固本平喘汤对慢阻肺合并肺纤维化大鼠细胞外基质影响的研究
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摘要
研究目的
在临床应用固本平喘汤防治慢性阻塞性肺疾病(COPD)合并肺纤维化(PF)并取得良好疗效的基础上,进一步研究该方对COPD合并PF动物模型细胞外基质Ⅲ型前胶原(PCⅢ)、透明质酸(HA)及层粘连蛋白(LN)含量的影响,探讨固本平喘汤防治COPD合并PF的作用机制,为临床应用提供理论依据。
研究方法
1.动物和分组：将体重180-200g大鼠60只,雌雄各30只,随机分为正常组对照组、模型组、西药组、固本平喘汤小剂量组、大剂量组,每组12只。
2.动物模型制作：将12只大鼠置于特制熏烟箱(60×55×70cm3)内,每9只香烟相接,一端固定于熏箱顶,点燃下端,燃毕换另12只,每日48只,每周7d,连续4周。第5周将除正常组外大鼠以10%水合氯醛(0.36ml/100g)腹腔注射麻醉,颈前剪毛后碘伏常规消毒,行颈正中切口,钝性分离暴露气管,经气管软骨环间隙向心端穿刺一次性注射器注入博莱霉素生理盐水溶液0.2ml(约博莱霉素4mg/kg),立即将动物直立并旋转鼠板,使药液在肺内均匀分布,缝合皮肤,常规消毒,待动物自然苏醒后置笼内常规饲养。造模后第2天开始用药,正常组及模型组均予等体积生理盐水灌胃,2次/日,西药组给予腹腔注射地塞米松0.3mg/100g,1次/日,固本平喘汤(炙麻黄、苦杏仁、甘草、紫苏子、白芥子、莱菔子、黄芩、地龙、淫羊藿、补骨脂、巴戟天、黄芪、丹参、川芎)大、小剂量组分别给予免煎冲剂4m1(根据人与大鼠药物剂量换算公式,设计药物剂量,浓缩成含生药2.7g/ml的免煎冲剂)、1ml(药物浓度同上)灌胃,2次／日,以上各组均给药28天。
3.检测方法和观测指标：①一般情况观测大鼠的活动度、对外界反应的灵敏度、皮毛光泽、饮食、饮水、体重、死亡情况。②HA、LN、PCⅢ检测于第8周,经10%的水合氯醛0.36ml/100g腹腔麻醉后,抽取外周血,严格按照试剂盒说明书采用酶联免疫吸附法检测大鼠血浆中HA、LN、PCⅢ水平。③病理切片形态学观察分批处死大鼠后,迅速取出右肺中叶,10%中性福尔马林溶液中固定,乙醇梯度脱水,二甲苯透明标本,浸蜡,包埋,切片,分别行HE染色,观察肺泡炎、肺纤维化程度以及在3个高倍镜下统计支气管黏膜下成纤维细胞、淋巴细胞、巨噬细胞数。
4.所有结果采用均数±标准差(x±s)表示,采用SPSS10.0软件进行均数单因素方差分析,P<0.05即认为有统计学意义,P<0.01有显著性差异。
研究结果
1.正常对照组皮毛光滑,身体健壮,饮食、活动、体质量增长正常。模型对照组皮毛干枯稀疏,精神不振,反应迟钝,进食、活动均减少,剧烈运动后出现喘息症状,体重较造模前略有增加,个别体重下降。西药组开始尚可,后期出现皮毛干枯,活动及饮食均减少。固本平喘汤大小剂量组皮毛尚可,身体较壮,活动及饮食较正常,体重明显增加。
2.正常组大鼠肺内结构均正常,细支气管周围炎性细胞浸润不明显,血管壁肌层无增厚,肺泡间隔无水肿、炎症及纤维化表现,肺泡内无明显渗出。模型组可见急性渗出性炎,有浆液性物质渗出,充满肺泡间隔内,部分肺泡结构破坏,肺泡融合,成纤维细胞、淋巴细胞、胶原纤维大量聚集。西药组与中药大、小剂量组病理过程同模型组,但纤维化病变程度及范围均较模型组小(见附图1至5)。
3.模型组第8周外周血中PCⅢ、HA及LN含量明显高与同期正常组,P<0.01。西药组、中药小剂量组、中药大剂量组第8周外周血中PCⅢ、HA及LN含量高与同期模型组,均P<0.05。西药组与中药大小剂量组相比无明显差异性,均P>0.05。
4.模型组支气管黏膜下成纤维细胞、淋巴细胞、巨噬细胞计数明显高与正常组,P<0.01。西药组支气管黏膜下成纤维细胞、淋巴细胞、巨噬细胞计数明显小与模型组,P<0.01。西药组与中药小剂量组、中药大剂量组相比较无明显差异性,P>0.05。符合COPD慢性气道炎症以及PF进程中成纤维细胞大量增生的病理特点。
结论
固本平喘汤能有效减轻大鼠肺纤维化,降低肺纤维化大鼠细胞外基质中UA、PCⅢ、LN含量是其作用机制之一。
Research Purpose:
On the basis of good efficacy in the clinical application of the strengths of treating antiasthmatic decoction to prevent chronic obstructive pulmonary disease with pulmonary fibrosis, do further study about influence of this medicament on chronic obstructive pulmonary disease with pulmonary fibrosis in animal models of extracellular matrix procollagen type III (PC III), and transparent quality acid (HA) and laminin (LN) content, explore the mechanism action of treating antiasthmatic decoction to treatment of pulmonary fibrosis, and provide a theoretical basis for clinical application.
Research Approach:
