麻杏石甘汤对哮喘模型小鼠气道重塑及肺组织MMP-9和TIMP-1表达的影响
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摘要
目的:观察麻杏石甘汤对哮喘模型小鼠气道重塑及肺组织基质金属蛋白酶9(MMP-9)和金属蛋白酶组织抑制物1(TIMP-1)表达的影响,探讨其治疗哮喘的可能机制。方法:将72只健康雌性BALB/c小鼠随机分为:空白对照组,模型组,麻杏石甘汤低剂量组、中剂量组和高剂量组,阳性对照组。采用卵清蛋白致敏激发建立小鼠哮喘模型。空白对照组和模型组于激发前30 min以生理盐水灌胃;麻杏石甘汤低剂量组、中剂量组和高剂量组分别于激发前30 min以麻杏石甘汤按5.0 g/kg、10.0 g/kg和20.0 g/kg剂量灌胃;阳性对照组于激发前30 min以地塞米松按0.005 g/kg剂量灌胃。连续给药7 d后,观察气道反应性、支气管肺泡冲洗液(BALF)中嗜酸性粒细胞(EOS)计数、杯状细胞百分比和胶原沉积的变化;ELISA法检测MMP-9和TIMP-1水平;Western blot法检测MMP-9和TIMP-1的蛋白表达;RT-qPCR法分析检测MMP-9和TIMP-1的mRNA表达。结果:与空白对照组比较,模型组的气道反应性、杯状细胞百分比、胶原沉积、BALF中EOS计数及肺组织MMP-9和TIMP-1的mRNA和蛋白水平均显著升高(P <0.01);与模型组比较,麻杏石甘汤低剂量组、中剂量组和高剂量组及阳性对照组上述指标则明显降低(P <0.05或P <0.01)。结论:麻杏石甘汤可能通过降低MMP-9和TIMP-1的表达改善哮喘模型小鼠气道重塑状态。
AIM:To investigate the effects of Maxing-Shigan decoction on airway remodeling and expression of matrix metalloproteinase-9(MMP-9)and tissue inhibitor of metalloproteinase-1(TIMP-1)in the lung tissues of asthmatic mice,and to explore its possible mechanism in treatment of asthma.METHODS:The BALB/c mice were divided into blank control group,model group,low-dose Maxing-Shigan decoction group,middle-dose Maxing-Shigan decoction group,high-dose Maxing-Shigan decoction group and positive control group.The mice were sensitized and challenged with ovalbumin to establish asthma model.The mice in blank control group and model group were given saline by oral administration before 30 min of suscitation.The mice in low-dose,middle-dose and high-dose Maxing-Shigan decoction groups were given Maxing-Shigan decoction at 5.0 g/kg,10.0 g/kg and 20.0 g/kg,respectively,by oral administration before 30 min of suscitation.The mice in positive control group was given dexamethasone at 0.005 g/kg by oral administration before 30 min of suscitation.After consecutive administration for 7 d,the variations of airway responsiveness,the percentage of the goblet cells,the collagen deposition,and the eosinophil(EOS)counts in bronchoalveolar lavage fluid(BALF)of each group were observed.The protein levels of MMP-9 and TIMP-1 in the lung tissues were determined by ELISA and Western blot.The mRNA expression of MMP-9 and TIMP-1 was detected by RT-qPCR.RESULTS:Compared with blank control group,the airway responsiveness,the goblet cell percentage,the collagen deposition,the EOS counts in BALF,the protein levels of MMP-9 and TIMP-1,and the mRNA expression of MMP-9 and TIMP-1 were significantly increased in model group(P<0.01).Compared with model group,all of the indexes were reversed in low-dose,middle-dose and high-dose Maxing-Shigan decoction groups and positive control group(P<0.05 or P<0.01).CONCLUSION:Maxing-Shigan decoction improves airway remodeling in asthma model mice by down-regulating the expression of MMP-9 and TIMP-1.
