清金化痰颗粒对COPD急性期(痰热郁肺型)大鼠肺组织STAT1,STAT3的调控作用
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摘要
目的:研究清金化痰颗粒对慢性阻塞性肺疾病(COPD)急性期(痰热郁肺型)大鼠肺组织中信号转导与转录激活因子1,3(STAT1,STAT3)的调控作用。方法:采用烟熏+脂多糖(LPS)气管注射方法建立COPD痰热郁肺证大鼠模型(正常组除外),随机分为正常组,模型组,罗红霉素组(0.031 5 g·kg~(-1)),清金化痰颗粒低、高剂量组(9.4,37.6 g·kg-1),每组8只。每日ig给药1次,连续给药2周。观察各组大鼠的一般行为活动和病理学变化特点,采用大鼠肺功能检测仪测定各组大鼠相关肺功能指标:0.3 s用力肺活量(FEV0.3),用力肺活量(FVC),FEV0.3/FVC,肺活量(VC),采用实时荧光定量聚合酶链式反应(Real-time PCR)法测定肺组织白细胞介素-6(IL-6),STAT1及STAT3 mRNA表达,免疫组化法检测肺组织IL-6,STAT1及STAT3蛋白表达。结果:与正常组比较,模型组肺组织中IL-6,STAT1及STAT3 mRNA表达显著升高(P<0.01);与模型组比较,罗红霉素组、清金化痰颗粒低、高剂量组均能显著降低大鼠肺组织IL-6,STAT1及STAT3 mRNA表达(P<0.01),罗红霉素组和清金化痰高剂量组减低更明显,二者之间无明显差异。各组大鼠肺组织中IL-6,STAT1及STAT3蛋白的表达中,与正常组比较,模型大鼠肺组织中的IL-6,STAT1及STAT3平均积分吸光度IA值显著升高(P<0.01);与正常组比较,模型组大鼠肺功能参数FEV0.3,FVC,FEV0.3/FVC,VC值均降低(P<0.01);与模型组比较,各给药组肺功能参数FEV0.3,FVC,VC值均有升高(P<0.01,P<0.05);与模型组比较,罗红霉素组、清金化痰颗粒低、高剂量组能显著降低大鼠肺组织IL-6,STAT1及STAT3平均IA值(P<0.01),罗红霉素组和清金化痰颗粒高剂量组减低更明显,二者之间差异无统计学意义。结论:清金化痰颗粒可下调JAK/STAT信号通路中STAT1,STAT3的过度表达和持续活化,来抑制IL-6的水平的升高,减轻气道炎症,抑制COPD急性发作与加重。
Objective: To explore the effect of Qingjin Huatan granule( QJHTG) on regulation of transcriptional activator and signal transducer STAT1 and STAT3 in lung tissues of rat models with acute exacerbation of chronic obstructive pulmonary disease( COPD)( phlegm-heat stagnating in lung). Method: COPD rat models with phlegm-heat stagnating in lung were established by smoking and intratracheal injection of LPS( except normal group). The rats were randomly divided into normal group,model group,roxithromycin( ROX)group( 0. 031 5 g·kg~(-1)),QJHTG low dose group and high dose group( 9. 40,37. 6 g·kg~(-1)),n = 8 in each group. The medicines were given once a day by ig administration for 2 weeks. General activities and pathological change features were observed in all groups. The lung function indexes of rats were measured by using lung function tester: forced expiratory volume in 0. 3 second( FEV0. 3),force vital capacity( FVC),FEV0. 3/ FVC and vital capacity( VC). The mRNA expression levels of interleukin-6( IL-6),STAT1 and STAT3 in lung tissues were determined by Real-time PCR method and the protein expression levels of IL-6,STAT1 and STAT3 in lung tissues were determined by immunohistochemical method. Result: As compared with the normal group,the mRNA expression levels of IL-6,STAT1 and STAT3 in lung tissues of model group were significantly increased( P <0. 01). As compared with the model group,the mRNA expression levels of IL-6,STAT1 and STAT3 in lung tissues were significantly decreased in ROX group,QJHTG low and high dose groups( P < 0. 01),and the decrease was more obvious in ROX group and QJHTG high dose group,with no significant difference between these two groups.The average optical density of IL-6,STAT1 and STAT3 protein expression in the model group was significantly higher than that in the normal group( P < 0. 01); the pulmonary function parameters of rats in model group,including FEV0. 3,FVC,FEV0. 3/ FVC and VC were lower than those in normal group( P < 0. 01). As compared with the model group,FEV0. 3,FVC,and VC values were increased in various treatment groups( P < 0. 01,P <0. 05); the average optical density of IL-6,STAT1 and STAT3 was significantly decreased in ROX group,QJHTG low and high dose groups( P < 0. 01),and the decrease was more obvious in ROX group and QJHTG high dose group,with no statistical differences between these two groups. Conclusion: QJHTG can inhibit the increase of IL-6,reduce airway inflammation,and inhibit COPD acute attack and exacerbation by down-regulating excessive expression and sustained activation of STAT1 and STAT3 in the JAK / STAT signaling pathway.
