清金化痰颗粒对慢性阻塞性肺疾病急性加重期痰热郁肺型大鼠肺组织JAK/STAT信号通路的影响
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摘要
目的探讨清金化痰颗粒治疗慢性阻塞性肺疾病急性加重期(AECOPD)痰热郁肺型的可能作用机制。方法将40只大鼠随机分成空白组、模型组、西药组及中药低、高剂量组各8只。除正常组外,其余大鼠均采用烟熏+脂多糖气管滴入方法建立AECOPD痰热郁肺型大鼠模型。造模后西药组给予罗红霉素0.0315 g/(kg·d)灌胃,中药低、高剂量组分别给予清金化痰颗粒9.4、37.6 g/(kg·d)灌胃,空白组及模型组分别给予10 ml/(kg·d)生理盐水灌胃。给药2周后观察各组大鼠肺泡灌洗液(BALF)中白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)含量,肺组织中酪氨酸激酶2(JAK2)、细胞因子信号抑制物3(SOCS3)mRNA表达,检测磷酸化信号转导转录激活因子1(p-STAT1)、磷酸化信号转导转录激活因子3(p-STAT3)、JAK2、磷酸化酪氨酸激酶2(p-JAK2)、SOCS3蛋白表达。结果与模型组比较,各给药组大鼠BALF中IL-1β、TNF-α含量明显降低,肺组织中JAK2 mRNA、蛋白表达及p-STAT1、p-STAT3、pJAK2蛋白表达明显下降,SOCS3 mRNA及蛋白表达明显升高(P<0.05或P<0.01),且中药高剂量组与西药组相当(P>0.05),中药低剂量组效果低于中药高剂量组和西药组(P<0.05或P<0.01)。结论清金化痰颗粒治疗AECOPD痰热郁肺型的作用机制之一,可能通过下调p-STAT1、p-STAT3、p-JAK2蛋白及JAK2蛋白及基因表达,上调SOCS3蛋白及基因表达从而抑制AECOPD炎症反应,减轻肺组织损伤,且高剂量效果更好。
Objective To discuss the potential mechanism of Qingjin Huatan Granule( QJHTG)( 清金化痰颗粒) in treating acute exacerbation of chronic obstructive pulmonary disease( AECOPD) with phlegm-heat stagnating in lung. Methods A total of 40 Sprague Dawley rats were randomized into blank group,model group,western medicine group and the low,high dose QJHTG groups,with 8 rats in each group. The rat model of phlegm-heat stagnating in lung of AECOPD was established by smoking and dripping the LPS into the trachea except those in the blank group. After modeling,the western medicine group was intragastrical administration 0. 0315 g/( kg·d) of roxithromycin( ROX). The low,high dose QJHTG groups were respectively intragastrical administration 9. 4 g/( kg·d),and37. 6 g/( kg · d) of QJHTG. The blank group and the model group were respectively intragastrical administration10 ml/( kg·d) of normal saline. After 2 weeks' drug administration,the inflammatory mediators contents of interleukin-1β( IL-1β),tumor necrosis factor-α( TNF-α) in bronchoalveolar lavage fluid( BALF) in each group were observed,and the expression of Janus kinase 2( JAK2) and suppressors-of-cytokine-signaling 3( SOCS3) mRNA in lung tissues was detected. The phosphorylation signal transducer and activator of transcription 1( p-STAT1),phosphorylation signal transducer and activator of transcription 3( p-STAT3),JAK2,p-JAK2,SOCS3 protein expression were detected by Western Blot. Results After drug administration,compared with the model group,the inflammatory mediators contents of IL-1β,TNF-α in BALF in the QJHTG groups were significantly decreased,and JAK2 mRNA and protein expression,p-STAT1,p-STAT3,p-JAK2 protein expression decreased obviously in the lung tissues. SOCS3 mRNA and protein expression significantly increased in the lung tissues( P < 0. 05 or P < 0. 01). The effects of high dose QJHTG group and western medicine group was similar( P > 0. 05). The effects of low dose QJHTG group were lower than that of high dose QJHTG group and Western medicine group( P < 0. 05 or P < 0. 01).Conclusion One of the mechanisms for QJHTG treatment phlegm-heat stagnating in the lung of AECOPD may be by down-regulated the expression of p-STAT1,p-STAT3,JAK2,p-JAK2 and up-regulated the expression of SOCS3,consequently inhibiting the inflammatory response of AECOPD and alleviating the lung tissue injury. The high dose shows better effects.
