保肺定喘汤对慢性阻塞性肺疾病大鼠肺动脉JAK/STAT信号传导通路的影响
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摘要
目的探讨保肺定喘汤治疗慢性阻塞性肺疾病(COPD)的可能作用机制。方法 50只大鼠随机分为正常组、模型组、中药组、西药组和中西医结合组,每组10只。除正常组外其余各组采用气道滴注脂多糖联合香烟烟雾暴露的方法制备大鼠COPD模型。造模后第29~70天中药组给予保肺定喘汤12.4 g/(kg·d)灌胃,西药组雾化吸入布地奈德混悬液50μg/(kg·d),中西医结合组同时予保肺定喘汤灌胃及雾化吸入布地奈德混悬液,正常组、模型组用4 ml(kg·d)生理盐水灌胃。干预后对各组大鼠肺小动脉进行Masson染色并检测管壁内胶原纤维及α-平滑肌肌动蛋白(α-SMA)表达变化,检测血清白细胞介素6(IL-6)含量及大鼠肺动脉磷酸化Janus激酶2(p-JAK2)和磷酸化信号转导及转录激活因子1(p-STAT1)表达情况。结果与正常组比较,模型组大鼠肺小动脉胶原纤维百分比、α-SMA阳性细胞表达显著增加,血清IL-6含量升高,肺动脉p-JAK2和p-STAT1表达显著增加(P<0.05或P<0.01)。西药组、中药组与中西医结合组肺小动脉胶原纤维百分比、α-SMA阳性细胞表达、血清IL-6含量及肺动脉p-JAK2和p-STAT1表达均较模型组显著减少(P<0.05或P<0.01);中西医结合组较中药组p-JAK2表达显著下降(P<0.05)。结论保肺定喘汤能有效减轻COPD大鼠肺血管重构,其机制可能与抑制JAK/STAT信号传导通路、调节大鼠肺动脉胶原纤维和α-SMA表达有关。
Objective To explore the possible mechanism of Baofei Dingchuan Decoction( 保肺定喘汤) in treating chronic obstructive pulmonary diseases( COPD). Methods Fifty rats were randomized into normal group,model group,herb group,western medicine group and integration group,with 10 in each group. Except the normal group,rats in other groups were made COPD models by lipopolysaccharide airway instillation combined with cigarette smoking exposure. Since the 29 th day to the 70 th day of modeling,the herb group was given Baofei Dingchuan Decoction 12. 4 g/( kg·d) by gavage,the western medicine group was given budesonide suspension 50 μg/( kg·d) by atomizing inhalation. The integration group was given Baofei Dingchuan Decoction by gavage and budesonide suspension by atomizing inhalation. The normal group and the model group were given normal saline 4 ml( kg ·d) by gavage.After intervention,rats' pulmonary arterioles were made Masson staining in each group. The changes of collagen fiber within the vascular wall and the expression ofα-smooth muscle actin( α-SMA) were detected. Serum interleukin 6( IL-6) level,as well as the expressions of rats' pulmonary artery phosphorylated Janus Kinase 2( p-JAK2) and phosphorylated signal transducersand activators of transcription 1( p-STAT1) were detected. Results Compared with the normal group,rats' pulmonary arterioles collagen fiber percentage and α-SMA positive cell expression in the model group decreased,with serum IL-6 level increasing and the expressions of pulmonary artery p-JAK2 and p-STAT1 increasing( P < 0. 05 or P < 0. 01). Compared with the model group,pulmonary arterioles collagen fiber percentage,α-SMA positive cell expression,serum IL-6 level and the expressions of pulmonary artery p-JAK2 and p-STAT1 in the Western medicine group,the herb group and the integration group decreased( P < 0. 05 or P < 0. 01). The expression of p-JAK2 in the integration group was lower than that in the herb group( P < 0. 05). Conclusion Baofei Dingchuan Decoction might reduce the vascular remodeling of COPD rats; the mechanism might relate to inhibiting JAK/STAT signaling pathways and regulating rats' pulmonary arterioles collagen fiber andα-SMA expression.
