健脾补肾方对吸烟诱导COPD小鼠NEDD4-PTEN-PI3k/AKT信号通路的影响
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摘要
目的评价健脾补肾方对吸烟诱导的慢性阻塞性肺疾病小鼠模型及对其NEDD4-PTEN-PI3k/AKT信号通路的作用。方法 60只C57BL/6J小鼠随机分为空白组(WT),造模组(CS)以及中药组(CH),每组20只。运用H&E染色对COPD小鼠肺脏形态学进行观察,评价中药治疗效果;采用Western blot方法检测小鼠肺组织NEDD4、PTEN、PI3k、AKT蛋白表达水平,qPCR检测小鼠肺组织NEDD4、PTEN、PI3k、mTOR、AKT mRNA表达水平。结果与模型组比较,健脾补肾方能改善小鼠整体状态,可降低COPD模型小鼠气道阻力(P<0.05),降低准静态肺顺应性和功能残气量;降低肺泡炎性因子(P<0.05),下调NEDD4、上调PTEN、下调PI3k与AKT的表达(P<0.05)。结论健脾补肾方改善COPD模型小鼠一般状态,保护肺功能,减轻肺组织结构损害的机制可能是通过调控NEDD4-PTEN-PI3k/AKT信号通路来实现的。
Objective To investigate the effects of Jianpibushen decoction on smoking-induced mice with chronic obstructive pulmonary disease(COPD) and its role in NEDD4-PTEN-PI3k/AKT signaling pathway.Methods Sixty C57BL/6J mice were randomly divided into three groups:blank group,modeling group and TCM group,with 20 mice in each group.The HE staining was used to observe the morphological changes of lungs in mice with COPD and to evaluate the therapeutic effect of TCM.Moreover the Western Blot was used to detect the expression levels of NEDD4,PTEN,PI3k and AKT protein in lung tissue of mice,and qPCR was used to detect the expression levels of NEDD4, PTEN, PI3k,mTOR,AKT mRNA in lung tissue of mice.Results As compared with model group,Jianpibushen decoction could improve the overall state of the mice,reduce airway resistance of mice with COPD in model group(P<0.05),moreover,which could decrease quasi-static lung compliance and functional residual capacity,and reduce alveolar inflammatory factors(P<0.05).In addition Jianpibushen decoction could down-regulate the expression levels of NEDD4 and up regulate the expression levels of PTEN,which could down regulate expression levels of PI3k and AKT(P<0.05).Conclusion The Jianpibushen decoction can improve the general state of mice with COPD,protect lung function,and alleviate the structural damage of lung tissue,and the action mechanism may be correlated with the regulation of NEDD4-PTEN-PI3k/AKT signaling pathway.
Objective To investigate the effects of Jianpibushen decoction on smoking-induced mice with chronic obstructive pulmonary disease(COPD) and its role in NEDD4-PTEN-PI3k/AKT signaling pathway.Methods Sixty C57BL/6J mice were randomly divided into three groups:blank group,modeling group and TCM group,with 20 mice in each group.The HE staining was used to observe the morphological changes of lungs in mice with COPD and to evaluate the therapeutic effect of TCM.Moreover the Western Blot was used to detect the expression levels of NEDD4,PTEN,PI3k and AKT protein in lung tissue of mice,and qPCR was used to detect the expression levels of NEDD4, PTEN, PI3k,mTOR,AKT mRNA in lung tissue of mice.Results As compared with model group,Jianpibushen decoction could improve the overall state of the mice,reduce airway resistance of mice with COPD in model group(P<0.05),moreover,which could decrease quasi-static lung compliance and functional residual capacity,and reduce alveolar inflammatory factors(P<0.05).In addition Jianpibushen decoction could down-regulate the expression levels of NEDD4 and up regulate the expression levels of PTEN,which could down regulate expression levels of PI3k and AKT(P<0.05).Conclusion The Jianpibushen decoction can improve the general state of mice with COPD,protect lung function,and alleviate the structural damage of lung tissue,and the action mechanism may be correlated with the regulation of NEDD4-PTEN-PI3k/AKT signaling pathway.
引文
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8 景璇,郭洁,赵小静,等.慢性阻塞性肺疾病急性加重期的中医治疗研究进展.世界中医药,2016,11:1384-1388.
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13 John G,Kohse K,Orasche J,et al.The composition of cigarette smoke determines inflammatory cell recruitment to the lung in COPD mouse models.Clinical Science,2014,126:207-221.
14 周奎龙,解庆凡,王志恒,等.健脾补肾法在慢性阻塞性肺疾病中的临床应用.河北中医杂志,2016,38:1883-1886.
