阿米福汀对小鼠放射性肺损伤的防护作用研究  

Protective effect of Amifostine against radiation-induced lung injury in mice and its mechanism

作　　者：宣伟[1] 李帅[1] 吴秀艳[2] 耿艳[1] 张卓[1] Wei Xuan;Shuai Li;Xiu-yan Wu;Yan Geng;Zhuo Zhang(Department of Radiotherapy the Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116023, China;Department of Medical Oncology, the Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116023, China)

机构地区：[1]大连医科大学附属第二医院肿瘤放疗科,辽宁大连116023 [2]大连医科大学附属第二医院肿瘤科,辽宁大连116023

出　　处：《中国现代医学杂志》2018年第2期26-32,共7页China Journal of Modern Medicine

基　　金：辽宁省科学技术计划项目(No:2014225003)

摘　　要：目的探讨阿米福汀(Amifostine,AMI)对小鼠放射性肺损伤(RILI)的防护作用机制。方法将24只雌性C57BL/6J小鼠随机分为对照组、单纯照射组、AMI组。用直线加速器6MV-X射线单次12Gy照射小鼠全肺,照射前30 min腹腔注射AMI,对照组和单纯照射组注射同等剂量的生理盐水。照射14 d后,留取小鼠肺组织标本,采用苏木精-伊红染色法(HE)观察病理改变;酶联免疫吸附法(ELISA)检测支气管肺泡灌洗液中白介素6(IL-6)、肿瘤坏死因子-α(TNF-α)、转化生长因子-β_1(TGF-β_1)表达水平;凝胶电泳迁移实验检测核转录因子-κB(NF-κB)活性;免疫组织化学法观察NLRP3蛋白的表达和定位;实时荧光定量聚合酶链反应检测肺组织中NLRP 3和IL-1βm RNA的表达;Western blot检测NF-κB p65、NLRP3、IL-1β蛋白的表达。结果照射后第14天,AMI组与单纯照射组比较,AMI组肺组织急性炎症反应减轻;AMI组支气管肺泡灌洗液中IL-6、TNF-α水平下降,TGF-β_1水平升高(P<0.05);AMI组肺组织中NF-κB活性下降(P<0.05);AMI组肺组织中NLRP3蛋白表达下降(P<0.05);AMI组NLRP3和IL-1βm RNA表达下降(P<0.05);AMI组NF-κB p65、NLRP3、IL-1β蛋白表达下降(P<0.05)。结论 AMI可能通过抑制辐射引起的NF-κB激活,进而抑制NLRP3基因的转录和表达,抑制炎症因子的释放,减轻RILI。Objective To investigate the radioprotective function of Amifostine(AMI)in mice with acute radiation-induced lung injury and its mechanism.Methods Totally24female C57BL/6J mice were randomized into3groups as AMI group(treated by AMI200mg/kg plus radiation),radiation group and solvent control group.The mouse lungs in the radiation group and the AMI group were irradiated with linear accelerator6MV X-ray at a single dose of12Gy,and those in the solvent control group received sham radiation.The mice in the AMI group were intraperitoneally injected with Amifostine30minutes before irradiation,while the same volume of solvent was given to the control and radiation groups.The mice were sacrified and the mouse lung tissue was collected14days after irradiation.The pathological changes in the lung tissue were observed after HE staining.The levels of IL-6,TNF-αand TGF-β1in the bronchoalveolar lavage fluid(BALF)were measured by ELISA.The NF-κB activity was detected by EMSA.The expression and positioning of nucleotide-binding domain and leucine-rich repeat containing protein3(NLRP3)in the lung tissue were observed by immunohistochemical method.The expressions of NLRP3and IL-1βmRNAs in the lung tissue were assayed by qRT-PCR.And the expressions of NF-κB p65,NLRP3and IL-1βproteins in the lung tissue were assayed by Western blot.Results On the14th day after irradiation,compared with the radiation group,acute inflammatory reaction of the lung tissue was alleviated,the IL-6and TNF-αlevels in BALF were markedly decreased(P<0.05),the level of TGF-β1in BALF rose slightly(P<0.05),and NF-κB activity in the lung tissue was obviously reduced(P<0.05)in the AMI group.Immunohistochemical results showed that the expressions of NLRP3and IL-1βmRNAs were obviously reduced(P<0.05),and the expressions of NF-κB p65,NLRP3and IL-1βproteins in the lung tissue were obviously reduced(P<0.05)in the AMI group compared to the radiation group.Conclusions Amifostine can down-regulate the activitity of NF–κB,then inhibit transcription and expre

关 键 词：阿米福汀 放射性肺损伤 NF-ΚB NLRP3 白介素-1Β 

分 类 号：R818.74[医药卫生—放射医学]