Objective To explore the possible mechanism of Baofei Dingchuan Decoction( 保肺定喘汤) in treating chronic obstructive pulmonary diseases( COPD). Methods Fifty rats were randomized into normal group,model group,herb group,western medicine group and integration group,with 10 in each group. Except the normal group,rats in other groups were made COPD models by lipopolysaccharide airway instillation combined with cigarette smoking exposure. Since the 29 th day to the 70 th day of modeling,the herb group was given Baofei Dingchuan Decoction 12. 4 g/( kg·d) by gavage,the western medicine group was given budesonide suspension 50 μg/( kg·d) by atomizing inhalation. The integration group was given Baofei Dingchuan Decoction by gavage and budesonide suspension by atomizing inhalation. The normal group and the model group were given normal saline 4 ml( kg ·d) by gavage.After intervention,rats' pulmonary arterioles were made Masson staining in each group. The changes of collagen fiber within the vascular wall and the expression ofα-smooth muscle actin( α-SMA) were detected. Serum interleukin 6( IL-6) level,as well as the expressions of rats' pulmonary artery phosphorylated Janus Kinase 2( p-JAK2) and phosphorylated signal transducersand activators of transcription 1( p-STAT1) were detected. Results Compared with the normal group,rats' pulmonary arterioles collagen fiber percentage and α-SMA positive cell expression in the model group decreased,with serum IL-6 level increasing and the expressions of pulmonary artery p-JAK2 and p-STAT1 increasing( P < 0. 05 or P < 0. 01). Compared with the model group,pulmonary arterioles collagen fiber percentage,α-SMA positive cell expression,serum IL-6 level and the expressions of pulmonary artery p-JAK2 and p-STAT1 in the Western medicine group,the herb group and the integration group decreased( P < 0. 05 or P < 0. 01). The expression of p-JAK2 in the integration group was lower than that in the herb group( P < 0. 05). Conclusion Baofei Dingchuan Decoction might reduce the vascular remodeling of COPD rats; the mechanism might relate to inhibiting JAK/STAT signaling pathways and regulating rats' pulmonary arterioles collagen fiber andα-SMA expression.
引文
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[10]HARKNESS LM,KANABAR V,SHARMA HS,et al.Pulmonary vascular changes in asthma and COPD[J].Pulm Pharmacol Ther,2014,29(2):144-155.
[11]BARBERJA.Mechanisms of development of chronic obstructive pulmonary disease-associated pulmonary hypertension[J].Pulm Circ,2013,3(1):160-164.
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[13]何飞.运用肺络理论治疗肺系疾病刍议[J].浙江中医杂志,2011,46(8):549-550.
[14]何飞,汝触会,陈爱凤,等.益气活血通络法治疗慢性阻塞性肺疾病D组稳定期的临床观察[J].中华中医药学刊,2017,35(6):1466-1468.
[2]LEVY O,TY G.Arginine-vasopressin activates the JAKSTAT pathway in vascular smooth muscle cells[J].J Biol Chem,2006,281(23):15597-15604.
[3]张琼,樊长征,苗青,等.慢性阻塞性肺疾病继发肺动脉高压的中医发病机制及治疗思路[J].中医杂志,2013,54(4):290-292.
[4]何飞,徐俭朴,周忠辉,等.保肺定喘汤对小牛肺动脉平滑肌细胞增殖的干预作用[J].中国中医药科技,2009,16(5):355-356.
[5]方苏榕,谷伟,谭焰,等.烟熏联合内毒素构建COPD大鼠模型[J].南京医科大学学报(自然科学版),2013,33(9):1226-1230.
[6]章元沛.药理实验方法学[M].2版.北京:人民卫生出版,1996:238.
[7]O'SULLIVAN LA,LIONGUE C,LEWIS RS,et al.Cytokine receptor signaling through the Jak-Stat-Socs pathway in disease[J].Mol Immunol,2007,44(10):2497-2506.
[8]GHORESCHI K,LAURENCE A,O'SHEA JJ.Janus kinases in immune cell signaling[J].Immunol Rev,2009,228(1):273-287.
[9]WATANABE S,MU W,KAHN A,et al.Role of JAK/STAT pathway in IL-6-induced activation of vascular smooth muscle cells[J].Am J Nephrol,2004,24(4):387-392.
[10]HARKNESS LM,KANABAR V,SHARMA HS,et al.Pulmonary vascular changes in asthma and COPD[J].Pulm Pharmacol Ther,2014,29(2):144-155.
[11]BARBERJA.Mechanisms of development of chronic obstructive pulmonary disease-associated pulmonary hypertension[J].Pulm Circ,2013,3(1):160-164.
[12]ARCINIEGAS E,FRID MG,DOUGLAS IS,et al.Perspectives on endothelial-to-mesenchymal transition:potential contribution to vascular remodeling in chronic pulmonary hypertension[J].Am J Physiol Lung Cell Mol Physiol,2007,293(1):L1-L8.
[13]何飞.运用肺络理论治疗肺系疾病刍议[J].浙江中医杂志,2011,46(8):549-550.
[14]何飞,汝触会,陈爱凤,等.益气活血通络法治疗慢性阻塞性肺疾病D组稳定期的临床观察[J].中华中医药学刊,2017,35(6):1466-1468.