AIM:To investigate the effects of Maxing-Shigan decoction on airway remodeling and expression of matrix metalloproteinase-9(MMP-9)and tissue inhibitor of metalloproteinase-1(TIMP-1)in the lung tissues of asthmatic mice,and to explore its possible mechanism in treatment of asthma.METHODS:The BALB/c mice were divided into blank control group,model group,low-dose Maxing-Shigan decoction group,middle-dose Maxing-Shigan decoction group,high-dose Maxing-Shigan decoction group and positive control group.The mice were sensitized and challenged with ovalbumin to establish asthma model.The mice in blank control group and model group were given saline by oral administration before 30 min of suscitation.The mice in low-dose,middle-dose and high-dose Maxing-Shigan decoction groups were given Maxing-Shigan decoction at 5.0 g/kg,10.0 g/kg and 20.0 g/kg,respectively,by oral administration before 30 min of suscitation.The mice in positive control group was given dexamethasone at 0.005 g/kg by oral administration before 30 min of suscitation.After consecutive administration for 7 d,the variations of airway responsiveness,the percentage of the goblet cells,the collagen deposition,and the eosinophil(EOS)counts in bronchoalveolar lavage fluid(BALF)of each group were observed.The protein levels of MMP-9 and TIMP-1 in the lung tissues were determined by ELISA and Western blot.The mRNA expression of MMP-9 and TIMP-1 was detected by RT-qPCR.RESULTS:Compared with blank control group,the airway responsiveness,the goblet cell percentage,the collagen deposition,the EOS counts in BALF,the protein levels of MMP-9 and TIMP-1,and the mRNA expression of MMP-9 and TIMP-1 were significantly increased in model group(P<0.01).Compared with model group,all of the indexes were reversed in low-dose,middle-dose and high-dose Maxing-Shigan decoction groups and positive control group(P<0.05 or P<0.01).CONCLUSION:Maxing-Shigan decoction improves airway remodeling in asthma model mice by down-regulating the expression of MMP-9 and TIMP-1.
引文
[1]张方琪,韩新鹏,张芳,等.动态观察过敏原诱导的哮喘小鼠模型从急性炎症到慢性重塑的发展[J].中国病理生理杂志,2017,33(2):380-384.
[2]Coleman SL,Shaw OM.Progress in the understanding of the pathology of allergic asthma and the potential of fruit proanthocyanidins as modulators of airway inflammation[J].Food Funct,2017,8(12):4315-4324.
[3]朱金凤.支气管哮喘的中医辨治思维与方法研究[J].中国中医基础医学杂志,2016,22(2):284-286.
[4]王盛隆,杨爽,王强,等.支气管哮喘中医药全病程治疗方案研究进展[J].时珍国医国药,2015,26(3):699-701.
[5]李献超.麻杏石甘汤治疗支气管哮喘急发期的临床观察[J].中药药理与临床,2015,31(1):292-293.
[6]刘春雨.麻杏石甘汤治疗支气管哮喘急发期的临床疗效分析[J].中国医学创新,2012,9(35):47-48.
[7]虞琳,朱丽君,张怡静,等.麻杏石甘汤对哮喘大鼠气道上皮STAT6和黏蛋白表达的调控[J].医学研究杂志,2014,43(5):40-44.
[8]高尚泽,杨林伟,王小莹,等.麻杏石甘汤对哮喘大鼠肺组织中Sec14l3表达的影响[J].天津中医药,2013,30(12):745-748.
[9]韩凤芹,孙雪文,张新国,等.麻杏石甘汤对哮喘模型小鼠支气管黏膜和肺组织免疫的影响[J].时珍国医国药,2012,23(6):1398-1399.
[10]黄丰,童晓云,张荣华.麻杏石甘汤调节哮喘模型小鼠Th1/Th2反应的机制初探[J].中药材,2008,31(10):1519-1522.
[11]李厚忠,高照渝,黄伟,等.中药川贝母对哮喘模型小鼠MMP-2,MMP-9和TIMP-1的影响[J].中国中药杂志,2017,42(21):4180-4186.
[12]杨汀,王辰,庞宝森,等.豚鼠哮喘模型气道重建对气道反应性的影响[J].中国病理生理杂志,2004,20(11):2033-2036.
[13]Liccardi G,Salzillo A,Piccolo A,et al.Dysfunction of small airways and prevalence,airway responsiveness and inflammation in asthma:much more than small particle size of pet animal allergens[J].Ups J Med Sci,2016,121(3):196-197.
[14]张方琪,韩新鹏,张芳,等.动态观察过敏原诱导的哮喘小鼠模型从急性炎症到慢性重塑的发展[J].中国病理生理杂志,2017,33(2):380-384.
[15]Grenier PA,Fetita CI,Brillet PY.Quantitative computed tomography imaging of airway remodeling in severe asthma[J].Quant Imaging Med Surg,2016,6(1):76-83.
[16]王爱华,赵霞,董盈妹,等.固本防哮饮对支气管哮喘缓解期小鼠肺组织MMP-2、MMP-9、TIMP-1、TIMP-2蛋白的影响[J].辽宁中医杂志,2018,45(1):183-186,229.
[17]Kim JS,Kang JY,Ha JH,et al.Expression of nerve growth factor and matrix metallopeptidase-9/tissueinhibitor of metalloproteinase-1 in asthmatic patients[J].J Asthma,2013,50(7):712-717.
[18]田金娜,李建保,刘小凡.丹龙定喘汤对哮喘小鼠气道重塑及MMP-9、TIMP-1的影响[J].中国实验方剂学杂志,2012,18(5):164-166.
