摘要
目的:探讨敲除糖基转移酶N-乙酰氨基葡萄糖转移酶(Ogt)加重小鼠缺血再灌注诱导肺损伤及其背后机制。方法:取6~8周龄Ogt条件性敲除的小鼠(Ogt-CKO)及未使用他莫昔芬诱导杂交同窝鼠(Ogtfl/fl)各12只,根据处理方式不同——假手术(sham)和缺血再灌注(I/R),分成4组(n=6):sham+Ogtfl/fl;I/R+Ogtfl/fl;sham+OgtCKO;I/R+Ogt-CKO。建立I/R模型后,光镜下观察HE染色后肺组织病理改变;电镜下观察肺上皮细胞超微结构改变;免疫荧光染色后分析活性氧(ROS);采用Western Blot测定肺组织中SLC7A11、G6PDH及Nrf2表达。结果:与sham+Ogtfl/fl组相比,肺损伤在I/R+Ogtfl/fl组小鼠加重,电镜下表现出细胞缩小、核固缩,ROS增多,SLC7A11表达减少,G6PDH及Nrf2表达增多。敲除Ogt进一步加重I/R造成的肺组织损伤和铁死亡。结论:Ogt的敲除加重了缺血再灌注诱导的铁死亡,这一过程可能与Nrf2/G6PDH有关。
Objective:To investigate the influence of O⁃GlcNAc transferase(Ogt)deficiency on ischemia/reperfusion(I/R)⁃induced ferroptosis and the mechanism behind it.Methods:Twelve Ogt conditional knockout mice with tamoxifen treatment(Ogt⁃CKO)and 12 Ogt⁃conditional knockout mice without tamoxifen treatment(Ogtfl/fl),all aging from 6 to 8 weeks,were divided into four groups according to different treatments(sham treatment or I/R treatment):sham+Ogtfl/fl,I/R+Ogtfl/fl,sham+Ogt⁃CKO,and I/R+Ogt⁃CKO.The model of lung I/R was induced by occluding the left pulmonary hi⁃lum for 1 h followed by 4 h reperfusion.In the sham group,mice underwent left thoracotomies with⁃out I/R followed by ventilation for 5 h.The mice were sacrificed after 4 h reperfusion or 5 h ventilation.By using HE staining,lung tissue sections were observed under the light microscope,and the pathological injury scores were calculated.The morphology of cells was observed under the transmis⁃sion electron microscope.The level of ROS was identified by immunofluorescent staining.By using Western Blot,the expressions of SLC7A11,G6PDH,and Nrf2 were detected.Results:Compared with the group of sham+Ogtfl/fl,the pathological injury scores of lung tissue,the expression of G6PDH,Nrf2,and the production of ROS were increased,and the expression of SLC7A11 was de⁃creased.Moreover,smaller pulmonary epithelial cells and karyopyknosis occurred in I/R+Ogtfl/fl group.Compared with the I/R+Ogtfl/fl group,the pathological injury scores of lung tissue and the lev⁃els of ROS were markedly increased,the damages of epithelial cells were more obvious,and the ex⁃pression of SLC7A11,G6PDH,and Nrf2 were significantly decreased in I/R+Ogt⁃CKO group.Conclusion:Ogt deficiency aggravated pulmonary ischemia⁃reperfusion induced ferroptosis via Nrf2/G6PDH pathway.
作者
杨柳青
彭君
桑阿明
张静
李心怡
YANG Liuqing;PENG Jun;SANG Aming;ZHANG Jing;LI Xinyi(Wuhan University School of Medicine,Wuhan 430071,Hubei,China;Dept.of Anesthesiology,Zhongnan Hospital of Wuhan University,Wuhan 430071,Hubei,China)
出处
《武汉大学学报（医学版）》
CAS
2024年第3期253-258,共6页
Medical Journal of Wuhan University
基金
国家自然科学基金资助项目(编号:82002033)。