1. Animals and grouping:Total 60 rats all with weight 180-200g—both males and females are 30. Randomly divide them into normal control group, model group, western medicine group, treating antiasthmatic decoction low-does group, high-dose group. There are 12 rats in each group.
2. Animal modeling:The 12 rats are placed in a special fume box (60×55×70cm3), each nine cigarettes phased, one end fixed on the smoke box top and bottom lighting, after complete combustion change another 12 rats, daily 48 rats, weekly 7d, and act 4 weeks continually. In the 5th week, injecting of anesthesia for all rats except in normal group at 10% chloral hydrate (0.36ml/100g) intraperitoneal, shearing conventional iodophor disinfection after the anterior, Line neck incision, blunt separation of the trachea exposed, a one-time injection of bleomycin-side saline solution 0.2ml (bleomycin 4mg/kg body weight) via tracheal cartilage ring gap to the heart puncture. Then let animals upright immediately and rotate mouse plate so that the liquid uniformly distributed in the lungs, skin suture, and routine disinfection. After the animals regained consciousness,put them in cages to breed them conventionally. After modeling from the second day starting medication, the normal group and model group are given normal saline 1ml/150g gavage,2times/day, the western medicine group are given intraperitoneal injection of dexamethasone 0.3mg/100g,1 times/day, treating antiasthmatic decoction low-does group, high-dose group are given free fried granules 4ml, 1ml gavage,2 times/day. All above groups are administered for 28 days.
3. Detection methods and observation standard:①generally observe activity, the sensitivity to external reactions, shiny fur, food, water, weight, and deaths of rats.②HA, LN, PCⅢdetected,in the 8th week,after intraperitoneal anesthesia by a 10% chloral hydrate 0.36ml/100g, extract blood strictly according to kit instructions using enzyme-linked immunosorbent assay in rat plasma HA, LN, the level of PCⅢ.③Pathological morphology, kill rats in batches, promptly remove the right middle lobe and fix in 10% neutral formalin. Gradient ethanol dehydration, xylene and transparent specimens, dipping wax, embedding, slicing, and treated by HE, Masson staining to observe the extent of alveolitis and pulmonary fibrosis and count bronchial mucosa fibroblasts, lymphocytes and macrophages under thress high magnifications.
4. All results using mean±standard deviation (x±s) to denote, using SPSS10.0 software mean one-way ANOVA to analyze. P<0.05 is considered to be with statistically significant, P<0.01 means significant differences.