Objective To discuss the potential mechanism of Qingjin Huatan Granule( QJHTG)( 清金化痰颗粒) in treating acute exacerbation of chronic obstructive pulmonary disease( AECOPD) with phlegm-heat stagnating in lung. Methods A total of 40 Sprague Dawley rats were randomized into blank group,model group,western medicine group and the low,high dose QJHTG groups,with 8 rats in each group. The rat model of phlegm-heat stagnating in lung of AECOPD was established by smoking and dripping the LPS into the trachea except those in the blank group. After modeling,the western medicine group was intragastrical administration 0. 0315 g/( kg·d) of roxithromycin( ROX). The low,high dose QJHTG groups were respectively intragastrical administration 9. 4 g/( kg·d),and37. 6 g/( kg · d) of QJHTG. The blank group and the model group were respectively intragastrical administration10 ml/( kg·d) of normal saline. After 2 weeks' drug administration,the inflammatory mediators contents of interleukin-1β( IL-1β),tumor necrosis factor-α( TNF-α) in bronchoalveolar lavage fluid( BALF) in each group were observed,and the expression of Janus kinase 2( JAK2) and suppressors-of-cytokine-signaling 3( SOCS3) mRNA in lung tissues was detected. The phosphorylation signal transducer and activator of transcription 1( p-STAT1),phosphorylation signal transducer and activator of transcription 3( p-STAT3),JAK2,p-JAK2,SOCS3 protein expression were detected by Western Blot. Results After drug administration,compared with the model group,the inflammatory mediators contents of IL-1β,TNF-α in BALF in the QJHTG groups were significantly decreased,and JAK2 mRNA and protein expression,p-STAT1,p-STAT3,p-JAK2 protein expression decreased obviously in the lung tissues. SOCS3 mRNA and protein expression significantly increased in the lung tissues( P < 0. 05 or P < 0. 01). The effects of high dose QJHTG group and western medicine group was similar( P > 0. 05). The effects of low dose QJHTG group were lower than that of high dose QJHTG group and Western medicine group( P < 0. 05 or P < 0. 01).Conclusion One of the mechanisms for QJHTG treatment phlegm-heat stagnating in the lung of AECOPD may be by down-regulated the expression of p-STAT1,p-STAT3,JAK2,p-JAK2 and up-regulated the expression of SOCS3,consequently inhibiting the inflammatory response of AECOPD and alleviating the lung tissue injury. The high dose shows better effects.
引文
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[2]田燕歌,李亚,李建生,等.调补肺肾法对COPD大鼠JAK/STAT信号转导的影响及远后效应[J].中国中药杂志,2013,38(1):75-80.
[3]YEW-BOOTH L,BIRRELL MA,LAU MS,et al.JAKSTAT pathway activation in COPD[J].Eur Respir J,2015,46(3):843-845.
[4]FRIDMAN JS,SCHERLE PA,COLLINS R,et al.Selective inhibition of JAK1 and JAK2 is efficacious in rodent models of arthritis:preclinical characterization of INCB028050[J].J Immunol,2010,184(9):5298-5307.
[5]李建生,张海龙,王海峰,等.慢性阻塞性肺疾病证候演变特点临床调查[J].中医杂志,2017,58(9):772-776.
[6]赵媚.清金化痰汤对慢性阻塞性肺疾病急性加重期痰热郁肺证患者临床疗效及相关炎症因子的影响[D].南宁:广西中医药大学,2016.
[7]宋小莲,王昌惠,白冲.脂多糖结合熏烟法和单纯熏烟法建立慢性阻塞性肺病大鼠模型的比较[J].第二军医大学学报,2010,31(3):246-249.
[8]许光兰,赵媚,钟云青,等.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[J].中国实验方剂学杂志,2017,23(2):91-97.
[9]徐淑云,卞如濂,陈修.药理实验方法学[M].3版.北京:人民卫生出版社,2001:202-203.
[10]VESTBO J,HOGG JC.Convergence of the epidemiology and pathology of COPD[J].Thorax,2006,61(1):86-88.