[19]张玉英,何云义,惠朋利,等.姜辛夏颗粒对支气管哮喘大鼠肺组织基质金属蛋白酶及其抑制剂含量的影响[J].中国中医药信息杂志,2012,19(2):30-32.
[20]马小娟,孙湛,于文燕,等.维药神香草对哮喘小鼠气道高反应性的抑制作用[J].免疫学杂志,2013,29(8):650-653,658.
[21]张益谋.黄芪注射液对支气管哮喘患儿免疫及气道重塑状态的影响[J].中国临床研究,2014,27(8):994-995.
[22]董雷,蔡宛如.芍药甘草汤对哮喘大鼠气道重塑的影响及相关机制研究[J].中药材,2016,93(4):887-890.
[2]Coleman SL,Shaw OM.Progress in the understanding of the pathology of allergic asthma and the potential of fruit proanthocyanidins as modulators of airway inflammation[J].Food Funct,2017,8(12):4315-4324.
[3]朱金凤.支气管哮喘的中医辨治思维与方法研究[J].中国中医基础医学杂志,2016,22(2):284-286.
[4]王盛隆,杨爽,王强,等.支气管哮喘中医药全病程治疗方案研究进展[J].时珍国医国药,2015,26(3):699-701.
[5]李献超.麻杏石甘汤治疗支气管哮喘急发期的临床观察[J].中药药理与临床,2015,31(1):292-293.
[6]刘春雨.麻杏石甘汤治疗支气管哮喘急发期的临床疗效分析[J].中国医学创新,2012,9(35):47-48.
[7]虞琳,朱丽君,张怡静,等.麻杏石甘汤对哮喘大鼠气道上皮STAT6和黏蛋白表达的调控[J].医学研究杂志,2014,43(5):40-44.
[8]高尚泽,杨林伟,王小莹,等.麻杏石甘汤对哮喘大鼠肺组织中Sec14l3表达的影响[J].天津中医药,2013,30(12):745-748.
[9]韩凤芹,孙雪文,张新国,等.麻杏石甘汤对哮喘模型小鼠支气管黏膜和肺组织免疫的影响[J].时珍国医国药,2012,23(6):1398-1399.
[10]黄丰,童晓云,张荣华.麻杏石甘汤调节哮喘模型小鼠Th1/Th2反应的机制初探[J].中药材,2008,31(10):1519-1522.
[11]李厚忠,高照渝,黄伟,等.中药川贝母对哮喘模型小鼠MMP-2,MMP-9和TIMP-1的影响[J].中国中药杂志,2017,42(21):4180-4186.
[12]杨汀,王辰,庞宝森,等.豚鼠哮喘模型气道重建对气道反应性的影响[J].中国病理生理杂志,2004,20(11):2033-2036.
[13]Liccardi G,Salzillo A,Piccolo A,et al.Dysfunction of small airways and prevalence,airway responsiveness and inflammation in asthma:much more than small particle size of pet animal allergens[J].Ups J Med Sci,2016,121(3):196-197.
[14]张方琪,韩新鹏,张芳,等.动态观察过敏原诱导的哮喘小鼠模型从急性炎症到慢性重塑的发展[J].中国病理生理杂志,2017,33(2):380-384.
[15]Grenier PA,Fetita CI,Brillet PY.Quantitative computed tomography imaging of airway remodeling in severe asthma[J].Quant Imaging Med Surg,2016,6(1):76-83.
[16]王爱华,赵霞,董盈妹,等.固本防哮饮对支气管哮喘缓解期小鼠肺组织MMP-2、MMP-9、TIMP-1、TIMP-2蛋白的影响[J].辽宁中医杂志,2018,45(1):183-186,229.
[17]Kim JS,Kang JY,Ha JH,et al.Expression of nerve growth factor and matrix metallopeptidase-9/tissueinhibitor of metalloproteinase-1 in asthmatic patients[J].J Asthma,2013,50(7):712-717.
[18]田金娜,李建保,刘小凡.丹龙定喘汤对哮喘小鼠气道重塑及MMP-9、TIMP-1的影响[J].中国实验方剂学杂志,2012,18(5):164-166.
[19]张玉英,何云义,惠朋利,等.姜辛夏颗粒对支气管哮喘大鼠肺组织基质金属蛋白酶及其抑制剂含量的影响[J].中国中医药信息杂志,2012,19(2):30-32.
[20]马小娟,孙湛,于文燕,等.维药神香草对哮喘小鼠气道高反应性的抑制作用[J].免疫学杂志,2013,29(8):650-653,658.
[21]张益谋.黄芪注射液对支气管哮喘患儿免疫及气道重塑状态的影响[J].中国临床研究,2014,27(8):994-995.
[22]董雷,蔡宛如.芍药甘草汤对哮喘大鼠气道重塑的影响及相关机制研究[J].中药材,2016,93(4):887-890.