Research Result:
1. The normal control group:smooth fur, able-bodied, diet, activity, body in normal growth. The model control group:dry sparse fur, lack of energy, the reaction retardation, eating, and activities reduced. Wheezing symptoms after strenuous exercise, weight increasing slightly compared with pre-made model, individual ones losing weight. Western Group: adequate at the beginning, dry fur in later period, activities and diet reducing. Treating antiasthmatic decoction group:adequate fur, strong body, normal activity and diet, and the weight increased significantly.
2. Rats in normal group:normal structure of lungs, unobvious infiltration around the bronchial inflammatory cell, no thickening of vessel wall muscle, no edema, inflammation, fibrosism and no clear alveolar exudates in alveolar septum. For the model group:acute exudative inflammation, with serous exudation of material full of alveolar septum, the part of the alveolar structural damaged, alveolar fusion, fibroblasts, lymphocytes and collagen fibers gathering in large numbers. Traditional Chinese medicine, the large and low-dose groups were with the same pathologic process as the model group, but the degree and extent of fibrosis were lower than the model group (see the attached 1 to 5).
3. In the 8th week PCⅢ, HA and LN level in peripheral blood of model group are higher than the same perioa in the normal group, western medicine and Chinese medicine large and small dose group, all P<0.01. In the 8th week PCⅢ, HA and LN level in peripheral blood of western medicine, Chinese medicine large and small dose group is higher than the normal group in the same period, P<0.05. Western medicine group is no difference from Chinese medicine large and small dose group, P>0.05.
4. Counts of fibroblasts, lymphocytes and macrophages in bronchial mucosa of the model group was higher than the normal group, P<0.01. Counts of fibroblasts, lymphocytes and macrophages in bronchial mucosa of the western medicine was lower than the model group, P<0.05. Western medicine group is no difference from Chinese medicine large and small dose group, P>0.05. Consistent with chronic airway inflammation in COPD, and fiber cells in the PF process proliferated into the pathological features.
Conclusion:
Treating antiasthmatic decoction can effectively ease pulmonary fibrosis of rats. One of the possible role of its mechanisms is to reduce the content of UA, PCⅢ, LN in extracellular matrix of pulmonary fibrosis rats.