[11]JENKINS BJ.Transcriptional regulation of pattern recognition receptors by Jak/STAT signaling,and the implications for disease pathogenesis[J].J Interferon Cytokine Res,2014,34(10):750-758.
[12]YANG L,LIAN X,COWEN A,et al.Synergy between signal transducer and activator of transcription 3 and retinoic acid receptor-alpha in regulation of the surfactant protein B gene in the lung[J].Mol Endocrinol,2004,18(6):1520-1532.
[13]许光兰,赵媚,李娇,等.JAK/STAT信号通路在慢性呼吸系统疾病中的研究进展[J].辽宁中医杂志,2016,43(7):1549-1552.
[14]FERRARI L,KRANE MK,FICHERA A.Inflammatory bowel disease surgery in the biologic era[J].World J Gastrointest Surg,2016,8(5):363-370.
[15]SHAW JG,VAUGHAN A,DENT AG,et al.Biomarkers of progression of chronic obstructive pulmonary disease(COPD)[J].J Thorac Dis,2014,6(11):1532-1547.
[16]CARAMORI G,ADCOCK IM,DI STEFANO A,et al.Cytokine inhibition in the treatment of COPD[J].Int J Chron Obstruct Pulmon Dis,2014,9:397-412.doi:10.2147/COPD.S42544.
[2]田燕歌,李亚,李建生,等.调补肺肾法对COPD大鼠JAK/STAT信号转导的影响及远后效应[J].中国中药杂志,2013,38(1):75-80.
[3]YEW-BOOTH L,BIRRELL MA,LAU MS,et al.JAKSTAT pathway activation in COPD[J].Eur Respir J,2015,46(3):843-845.
[4]FRIDMAN JS,SCHERLE PA,COLLINS R,et al.Selective inhibition of JAK1 and JAK2 is efficacious in rodent models of arthritis:preclinical characterization of INCB028050[J].J Immunol,2010,184(9):5298-5307.
[5]李建生,张海龙,王海峰,等.慢性阻塞性肺疾病证候演变特点临床调查[J].中医杂志,2017,58(9):772-776.
[6]赵媚.清金化痰汤对慢性阻塞性肺疾病急性加重期痰热郁肺证患者临床疗效及相关炎症因子的影响[D].南宁:广西中医药大学,2016.
[7]宋小莲,王昌惠,白冲.脂多糖结合熏烟法和单纯熏烟法建立慢性阻塞性肺病大鼠模型的比较[J].第二军医大学学报,2010,31(3):246-249.
[8]许光兰,赵媚,钟云青,等.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[J].中国实验方剂学杂志,2017,23(2):91-97.
[9]徐淑云,卞如濂,陈修.药理实验方法学[M].3版.北京:人民卫生出版社,2001:202-203.
[10]VESTBO J,HOGG JC.Convergence of the epidemiology and pathology of COPD[J].Thorax,2006,61(1):86-88.
[11]JENKINS BJ.Transcriptional regulation of pattern recognition receptors by Jak/STAT signaling,and the implications for disease pathogenesis[J].J Interferon Cytokine Res,2014,34(10):750-758.
[12]YANG L,LIAN X,COWEN A,et al.Synergy between signal transducer and activator of transcription 3 and retinoic acid receptor-alpha in regulation of the surfactant protein B gene in the lung[J].Mol Endocrinol,2004,18(6):1520-1532.
[13]许光兰,赵媚,李娇,等.JAK/STAT信号通路在慢性呼吸系统疾病中的研究进展[J].辽宁中医杂志,2016,43(7):1549-1552.
[14]FERRARI L,KRANE MK,FICHERA A.Inflammatory bowel disease surgery in the biologic era[J].World J Gastrointest Surg,2016,8(5):363-370.
[15]SHAW JG,VAUGHAN A,DENT AG,et al.Biomarkers of progression of chronic obstructive pulmonary disease(COPD)[J].J Thorac Dis,2014,6(11):1532-1547.
[16]CARAMORI G,ADCOCK IM,DI STEFANO A,et al.Cytokine inhibition in the treatment of COPD[J].Int J Chron Obstruct Pulmon Dis,2014,9:397-412.doi:10.2147/COPD.S42544.