川麦冬须根提取物对烟气致小鼠气道损伤的保护作用及机制初探
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摘要
目的:研究川麦冬须根提取物对烟气引起小鼠气道损伤的保护作用并初步探讨其作用机制。方法:将SPF级KM小鼠随机分为空白对照组、模型组及给药组。给药组提前3天给药,模型组与对照组分别予以等体积纯净水。实验采用模拟被动吸烟法建立小鼠肺损伤模型,在使用烟气对模型组和给药组染毒4周后,分别测定小鼠氨水引咳潜伏期及咳嗽次数和酚红排泌量,对气管和肺组织进行组织病理学检查。采用分光光度法检测川麦冬须根皂苷组分、皂苷与黄酮组分对DPPH自由基、超氧阴离子和羟自由基的清除作用。结果:川麦冬须根提取物样品A(黄酮+皂苷)及样品B(皂苷)可促进烟气染毒小鼠体重增长,在给药3周末和给药4周末,样品A 2g/kg组、4g/kg组及样品B 4g/kg组体重均显著高于模型组。样品A 2g/kg组、4g/kg组及样品B 2g/kg组、4g/kg组的小鼠咳嗽次数和酚红排泌量均显著低于模型小鼠;另外,样品A和样品B还可改善烟气所致小鼠气管柱状上皮细胞纤毛消失,细胞扁平变薄或细胞层数增加变厚,粘膜下层可偶见少量炎细胞浸润的病理改变。同时,体外抗氧化活性研究显示,川麦冬须根皂苷提取组分清除DPPH自由基、羟自由基和超氧阴离子自由基的IC50分别为0. 8067g/L、0. 7899 g/L和1. 357 g/L;川麦冬须根黄酮皂苷混合提取组分清除DPPH自由基、羟自由基和超氧阴离子自由基的IC50分别为0. 7975 g/L、0. 5413 g/L和1. 902 g/L。结论:川麦冬须根提取物对烟气所致小鼠气道损伤具有明显的保护作用,推测其可能与其具有一定的抗氧化能力有关,且黄酮与皂苷混合提取组分的抗氧化能力略强于总皂苷组分。其详细机理有待进一步深入探讨。
Objective: To study the protective effect and mechanism of extracts from Fibrous Root of Sichuan Ophiopogon japonicus on tobacco smoke induced airway injury mice. Methods: The KM mice were randomly divided into four groups,including the control group,the model group,OJ group A( Saponin + flavone) and OJ group B( Saponin). All mice were administered for 3 days,while equal volume of pure water was given to mice in the model group and the control group. Mice in the model group and OJ group were exposed to the tobacco smoke for4 weeks to establish airway injury model,then the incubation period of cough,cough frequency and the phenol red excretion were measured,and the histopathological examination of the trachea and lung tissue was carried out. In addition,the scavenging effects of OJ group A and OJ group B on DPPH free radical,superoxide anion and hydroxyl radical were detected by spectrophotometric method. Results: The body weights in 2g/kg A group,4g/kg A and 4g/kg B were higher than those in the model group at the end of 3 weeks and 4 weeks. Cough frequency and phenol red excretion in A group and B group were significantly decreased than those in the model group. Furthermore,in OJ group,cilia of tracheal columnar epithelial cells in mice exposed to the tobacco smoke was improved. It was observed that the cells were flattened,the cell layers became thick,or a small number of inflammatory cells were infiltrated. In vitro,IC 50 values of Saponin for the ability of scavenging DPPH radical,hydroxyl radical and superoxide radical anion were 0. 8067g/L,0. 7899 g/L,1. 357 g/L respectively,IC50 values of Flavone saponins mixture were 0. 7975 g/L,0. 5413 g/L,1. 902 g/L. Conclusion: Fibrous Root of Sichuan Ophiopogon japonicus has the protective effect on airway injury induced by smoke in mice,which may be related to its antioxidant ability. Combination of saponins and flavone shows stronger scavenging ability than single use of saponins,and the mechanism needs further research.
Objective: To study the protective effect and mechanism of extracts from Fibrous Root of Sichuan Ophiopogon japonicus on tobacco smoke induced airway injury mice. Methods: The KM mice were randomly divided into four groups,including the control group,the model group,OJ group A( Saponin + flavone) and OJ group B( Saponin). All mice were administered for 3 days,while equal volume of pure water was given to mice in the model group and the control group. Mice in the model group and OJ group were exposed to the tobacco smoke for4 weeks to establish airway injury model,then the incubation period of cough,cough frequency and the phenol red excretion were measured,and the histopathological examination of the trachea and lung tissue was carried out. In addition,the scavenging effects of OJ group A and OJ group B on DPPH free radical,superoxide anion and hydroxyl radical were detected by spectrophotometric method. Results: The body weights in 2g/kg A group,4g/kg A and 4g/kg B were higher than those in the model group at the end of 3 weeks and 4 weeks. Cough frequency and phenol red excretion in A group and B group were significantly decreased than those in the model group. Furthermore,in OJ group,cilia of tracheal columnar epithelial cells in mice exposed to the tobacco smoke was improved. It was observed that the cells were flattened,the cell layers became thick,or a small number of inflammatory cells were infiltrated. In vitro,IC 50 values of Saponin for the ability of scavenging DPPH radical,hydroxyl radical and superoxide radical anion were 0. 8067g/L,0. 7899 g/L,1. 357 g/L respectively,IC50 values of Flavone saponins mixture were 0. 7975 g/L,0. 5413 g/L,1. 902 g/L. Conclusion: Fibrous Root of Sichuan Ophiopogon japonicus has the protective effect on airway injury induced by smoke in mice,which may be related to its antioxidant ability. Combination of saponins and flavone shows stronger scavenging ability than single use of saponins,and the mechanism needs further research.
