C-EPO预处理对创伤性脑损伤合并海水淹溺大鼠肺组织Nrf2/GPX4通路及铁死亡的影响  

Effect of carbamylated erythropoietin on Nrf2/GPX4 pathway and ferroptosis in lung tissue of rats with traumatic brain injury combined with seawater drowning

作　　者：张灏 叶丹[2] 潘振洪 魏梁锋 徐永君 吴再新 王守森 Zhang Hao;Ye Dan;Pan Zhenhong;Wei Liangfeng;Xu Yongjun;Wu Zaixin;Wang Shousen(Department of Neurosurgery,Fuzong Clinical Medical college of Fujian Medical University(No.900 hospital of PLA),Fuzhou 350025,China;Department of Neurosurgery,Fuzong Teaching Hospital of Fujian University of Traditional Chinese Medicine(No.900 hospital of PLA),Fuzhou 350025,China;Laboratory of Basic Medicine,No.900 hospital of PLA,Fuzhou 350025,China;Department of Military Medical Service,No.900 hospital of PLA,Fuzhou 350025,China)

机构地区：[1]福建医科大学福总临床医学院(联勤保障部队第九〇〇医院)神经外科,福州350025 [2]福建中医药大学福总教学医院(联勤保障部队第九〇〇医院)神经外科,福州350025 [3]联勤保障部队第九〇〇医院基础医学实验室,福州350025 [4]联勤保障部队第九〇〇医院卫勤部,福州350025

出　　处：《中华航海医学与高气压医学杂志》2023年第6期709-714,共6页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：全军后勤科技重大项目子课题(AHJ14J001);联勤保障部队第九〇〇医院院内课题(2022ZL01,2022ZL03)。

摘　　要：目的探究氨甲酰化促红细胞生成素(C-EPO)对创伤性脑损伤(TBI)合并海水淹溺(SWD)大鼠肺组织核因子E2相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)通路及铁死亡的影响。方法选取72只SD大鼠按随机数字表法分为假手术(sham)组、TBI+SWD组、C-EPO组和C-EPO+ML385(Nrf2抑制剂)组各18只(其中每组6只大鼠为备用)。采用控制性皮质撞击(CCI)颅脑致伤法+气管内泵入海水法(3 ml/kg)建立TBI+SWD大鼠模型。每24 h腹腔注射50 μg/kg的C-EPO。Nrf2抑制剂采用ML385, 30 mg/kg, 1次/d。对各组大鼠进行神经功能评分与肺组织含水量测定, HE染色观察肺形态, 普鲁士蓝染色观察肺组织中铁沉积情况, 分光光度计法测定肺组织中铁、丙二醛(MDA)、谷胱甘肽(GSH)含量及GPX4活性, qPCR法检测肺组织中Nrf2 mRNA和GPX4 mRNA表达水平。结果 4组大鼠造模后24 h神经功能评分差异有统计学意义(F=21.30, P<0.001), TBI+SWD组、C-EPO+ML385组大鼠神经功能评分均低于sham组、C-EPO组(P<0.05)。4组大鼠造模后24 h肺组织含水量差异有统计学意义(F=31.21, P<0.001), TBI+SWD组、C-EPO组、C-EPO+ML385组大鼠含水量高于sham组, TBI+SWD组、C-EPO+ML385组高于C-EPO组(P<0.05)。4组大鼠造模后24 h肺组织中铁沉积量差异有统计学意义(F=872.1, P<0.001), TBI+SWD组大鼠肺组织中铁沉积量多于sham组和C-EPO组(P<0.05)。4组大鼠造模后24 h肺组织中铁、MDA、GSH含量及GPX4活性差异均有统计学意义(F =748.18、453.24、121.37、1 042.31, 均P<0.001);TBI+SWD组、C-EPO+ML385组大鼠肺组织中铁和MDA含量均高于sham组、C-EPO组, GSH含量和GPX4活性均低于sham组、C-EPO组(P<0.05)。4组大鼠造模后24 h肺组织中Nrf2 mRNA和GPX4 mRNA表达水平差异均有统计学意义(F=96.24、432.49, 均P<0.001);TBI+SWD组、C-EPO+ML385组大鼠肺组织中Nrf2 mRNA及GPX4 mRNA表达水平均低于sham组、C-EPO组(P<0.05)。结论 C-EPO可能通过调控Nrf2/GPX4信号轴抑制TBI+SWD大鼠肺组织�Objective To investigate the effects of carbamylated erythropoietin(C-EPO)on the nuclear factor E2-related factor 2(NRF2)/glutathione peroxidase 4(GPX4)pathway and ferroptosis in the lung tissue of rats with traumatic brain injury(TBI)combined with seawater drowning(SWD).Methods A total of 48 Sprague-Dawley(SD)rats were randomly divided into four groups,according to random number table method,i.e.,sham group,TBI+SWD group,C-EPO group,and C-EPO+ML385 group,with 18 rats in each group(each group has six spare rats).The TBI+SWD rat model was established using the controlled cortical impact(CCI)method followed by seawater instillation(3 ml/kg)through the trachea.C-EPO was administered by intraperitoneal injection at a dose of 50μg/kg every 24 hours;and ML385,the Nrf2 inhibitor,was administered at a dose of 30 mg/kg once daily.The neurological function scores and lung tissue water contents were assessed;lung morphology was observed by HE staining;iron deposition in lung tissue was evaluated by Prussian blue staining;and the contents of iron,malondialdehyde(MDA),and glutathione(GSH)and GPX4 activity in lung tissue were measured by spectrophotometric assay.The expression levels of Nrf2 mRNA and GPX4 mRNA in the lung tissue were assessed by quantitative PCR(qPCR).Results The differences in neurological function scores at 24 hours post-injury were statistically significant among the four groups(F=21.30,P<0.001),with the TBI+SWD group and the C-EPO+ML385 group showing lower scores compared with the sham group and the C-EPO group(P<0.05).Lung tissue water contents at 24 hours post-injury also exhibited significant differences among the four groups(F=31.21,P<0.001),with the TBI+SWD group,the C-EPO group,and the C-EPO+ML385 group displaying higher water contents than the sham group,and the TBI+SWD group and the C-EPO+ML385 group displaying higher water contents than the C-EPO group(P<0.05).Iron deposition in the lung tissue at 24 hours post-injury showed significant differences among the four groups(F=872.1,P<0.001),with the

关 键 词：创伤性脑损伤 海水淹溺 氨甲酰化促红细胞生成素 铁死亡 大鼠 

分 类 号：R651.15[医药卫生—外科学] R649.3[医药卫生—临床医学]