15 Malaviya R,Laskin JD,Laskin DL.Anti-TNF-α therapy in inflammatory lung diseases.Pharmacology & therapeutics,2017,41:417-425.
16 Rabe KF.Global Initiative for Chronic Obstructive Lung Disease.Global strategy for the diagnosis,management,and prevention of chronic obstructive pulmonary disease.GOLD executive summary.Am J Respir Crit Care Med,2007,176:532-555.
17 Sapey E,Stockley RA.COPD exacerbations:Aetiology.Thorax,2006,61:250-258.
18 Bozinovski S,Vlahos R,Hansen M,et al.Akt in the pathogenesis of COPD.International journal of chronic obstructive pulmonary disease,2006,1:31.
19 Jiang Y,Wang X,Hu D.Mitochondrial alterations during oxidative stress in chronic obstructive pulmonary disease.International journal of chronic obstructive pulmonary disease,2017,12:1153.
20 Huang ZJ,Zhu JJ,Yang XY,et al.NEDD4 promotes cell growth and migration via PTEN/PI3K/AKT signaling in hepatocellular carcinoma.Oncology letters,2017,14:2649-2656.
2 Barnes PJ,Shapiro SD,Pauwels RA.Chronic obstructive pulmonary disease:molecular and cellularmechanisms.European Respiratory Journal,2003,22:672-688.
3 Schikowski T,Mills IC,Anderson HR,et al.Ambient air pollution:a cause of COPD.European Respiratory Journal,2014,43:250-263.
4 Salvi S.Tobacco smoking and environmental risk factors for chronic obstructive pulmonary disease.Clinics in chest medicine,2014,35:17-27.
5 Pérez-Padilla R,Ramirez-Venegas A,Sansores-Martinez R.Clinical characteristics of patients with biomass smoke-associated COPD and chronic bronchitis,2004-2014.Chronic Obstructive Pulmonary Diseases:Journal of the COPD Foundation,2014,1:23.
6 Capistrano S J,van Reyk D,Chen H,et al.Evidence of Biomass Smoke Exposure as a Causative Factor for the Development of COPD.Toxics,2017,5:36.
7 Decramer M,Janssens W,Miravitlles M.Chronic obstructive pulmonary disease.Lancet,2012,379:1341-1351.
8 景璇,郭洁,赵小静,等.慢性阻塞性肺疾病急性加重期的中医治疗研究进展.世界中医药,2016,11:1384-1388.
9 吴健广,郭思佳,封继宏,等.中医辨证论治慢性阻塞性肺疾病稳定期研究进展.中华中医药学刊,2015,33:1104-1106.
10 乐一萍,孙燕妮.中医防治慢性阻塞性肺疾病的机制研究进展.环球中医药,2013,6:226-229.
11 俞宁宁,王真,杨珺超.COPD稳定期中医证治研究进展.浙江中医药大学学报,2012,36:467-470.
12 Eltom S,Stevenson C,Birrell MA.Cigarette smoke exposure as a model of inflammation associated with COPD.Current protocols in pharmacology,2013,24:11-18.
13 John G,Kohse K,Orasche J,et al.The composition of cigarette smoke determines inflammatory cell recruitment to the lung in COPD mouse models.Clinical Science,2014,126:207-221.
14 周奎龙,解庆凡,王志恒,等.健脾补肾法在慢性阻塞性肺疾病中的临床应用.河北中医杂志,2016,38:1883-1886.
15 Malaviya R,Laskin JD,Laskin DL.Anti-TNF-α therapy in inflammatory lung diseases.Pharmacology & therapeutics,2017,41:417-425.
16 Rabe KF.Global Initiative for Chronic Obstructive Lung Disease.Global strategy for the diagnosis,management,and prevention of chronic obstructive pulmonary disease.GOLD executive summary.Am J Respir Crit Care Med,2007,176:532-555.
17 Sapey E,Stockley RA.COPD exacerbations:Aetiology.Thorax,2006,61:250-258.
18 Bozinovski S,Vlahos R,Hansen M,et al.Akt in the pathogenesis of COPD.International journal of chronic obstructive pulmonary disease,2006,1:31.
19 Jiang Y,Wang X,Hu D.Mitochondrial alterations during oxidative stress in chronic obstructive pulmonary disease.International journal of chronic obstructive pulmonary disease,2017,12:1153.
20 Huang ZJ,Zhu JJ,Yang XY,et al.NEDD4 promotes cell growth and migration via PTEN/PI3K/AKT signaling in hepatocellular carcinoma.Oncology letters,2017,14:2649-2656.