁死亡的影响。方法 本实验时间为2022年6月—2023年6月。选取2月龄SPF级雄性Wistar大鼠40只,采用抽签法将大鼠分为空白组、模型组、罗红霉素组、GBE组,每组10只。模型组、罗红霉素组、GBE组大鼠采用脂多糖联合香烟烟雾构建COPD大鼠模型,造模成功后,空白组和模型组大鼠给予0.9%氯化钠溶液灌胃,罗红霉素组大鼠给予罗红霉素分散片0.017 5 g·kg^(-1)·d^(-1)灌胃,GBE组大鼠给予GBE 14 mg·kg^(-1)·d^(-1)腹腔注射。麻醉并处死各组大鼠,取腹主动脉血液和肺组织。观察各组大鼠肺组织HE染色结果,采用酶联免疫吸附试验(ELISA)检测各组大鼠血清肿瘤坏死因子α(TNF-α)、白介素6(IL-6)水平,采用qRT-PCR法检测各组大鼠肺组织GPX4、铁蛋白重链1(FTH1)、转铁蛋白(TF)mRNA相对表达量,采用Western blot法检测各组大鼠肺组织GPX4、FTH1、TF表达量。结果 HE染色结果显示,与空白组相比,模型组大鼠肺组织中可见较多充血以及水肿,炎性细胞浸润,支气管变形,肺泡破裂,较多肺大疱生成;与模型组相比,罗红霉素组与GBE组大鼠肺组织中炎性细胞减少,肺泡结构完整,肺组织结构相对完整。模型组大鼠血清TNF-α、IL-6水平高于空白组(P<0.05);罗红霉素组、GBE组大鼠血清TNF-α、IL-6水平低于模型组(P<0.05);罗红霉素组与GBE组大鼠血清TNF-α、IL-6水平比较,差异无统计学意义(P>0.05)。模型组大鼠肺组织GPX4、FTH1 mRNA相对表达量低于空白组,TF mRNA相对表达量高于空白组(P<0.05);罗红霉素组、GBE组大鼠肺组织GPX4、FTH1 mRNA相对表达量高于模型组,TF mRNA相对表达量低于模型组(P<0.05);罗红霉素组与GBE组大鼠肺组织GPX4、FTH1、TF mRNA相对表达量比较,差异无统计学意义(P>0.05)。模型组大鼠肺组织GPX4、FTH1表达量低于空白组,TF表达量高于空白组(P<0.05);罗红霉素组、GBE组大鼠肺组织GPX4、FTH1表达量高于模型组,TF表达量低于模型组(P<0.05);罗红霉素组与GBE组大鼠肺组织GPX4、FTH1、TF表达量比较,差异无统计学意义(P>0.05)。结论 COPD的发生与肺组织铁死亡有关。GBE可通过GPX4信号通路降低炎症因子水平,提高FTH1基因及蛋白表达水平,降低TF基因及蛋白表达水平,从而抑制COPD大鼠肺组织铁死亡。
Objective To explore the effect of ginkgo biloba extract(GBE)on ferroptosis in lung tissue of rats with chronic obstructive pulmonary disease(COPD)based on glutathione peroxidase 4(GPX4)signaling pathway.Methods The experiment was conducted from June 2022 to June 2023.Forty male Wistar rats of SPF grade at 2 months of age were selected and divided into blank group,model group,roxithromycin group,and GBE group by drawing method,with 10 rats in each group.The COPD rat model was constructed in the rats in the model group,roxithromycin group,and GBE group through lipopolysaccharide combined with cigarette smoke.After successful modeling,the rats in the blank group and model group were given 0.9%sodium chloride solution by gavage,while the rats in the roxithromycin group were given roxithromycin dispersible tablets 0.0175 g·kg^(-1)·d^(-1) by gavage,and the rats in the GBE group were given GBE 14 mg·kg^(-1)·d^(-1) by intraperitoneal injection.The rats in each group were anesthetically sacrificed and the abdominal aorta blood and lung tissues were collected.The results of HE staining of lung tissues of rat in each group were observed,and the levels of serum tumor necrosis factor alpha(TNF-α)and interleukin-6(IL-6)of rats in each group were detected by enzyme-linked immunosorbent assay(ELISA).The relative expression levels of GPX4,ferritin heavy chain 1(FTH1),and transferrin(TF)mRNA in lung tissues of rats in each group were detected by qRT-PCR,and the expression levels of GPX4,FTH1,and TF in lung tissues of rats in each group were detected by Western blot.Results The results of HE staining showed that,compared with the blank group,more congestion and edema,inflammatory cell infiltration in the lung tissue,bronchial deformation,alveolar rupture,and more pulmonary bullae were observed in the model group;compared with the model group,the inflammatory cells in the lung tissue of rats decreased,the alveolar structure was intact,and the lung tissue structure was relatively complete in the roxithromycin group and GBE group.The levels of serum TNF-αand IL-6 in model group were higher than those in blank group(P<0.05);the levels of serum TNF-αand IL-6 in roxithromycin group and GBE group were lower than those in model group(P<0.05);there was no statistically significant difference in the levels of serum TNF-αand IL-6 between erythromycin group and GBE group(P>0.05).The relative expression levels of GPX4 and FTH1 mRNA in model group were lower than those in blank group,relative expression level of TF mRNA was higher than that in blank group(P<0.05);the relative expression levels of GPX4 and FTH1 mRNA in roxithromycin group and GBE group were higher than those in model group,relative expression level of TF mRNA was lower than that in model group(P<0.05);there was no statistically significant difference in the relative expression levels of GPX4,FTH1 and TF mRNA between erythromycin group and GBE group(P>0.05).The expression levels of GPX4 and FTH1 in model group were lower than those in blank group,expression level of TF was higher than that in blank group(P<0.05);the expression levels of GPX4 and FTH1 in roxithromycin group and GBE group were higher than those in model group,expression level of TF was lower than that in model group(P<0.05);there was no statistically significant difference in the expression levels of GPX4,FTH1 and TF between erythromycin group and GBE group(P>0.05).Conclusion The occurrence of COPD is related to ferroptosis in lung tissues.GBE can reduce the level of inflammatory factors,increase the expression level of FTH1 gene and protein,and decrease the expression level of TF gene and protein through the GPX4 signaling pathway,thereby regulating ferroptosis in lung tissues of COPD rats.
作者
黄海屏
裴凯
邱璇
陈佳民
方勇
韩晓瑜
李苏舒
HUANG Haiping;PEI Kai;QIU Xuan;CHEN Jiamin;FANG Yong;HAN Xiaoyu;LI Sushu(Respiratory and Critical Care Medicine Department,Liuzhou Traditional Chinese Medicine Hospital,Liuzhou 545001,China)
出处
《实用心脑肺血管病杂志》
2024年第7期64-70,共7页
Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease
基金
广西自然科学基金资助项目(2020GXNSFBA297022)
柳州市科技计划项目(2022SB029)
广西中医药大学联合基金项目(2023L2007)。
关键词
肺疾病
慢性阻塞性
银杏叶
铁死亡
谷胱甘肽过氧化物酶4
Pulmonary disease,chronic obstructive
Ginkgo folium
Ferroptosis
Glutathione peroxidase 4
作者简介
通信作者:李苏舒,E-mail:Lisushu1025@163.com。
