肌苷单磷酸脱氢酶在卵清蛋白诱导过敏性哮喘中的作用  

The Role of Inosine Monophosphate Dehydrogenase in Ovalbumin-induced Allergic Asthma

作　　者：桑碧敏 孔春雪 曹静蕾 梁景仪 卢芳菲 张国薇 李金泉 李平 周婷[1] Sang Bimin;Kong Chunxue;Cao Jinglei(Environmental Toxicology Laboratory,Hubei Province Key Laboratory of Occupational Hazard Identification and Control,School of Public Health,Wuhan University of Science and Technology,Wuhan 430065,China)

机构地区：[1]武汉科技大学公共卫生学院,职业危害识别与控制湖北省重点实验室,环境毒理学研究室,武汉430065 [2]武汉科技大学创新创业学院,武汉430065 [3]湖北医药学院附属襄阳市第一人民医院呼吸与危重医学科,襄阳441000

出　　处：《华中科技大学学报（医学版）》2024年第5期578-585,共8页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81903278);武汉科技大学研究生创新创业基金项目(No.JCX2023120);武汉科技大学“十四五”湖北省优势特色学科(群)项目(No.2023C0101)。

摘　　要：目的探讨肌苷单磷酸脱氢酶(inosine monophosphate dehydrogenase,IMPDH)在卵清蛋白(ovalbumin,OVA)诱导过敏性哮喘发生发展中的作用。方法将36只BALB/c雌性小鼠随机分为健康对照组、OVA哮喘组和IMPDH抑制剂(利巴韦林)干预组,每组12只。收集每只小鼠的肺泡灌洗液(bronchoalveolar lavage fluid,BALF)、肺组织、外周血及脾脏,采用苏木精-伊红染色分析肺组织病理改变,免疫组织化学法检测肺组织内CD3的表达情况,血球分析仪检测BALF和外周血中白细胞总数及分类计数,qRT-PCR和Western blot分别检测肺组织IMPDH mRNA和蛋白表达水平,免疫荧光法检测肺组织IMPDH蛋白定位和表达情况,流式细胞术检测外周血和脾脏淋巴细胞亚群及其细胞内IMPDH蛋白表达水平。结果与健康对照组相比,OVA哮喘组小鼠肺内小气道周围及肺泡间隔大量炎症细胞浸润,BALF和外周血中白细胞总数均显著增加,以中性粒细胞、嗜酸性粒细胞和单核细胞的数量增多为主(均P<0.05),且肺组织内IMDPH蛋白水平显著升高(P<0.05),炎症细胞胞质内明显增多的IMPDH蛋白呈分散模式。给予利巴韦林干预后哮喘小鼠肺部炎症明显减轻,尤以BALF中嗜酸性粒细胞和中性粒细胞显著减少为特征(均P<0.05),同时肺部IMPDH蛋白水平降低(P<0.05),炎症细胞胞质内IMPDH表达减少,但细胞核内出现较多IMPDH丝状结构。此外,OVA哮喘组小鼠脾脏中B细胞占比明显增多(P<0.05),但利巴韦林干预后哮喘小鼠脾脏中B细胞占比显著减少,而NK、CD3^(+)T和CD8^(+)T细胞占比较OVA哮喘组明显增高(均P<0.05)。同时利巴韦林干预组脾脏NK细胞内IMPDH蛋白表达水平较健康对照组升高,而CD8^(+)T细胞内IMPDH蛋白表达水平降低。结论抑制IMPDH可有效缓解以啫酸性粒细胞为主的气道及全身炎症反应,同时还能降低脾脏B细胞免疫应答,增强NK及CD8^(+)T细胞等杀伤细胞的免疫应答效应,其潜在的机制可能与炎症细Objective To investigate the role of inosine monophosphate dehydrogenase(IMPDH)in the development of ovalbumin(OVA)-induced allergic asthma.Methods Thirty-six BALB/c female mice were randomly divided into a healthy control group,an OVA asthma group,and an IMPDH inhibitor(ribavirin)intervention group,with 12 mice in each group.The bronchoalveolar lavage fluid(BALF),lung,peripheral blood and spleen were collected from each mouse.HE staining was used to analyze the pathological changes in the lung tissue,and immunohistochemistry was used to detect the expression of CD3 in the lung tissue.A hemocyte analyzer was used to determine the total number and classified counts of leukocytes in the BALF and peripheral blood.RT-PCR and Western blotting were used to detect IMPDH mRNA and protein levels,respectively,in the lung tissue.Immunofluorescence was used to detect IMPDH protein localization and expression in lung tissue,and flow cytometry was used to analyze lymphocyte subpopulations and their intracellular IMPDH protein levels in peripheral blood and spleen tissue.Results Compared with those in the healthy control group,the number of inflammatory cells around small airways and in alveolar spaces in the lungs of the OVA asthma group was greater,and the total number of leukocytes in both the BALF and peripheral blood was significantly higher,with significant increases in the numbers of neutrophils,eosinophils,and monocytes(P<0.05).There was also a significant increase in the level of IMPDH protein in lung tissue(P<0.05),and immunostaining revealed increased expression of IMPDH protein in the cytoplasm of inflammatory cells in a dispersed pattern.Subsequently,lung inflammation was significantly reduced in OVA-induced asthmatic mice after ribavirin intervention,as characterized specifically by a significant reduction in eosinophils and neutrophils in the BALF(P<0.05)and a decrease in the level of IMPDH protein in the lungs(P<0.05).Furthermore,there was also a decrease in the expression of IMPDH in the cytoplasm of inflammato

关 键 词：肌苷单磷酸脱氢酶 过敏性哮喘 气道炎症 淋巴细胞 利巴韦林 

分 类 号：R562.25[医药卫生—呼吸系统] R135.1[医药卫生—内科学]