细粒棘球蚴囊液致敏过程中IL⁃1β受体阻滞剂对肺损伤的治疗作用  

Therapeutic effects of IL⁃1βreceptor blocker on lung injury during the sensitization process of Echinococcus granulosus cyst fluid

作　　者：王春生 乌尔格力 西利扎提·库来西 李玉倩 先依旦·阿布拉 苏比·泰来提 蒲雪莉 王佳玲 李孟 房志远 叶建荣[1] WANG Chunsheng;WU Ergeli;XILIZATI Kulaixi;LI Yuqian;XIANYIDAN Abula;SUBI Tailaiti;PU Xueli;WANG Jialing;LI Meng;FANG Zhiyuan;YE Jianrong(Department of Anesthesiology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830000,China;Department of Anesthesia and Surgery,Changji People’s Hospital,Changji 831100,Xinjiang,China;Department of Anesthesiology,Dongguan First Hospital Affiliated to Guangdong Medical University,Dongguan 523710,China)

机构地区：[1]新疆医科大学第一附属医院麻醉科,乌鲁木齐830000 [2]昌吉市人民医院麻醉手术部,新疆昌吉831100 [3]广东医科大学附属东莞第一医院麻醉科,东莞523710

出　　处：《中国寄生虫学与寄生虫病杂志》2024年第2期199-203,210,共6页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(82060581);新疆围术期器官保护实验室(XJDX1411);自治区重点实验室开放课题(2020D04025)。

摘　　要：目的探究细粒棘球蚴囊液外溢致敏过程中白细胞介素-1β(IL-1β)受体阻滞剂对肺损伤的治疗作用机制。方法细粒棘球蚴包囊采集自新疆乌鲁木齐华凌屠宰场感染细粒棘球蚴的新鲜羊肝,收集包囊囊液,消化沉淀获得原头节,上清经内毒素处理获得致敏囊液。将24只小鼠随机分为对照组、致敏组、阻滞剂治疗组和阻滞剂预防组,每组6只。致敏组、阻滞剂治疗组和阻滞剂预防组小鼠分别腹腔注射约2000个细粒棘球蚴原头节,对照组小鼠腹腔注射等体积生理盐水(约1 ml)。3个月后致敏组小鼠腹腔注射致敏囊液(0.1 ml/g体质量),阻滞剂治疗组小鼠先腹腔注射致敏囊液、15 min后尾静脉注射IL-1β受体阻滞剂(4μg/g体质量),阻滞剂预防组小鼠先尾静脉注射阻滞剂、15 min后腹腔注射致敏囊液,对照组小鼠腹腔注射等体积生理盐水。取小鼠肺组织,制备石蜡切片后苏木精-伊红(HE)染色,显微镜下观察肺组织的炎症损伤情况。提取小鼠肺组织总RNA,qRT-PCR检测肿瘤坏死因子α(TNF-α)、IL-6、半胱氨酸天冬氨酸蛋白酶-8(caspase-8)、磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(Akt)、核因子-κB(NF-κB)的mRNA相对转录水平。提取小鼠肺组织蛋白,以β肌动蛋白(β-actin)为内参,蛋白质免疫印迹(Western blotting)检测PI3K、Akt、NF-κB的蛋白相对表达水平。应用GraphPad Prism软件对数据进行统计学分析,组间比较采用单因子方差分析。结果HE染色结果显示,致敏组小鼠肺组织局部充血,充血部位周围炎症细胞游出、淋巴细胞聚集;阻滞剂治疗组、阻滞剂预防组和对照组肺组织均未见明显炎症反应。qRT-PCR结果显示,对照组小鼠肺组织中IL-6、TNF-α、caspase-8、PI3K、Akt和NF-κB的mRNA相对转录水平分别为1.057±0.363、1.020±0.217、1.004±0.097、1.000±0.031、1.035±0.312、1.029±0.304,致敏组分别为2.013±0.514、2.189±0.194、6.433±0.340、1.594±0.117、2.902±0.Objective To investigate the therapeutic mechanism of interleukin-1β(IL-1β)receptor blocker on lung injury in the sensitization process of Echinococcus granulosus cyst extravasated fluid.Methods The cysts of E.granulosus were collected from fresh sheep livers infected with E.granulosus from Hualing Slaughterhouse of Urumqi,Xinjiang.The cyst fluid was collected,digested and sedimented to obtain protoscoleces,and the supernatant was treated with endotoxin to obtain allergenic cyst fluid.Twenty-four mice were randomly divided into control group,sensitized group,blocker treatment group and blocker prevention group,with 6 mice in each group.Mice in the sensitized group,the blocker treatment group and the blocker prevention group were intraperitoneally injected with about 2000 protoscoleces,respectively,while mice in the control group were injected with about the same volume(1 ml)of normal saline.Three months later,mice in the sensitized group were intraperitoneally injected with allergenic cyst fluid(0.1 ml per g weight);mice in the blocker treatment group were injected with allergenic cyst fluid prior to intravenous injection with IL-1βblocker(4μg per g weight)15 minutes later;mice in the blocker prevention group were injected with IL-1βblocker prior to intraperitoneal injection with allergenic cyst fluid 15 minutes later;mice in the control group were injected with the same volume of normal saline.The paraffin sections of mice lung tissues were stained with hematoxylin-eosin(HE)to observe the inflammatory damage under microscope.Total RNA was extracted from lung tissue and the mRNA relative transcription levels of tumor necrosis factorα(TNF-α),IL-6,cysteinyl aspartate specific proteinase-8,phosphatidylinositol 3 kinase(PI3K),protein kinase B(Akt)and nuclear factor kappa-B(NF-κB)were detected by qRT-PCR.Western blotting was used to detect the protein relative expression levels of PI3K,Akt and NF-κB,withβ-actin as an internal reference.Data was analyzed by GraphPad Prism software.One-way analysis of variance

关 键 词：细粒棘球绦虫 囊液致敏 IL⁃1β受体阻滞剂 肺损伤治疗 

分 类 号：R532.32[医药卫生—内科学]