在临床应用固本平喘汤防治慢性阻塞性肺疾病(COPD)合并肺纤维化(PF)并取得良好疗效的基础上,进一步研究该方对COPD合并PF动物模型细胞外基质Ⅲ型前胶原(PCⅢ)、透明质酸(HA)及层粘连蛋白(LN)含量的影响,探讨固本平喘汤防治COPD合并PF的作用机制,为临床应用提供理论依据。
研究方法
1.动物和分组：将体重180-200g大鼠60只,雌雄各30只,随机分为正常组对照组、模型组、西药组、固本平喘汤小剂量组、大剂量组,每组12只。
2.动物模型制作：将12只大鼠置于特制熏烟箱(60×55×70cm3)内,每9只香烟相接,一端固定于熏箱顶,点燃下端,燃毕换另12只,每日48只,每周7d,连续4周。第5周将除正常组外大鼠以10%水合氯醛(0.36ml/100g)腹腔注射麻醉,颈前剪毛后碘伏常规消毒,行颈正中切口,钝性分离暴露气管,经气管软骨环间隙向心端穿刺一次性注射器注入博莱霉素生理盐水溶液0.2ml(约博莱霉素4mg/kg),立即将动物直立并旋转鼠板,使药液在肺内均匀分布,缝合皮肤,常规消毒,待动物自然苏醒后置笼内常规饲养。造模后第2天开始用药,正常组及模型组均予等体积生理盐水灌胃,2次/日,西药组给予腹腔注射地塞米松0.3mg/100g,1次/日,固本平喘汤(炙麻黄、苦杏仁、甘草、紫苏子、白芥子、莱菔子、黄芩、地龙、淫羊藿、补骨脂、巴戟天、黄芪、丹参、川芎)大、小剂量组分别给予免煎冲剂4m1(根据人与大鼠药物剂量换算公式,设计药物剂量,浓缩成含生药2.7g/ml的免煎冲剂)、1ml(药物浓度同上)灌胃,2次／日,以上各组均给药28天。
3.检测方法和观测指标：①一般情况观测大鼠的活动度、对外界反应的灵敏度、皮毛光泽、饮食、饮水、体重、死亡情况。②HA、LN、PCⅢ检测于第8周,经10%的水合氯醛0.36ml/100g腹腔麻醉后,抽取外周血,严格按照试剂盒说明书采用酶联免疫吸附法检测大鼠血浆中HA、LN、PCⅢ水平。③病理切片形态学观察分批处死大鼠后,迅速取出右肺中叶,10%中性福尔马林溶液中固定,乙醇梯度脱水,二甲苯透明标本,浸蜡,包埋,切片,分别行HE染色,观察肺泡炎、肺纤维化程度以及在3个高倍镜下统计支气管黏膜下成纤维细胞、淋巴细胞、巨噬细胞数。
4.所有结果采用均数±标准差(x±s)表示,采用SPSS10.0软件进行均数单因素方差分析,P<0.05即认为有统计学意义,P<0.01有显著性差异。
研究结果
1.正常对照组皮毛光滑,身体健壮,饮食、活动、体质量增长正常。模型对照组皮毛干枯稀疏,精神不振,反应迟钝,进食、活动均减少,剧烈运动后出现喘息症状,体重较造模前略有增加,个别体重下降。西药组开始尚可,后期出现皮毛干枯,活动及饮食均减少。固本平喘汤大小剂量组皮毛尚可,身体较壮,活动及饮食较正常,体重明显增加。
2.正常组大鼠肺内结构均正常,细支气管周围炎性细胞浸润不明显,血管壁肌层无增厚,肺泡间隔无水肿、炎症及纤维化表现,肺泡内无明显渗出。模型组可见急性渗出性炎,有浆液性物质渗出,充满肺泡间隔内,部分肺泡结构破坏,肺泡融合,成纤维细胞、淋巴细胞、胶原纤维大量聚集。西药组与中药大、小剂量组病理过程同模型组,但纤维化病变程度及范围均较模型组小(见附图1至5)。
3.模型组第8周外周血中PCⅢ、HA及LN含量明显高与同期正常组,P<0.01。西药组、中药小剂量组、中药大剂量组第8周外周血中PCⅢ、HA及LN含量高与同期模型组,均P<0.05。西药组与中药大小剂量组相比无明显差异性,均P>0.05。
4.模型组支气管黏膜下成纤维细胞、淋巴细胞、巨噬细胞计数明显高与正常组,P<0.01。西药组支气管黏膜下成纤维细胞、淋巴细胞、巨噬细胞计数明显小与模型组,P<0.01。西药组与中药小剂量组、中药大剂量组相比较无明显差异性,P>0.05。符合COPD慢性气道炎症以及PF进程中成纤维细胞大量增生的病理特点。
结论
固本平喘汤能有效减轻大鼠肺纤维化,降低肺纤维化大鼠细胞外基质中UA、PCⅢ、LN含量是其作用机制之一。
Research Purpose:
On the basis of good efficacy in the clinical application of the strengths of treating antiasthmatic decoction to prevent chronic obstructive pulmonary disease with pulmonary fibrosis, do further study about influence of this medicament on chronic obstructive pulmonary disease with pulmonary fibrosis in animal models of extracellular matrix procollagen type III (PC III), and transparent quality acid (HA) and laminin (LN) content, explore the mechanism action of treating antiasthmatic decoction to treatment of pulmonary fibrosis, and provide a theoretical basis for clinical application.
Research Approach:
1. Animals and grouping:Total 60 rats all with weight 180-200g—both males and females are 30. Randomly divide them into normal control group, model group, western medicine group, treating antiasthmatic decoction low-does group, high-dose group. There are 12 rats in each group.