引文
[1]中华人民共和国药典第一部,中国医药科技出版社,2015∶155.
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[4]Chen J,Yuan J,Zhou L,et al. Regulation of different components from Ophiopogon japonicus on autophagy in human lung adenocarcinoma A549Cells through PI3K/Akt/m TOR signaling pathway. Biomedicine&Pharmacotherapy,2017,87∶118~126.
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[7]熊静悦,张娅莉,陈慧泼,等.健肺丸补气润肺和镇咳祛痰功效的研究.华西药学杂志,2012,27(6)∶655~658.
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[9]吴兑.灰霾天气的形成与演化.环境科学与技术,2011; 34(3)∶157~161.
[10]李建强,赵卉,刘卓拉,等.胆红素对烟雾暴露大鼠肺泡巨噬细胞核因子κB及白细胞介素8表达的影响.中华结核和呼吸杂志,2005,28(1)∶59~60.
[11]袁峰,杜永成,胡晓芸,等.被动吸烟对致敏大鼠气道5-脂氧合酶及其mRNA表达的影响.中华哮喘杂志(电子版),2010,30(4)∶527~530.
[12]Scesnaite A,Jarmalaite S,Mutanen P,et al. Similar DNA methylation pattern in lung tumours from smokers and never-smokers with second-hand tobacco smoke exposure. Mutagenesis,2012,27(4)∶423~429.
[13]Malinovschi A,Janson C,Holmkvist T,et al. Effect of smoking on exhaled nitric oxide and flow-independent nitric oxide exchange parameters. European Respiratory Journal,2006,28(2)∶339~345.
[14]Slater T F. Free Radical Mechanism in Tissue Injury. Biochemical Journal,1984,222(1)∶1~15.
[15]赵保路.吸烟、自由基与健康.生物物理学报,2012,28(4)∶332~340.
[16]吴彦,杨鸿,刘红,等.中药组分清除烟气有害物质及降低细胞毒性的研究,中国中药杂志,2011,36(22)∶3184~3188.
[2]Kitahiro Y,Koike A,Sonoki A,et al. Anti-inflammatory activities of Ophiopogonis Radix on hydrogen peroxide-induced cellular senescence of normal human dermal fibroblasts. Journal of Natural Medicines,2018,72(4)∶905~914.
[3]彭婉,马骁,王建,等.麦冬化学成分及药理作用研究进展.中草药,2018,(2)∶477~488.
[4]Chen J,Yuan J,Zhou L,et al. Regulation of different components from Ophiopogon japonicus on autophagy in human lung adenocarcinoma A549Cells through PI3K/Akt/m TOR signaling pathway. Biomedicine&Pharmacotherapy,2017,87∶118~126.
[5]何礼,金虹,梁雪兰,等. HPLC测定川麦冬块根及须根中的麦冬皂苷.华西药学杂志,2014,29(5)∶583~584.
[6]金虹,王化东,何礼,等.川产麦冬及其须根组织学与麦冬皂苷的含量对比研究.中草药,2014,(4)∶1002~1005.
[7]熊静悦,张娅莉,陈慧泼,等.健肺丸补气润肺和镇咳祛痰功效的研究.华西药学杂志,2012,27(6)∶655~658.
[8]钟小宁,白晶,施焕中,等.慢性支气管炎与肺气肿大鼠气道炎症与重塑的实验研究.中华结核和呼吸杂志,2003,26(12)∶750~755.
[9]吴兑.灰霾天气的形成与演化.环境科学与技术,2011; 34(3)∶157~161.
[10]李建强,赵卉,刘卓拉,等.胆红素对烟雾暴露大鼠肺泡巨噬细胞核因子κB及白细胞介素8表达的影响.中华结核和呼吸杂志,2005,28(1)∶59~60.
[11]袁峰,杜永成,胡晓芸,等.被动吸烟对致敏大鼠气道5-脂氧合酶及其mRNA表达的影响.中华哮喘杂志(电子版),2010,30(4)∶527~530.
[12]Scesnaite A,Jarmalaite S,Mutanen P,et al. Similar DNA methylation pattern in lung tumours from smokers and never-smokers with second-hand tobacco smoke exposure. Mutagenesis,2012,27(4)∶423~429.
[13]Malinovschi A,Janson C,Holmkvist T,et al. Effect of smoking on exhaled nitric oxide and flow-independent nitric oxide exchange parameters. European Respiratory Journal,2006,28(2)∶339~345.
[14]Slater T F. Free Radical Mechanism in Tissue Injury. Biochemical Journal,1984,222(1)∶1~15.
[15]赵保路.吸烟、自由基与健康.生物物理学报,2012,28(4)∶332~340.
[16]吴彦,杨鸿,刘红,等.中药组分清除烟气有害物质及降低细胞毒性的研究,中国中药杂志,2011,36(22)∶3184~3188.