Objective: To explore the effect of Qingjin Huatan granule( QJHTG) on regulation of transcriptional activator and signal transducer STAT1 and STAT3 in lung tissues of rat models with acute exacerbation of chronic obstructive pulmonary disease( COPD)( phlegm-heat stagnating in lung). Method: COPD rat models with phlegm-heat stagnating in lung were established by smoking and intratracheal injection of LPS( except normal group). The rats were randomly divided into normal group,model group,roxithromycin( ROX)group( 0. 031 5 g·kg~(-1)),QJHTG low dose group and high dose group( 9. 40,37. 6 g·kg~(-1)),n = 8 in each group. The medicines were given once a day by ig administration for 2 weeks. General activities and pathological change features were observed in all groups. The lung function indexes of rats were measured by using lung function tester: forced expiratory volume in 0. 3 second( FEV0. 3),force vital capacity( FVC),FEV0. 3/ FVC and vital capacity( VC). The mRNA expression levels of interleukin-6( IL-6),STAT1 and STAT3 in lung tissues were determined by Real-time PCR method and the protein expression levels of IL-6,STAT1 and STAT3 in lung tissues were determined by immunohistochemical method. Result: As compared with the normal group,the mRNA expression levels of IL-6,STAT1 and STAT3 in lung tissues of model group were significantly increased( P <0. 01). As compared with the model group,the mRNA expression levels of IL-6,STAT1 and STAT3 in lung tissues were significantly decreased in ROX group,QJHTG low and high dose groups( P < 0. 01),and the decrease was more obvious in ROX group and QJHTG high dose group,with no significant difference between these two groups.The average optical density of IL-6,STAT1 and STAT3 protein expression in the model group was significantly higher than that in the normal group( P < 0. 01); the pulmonary function parameters of rats in model group,including FEV0. 3,FVC,FEV0. 3/ FVC and VC were lower than those in normal group( P < 0. 01). As compared with the model group,FEV0. 3,FVC,and VC values were increased in various treatment groups( P < 0. 01,P <0. 05); the average optical density of IL-6,STAT1 and STAT3 was significantly decreased in ROX group,QJHTG low and high dose groups( P < 0. 01),and the decrease was more obvious in ROX group and QJHTG high dose group,with no statistical differences between these two groups. Conclusion: QJHTG can inhibit the increase of IL-6,reduce airway inflammation,and inhibit COPD acute attack and exacerbation by down-regulating excessive expression and sustained activation of STAT1 and STAT3 in the JAK / STAT signaling pathway.
引文
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[5]宋一平,崔德健,茅培英,等.慢性阻塞性肺病大鼠模型的建立及药物干预的影响[J].中华内科杂志,2000,39(8):556-559.
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[9]Samavati L,Rastogi R,DU W,et al.STAT3 tyrosine phosphorylation is critical for interleukin 1 beta and interleukin-6 production in response to lipopolysaccharide and live bacteria[J].Mol Immunol,2009,46(8):1867-1877.
[10]岳圆圆,李永春.丹参对慢性阻塞性肺疾病大鼠模型气道保护作用的实验研究[J].临床合理用药,2011,10(32):15-17.
[11]冀兰鑫,黄浩,李长志,等.赤芍药理作用的研究进展[J].药物评价研究,2010,33(3):233-236.
[12]张翠英,章洪,戚琼华.川芎的有效成分及药理研究进展[J].辽宁中医杂志,2014,41(10):2264-2265.
[13]张桂菊,卢立伟,吴金勇,等.加味茵陈蒿汤对湿热哮喘大鼠气道白细胞介素-6、信号转导因子-3表达的影响及与气道炎症的关系[J].中国实验方剂学杂志,2014,21(15):133-137.
[2]许光兰,李娇,韦艾凌.慢性阻塞性肺疾病中医证型相关文献统计分析[J].山东中医杂志,2012,31(1):11-12.
[3]熊瑛,李国平,扬礼腾,等.白介素5与转录激活因子和信号转导子1表达水平及其信号转导对支气管哮喘豚鼠气道炎症影响的研究[J].中华哮喘杂志,2008,2(1):41-45.
[4]Quinton L J,Jones M R,Robson B E,et al.Alveolar epithelial STAT3,IL-6 family cytokines,and host defense during Escherichia coli pneumonia[J].Am J Respir Cell Mol Biol,2008,38(6):699-706.
[5]宋一平,崔德健,茅培英,等.慢性阻塞性肺病大鼠模型的建立及药物干预的影响[J].中华内科杂志,2000,39(8):556-559.
[6]李雯,徐永健,魏华平.白介素-4和白介素-6在慢性阻塞性肺病肺组织中的表达[J].中国组织化学与细胞化学杂志,2001,10(4):425-428.
[7]何智辉,陈平,伍国宝,等.白细胞介素6在慢性阻塞性疾病患者痰和血清中的浓度变化[J].中国现代医学杂志,2005,15(2):1768-1770.
[8]GONG J H,Shin D,HAN S Y,et al.Blockade of airway inflammation by kaempferol via disturbing Tyk-STAT signaling in airway epithelial cells and in asthmatic mice[J].Evid Based Complement Alternat Med,2013,doi:10.1155/2013/250725.
[9]Samavati L,Rastogi R,DU W,et al.STAT3 tyrosine phosphorylation is critical for interleukin 1 beta and interleukin-6 production in response to lipopolysaccharide and live bacteria[J].Mol Immunol,2009,46(8):1867-1877.
[10]岳圆圆,李永春.丹参对慢性阻塞性肺疾病大鼠模型气道保护作用的实验研究[J].临床合理用药,2011,10(32):15-17.
[11]冀兰鑫,黄浩,李长志,等.赤芍药理作用的研究进展[J].药物评价研究,2010,33(3):233-236.
[12]张翠英,章洪,戚琼华.川芎的有效成分及药理研究进展[J].辽宁中医杂志,2014,41(10):2264-2265.
[13]张桂菊,卢立伟,吴金勇,等.加味茵陈蒿汤对湿热哮喘大鼠气道白细胞介素-6、信号转导因子-3表达的影响及与气道炎症的关系[J].中国实验方剂学杂志,2014,21(15):133-137.