2. Animal modeling:The 12 rats are placed in a special fume box (60×55×70cm3), each nine cigarettes phased, one end fixed on the smoke box top and bottom lighting, after complete combustion change another 12 rats, daily 48 rats, weekly 7d, and act 4 weeks continually. In the 5th week, injecting of anesthesia for all rats except in normal group at 10% chloral hydrate (0.36ml/100g) intraperitoneal, shearing conventional iodophor disinfection after the anterior, Line neck incision, blunt separation of the trachea exposed, a one-time injection of bleomycin-side saline solution 0.2ml (bleomycin 4mg/kg body weight) via tracheal cartilage ring gap to the heart puncture. Then let animals upright immediately and rotate mouse plate so that the liquid uniformly distributed in the lungs, skin suture, and routine disinfection. After the animals regained consciousness,put them in cages to breed them conventionally. After modeling from the second day starting medication, the normal group and model group are given normal saline 1ml/150g gavage,2times/day, the western medicine group are given intraperitoneal injection of dexamethasone 0.3mg/100g,1 times/day, treating antiasthmatic decoction low-does group, high-dose group are given free fried granules 4ml, 1ml gavage,2 times/day. All above groups are administered for 28 days.
3. Detection methods and observation standard:①generally observe activity, the sensitivity to external reactions, shiny fur, food, water, weight, and deaths of rats.②HA, LN, PCⅢdetected,in the 8th week,after intraperitoneal anesthesia by a 10% chloral hydrate 0.36ml/100g, extract blood strictly according to kit instructions using enzyme-linked immunosorbent assay in rat plasma HA, LN, the level of PCⅢ.③Pathological morphology, kill rats in batches, promptly remove the right middle lobe and fix in 10% neutral formalin. Gradient ethanol dehydration, xylene and transparent specimens, dipping wax, embedding, slicing, and treated by HE, Masson staining to observe the extent of alveolitis and pulmonary fibrosis and count bronchial mucosa fibroblasts, lymphocytes and macrophages under thress high magnifications.
4. All results using mean±standard deviation (x±s) to denote, using SPSS10.0 software mean one-way ANOVA to analyze. P<0.05 is considered to be with statistically significant, P<0.01 means significant differences.
Research Result:
1. The normal control group:smooth fur, able-bodied, diet, activity, body in normal growth. The model control group:dry sparse fur, lack of energy, the reaction retardation, eating, and activities reduced. Wheezing symptoms after strenuous exercise, weight increasing slightly compared with pre-made model, individual ones losing weight. Western Group: adequate at the beginning, dry fur in later period, activities and diet reducing. Treating antiasthmatic decoction group:adequate fur, strong body, normal activity and diet, and the weight increased significantly.
2. Rats in normal group:normal structure of lungs, unobvious infiltration around the bronchial inflammatory cell, no thickening of vessel wall muscle, no edema, inflammation, fibrosism and no clear alveolar exudates in alveolar septum. For the model group:acute exudative inflammation, with serous exudation of material full of alveolar septum, the part of the alveolar structural damaged, alveolar fusion, fibroblasts, lymphocytes and collagen fibers gathering in large numbers. Traditional Chinese medicine, the large and low-dose groups were with the same pathologic process as the model group, but the degree and extent of fibrosis were lower than the model group (see the attached 1 to 5).
3. In the 8th week PCⅢ, HA and LN level in peripheral blood of model group are higher than the same perioa in the normal group, western medicine and Chinese medicine large and small dose group, all P<0.01. In the 8th week PCⅢ, HA and LN level in peripheral blood of western medicine, Chinese medicine large and small dose group is higher than the normal group in the same period, P<0.05. Western medicine group is no difference from Chinese medicine large and small dose group, P>0.05.
4. Counts of fibroblasts, lymphocytes and macrophages in bronchial mucosa of the model group was higher than the normal group, P<0.01. Counts of fibroblasts, lymphocytes and macrophages in bronchial mucosa of the western medicine was lower than the model group, P<0.05. Western medicine group is no difference from Chinese medicine large and small dose group, P>0.05. Consistent with chronic airway inflammation in COPD, and fiber cells in the PF process proliferated into the pathological features.
