胆木提取物对哮喘小鼠肺泡灌洗液中炎性细胞及细胞因子的影响
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摘要
目的探讨通过预先应用胆木提取物的干预,了解其对哮喘小鼠肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中炎性细胞及细胞因子的影响以及剂量相关性。方法将实验动物随机分为6组(正常对照组,哮喘模型组,胆木提取物低剂量组、中剂量组、高剂量组,地塞米松组),每组10只,实验组通过卵白蛋白(ovalbumin,OVA)致敏,随后每日一次雾化吸入OVA激发复制小鼠支气管哮喘模型,除正常对照组外,每次哮喘激发前30 min腹腔注射给药(低剂量组1.0 mg/kg,中剂量组2.0 mg/kg,高剂量组4.0 mg/kg,地塞米松组2 mg/kg)。在连续雾化激发28 d后断颈处死小鼠,留取BALF,用ELISA方法对IL-2、IL-4、IL-5、IL-10、IFN-γ浓度进行检测;瑞氏染色计数白细胞(white blood cell,WBC)、嗜酸性粒细胞(eosinophile granulocyte,EOS)、中性粒细胞(neutrophile granulocyte,NEU)。结果与哮喘模型组对比,经胆木提取物干预处理,哮喘小鼠BALF中WBC、EOS、NEU均降低,其数值与胆木提取物的干预剂量呈负相关;而细胞因子IL-2、IL-10、IFN-γ的浓度则比哮喘模型组升高,其升高幅度与干预剂量呈正相关,细胞因子IL-4、IL-5浓度比哮喘模型组降低,降低幅度与干预剂量呈负相关。结论胆木提取物可能通过影响细胞因子(IL-2、IL-4、IL-5、IL-10、IFN-γ)的分泌及炎性细胞在气道中的浸润,对支气管哮喘小鼠的气道炎症进行调控。
Objective Through the advance intervention of Danmu extract, to understand the effect of inflammatorycells and cytokines in bronchoalveolar lavage fluid in asthma mice and its dose correlation. Methods The experimentalanimals were randomly divided into 6 groups(normal control group, asthma model group, low-dose group of Danmu extract,medium-dose group, high-dose group, dexamethasone group), 10 of each group. The experimental groups were sensitized byovalbumin(OVA), and then the inhalation of OVA induced the mouse model of bronchial asthma per day. Except the normalcontrol group, the experimental animals were injected by intraperitoneal 30 minutes before every induction(low dose group: 1.0 mg/kg, medium dose group: 2.0 mg/kg, high dose group: 4.0 mg/kg, dexamethasone group 2 mg/kg). After 28 days of continuousatomization, killing the mouse by the neck and take the BALF, the concentration of IL-2, IL-4, IL-5, IL-10 and IFN-gammawere detected by ELISA method and count white blood cell(WBC), eosinophil(EOS), neutrophil(NEU) by Wright's staining.Results Comparison with asthma model group, by the intervention of Danmu extract, the WBC, EOS, and NEU in BALF wereall reduced, and the quantity was negatively correlated with the dose of Danmu extract. The concentration of cytokine IL-2, IL-10, IFN-gamma were higher than the asthma model group, and its rising were positively correlated with the dose of Danmuextract. The concentration of cytokines IL-4, IL-5 were lower than the asthma model group, and the reduction were negativelycorrelated with intervention dose. Conclusion Danmu extract may regulate the airway inflammation in bronchial asthma miceby influencing cytokines(IL-2, IL-4, IL-5, IL-10 and IFN-gamma) secretion and inflammatory cells infiltration in airway.
