益气消癥通络方对慢性阻塞性肺疾病血管重塑模型大鼠VEGF、ET-1和CTGF表达的影响
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摘要
目的:探讨益气消癥通络方对慢性阻塞性肺疾病(COPD)血管重塑模型大鼠血清血管内皮细胞生长因子(VEGF)、内皮素-1(ET-1)、结缔组织生长因子(CTGF)表达的影响。方法:60只Wistar大鼠随机分为6组:空白组(n=10)、模型组(n=10)、西药组(n=10)、益气消癥通络方低剂量组(n=10)、中剂量组(n=10)及高剂量组(n=10)。造模采用香烟烟雾暴露联合气道注入脂多糖(LPS)法制备COPD血管重塑大鼠模型。实验第4天起,模型组大鼠按10 mL/(kg·d)给予生理盐水灌胃;西药组大鼠雾化吸入布地奈德混悬液(剂量按体表面积换算为成人量的6.17倍),1次/d;中药组干预分别按照益气消癥通络方低剂量组[17.89 g/(kg·d)]、中剂量组[35.78 g/(kg·d)]、高剂量组[53.67 g/(kg·d)]灌胃给药,共12周。用酶联免疫吸附法(ELISA)检测大鼠血清VEGF、ET-1、CTGF水平。结果:大鼠肺血管病理改变显示COPD血管重塑大鼠模型造模成功。ELISA检测结果:与正常组大鼠血清VEGF、ET-1、CTGF表达水平相比,其余各组均升高,差异有统计学意义(P<0.01);与模型组VEGF、ET-1、CTGF表达水平相比,西药组的VEGF、ET-1含量表达下降,差异有统计学意义(P<0.01),益气消癥通络方低剂量组、中剂量组及高剂量组的VEGF、ET-1、CTGF表达水平明显均有下降趋势,差异有统计学意义(P<0.01);与西药组大鼠血清VEGF、ET-1、CTGF水平比较,益气消癥通络方低剂量组ET-1、CTGF表达水平均下降,差异有统计学意义(P<0.05),中剂量组及高剂量组VEGF、ET-1、CTGF含量均下降,差异均有统计学意义(P<0.05);与益气消癥通络方高剂量组比较,西药组、低剂量及中剂量组差异均具有统计学意义(P<0.05)。结论:益气消癥通络方能够通过降低VEGF、ET-1、CTGF表达水平,干预COPD血管重塑,其中益气消癥通络方高剂量效果更为显著。
Obective: To obeserve the effect of Yiqi Xiaozheng Tongluo Recipe on expressions of VEGF,ET-1 and CTGF in rats with COPD vascular remodeling. Methods: Sixty Wistar rats were randomly divided into the normal group,the model group,the western medicine group,the low,middle and high dose groups of Yiqi Xiaozheng Tongluo Recipe,10 rats in each group. Cigarette smoking exposure combined with airway injection of lipopolysaccharide( LPS) was used to prepare rats of COPD vascular remodeling. On the fourth day of the experiment,the rats in model group were given the normal saline by 10 mL/( kg· d). The group of western medicine received nebulized inhalation of budesonide suspension ( the dose was 6. 17 times the body size),1 time/d and the intervented groups of traditional Chinese medicine were administered according to the low dose[17. 89 g/( kg· d) ],middle dose[( 35. 78 g/( kg·d) ],and high dose[( 53. 67 g/( kg·d) ],totally 12 weeks. Enzyme-linked immunosorbent assay( ELISA) was used to detect VEGF,ET-1 and CTGF levels in rats. Results: The pathological changes of pulmonary vessels in rats showed that models of COPD vascular remodeling rats were successful. ELISA test results: Compared with the normal group,the expression levels of VEGF,ET-1 and CTGF in all other groups increased and the difference was statistically significant( P < 0. 01). Compared with the expression levels of VEGF,ET-1 and CTGF in the model group,the expressions of VEGF and ET-1 in the western medicine group decreased and the difference was statistically significant( P < 0. 01). The expressions of VEGF,ET-1 and CTGF in the low dose group,the middle dose group and the high dose group were obviously decreased and the difference was statistically significant( P < 0. 01). Compared with the levels of serum VEGF,ET-1 and CTGF in the western medicine group,the expression levels of ET-1 and CTGF decreased in the low dose group of Yiqi Xiaozheng Tongluo Recipe group and the difference was statistically significant( P < 0. 05). The contents of VEGF,ET-1 and CTGF decreased in both middle and high dose groups and the difference was statistically significant( P < 0. 05). Compared with the high dose group,western medicine group,the low dose group and the middle dose group were statistically significant( P < 0. 05). Conclusion: Yiqi Xiaozheng Tongluo Recipe can interfere COPD vascular remodeling by reducing the expression levels of VEGF,ET-1 and CTGF,in which the effect of high dose is more obvious.
