支气管哮喘小鼠细胞免疫因子及Th_(17)细胞水平的变化及发病机制研究
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摘要
目的探讨支气管哮喘小鼠细胞免疫因子及辅助性T细胞(Th)_(17)细胞水平的变化及发病机制,旨在为支气管哮喘的临床治疗提供参考。方法选择SPF级昆明种小鼠24只,采用随机数字表法分为正常对照组和哮喘模型组各12只。哮喘模型组小鼠分别于实验开始当天和第8天腹腔内注入卵清蛋白(OVA)混悬液[含OVA/Al(OH)_3 10mg/g]50μg/次致敏,并于第16天给予雾化吸入含1%OVA的0.9%氯化钠溶液30~40ml进行激发,持续30~40min,1次/d,连续7d。正常对照组小鼠则给予等量的0.9%氯化钠溶液。比较两组小鼠行为变化情况、肺组织病理切片HE染色结果、肺泡灌洗液(BALF)中炎性细胞分类计数、BALF和血清中细胞免疫因子和(Th)_(17)细胞水平。结果哮喘模型组小鼠肺组织内可见大量中性粒细胞和嗜酸性粒细胞,且相对阳性着色面积增大。与正常对照组小鼠相比,哮喘模型组小鼠BALF中细胞总数、嗜酸性粒细胞、中性粒细胞和淋巴细胞等炎性细胞分类计数均明显升高,BALF和血清中IL-4、IL-5、IFN-γ和IL-17水平均明显升高,而IL-10水平明显降低,差异均有统计学意义(均P<0.05)。结论支气管哮喘小鼠BALF中细胞总数、嗜酸性粒细胞、中性粒细胞和淋巴细胞等炎性细胞分类计数均明显升高,可能与BALF和血清中IL-4、IL-5、IFN-γ和IL-17水平明显升高,而IL-10水平明显降低密切相关。
Objective To investigate Th_(17) lymphocyte and inflammation-related cytokine levels in asthmatic mice and the mechanisms. Methods Twenty-four SPF Kunming mice were divided into normal control group and asthma model group with 12 in each group. The asthma model was induced by intraperitoneal injection with OVA suspension, the mice in the control group were given the same volume of normal saline. The morphological changes of lung tissue were observed, the inflammatory cells in brochoalveolar lavage fluid(BALF) were examined, inflammatory cytokines and Th_(17) cells in serum and BALF were determined in two groups. Results Histological examination showed a large number of neutrophils and eosinophils and increased positive staining area in lung tissue of asthma model group. Compared with control group, the total number of eosinophils, neutrophils,lymphocytes and other inflammatory cells in BALF of asthma model group were increased significantly; IL-4, IL-5, IFN-γand IL-17 levels in BALF and serum were increased significantly; and IL-10 levels in BALF and serum were decreased significantly(all P<0.05). Conclusion The increased inflammatory cells in BALF of asthma mice are correlated with the increased IL-4, IL-5, IFN-γ,IL-17 levels and decreased IL-10 levels in BALF and serum.
Objective To investigate Th_(17) lymphocyte and inflammation-related cytokine levels in asthmatic mice and the mechanisms. Methods Twenty-four SPF Kunming mice were divided into normal control group and asthma model group with 12 in each group. The asthma model was induced by intraperitoneal injection with OVA suspension, the mice in the control group were given the same volume of normal saline. The morphological changes of lung tissue were observed, the inflammatory cells in brochoalveolar lavage fluid(BALF) were examined, inflammatory cytokines and Th_(17) cells in serum and BALF were determined in two groups. Results Histological examination showed a large number of neutrophils and eosinophils and increased positive staining area in lung tissue of asthma model group. Compared with control group, the total number of eosinophils, neutrophils,lymphocytes and other inflammatory cells in BALF of asthma model group were increased significantly; IL-4, IL-5, IFN-γand IL-17 levels in BALF and serum were increased significantly; and IL-10 levels in BALF and serum were decreased significantly(all P<0.05). Conclusion The increased inflammatory cells in BALF of asthma mice are correlated with the increased IL-4, IL-5, IFN-γ,IL-17 levels and decreased IL-10 levels in BALF and serum.
引文
[1]Paneliya AM,Patgiri B,Galib R,et al.Efficacy of Vasa Avaleha and its granules on Tamaka Shwasa(bronchial asthma):Open-label randomized clinical study[J].Ayu,2015,36(3):271-277.doi:10.4103/0974-8520.182760.
[2]Huang HR,Zhong YQ,Wu JF.The association between IFN-γand IL-4 genetic polymorphisms and childhood susceptibility tobronchial asthma[J].Gene,2012,494(1):96-101.doi:10.1016/j.gene.2011.09.027.
[3]窦金丽,陈静,杨青慧.健康教育对支气管哮喘患者吸入糖皮质激素口咽部不良反应的影响[J].世界最新医学信息文摘,2016,16(58):172.doi:10.3969/j.issn.1671-3141.2016.58.152.
[4]Zusska-Prot M,Jaroszewski JJ,Maslanka T.Involvement of regulatory T cells and selected cytokines in the pathogenesis of bronchial asthma[J].Postepy Hig Med Dosw(Online),2016,70(0):668-677.doi:10.5604/17322693.1207511.
[5]金华良,王利民,王娇莉,等.雷帕霉素对哮喘缓解期小鼠模型气道炎症的作用[J].浙江医学,2017,39(11):860-864,870.doi:10.12056/j.issn.1006-2785.2017.39.11.2017-19.
