一氧化氮合酶抑制剂减轻哮喘模型小鼠气道炎性反应

Nitric oxide synthase inhibitor reduces airway inflammation in mouse models with asthma

目的 探究一氧化氮合酶(NOS)抑制剂对哮喘小鼠的治疗效果。方法 将雌性BALB/c 5~6周小鼠50只,随机分为5组,每组10只。1)对照组;2)哮喘组:用卵清蛋白(OVA)致敏小鼠建立哮喘模型;3)S-甲基异硫脲硫酸盐(SMT)组:给予SMT干预;4)N-硝基-L-精氨酸甲酯(L-NAME)组:给予L-NAME干预;5)NG-单甲基-L-精氨酸(L-NMMA)组:给予L-NMMA干预。用HE染色进行组织病理学分析。使用乙酰甲胆碱激发试验评估气道反应性。用ELISA检测血清及肺泡灌洗液(BALF)中一氧化氮(NO)及细胞因子的水平。结果 哮喘组与对照组相比,可见杯状细胞增生、黏液分泌增加,气管血管周围大量的炎性细胞浸润,气道阻力明显升高,给予NOS抑制剂后好转,尤其是SMT组。哮喘组与对照组相比,血浆、肺泡灌洗液及肺组织匀浆中NO水平明显升高(P<0.05),血浆、肺泡灌洗液IL-2、IL-4、IL-17、TNF-α水平均明显升高(P<0.05)。SMT组与哮喘组相比,血浆、BALF及肺组织匀浆中NO水平明显下降(P<0.05),血浆、BALF IL-2、IL-4、IL-17、TNF-α水平均明显下降(P<0.05)。L-NAME组与哮喘组相比,血浆NO、IL-2、IL-17、TNF-α水平明显下降(P<0.05),BALF NO、IL-2、IL-17、TNF-α水平明显下降(P<0.05)。L-NMMA组与哮喘组相比,血浆NO、IL-2、IL4、IL-17、TNF-α水平明显下降(P<0.05),BALF NO、IL-2、IL4、IL-17、TNF-α水平明显下降(P<0.05),肺组织匀浆中NO水平明显下降(P<0.05)。结论 一氧化氮合酶抑制剂,特别是SMT能降低哮喘小鼠气道炎性反应。

Objective To explore the therapeutic effect of nitric oxide synthase(NOS) inhibitor in asthmatic mice.Methods BALB/c mice of 5 to 6 weeks were randomly divided into five groups with 10 in each:1)control group;2)asthma group:The asthma model was established by ovalbumin(OVA)-sensitized mice;3)S-methyl isothiourea sulfate(SMT) group:OVA-sensitized asthmatic mice were stimulated and then given with SMT;4)N-nitro-L-arginine methyl ester(L-NAME) group:OVA-sensitized asthmatic mice were stimulated and then given with L-NAME;5) NG-monomethyl-L-arginine(L-NMMA) group:OVA-sensitized asthmatic mice were stimulated and then given with L-NMMA.HE staining was used for histopathological analysis.Airway reactivity was assessed usinga methacholine challenge test.The levels of nitric oxide(NO) and cytokines in serum and bronchoalveolar lavage fluid(BALF) were detected by ELISA.Results Compared with the control group,goblet cell proliferation and increased mucus secretion were found in the asthma group,together with a large number of inflammatory cell infiltration around the trachea and blood vessels and significantly increased airway resistance,which were improved after the administration of NOS inhibitors,especially in the SMT group.Compared with the control group,the levels of NO in plasma,BALF and lung tissue homogenate were significantly increased in the asthma group(P<0.05),while the level of IL-2,IL-4,IL-17 and TNF-α in plasma and BALF was significantly increased(P<0.05).Compared with the asthma group,the level of NO in plasma,BALF and lung tissue homogenate was significantly decreased in SMT group(P<0.05),while the level of IL-2,IL-4,IL-17 and TNF-α in plasma and BALF was significantly decreased in SMT group(P<0.05).Compared with the asthma group,the level of plasma NO,IL-2,IL-17,and TNF-α in the L-NAME group was significantly decreased(P<0.05).The level of NO,IL-2,IL-17,and TNF-α in BALF was significantly decreased(P<0.05).Compared with the asthma group,the plasma level of NO,IL-2,IL-4,IL-17 and TNF-α in the L-NMMA group was significantly decreased(P<0.05).The level of NO,IL-2,IL-4,IL-17 and TNF-α in BALF was significantly decreased(P<0.05),and the level of NO in lung tissue homogenate was significantly decreased(P<0.05).Conclusions NOS inhibitor,especially SMT may reduce airway inflammation of asthmatic mice.