哺乳期全氟辛烷磺酸暴露诱导子代青春期大鼠慢性肺损伤及其机制  

Lactational perfluorooctane sulfonate exposure induces chronic lung injury in adolescent offspring rats and its mechanism

作　　者：覃珊 莫佳丽 张慧珊 叶乐平 QIN Shan;MO Jia-li;ZHANG Hui-shan;YE Le-ping(Department of Pediatrics,Peking University First Hospital,Beijing 100034,China;Department of Pediatrics,Shanghai Children's Medical Center,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China)

机构地区：[1]北京大学第一医院儿科,北京100034 [2]上海交通大学医学院附属上海儿童医学中心,上海200127

出　　处：《中国病理生理杂志》2022年第4期658-664,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81971424;No.81771635)。

摘　　要：目的:探讨gasdermin E(GSDME)介导的细胞焦亡及其引起的细胞因子风暴在哺乳期全氟辛烷磺酸(PFOS)诱导的子代青春期大鼠慢性肺损伤中的作用。方法:24只产后SD大鼠随机分成4组(每组6只):对照组(0 mg·kg^(−1)·d^(−1) PFOS)、低剂量(5 mg·kg^(−1)·d^(−1))PFOS(PFOS-L)组、中剂量(10 mg·kg^(−1)·d^(−1))PFOS(PFOS-M)组和高剂量(15 mg·kg^(−1)·d^(−1))PFOS(PFOS-H)组。从产后1 d(P1)至产后21 d(P21)给予染毒组大鼠相应剂量的PFOS灌胃,仔鼠经哺乳染毒。于出生后35 d(P35)处死仔鼠,收集支气管肺泡灌洗液(BALF)和肺组织;采用质谱法检测肺组织中PFOS含量;肺组织石蜡切片进行HE染色;免疫组化法检测肺组织中GSDME蛋白表达情况;Western blot法检测肺组织中caspase-3、cleaved caspase-3和GSDME蛋白水平;ELISA法检测肺组织白细胞介素18(IL-18)、髓过氧化物酶(MPO)和可溶性白细胞介素2受体(sIL-2R)含量;使用乳酸脱氢酶(LDH)试剂盒检测BALF中LDH活性。结果:PFOS染毒组仔鼠的体重显著低于对照组(P<0.01)。各染毒组仔鼠肺组织内PFOS含量均显著高于对照组(P<0.01)。HE染色结果显示,各染毒组肺组织结构紊乱,支气管内膜和血管壁增厚,肺泡出血,巨噬细胞和淋巴细胞浸润,PFOS-M组和PFOS-H组的肺泡结构被破坏,PFOS-H组的肺泡壁可见胶原沉积。免疫组化结果显示,PFOS-M组和PFOS-H组仔鼠肺组织中GSDME蛋白表达量较对照组显著升高(P<0.01)。Western blot结果显示,与对照组相比,PFOS-M组和PFOS-H组仔鼠肺组织GSDME和caspase-3蛋白表达量显著增高(P<0.01)。ELISA结果显示,PFOS-M组和PFOS-H组仔鼠肺组织中MPO、IL-18和sIL-2R含量较对照组显著升高(P<0.01)。此外,与对照组相比,各染毒组BALF中LDH活性均显著升高(P<0.05)。结论:哺乳期PFOS暴露抑制子代青春期大鼠体重增长,并通过GSDME介导的细胞焦亡及其引起的细胞因子风暴导致肺慢性炎症和慢性肺损伤。AIM:To investigate the effect of gasdermin E(GSDME)-mediated pyroptosis and cytokine storm on chronic lung injury in adolescent offspring rats induced by lactational perfluorooctane sulfonate(PFOS)exposure.METHODS:Twenty-four postpartum SD rats were randomly divided into 4 groups(6 dams per group):control group(0 mg·kg^(−1)·d^(−1) PFOS),low-dose(5 mg·kg^(−1)·d^(−1))PFOS(PFOS-L)group,middle-dose(10 mg·kg^(−1)·d^(−1))PFOS(PFOS-M)group and high-dose(15 mg·kg^(−1)·d^(−1))PFOS(PFOS-H)group.The dams in exposure groups were given PFOS by gavage from postpartum day 1(P1)to 21(P21),and the pups were exposed by breast feeding.The pups were sacrificed at P35,and then the bronchoalveolar lavage fluid(BALF)and lung tissues were collected.The content of PFOS in lung tissues was detected by HPLC-MS.Pathological sections of lung tissues were made for HE staining and lung injury scoring.The protein expression of GSDME in lung tissues was detected by immunohistochemical staining.The protein levels of caspase-3,cleaved caspase-3 and GSDME in lung tissues were detected by Western blot.The content of interleukin-18(IL-18),myeloperoxidase(MPO)and soluble interleukin-2 receptor(sIL-2R)in lung tissues was detected by ELISA,and the activity of lactate dehydrogenase(LDH)in BALF was measured by LDH assay kit.RESULTS:The body weight of off⁃spring rats in PFOS exposure groups was significantly lower than that in control group(P<0.01).The content of PFOS in the lung tissues of offspring rats in PFOS exposure groups was significantly higher than that in control group(P<0.01).The results of HE staining showed that the lung structure in PFOS exposure groups was disordered,accompanied by bron⁃chial intima and vascular wall thickening,alveolar hemorrhage,and macrophage and lymphocyte infiltration.The alveoli of PFOS-M and PFOS-H groups were damaged,and collagen deposition was seen in the alveolar wall of PFOS-H group.In addition,the lung injury score in PFOS exposure groups was significantly higher than that in control

关 键 词：全氟辛烷磺酸 青春期大鼠 肺损伤 Gasdermin E 细胞因子风暴 

分 类 号：R363.2[医药卫生—病理学] R135.2[医药卫生—基础医学]