利巴韦林对呼吸道合胞病毒感染哮喘加重小鼠胸腺基质淋巴细胞生成素分泌的影响

目的 明确利巴韦林(RIB)在呼吸道合胞病毒(RSV)感染哮喘加重治疗中的作用.方法 (1)细胞试验:人支气管上皮细胞16HBE随机分为RSV组、RSV/RIB组、RIB组和对照组,每组8瓶.感染细胞用RSV病毒液的感染复数(MOI)为2;RIB浓度为50 μg/ml.蛋白质印迹法检测各组细胞胸腺基质淋巴细胞生成素(TSLP)蛋白表达.(2)动物实验:雌性BALB/c小鼠随机分为卵清蛋白(OVA)组、OVA/RSV组、OVA/RSV/RIB组和对照组,每组8只.应用OVA腹腔注射致敏、OVA雾化吸入结合RSV病毒液滴鼻激发哮喘,RIB 10 mg/kg肌内注射.动物肺功能分析系统检测各组小鼠气道反应性;酶联免疫吸附试验检测小鼠血清白细胞介素4(IL-4)、IL-5、IL-13、干扰素γ(IFN-γ)和支气管肺泡灌洗液(BALF)TSLP浓度;取肺组织观察炎症反应和气道上皮细胞TSLP表达水平.结果 细胞试验显示RSV、RSV/RIB、RIB和对照组TSLP蛋白表达量分别为1.97±0.22、1.16±0.19、0.99±0.17和0.89±0.08,RSV/RIB组明显低于RSV组(P＜0.01).动物实验显示OVA/RSV/RIB组小鼠气道反应性明显低于OVA/RSV组(P＜0.01);血清IL-4、IL-5、IFN-γ和BALF中TSLP浓度分别为(109.7±41.9)、(220.8±30.9)、(13.0±3.4)和(1945±82)pg/ml,均明显低于OVA/RSV组分别为(274.2±103.7)、(293.3±46.1)、(30.1±5.7)、(2127±46)pg/ml,均P＜0.01];气道炎症细胞浸润和气道上皮细胞TSLP表达均明显少于OVA/RSV组.结论 RIB可以抑制RSV感染引起的TSLP分泌增加和哮喘加重.

Effect of ribavirin upon secretion of thymic stromal lymphopoietin in respiratory syncytial virus-induced asthma exacerbation of mice

Objective To investigate the role of ribavirin (RIB) in treating respiratory syncytial virus (RSV)-induced asthma exacerbation of mice.Methods (1)Cell experiment: 32 flasks of human airway epithelial cell 16HBEs were randomly divided into four groups: RSV group, RSV/RIB group, RIB group and control group. 16HBEs were infected with RSV at a multiplicity of infection (MOI) of 2. RIB 50 μg/ml was added in culture medium and Western blot used to detect the production of thymic stromal lymphopoietin (TSLP) protein; (2) Animal experiment: 32 female BALB/c mice were randomly divided into four groups: Ovalbumin (OVA) group, OVA/RSV group, OVA/RSV /RIB group and control group. Mice were sensitized by OVA and stimulated with nebulized OVA. RSV was inoculated into murine nasal cavity and RIB 10 mg/kg intramuscularly administered. BUXCO noninvasive murine lung function detection instrument was used to examine the airway response to metacholine; ELISA was used to detect IL-4, IL-5, IL-13 and IFN-γ in murine serum and TSLP in supernatants of bronchoalveolar lavage fluid (BALF);murine lung specimens were stained with HE to observe inflammation and immunohistochemical technique was employed to observe the production of TSLP in murine airway epithelial cells.Results The cell experiment demonstrated the productions of TSLP protein in RSV group, RSV/RIB group, RIB group and control group were 1.97±0.22, 1.16±0.19, 0.99±0.17 and 0.89±0.08 respectively, and the production of TSLP in RSV/RIB group was lower than that in RSV group(P＜0.01). The animal experiment demonstrated that the murine airway responsiveness in RSV/OVA/RIB group was lower than that in OVA/RSV group(P＜0.01). The levels of IL-4 (109.7±41.9) pg/ml], IL-5 (220.8±30.9) pg/ml], IFN-γ (13.0±3.4) pg/ml] in murine serum and TSLP (1945±82) pg/ml] in BALF of RSV/OVA/RIB group were significantly lower than those in OVA/RSV group (274.2±103.7), (293.3±46.1), (30.1±5.7) and (2127±46) pg/ml respectively, all P＜0.01]; less infiltration of airway inflammatory cells in OVA/RSV/RIB group was observed than that in OVA/RSV group. Immunohistochemical staining of TSLP also showed a lower production of TSLP in airway epithelial cells of OVA/RSV/RIB group than OVA/RSV group.ConclusionRibavirin can inhibit the elevated production of TSLP after RSV infection and relieve RSV-induced asthma exacerbation in mice.