Objective Through the advance intervention of Danmu extract, to understand the effect of inflammatorycells and cytokines in bronchoalveolar lavage fluid in asthma mice and its dose correlation. Methods The experimentalanimals were randomly divided into 6 groups(normal control group, asthma model group, low-dose group of Danmu extract,medium-dose group, high-dose group, dexamethasone group), 10 of each group. The experimental groups were sensitized byovalbumin(OVA), and then the inhalation of OVA induced the mouse model of bronchial asthma per day. Except the normalcontrol group, the experimental animals were injected by intraperitoneal 30 minutes before every induction(low dose group: 1.0 mg/kg, medium dose group: 2.0 mg/kg, high dose group: 4.0 mg/kg, dexamethasone group 2 mg/kg). After 28 days of continuousatomization, killing the mouse by the neck and take the BALF, the concentration of IL-2, IL-4, IL-5, IL-10 and IFN-gammawere detected by ELISA method and count white blood cell(WBC), eosinophil(EOS), neutrophil(NEU) by Wright's staining.Results Comparison with asthma model group, by the intervention of Danmu extract, the WBC, EOS, and NEU in BALF wereall reduced, and the quantity was negatively correlated with the dose of Danmu extract. The concentration of cytokine IL-2, IL-10, IFN-gamma were higher than the asthma model group, and its rising were positively correlated with the dose of Danmuextract. The concentration of cytokines IL-4, IL-5 were lower than the asthma model group, and the reduction were negativelycorrelated with intervention dose. Conclusion Danmu extract may regulate the airway inflammation in bronchial asthma miceby influencing cytokines(IL-2, IL-4, IL-5, IL-10 and IFN-gamma) secretion and inflammatory cells infiltration in airway.
引文
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[14]叶惠英,韩珊珊,柯琴剑.1252例过敏性疾病患者血清过敏原检测与分析[J].实用预防医学,2016,23(4):462-464.
[15]周敏芳,张倩.哮喘患儿血清25-羟维生素D_3水平检测的临床意义[J].中国热带医学,2016,16(2):165-168.
[2]朱粉霞,王静静,殷蓉,等.乌檀属植物化学成分研究进展[J].中华中医药杂志,2014,29(9):2877-2880.
[3]OLIN J T,WECHSLER M E.Asthma:pathogenesis and novel drugs for treatment[J].BMJ,2014,349:g5517.
[4]卢春亚,张国俊.难治性哮喘的研究新进展[J].河南医学研究,2016,25(4):656-658.
[5]程思,邵天波,郭晓静.哮喘患者外周血调节性T细胞和Th1/Th2的变化及与病情的相关性分析[J].河北医学,2017,23(4):699-702.
[6]PAYNE A S,FREISHTAT R J.Conserved steroid hormone homology converges on nuclear factorκB to modulate inflammation in asthma[J].J Investig Med,2012,60(1):13-17.
[7]BOULET L P,TURCOTT H,PLANTE S,et al.Airway function,inflammation and regulatory T cell function in subjects in asthma remission[J].Can Respir J,2012,19(1):19-25.
[8]SANTUS P,RADOVANOVIC D.Prostaglandin D2 receptor antagonists in early development as potential therapeutic options for asthma[J].Expert Opin Investig Drugs,2016,25(9):1083-1092.
[9]WEGMANN M.Th2 cells as targets for therapeutic intervention in allergic bronchial asthma[J].Expert Rev Mol Diagn,2009,9(1):85-100.
[10]ULLMANN N,BOSSLEY C J,FLEMING L,et al.Blood eosinophil counts rarely reflect airway eosinophilia in children with severe asthma[J].Allergy,2013,68(3):402-406.
[11]马雅銮,胡镜清.胆木的研究进展[J].中华中医药杂志,2017,32(7):3079-3082.
[12]沈存思,尹庆锋,王蔚,等.6种中药口服液抗炎、镇痛、解热作用比较实验研究[J].世界中医药,2016,11(9):1663-1666.
[13]田锋,戴银芳.肥胖症合并哮喘急性期儿童血液指标和炎性因子变化[J].热带医学杂志,2017,17(9):1181-1183.
[14]叶惠英,韩珊珊,柯琴剑.1252例过敏性疾病患者血清过敏原检测与分析[J].实用预防医学,2016,23(4):462-464.
[15]周敏芳,张倩.哮喘患儿血清25-羟维生素D_3水平检测的临床意义[J].中国热带医学,2016,16(2):165-168.