间充质干细胞通过抑制铁死亡减轻小鼠放射性肺损伤的作用机制研究
Study on the mechanism of the action of mesenchymal stem cells in relieving radiation-induced lung injury in mice by inhibiting ferroptosis
谭鑫 1张沛 2杜乐辉 2解传滨 2曲宝林2
作者信息
- 1. 解放军总医院第一医学中心放射治疗科 北京 100853;解放军医学院 北京 100853
- 2. 解放军总医院第一医学中心放射治疗科 北京 100853
- 折叠
摘要
目的:探讨间充质干细胞(MSCs)通过铁死亡途径对小鼠放射性肺损伤的调控作用.方法:将小鼠在照射前按随机数表法分为正常对照组,MSCs处理组(MSCs组),单纯照射(IR)组(IR组)和IR组联合MSCs组(IR+MSCs组),使用钴60(60Co)γ射线全胸照射(单次20 Gy)构建小鼠放射性肺损伤模型,采用酶联免疫吸附测定(ELISA)法检测小鼠肿瘤坏死因子(TNF-α)和白细胞介素-6(IL-6)水平;采用苏木精-伊红(HE)、Masson染色评估小鼠肺组织损伤程度.使用蛋白免疫印迹法检测小鼠肺组织铁死亡蛋白核因子红细胞相关因子2(Nrf2)、4-羟基壬烯醛(4-HNE)和谷胱甘肽过氧化物酶4(GPX4)表达水平;使用定量聚合酶链式反应(qPCR)检测小鼠肺组织前列腺素内过氧化物合酶2(PTGS2)表达水平;使用二氢乙锭(DHE)检测辐射后小鼠肺组织活性氧(ROS)水平;检测辐射后肺组织的丙二醛(MDA)含量,评估小鼠肺组织氧化损伤水平.结果:IR组照射后小鼠血清中的TNF-α和IL-6水平分别为(768.8±31.42)pg/ml和(161.29±5.75)pg/ml,与对照组相比明显升高,差异有统计学意义(F=53.60、10.65,P＜0.05).HE、Masson染色显示MSCs能够减轻辐射诱导的早期放射性肺炎和晚期肺纤维化.照射后小鼠肺组织中4-HNE的表达水平上调,Nrf2表达下调,MSCs能够降低4-HNE的表达,上调Nrf2的表达;铁死亡基因PTGS2的mRNA表达水平和MDA含量与对照组比较显著升高,而MSCs能够显著降低其表达,差异有统计学意义(F=105.8、7.693,P＜0.05).结论:MSCs能够显著减轻电离辐射诱导的肺上皮细胞铁死亡,进而减轻放射性肺损伤.
Abstract
Objective:To investigate the regulatory role of mesenchymal stem cells(MSCs)on radiation-induced lung injury in mice by the ferroptosis pathway.Methods:The mice were divided into normal control group,MSCs-treated group(MSCs group),single irradiation group(IR group)and IR combined with MSCs group(IR+MSCs group)according to random number table method before irradiation.A mouse model of radiation-induced lung injury was constructed using whole thorax irradiation with cobalt 60(60Co)(20Gy each time),and TNF-α and IL-6 levels of mouse were detected by enzyme-linked immunosorbent assay(ELISA).The injury of lung tissue in mice was assessed using hematoxylin eosin(HE)staining and Masson staining.Western Blot was used to examine the expression levels of ferroptosis-related proteins in lung tissue,including nuclear factor erythroid NF-E2 related factor 2(Nrf2),4-Hydroxynonenal(4-HNE)and glutathione peroxidase 4(GPX4).The expression level of prostaglandin-endoperoxide synthase 2 Gene(PTGS2)in the lung tissue of mice was detected by using quantitative polymerase chain reaction(qPCR),and the level of reactive oxygen species(ROS)in the lung tissue of mice was detected after radiation by using dihydroethidium(DHE),and the level of malondialdehyde(MDA)content in the lung tissue was detected after radiation.And then,the level of oxidative damage in the lung tissue of mice was assessed.Results:Elisa results showed the serum TNF-α and IL-6 levels in mice after irradiation in IR group were significantly higher than those in normal control group(F=53.60,10.65,P＜0.05),respectively.The HE and MSCs staining of pathological analysis showed that MSCs treatment could significantly relieve both early radiation-induced pneumonitis and advanced pulmonary fibrosis.After radiation,the 4-HNE expression level was upregulation and the Nrf2 expression level was downregulation in the lung tissues of mice,whereas MSCs were able to significantly reduce the 4-HNE expression and upregulate the Nrf2 expression.The mRNA expression level and MDA content of the ferroptosis gene PTGS2 were significantly increased,which were significantly higher than those of normal control group,while MSCs were able to significantly reduce its expression,and the differences were statistically significant(F=105.8,7.693,P＜0.05).Conclusion:MSCs is able to relieve significantly ionizing radiation-induced ferroptosis of lung epithelial cells,thereby relieve radiation-induced lung injury.
关键词
间充质干细胞(MSCs)/放射性肺损伤/铁死亡/氧化应激Key words
Mesenchymal stem cells(MSCs)/Radiation-induced lung injury/Ferroptosis/Oxidative stress引用本文复制引用
基金项目
军队后勤基础加强计划技术领域基金(2021-JCJQ-JJ-1082)
出版年
2024
国家科技期刊平台
NSTL
万方数据