Obective: To obeserve the effect of Yiqi Xiaozheng Tongluo Recipe on expressions of VEGF,ET-1 and CTGF in rats with COPD vascular remodeling. Methods: Sixty Wistar rats were randomly divided into the normal group,the model group,the western medicine group,the low,middle and high dose groups of Yiqi Xiaozheng Tongluo Recipe,10 rats in each group. Cigarette smoking exposure combined with airway injection of lipopolysaccharide( LPS) was used to prepare rats of COPD vascular remodeling. On the fourth day of the experiment,the rats in model group were given the normal saline by 10 mL/( kg· d). The group of western medicine received nebulized inhalation of budesonide suspension ( the dose was 6. 17 times the body size),1 time/d and the intervented groups of traditional Chinese medicine were administered according to the low dose[17. 89 g/( kg· d) ],middle dose[( 35. 78 g/( kg·d) ],and high dose[( 53. 67 g/( kg·d) ],totally 12 weeks. Enzyme-linked immunosorbent assay( ELISA) was used to detect VEGF,ET-1 and CTGF levels in rats. Results: The pathological changes of pulmonary vessels in rats showed that models of COPD vascular remodeling rats were successful. ELISA test results: Compared with the normal group,the expression levels of VEGF,ET-1 and CTGF in all other groups increased and the difference was statistically significant( P < 0. 01). Compared with the expression levels of VEGF,ET-1 and CTGF in the model group,the expressions of VEGF and ET-1 in the western medicine group decreased and the difference was statistically significant( P < 0. 01). The expressions of VEGF,ET-1 and CTGF in the low dose group,the middle dose group and the high dose group were obviously decreased and the difference was statistically significant( P < 0. 01). Compared with the levels of serum VEGF,ET-1 and CTGF in the western medicine group,the expression levels of ET-1 and CTGF decreased in the low dose group of Yiqi Xiaozheng Tongluo Recipe group and the difference was statistically significant( P < 0. 05). The contents of VEGF,ET-1 and CTGF decreased in both middle and high dose groups and the difference was statistically significant( P < 0. 05). Compared with the high dose group,western medicine group,the low dose group and the middle dose group were statistically significant( P < 0. 05). Conclusion: Yiqi Xiaozheng Tongluo Recipe can interfere COPD vascular remodeling by reducing the expression levels of VEGF,ET-1 and CTGF,in which the effect of high dose is more obvious.
引文
[1] 中华医学会呼吸病学会慢性阻塞性肺疾病学组.慢性阻塞性肺疾病诊治指南(2013年修订版)[S].中国医学前沿杂志(电子版),2014,6(2):67-80.
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[6] 张伟,谷明明,孙璐璐,等.活血化瘀中药对COPD血管重塑的干预作用[J].中国实验方剂学杂志,2013,19(8):254-257.
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[12] 车向前,林育红.慢性阻塞性肺疾病患者血清干扰素-γ、肿瘤坏死因子-α和内皮素-1水平与肺功能关系的临床研究[J].检验医学与临床,2013,10(21):2829-2830.
[13] 王卓.老年COPD患者血清MMP-9、SAA、SOD和ET-1水平与病情急性发作的相关性分析[J].放射免疫学杂志,2012,25(6):654-657.
[14] 宋本艳,胡强.刺五加注射液对COPD稳定期患者血浆NO和内皮素-1的影响[J].中国临床新医学,2017,10(7):630-632.
[15] 喻东,夏熙郑.COPD大鼠肺组织TGF-β1及CTGF的表达及其相关性研究[J].中国实用医药,2007(36):79-80.
[16] JIANG X,ZHANG,LI F,et al.Allicin as possibleadiunctive therapeutic drug for stage II oral submucous fibrosis preliminary clinical trial in a Chinese cohort[J].Int J Oral Maxillofac Surg,2015,44(12):1540-1546.
[17] 刘娴娴,张杰,崔世超,等.结缔组织生长因子在慢性阻塞性肺疾病血管重建中表达研究(英文)[J].现代生物医学进展,2011,11(9):1663-1666.
[18] 叶烨,周仙仕,王伟荣.从“肺失治节”论治慢性阻塞性肺疾病[J].中国中医急症,2016,25(7):1328-1330.
[19] 王琦,吴海斌,张永生,等.“肺络微型癥瘕”与COPD气道重构的相关性探讨[J].北京中医药大学学报,2012,35(2):130-133.