Conclusion:
Treating antiasthmatic decoction can effectively ease pulmonary fibrosis of rats. One of the possible role of its mechanisms is to reduce the content of UA, PCⅢ, LN in extracellular matrix of pulmonary fibrosis rats.
引文
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[10]李卫青,柯新桥,孙劲松,等.固本防喘胶囊对慢性阻塞性肺 疾病急性加重期患者治疗作用的研究[J].中国中西医结合急救杂志,2006,13(2)：76-79.
[11]赵兰才,武维屏,纪雅文.肺气虚证慢阻肺合并肺纤维化金黄地鼠模型的建立[J].中国中医基础医学杂志,2002,8(7)：55-56.
[12]刘文萍,董亚苒.慢性阻塞性肺疾病合并肺间质纤维化23例临床分析[J].北京医学报,2006,29(3)：185-186.
[13]周仲瑛,金实,李明富,等.中医内科学,北京：中国中医药出版社,2003,1：117.
[14]陈建,何冰,刘新民,等.银杏叶制剂治疗肺间质纤维化的实验研究[J].中国中西医结合杂志,2000,20(6)：441-443.
[15]刘卫敏,徐启勇,林宇辉,等.浓当归注射液对博莱霉素致鼠肺纤维化治疗作用的研究[J].医学新知杂志,2001,11(1)：20-22.
[16]茹永新,李桂萍,张华梅,等.HT合剂防治肺纤维化的实验研究[J].中国中医药科技,2001,8(4)：221.
[17]任正斌,补肾中药药理作用浅析[N].辽宁中医学院报,2002,4(4)：312.
[18]陆再英,钟南山.内科学,北京：人民卫生出版社,2008,7：98.
[19]Low RB, Giancola MS, King TE JR, et al. Serum and bronchoalveolar lavage of N-terminal type Ⅲ procollagen peptides in idiopathic pulmonary fibrosis. Am Rev Respir Dis, 1992,146(3):701
[20]孙达,苗淼,张玮.HA与IPF发病机制关系的研究进展[J],中国医药导报,200.8：18(5)：28-29
[21]柯新桥.柯新桥论文集,北京：中国医药出版社,2006,1：160。
[1]荆小莉,王东平,李欣,刘建博,熊艳.清肺化痰祛瘀方治疗慢性阻塞性肺疾病急性加重期的研究[J].中国中西医结合急救杂志,2006,13(5)：207-272.
[2]刘建博,杨海娟.健脾清肺化痰法对慢性阻塞性肺疾病合并全身炎症反应综合征疗效及核因子-κB表达的影响[J].广州中医药大学学报,2009,26(2)：120.123.
[3]陈宁,李慧,赵丽芸,等.清热化痰法治疗慢性阻塞性肺疾病急性加重期全身炎症反应综合征的随机对照临床试验[J].中西医结合学报,2009,7(2)：105.109.
[4]李希,黄河清,张川林.李希黄河清张川林[J].中国现代药物应用,2008,2(4)：83.84.
[5]常建锋,李素云.中西医结合治疗慢性阻塞性肺疾病急性加重期36例[J].四川中医,2008,26(11)：69.70.
[6]刘国华.中西医结合治疗慢性阻塞性肺疾病急性加重期38例[J].实用中医内科学,2007,21(6)：56.
[7]张顺利.中西医结合治疗慢性阻塞性肺疾病急性加重期痰热郁肺证30例临床观察[J].河北中医,2008,30(2)：164-165.
[8]张穗坚,方楚权.二陈汤合三子养亲汤治疗慢性阻塞性肺疾病48例疗效观察[J].新中医,2006,38(10)：24.25.
[9]赵菁华,杨广源,白燕,等.中西医结合治疗慢性阻塞性肺疾病 急性发作42例临床观察[J].内蒙古中医药,2007,10：19.20.
[10]彭波王桂珍李泽庚,等.化痰降气胶囊对慢性阻塞性肺疾病急性加重期患者肺功能的影响[J].北京中医,2006,25(11)：648.649.
