氢气通过靶向调控miR-141-3p/TXNIP对支气管哮喘幼年大鼠炎症反应和氧化应激的影响  

Effects of hydrogen targeted regulation of miR-141-3p/TXNIP on inflammatory response and oxidative stress from bronchial asthma in young rats

作　　者：戴吉成[1] 姜明宇[1] 张玉鑫 任明永[1] 张羽航 DAI Jicheng;JIANG Mingyu;ZHANG Yuxin;REN Mingyong;ZHANG Yuhang(Department of Pediatrics,the First Affiliated Hospital of Harbin Medical University,Harbin,Heilongjiang 150001,China;Department of Pediatrics,Boli County People’s Hospital,Qitaihe,Heilongjiang 154500,China)

机构地区：[1]哈尔滨医科大学附属第一医院儿科,黑龙江哈尔滨150001 [2]七台河市勃利县人民医院儿科,黑龙江七台河154500

出　　处：《中国优生与遗传杂志》2023年第10期1987-1993,共7页Chinese Journal of Birth Health & Heredity

基　　金：2019年度黑龙江省教育厅临床青年专项课题(省属高等学校基本科研业务费科研项目:2019-KYYWF-0332)。

摘　　要：目的 观察氢气是否通过靶向调控miR-141-3p/TXNIP参与支气管哮喘幼年大鼠的炎症反应和氧化应激效应。方法 将支气管哮喘幼年大鼠造模成功后,按照随机数字法分为模型组、氢气组、氢气+miR-141-3pNC组、氢气+miR-141-3p antagomir组,每组10只。另设对照组,对照组用生理盐水替代药物进行处理。取幼年大鼠肺组织,并收集大鼠肺泡灌洗液;通过ELISA法检测IL-4、IL-5和IL-13三个炎症因子的表达水平;幼年大鼠肺组织HE染色后,显微镜下观察病理学变化同时肺损伤评分,计数嗜酸性粒细胞、淋巴细胞和中性粒细胞的变化;对大鼠肺组织中活性氧、丙二醛和髓过氧化物酶及其miR-141-3p的表达水平进行qRT-PCR分析,通过蛋白质印迹法测定大鼠肺组织中TXNIP蛋白的表达水平;采用萤光素酶报告实验分析miR-141-3p与TXNIP的靶向关系。结果 与相应的对照组对比,模型组和氢气+miR-141-3pantagomir组幼年大鼠肺损伤评分、肺泡灌洗液中炎症因子水平显著升高(均P<0.05),肺组织气道上皮细胞排列不规则,支气管上皮细胞肿胀,部分黏膜脱落,支气管平滑肌显著增生伴有大量炎性细胞浸润,嗜酸性粒细胞、淋巴细胞及中性粒细胞数量在肺泡灌洗液中显著增加(均P<0.05),肺组织中miR-141-3p表达水平显著降低、TXNIP蛋白表达水平显著升高(均P<0.05)。与模型组相比,氢气组、氢气+miR-141-3p NC组上述指标呈相反样改变。相对萤光素酶报告提示miR-141-3p与TXNIP存在靶向关系。结论 吸入氢气后,通过靶向调控miR-141-3p和TXNIP的表达,在支气管哮喘炎症反应和氧化应激的治疗机制中发挥重要作用。Objective To examine whether hydrogen can take part in the inflammatory reaction and oxidative stress effect from young rats with bronchial asthma by targeting miR-141-3p/TXNIP.Methods Young rats with bronchial asthma,according to the random number method,were divided into:Model group,hydrogen group,hydrogen+miR-141-3p NC group,hydrogen+miR-141-3p antagomir group with 10 rats in each group.The control group was set up,treated with normal saline instead of drugs.The lung tissues and alveolar lavage fluid of of young rats were separately collected.The expression levels of three inflammatory factors:IL-4,IL-5,and IL-13 were detected using ELISA method.After HE staining from the lung tissue of young rats,pathological changes were observed under a microscope,along with lung injury scores,and eosinophils,lymphocytes,and neutrophils were counted.The expression levels of ROS,MDA,MPO and miR-141-3p in lung tissues were measured by qRT-PCR,and the protein expression level of TXNIP was determined by Western blot method.Analysis of the targeting relationship between miR-141-3p and TXNIP was used by luciferase reporting experiment.Results Compared with the corresponding control group,the lung injury score and the level of inflammatory factors in the alveolar lavage fluid in the model group and the hydrogen+miR-141-3p antagomir group were significantly increased(all P<0.05),the airway epithelial cells in the lung tissue were irregularly arranged,the bronchial epithelial cells were swollen,part of the mucosa was shed,the bronchial smooth muscle was thickened,and a large number of inflammatory cells were seen.The number of EOS,lymphocytes,and neutrophils in the alveolar lavage fluid was dramatically elevated(all P<0.05).The expression level of miR-141-3p in lung tissue decreased significantly,and the expression level of TXNIP protein increased substantially(P<0.05).The comparative analysis with the model group,the hydrogen group and the hydrogen+miR-141-3p NC group presented opposite changes.Relative luciferase report suggests t

关 键 词：氢气 支气管哮喘 幼年大鼠 炎症反应 氧化应激 

分 类 号：R562.25[医药卫生—呼吸系统]