[20] 白晓旭,王琦,张永生,等.慢性阻塞性肺疾病的肺络癥、瘕、聚、散理论[J].中医杂志,2012,53(20):1717-1719.
[21] 刘勇明,吕晓东,庞立健,等.基于肺络构效理论的肺脏生理功能发微[J].中华中医药学刊,2017,35(10):2518-2520.
[22] 冯淬灵,武维屏,武红莉,等.益气活血化痰法治疗慢性阻塞性肺疾病76例临床资料分析[J].北京中医药大学学报,2007(6):419-422.
[2] Prevalence and risk factors of chronic obstructive pulmonary disease in China(the China Pulmonary Health[CPH]study):a national cross-sectional study.Lancet Published Online April 9,2018.
[3] 王英,李亚,李建生.慢性阻塞性肺疾病肺血管重塑研究进展[J].中医学报,2012,27(2):143-145.
[4] Calabrese C,Bocchino V,Vatrella A,et al.Evidence of angiogenesis in bronchial biopsies of smokers with and without airway obstruction[J].Respir Med,2006,100(8):1415-1422.
[5] 王昌明,王会娟.高迁移率族蛋白B1与慢性阻塞性肺疾病继发肺动脉高压的关系[J].中国老年学杂志,2013,33(10):5221-5223.
[6] 张伟,谷明明,孙璐璐,等.活血化瘀中药对COPD血管重塑的干预作用[J].中国实验方剂学杂志,2013,19(8):254-257.
[7] Tripp DA,Curtis Nickel J,Landis LR,et al.Predictors of quality of life and pain in chronic prostatitis/chronic pelvic pain syndrome:finding from the National Institutes of Health Cohort Study[J].BJU Int,2004,94(9):1279-1982.
[8] 高云,罗凤鸣,范志航.血管内皮生长因子与肺部疾病的关系[J].中华哮喘杂志(电子版),2013,7(5):369-374.
[9] Ishida A,Murray J,Saito Y,et al.Expression of vascular endothelial growth factor receptors in smooth muscle cell[J].J Cell Physiol,2001,188(3):359-368.
[10] 廖艺璇,陈亚红,米文君.慢性阻塞性肺疾病肺血管重塑的研究进展[J].生理科学进展,2016,47(4):295-299.
[11] Salud S,Victor I,Peinado J,et al.Enhanced expression of vascular endothelial growth factor in pulmonary arteries of smokers and patients with moderate chronic obstructive pulmonary disease[J].Am J Respir Crit Care Med,2003,167(9):1250-1256.
[12] 车向前,林育红.慢性阻塞性肺疾病患者血清干扰素-γ、肿瘤坏死因子-α和内皮素-1水平与肺功能关系的临床研究[J].检验医学与临床,2013,10(21):2829-2830.
[13] 王卓.老年COPD患者血清MMP-9、SAA、SOD和ET-1水平与病情急性发作的相关性分析[J].放射免疫学杂志,2012,25(6):654-657.
[14] 宋本艳,胡强.刺五加注射液对COPD稳定期患者血浆NO和内皮素-1的影响[J].中国临床新医学,2017,10(7):630-632.
[15] 喻东,夏熙郑.COPD大鼠肺组织TGF-β1及CTGF的表达及其相关性研究[J].中国实用医药,2007(36):79-80.
[16] JIANG X,ZHANG,LI F,et al.Allicin as possibleadiunctive therapeutic drug for stage II oral submucous fibrosis preliminary clinical trial in a Chinese cohort[J].Int J Oral Maxillofac Surg,2015,44(12):1540-1546.
[17] 刘娴娴,张杰,崔世超,等.结缔组织生长因子在慢性阻塞性肺疾病血管重建中表达研究(英文)[J].现代生物医学进展,2011,11(9):1663-1666.
[18] 叶烨,周仙仕,王伟荣.从“肺失治节”论治慢性阻塞性肺疾病[J].中国中医急症,2016,25(7):1328-1330.
[19] 王琦,吴海斌,张永生,等.“肺络微型癥瘕”与COPD气道重构的相关性探讨[J].北京中医药大学学报,2012,35(2):130-133.
[20] 白晓旭,王琦,张永生,等.慢性阻塞性肺疾病的肺络癥、瘕、聚、散理论[J].中医杂志,2012,53(20):1717-1719.
[21] 刘勇明,吕晓东,庞立健,等.基于肺络构效理论的肺脏生理功能发微[J].中华中医药学刊,2017,35(10):2518-2520.
[22] 冯淬灵,武维屏,武红莉,等.益气活血化痰法治疗慢性阻塞性肺疾病76例临床资料分析[J].北京中医药大学学报,2007(6):419-422.