达泮舒腈(OLT1177)减轻急性放射性肺损伤及机制初探

Dapansutrile(OLT1177)alleviates acute radiation⁃induced lung injury

目的研究核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体抑制剂达泮舒腈(OLT1177)对急性放射性肺损伤的治疗作用及其机制。方法将小鼠分为对照组、注射OLT1177组、照射组及照射后注射OLT1177组,采用单次22 Gy全肺60Co射线放射建立急性放射性肺损伤模型。建模6 h后腹腔给药OLT1177(100 mg/kg,1次/d)。连续给药14 d后,取小鼠肺组织并称重,计算并统计小鼠肺系数;苏木素-伊红染色(HE)和F4/80免疫组化染色观察小鼠肺组织病理变化和炎性细胞浸润情况;实时定量PCR(qPCR)检测肺组织NLRP3、白细胞介素-1β(IL-1β)等mRNA的转录水平;流式微球阵列(CBA)蛋白定量技术检测小鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)等炎性因子的分泌;Western印迹检测含半胱氨酸的天冬氨酸蛋白水解酶-1(Caspase-1)和IL-18蛋白的激活。结果放射导致小鼠肺组织急性炎症的发生,表现为肺组织水肿以及肺泡结构破坏,炎性细胞浸润增多,肺组织炎症相关基因NLRP3、IL-1β、TNF-α、IL-6表达及血清TNF-α、IL-6等炎症因子水平增加。用OLT1177处理可以显著改善放射引起的上述症状。OLT1177抑制放射导致的NLRP3炎症小体下游Caspase-1及其底物IL-18的剪切活化。结论OLT1177显著减轻小鼠急性放射性肺损伤,可能是通过抑制放射导致的NLRP3炎症小体活化。

Objective To investigate the therapeutic effect and mechanism of NLRP3 inflammasome inhibitordapansutrile(OLT1177)-against acute radiation lung injury.Methods Mice were divided into the control group,OLT1177 injection group,irradiation group,and irradiation+OLT1177 injection group.A single dose of 22 Gy whole-lung 60Co radiation was used to establish a model of acute radiation lung injury.After 6 h of radiation,OLT1177(100 mg/kg,once daily)was administered intraperitoneally.After 14 consecutive days of administration,lung tissues were collected and weighed while the lung coefficient was calculated.Hematoxylin-eosin(HE)staining and F4/80 immunohistochemical staining were used to observe the pathological changes and inflammatory cell infiltration in lung tissues.Real-time quantitative PCR(qPCR)was used to detect the transcription levels of NLRP3,IL-1β,and other mRNAs in lung tissues.Serum cytokines such as TNF-αand IL-6 were measured by cytometric bead array(CBA).The activation of Caspase-1 and IL-18 was detected by Western blotting.Results Radiation caused acute inflammation in the lung tissues of mice,manifested as edema in the lung tissues and destruction of the alveolar structure,increased macrophage infiltration,and elevated expressions of inflammatory genes NLRP3,IL-1β,TNF-α,and IL-6 in the lung tissues and higher serum levels of TNF-α,IL-6.Treatment with OLT1177 significantly improved the above symptoms induced by radiation.OLT1177 inhibited the activation of NLRP3 inflammasome downstream Caspase-1 and IL-18 induced by radiation.Conclusion OLT1177 can significantly alleviate acute radiation lung injury in mice,which may be due to its inhibition of NLRP3 inflammasome activation induced by radiation.