X线辐照激活NLRP3炎性体引起损伤肺组织细胞焦亡(pyroptosis)  

NLRP3 inflammasome induces pyroptosis in lung tissues of radiation-induced lung injury in mice

作　　者：韩蓉[1] 吴东明[1] 邓仕华 刘腾[1] 张婷 许颖[1] 

机构地区：[1]成都医学院第一附属医院,四川成都610500

出　　处：《细胞与分子免疫学杂志》2017年第9期1206-1211,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：四川省教育厅科研创新项目(17TD0012);四川省卫生和计划生育委员会(16PJ112;16ZD040);四川省医学科研青年创新课题(Q16015);成都医学院科研创新团队(CYTD15-03)

摘　　要：目的建立放射性肺损伤模型,研究胱天蛋白酶1(caspase-1)依赖的程序性细胞死亡即细胞焦亡(pyroptosis)在放射性肺炎发病机制中的作用。方法 BALB/c小鼠胸腔15 Gy X线辐照5 d,处死小鼠。HE染色观察肺组织形态变化,原位末端转移酶标记技术(TUNEL)检测辐照后肺组织细胞凋亡情况,Western blot法检测肺组织γ组蛋白2AX(γ-H2AX)、抗原KI-67(ki67)、含pyrin结构域NOD样受体家族3(NLRP3)、caspase-1和含CARD结构域凋亡相关斑点样蛋白(ASC/TMS-1)的表达情况,实时荧光定量PCR检测肺组织白细胞介素6(IL-6)、IL-8、肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)、含pyrin结构域NOD样受体家族3(NLRP3)、caspase-1、白细胞介素1β(IL-1β)、IL-18的m RNA水平,采用免疫组织化学染色检测肺组织NLRP3、caspase-1和TMS1的表达情况,使用caspase-1活性检测试剂盒检测caspase-1活性;ELISA检测血清IL-1β和IL-18水平。结果辐照后小鼠肺泡壁毛细血管扩张充血,炎性细胞浸润,肺泡壁增厚;肺组织细胞TUNEL染色阳性指数升高,γ-H2AX和ki67的表达增强,提示细胞DNA遭到破坏且细胞增殖活性降低;肺组织细胞中IL-6、IL-8、TNF-α和MCP-1 m RNA水平增加;血清IL-1β和IL-18水平增加,肺组织NLRP3、caspase-1、ASC/TMS-1表达水平增加,caspase-1活性增强。结论辐照后,激活小鼠肺组织NLRP3炎性体诱导肺组织的细胞发生pyroptosis。Objective To establish a radiation-induced lung injury model and investigate the role of caspase-1-dependent programmed cell death（ pyroptosis） in the pathogenesis of radiation pneumonitis. Methods BALB/c mice were sacrificed after receiving 5-day 15 Gy X-ray irradiation at chest cavity. The pathological changes of pulmonary tissues were observed by HE staining. The apoptosis of lung tissues cells after irradiation was detected by TUNEL assay. The expressions of γ-H2 AX,ki67,NLR family pyrin domain containing 3（ NLRP3）,caspase-1,apoptosis-associated speck-like protein containing a CARD（ ASC/TMS-1） were detected by Western blot analysis. Real-time quantitative PCR was used to check m RNA levels of interleukin-6（ IL-6）,IL-8,tumor necrosis factor α（ TNF-α）,monocyte chemoattractant protein 1（ MCP-1）,NLRP3,caspase-1,IL-1β and IL-18. Immunohistochemical staining was used to determine the expressions of NLRP3,caspase-1 and TMS1 in lung tissues. The activity of caspase-1 was evaluated by caspase-1 assay kit,and the serum levels of IL-1β and IL-18 were detected by ELISA. Results After irradiation,the capillaries of the alveolar wall of the mice were dilated and congested,inflammatory cells infiltrated,the alveolar wall thickened. Positive rate of lung tissue cells was raised in TUNEL staining. The expressions of γ-H2 AX and ki67 were elevated,indicating that DNA damage and cell proliferation activity decreased in lung tissues. The m RNA levels of IL-6,IL-8,TNF-α and MCP-1 in lung cells increased; the serum levels of IL-1β and IL-18 increased; the expressions of IL-1β,IL-18,NLRP3,caspase-1 and ASC/TMS-1 in lung tissues were enhanced; and caspase-1 activity increased. Conclusion After irradiation,the pyroptosis caused by the activation of NLRP3 inflammatory body occurred in the lung tissue of mice.

关 键 词：放射性肺炎 胱天蛋白酶1依赖的程序性细胞死亡 细胞焦亡(pyroptosis) 含pyrin结构域NOD样受体家族3(NLRP3) 炎性体 胱天蛋白酶1(caspase-1) 

分 类 号：R563[医药卫生—呼吸系统] R392-33[医药卫生—内科学]