[11]谢加富曹锐彬陈仕章,等.小青龙合剂配合基础治疗对慢性阻塞性肺疾病患者排痰效果的影响[J].实用医学杂志,2009,25(1)：144.146.
[12]郑柳鹏.中西医结合治疗老年人慢性阻塞性肺疾病30例疗效观察[J].实用心脑肺血管病杂志,2008,16(6)：73.74.
[13]周晓宏,谷雨.中西医结合治疗慢性阻塞性肺疾病急性发作期气道黏液高分泌临床观察[J].中华结核和呼吸杂志,1999,22(1)：30.32.
[14]杨兵文.中西医结合治疗慢性阻塞性肺疾病急性加重期48例疗效观察[J].新中医,2008,40(2)：56.57.
[15]许有志,王仕汉,张文东,等.中西医结合治疗慢性阻塞性肺疾病急性加重期100例[J].中医药临床杂志,2007,19(6)：579.580.
[16]汪丙柱,等.补肺通腑汤为主治疗慢性阻塞性肺疾病稳定期36例[J].浙江中医杂志,2009,44(1)：30.31.
[17]范发才,赵凤达,惠萍,等.苈黄导痰汤治疗慢性阻塞性肺疾病急性加重期临床观察[J].中医药学刊,2006,24(3)：566.568.
[18]李士瑾.通瘀泻脯汤治疗62例慢性阻塞性肺疾病临床观察[J].中医中医药现代远程教育,2008,6(4)：327.328.
[19]林女燕钊.通下法配合无创通气治疗慢性阻塞性肺疾病急性加重 期呼吸衰竭15例[J].新中医,2005,37(1)：41.43.
[20]郭文新,张凤宇.宣白承气汤加味联合沐舒坦对慢性阻塞性肺疾病急性加重期祛痰疗效观察[J].河北中医,2008,30(5)：502.503.
[21]肖波,刘晖,陈康桂,等.参麦注射液对慢性阻塞性肺疾病急性加重期的干预作用[J].新中医,2007,39(6)：11.12.
[22]陈红华.中西医结合治疗COPD急性加重期45例[J].浙江中西医结合杂志,2008,18(3)：171.172.
[23]谭光波,胡学军,卜献春.中西医结合治疗慢性阻塞性肺疾病42例临床观察[J].湖南中医杂志,2008,24(4)：12.13.
[24]茅万里.补肾法治疗慢性阻塞性肺疾病的近况[J].安徽中医临床杂志,2003,15(6)：544.545.
[25]许林生.中西医结合治疗对COPD稳定期中、重度患者肺功能的影响[J].中华中西医学杂志,2007,5(2)：55.56.
[26]柯新桥.固本防喘胶囊对慢性阻塞性肺疾病急性加重期患者治疗作用的研究[J].中国中西医结合急救杂志,2006,13(2)：76.79.
[27]张凤宇,郭文新,刘亚辉,等.中西医结合治疗慢性阻塞性肺疾病.缓解期50例临床观察[J].河北中医,2007,29(7)：615.616.
[28]瞿梅,许文清.中西医结合治疗慢性阻塞性肺疾病急性加重期46例总结[J].湖南中医杂志,2007,23(4)：6.8.
[29]刘小虹.活血化痰法对慢性阻塞性肺疾病稳定期患者肺功能和生存质量的影响[J].广州中医药大学学报,2006,23(4)：275.278.
[30]梁建萍,许萍,李平.通络活血和益气补肾法治疗慢性阻塞性 肺疾病的临床研究[J].中华中西医学杂志,2007,5(2)：55-56.
[31]孟强梅,李炜,李立民,等.宣肺祛瘀通络中药对慢性阻塞性肺疾病肺动脉高压伴高粘血症的影响[J].中国中医药信息杂志,2009,16(2)：70.71.
[2]吴小燕,卫小红.慢性阻塞性肺疾病合并肺纤维化的研究现状[J].医学综述,2006,12(10)：635-636.