[6]赵燕,冉学梅,黄毅,等.构建以Th17应答为主致气道中性粒细胞增高的小鼠哮喘模型[J].第三军医大学学报,2013,35(13):1376-1378.doi:10.16016/j.1000-5404.2013.13.007.
[7]仕丽,李晓红,王檀,等.定喘止哮颗粒对支气管哮喘动物模型血清IFN-γ、IL-4、IL-5的影响[J].中国老年学杂志,2014,34(12):3394-3395.doi:10.3969/j.issn.1005-9202.2014.12.084.
[8]Arora P,Ansari SH,Najmi AK,et al.Investigation of anti-asthmatic potential of dried fruits of Vitis vinifera L.in animal model of bronchial asthma[J].Allergy Asthma Clin Immunol,2016,12:42.doi:10.1186/s13223-016-0145-x.eC ollection 2016.
[9]Kamimura M,Arimoto Y,Homma C,et al.The effects of daily bathing on symptoms of patients with bronchial asthma[J].Asia Pac Allergy,2016,6(2):112-119.doi:10.5415/apallergy.2016.6.2.112.
[10]高阳,杨帆.支气管哮喘患者外周血Th1、Th2与Th17细胞表达水平及临床意义[J].解放军医药杂志,2017,29(1):76-79.doi:10.3969/j.issn.2095-140X.2017.01.019.
[11]朱智全,时新慧,刘红升,等.Th17及Th1相关细胞因子与支气管哮喘的关系研究[J].现代生物医学进展,2017,17(20):3932-3935.doi:10.13241/j.cnki.pmb.2017.20.031.
[12]魏秀娟,宗艳杰,宋丽琴,等.血清IL-17与IL-23在支气管哮喘患者中的表达水平及其相关性[J].现代生物学进展,2016,16(26):5138-5140.doi:10.13241/j.cnki.pmb.2016.26.037.
[13]张艳丽,王秀芳,雷瑞瑞,等.哮喘、毛细支气管炎患儿外周血MDSCs、IL-10和IL-12水平及意义[J].西安交通大学学报(医学版),2013,34(4):503-507,550.doi:10.3969/j.issn.1671-8259.2013.04.020.
[2]Huang HR,Zhong YQ,Wu JF.The association between IFN-γand IL-4 genetic polymorphisms and childhood susceptibility tobronchial asthma[J].Gene,2012,494(1):96-101.doi:10.1016/j.gene.2011.09.027.
[3]窦金丽,陈静,杨青慧.健康教育对支气管哮喘患者吸入糖皮质激素口咽部不良反应的影响[J].世界最新医学信息文摘,2016,16(58):172.doi:10.3969/j.issn.1671-3141.2016.58.152.
[4]Zusska-Prot M,Jaroszewski JJ,Maslanka T.Involvement of regulatory T cells and selected cytokines in the pathogenesis of bronchial asthma[J].Postepy Hig Med Dosw(Online),2016,70(0):668-677.doi:10.5604/17322693.1207511.
[5]金华良,王利民,王娇莉,等.雷帕霉素对哮喘缓解期小鼠模型气道炎症的作用[J].浙江医学,2017,39(11):860-864,870.doi:10.12056/j.issn.1006-2785.2017.39.11.2017-19.
[6]赵燕,冉学梅,黄毅,等.构建以Th17应答为主致气道中性粒细胞增高的小鼠哮喘模型[J].第三军医大学学报,2013,35(13):1376-1378.doi:10.16016/j.1000-5404.2013.13.007.
[7]仕丽,李晓红,王檀,等.定喘止哮颗粒对支气管哮喘动物模型血清IFN-γ、IL-4、IL-5的影响[J].中国老年学杂志,2014,34(12):3394-3395.doi:10.3969/j.issn.1005-9202.2014.12.084.
[8]Arora P,Ansari SH,Najmi AK,et al.Investigation of anti-asthmatic potential of dried fruits of Vitis vinifera L.in animal model of bronchial asthma[J].Allergy Asthma Clin Immunol,2016,12:42.doi:10.1186/s13223-016-0145-x.eC ollection 2016.
[9]Kamimura M,Arimoto Y,Homma C,et al.The effects of daily bathing on symptoms of patients with bronchial asthma[J].Asia Pac Allergy,2016,6(2):112-119.doi:10.5415/apallergy.2016.6.2.112.
[10]高阳,杨帆.支气管哮喘患者外周血Th1、Th2与Th17细胞表达水平及临床意义[J].解放军医药杂志,2017,29(1):76-79.doi:10.3969/j.issn.2095-140X.2017.01.019.
[11]朱智全,时新慧,刘红升,等.Th17及Th1相关细胞因子与支气管哮喘的关系研究[J].现代生物医学进展,2017,17(20):3932-3935.doi:10.13241/j.cnki.pmb.2017.20.031.
[12]魏秀娟,宗艳杰,宋丽琴,等.血清IL-17与IL-23在支气管哮喘患者中的表达水平及其相关性[J].现代生物学进展,2016,16(26):5138-5140.doi:10.13241/j.cnki.pmb.2016.26.037.
[13]张艳丽,王秀芳,雷瑞瑞,等.哮喘、毛细支气管炎患儿外周血MDSCs、IL-10和IL-12水平及意义[J].西安交通大学学报(医学版),2013,34(4):503-507,550.doi:10.3969/j.issn.1671-8259.2013.04.020.