程序性死亡受体1抑制剂通过促进细胞焦亡加重放射性心肌损伤的研究  

Effect of PD-1 inhibitor on aggravating radiation-induced myocardial injury by catalyzing pyroptosis

作　　者：赵沙沙 吴碧波 柏杰 王刚 刘瑶[1,2,3] 王羽 胡银祥[1,2,3] 欧阳伟炜 卢冰[1,2,3] 苏胜发 Zhao Shasha;Wu Bibo;Bai Jie;Wang Gang;Liu Yao;Wang Yu;Hu Yinxiang(Department of Oncology,Afiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Department of Oncology,Afiliated Cancer Hospital of Guizhou Medical University,Guiyang 550004,China;Department of Oncology,School of Clinical Medicine,Guizhou Medical University,Guiyang)

机构地区：[1]贵州医科大学附属医院肿瘤科,贵阳550004 [2]贵州医科大学附属肿瘤医院肿瘤科,贵阳550004 [3]贵州医科大学肿瘤学教研室,贵阳550004

出　　处：《中华放射医学与防护杂志》2024年第4期255-261,共7页Chinese Journal of Radiological Medicine and Protection

基　　金：贵州省基础研究计划重点项目ZK((2022)重点040);国家自然科学基金(81960548)。

摘　　要：目的探讨程序性死亡受体1(PD-1)抑制剂通过细胞焦亡通路对放射性心肌损伤的影响。方法20只C57BL/6雄性小鼠按随机数字表法分为4组:健康对照组;PD-1抑制剂组;单纯照射组(胸部单次照射20 Gy);照射联合PD-1抑制剂组,每组5只。超声心动测试评价受照后1个月小鼠左室射血分数(LVEF)、每搏输出量(SV)、左室缩短率(FS);采用苏木精-伊红(HE)、Masson染色观察心肌组织病理学改变;蛋白免疫印迹、实时荧光定量反转录聚合酶链式反应(RT-qPCR)检测心肌组织焦亡关键因子半胱氨酸天冬氨酸蛋白水解酶1(Caspase-1)、消皮素D(GSDMD)、凋亡相关斑点样蛋白(ASC)表达水平;酶联免疫吸附实验(ELISA)检测白介素18(IL-18)、IL-1β表达;流式细胞术检测心肌淋巴细胞浸润情况。结果照射后1个月,照射联合PD-1抑制剂组的LVEF、FS、SV较单纯照射组下降(t=4.50、27.93、3.11,P<0.05);照射联合PD-1抑制剂组小鼠心肌损伤及纤维化较单纯照射组更明显,心肌胶原容积分数较单纯照射组升高[(2.88±0.27)%vs.(3.81±0.57)%,t=2.90,P<0.05];与单纯照射组比较,照射联合PD-1抑制剂组心肌Caspase-1、Caspase-1 p20、GSDMD、GSDMD-N、ASC蛋白表达量增高(t=3.14、3.22、8.83、20.29、2.79,P<0.05),Caspase-1、GSDMD、ASC的mRNA表达量也升高(t=3.09、2.91、2.53,P<0.05);照射联合PD-1抑制剂组心肌及血清IL-18、IL-1β表达较单纯照射组均升高(t=3.46、3.75、7.58、8.24,P<0.05),CD8+T淋巴细胞百分比高于单纯照射组[(38.33±7.92)%vs.(54.70±4.01)%,t=3.29,P<0.05],而CD4+T淋巴细胞在各组间的差异无统计学意义(P>0.05)。结论PD-1抑制剂可通过Caspase-1/GSDMD促进细胞焦亡,进而加重放射性心肌损伤。Objective To investigate the effects of programmed cell death protein-1(PD-1)inhibitor on radiation-induced myocardial injury through the pyroptosis pathway.Methods Twenty male C57BL/6 mice were randomized into four groups,namely the control group,the PD-1 inhibitor group,the cardiac radiation group(with a single dose of 20 Gy),and the combined group treated with both cardiac radiation and PD-1 inhibitor,with five mice in each group.At 1 month post-irradiation,echocardiography was performed to assess left ventricular ejection fraction(LVEF),stroke volume(SV),and left ventricular fractional shortening(LVFS);Hematoxylin-eosin(HE)and Masson staining were employed to observe the pathological changes in myocardial tissue;Western blot and real-time reverse transcription-quantitative polymerase chain reaction(RT-qPCR)were used to determine the expression of pyroptosis-related key factors in myocardial tissue such as cysteine aspartate specific proteinase-1(Caspase-1),gasdermin D(GSDMD),and apoptosis-associated speck-like protein containing CARD(ASC).Enzyme-linked immunosorbent assay(ELISA)was performed to determine the expression of interleukin-18(IL-18)and interleukin-1β(IL-1β).Furthermore,flow cytometry was used to evaluate the infiltration of myocardial lymphocytes.Results Compared to the radiation group,the mice in the combined group exhibited lower LVEF,FS and SV(t=4.50,27.93,3.11,P<0.05),more significant myocardial injury and fibrosis,elevated myocardial collagen volume fraction[(2.88±0.27)%vs.(3.81±0.57)%,t=2.90,P<0.05)],increased expression of Caspase-1,Caspase-1 p20,GSDMD,GSDMD-N,and ASC(t=3.14,3.22,8.83,20.29,2.79,P<0.05),increased mRNA expression of Caspase-1,GSDMD and ASC(t=3.09,2.91,2.53,P<0.05),increased expression of IL-18 and IL-1βin myocardium and serum(t=3.46,3.75,7.58,8.24,P<0.05),and higher percentage of CD8+T lymphocytes[(38.33±7.92)%vs.(54.70±4.01)%,t=3.29,P<0.05].However,there was no significant difference in the percentage of CD4+T lymphocytes among the groups(P>0.05).Conclusions PD-1 inhi

关 键 词：程序性死亡受体1抑制剂 焦亡 心肌损伤 放射 

分 类 号：R730.55[医药卫生—肿瘤]