氧化苦参碱对哮喘大鼠肺组织中IL-17表达的影响

Effects of oxymatrine on the expression of IL-17 in lung tissues of asthmatic rats

目的观察不同剂量的氧化苦参碱(Oxymatrine,Oxy)对哮喘大鼠肺组织中IL-17蛋白和IL-17mRNA表达的影响,探讨Oxy治疗哮喘的作用机制。方法清洁级40只SD雌性大鼠按照随机数字表分为5组,每组8只。对照组第1天和第8天各腹腔注射生理盐水2mL,第15天开始应用37℃生理盐水雾化吸入激发,1次/天,60min/次,共7d。哮喘组OVA10mg加生理盐水制成2mL悬液代替生理盐水腹腔注射,1%OVA代替生理盐水雾化吸入激发,余同对照组。Oxy低、中、高剂量组在每次激发前30min分别用Oxy按50、100、150mg/kg腹腔注射,余同哮喘组。末次刺激后24h处死大鼠,取右上肺组织置液氮溶液中速冻,置于-80℃冰箱保存,分别应用Westernblot和RT-PCR检测组织中IL-17蛋白和IL-17mRNA;收集支气管肺泡灌洗液,检测炎症细胞数。结果对照组、Oxy高剂量组、Oxy中剂量组、Oxy低剂量组和哮喘组肺泡灌洗液炎症细胞数依次增加,IL-17蛋白和IL-17mRNA表达依次升高。与哮喘组比较,Oxy低、中、高剂量组和对照组炎症细胞总数和嗜酸性细胞数均下降(P<0.05),IL-17蛋白和IL-17mRNA表达减少(P<0.05)。与对照组比较,Oxy中、低剂量组炎症细胞总数、嗜酸性细胞数、IL-17表达增加(P<0.05),Oxy高剂量组则无明显变化(P>0.05)。结论Oxy可能通过降低哮喘大鼠肺组织中炎症促进因子IL-17蛋白和IL-17mRNA的表达,抑制肺组织炎症反应,Oxy抑制肺组织炎症反应随剂量增加而增强。

Objective To observe the influence of the different dose of Oxymatrine on the IL-17protein and IL-17mRNA of lung from asthmatic rats and explore its mechanism of oxymatrine in the treatment of asthma. Methods 40 healthy female clean SD rats were randomly divided into five groups.The rats of control group were injected normal saline 2 mL in abdominal cavity on the first day and 8th day and were inhalated 37℃ normal saline since the 15th day(once a day,60 min/ time,7 d in total ).The rats of the asthma model group were injected 2 mL suspension made by 10 mg OVA and saline in abdominal cavity on the first day and the 8th day and were inhalated 1% OVA since the 15th day(once a day,60 min/ time,7 d in total).The rats of low-dose,medium-dose and high-dose Oxy group were injected into the abdominal cavity by 50 mg/kg,100 mg/kg and 150 mg/kg Oxy respectively 30 min before aerosol inhalation,while the others were the same as the asthma group.The rats were killed 24 h after the rats were nebulizated,and the right lung tissue in liquid nitrogen solution was stored in the fridge at -80 degree.The IL-17protein and IL-17mRNA in the tissue were extracted by Western blot and RT-PCR respectively.Bronchial alveolar lavage was performed to detect the number of inflammatory cells. Results The number of inflammatory cells in bronchial alveolar lavage of the control group,the high-dose,the medium-dose,the low-dose Oxy group and the asthma group were successively increased,and the expression of IL-17protein and IL-17mRNA were successively increased.Compared with the asthma model group,the total number of inflammatory cells and eosinophilic cells in Oxy low-dose,medium-dose and high-dose group and control group were decreased ( P <0.05),and the expression of IL-17protein and IL-17mRNA was decreased( P <0.05).Compared with the control group,the total number of inflammatory cells,the number of eosinophilic cells,and the expression of IL-17 were increased in the Oxy medium-dose and low-dose group( P <0.05),while there was no significant change in the Oxy high-dose group( P >0.05).Conclusion Oxy may inhibit the inflammatory reaction of asthmatic rats’ lung tissues by reducing the expression of IL-17protein and IL-17mRNA.The inhibitory effect of Oxy on inflammatory reaction of lung tissue was enhanced with increasing dose.