[3]中华医学会呼吸病学分会慢性阻塞性肺疾病学组.慢性阻塞性肺疾病诊治指南[S].2007；46(3)：254-261.
[4]雷丰丰,李惠菊,万毅新,等。中医药治疗肺纤维化及实验研究进展[J].现代中西医结合杂志,2003,12(10)：1117-1118.
[5]Gross TJ, Hunninghake GW. Idiopathic pulmonary fibrosis[J].N EnglMed,2001,345(7):517-525.
[6]康卫国,刘久山,蔡绍曦等,血清透明质酸等的变化COPD肺纤维化的关系[J].实用医药杂志,2002,11(19)：840-842.
[7]Entzian P, Hucksladt A, Kreipe H, et al. Determination of serum concentrations of type Ⅲ procollagen peptide inmec hanically ventilation patients. Am Rev Respir Dis,1990,142: 1079.
[8]詹爱霞,蔡文霞,钱定良.肺纤维化患者血清LN、HA、PCⅢ、CⅣ检测的临床意义[J].全科医学临床与教育,2006,4(4)：289-290.
[9]孙子凯,奚肇庆.慢性阻塞性肺疾病合并肺间质纤维化的中西医治疗现状与探讨[J].辽宁中医药大学报,2007,9(6)：34-36·
[10]李卫青,柯新桥,孙劲松,等.固本防喘胶囊对慢性阻塞性肺 疾病急性加重期患者治疗作用的研究[J].中国中西医结合急救杂志,2006,13(2)：76-79.
[11]赵兰才,武维屏,纪雅文.肺气虚证慢阻肺合并肺纤维化金黄地鼠模型的建立[J].中国中医基础医学杂志,2002,8(7)：55-56.
[12]刘文萍,董亚苒.慢性阻塞性肺疾病合并肺间质纤维化23例临床分析[J].北京医学报,2006,29(3)：185-186.
[13]周仲瑛,金实,李明富,等.中医内科学,北京：中国中医药出版社,2003,1：117.
[14]陈建,何冰,刘新民,等.银杏叶制剂治疗肺间质纤维化的实验研究[J].中国中西医结合杂志,2000,20(6)：441-443.
[15]刘卫敏,徐启勇,林宇辉,等.浓当归注射液对博莱霉素致鼠肺纤维化治疗作用的研究[J].医学新知杂志,2001,11(1)：20-22.
[16]茹永新,李桂萍,张华梅,等.HT合剂防治肺纤维化的实验研究[J].中国中医药科技,2001,8(4)：221.
[17]任正斌,补肾中药药理作用浅析[N].辽宁中医学院报,2002,4(4)：312.
[18]陆再英,钟南山.内科学,北京：人民卫生出版社,2008,7：98.
[19]Low RB, Giancola MS, King TE JR, et al. Serum and bronchoalveolar lavage of N-terminal type Ⅲ procollagen peptides in idiopathic pulmonary fibrosis. Am Rev Respir Dis, 1992,146(3):701
[20]孙达,苗淼,张玮.HA与IPF发病机制关系的研究进展[J],中国医药导报,200.8：18(5)：28-29
[21]柯新桥.柯新桥论文集,北京：中国医药出版社,2006,1：160。
[1]荆小莉,王东平,李欣,刘建博,熊艳.清肺化痰祛瘀方治疗慢性阻塞性肺疾病急性加重期的研究[J].中国中西医结合急救杂志,2006,13(5)：207-272.
[2]刘建博,杨海娟.健脾清肺化痰法对慢性阻塞性肺疾病合并全身炎症反应综合征疗效及核因子-κB表达的影响[J].广州中医药大学学报,2009,26(2)：120.123.
[3]陈宁,李慧,赵丽芸,等.清热化痰法治疗慢性阻塞性肺疾病急性加重期全身炎症反应综合征的随机对照临床试验[J].中西医结合学报,2009,7(2)：105.109.
[4]李希,黄河清,张川林.李希黄河清张川林[J].中国现代药物应用,2008,2(4)：83.84.
[5]常建锋,李素云.中西医结合治疗慢性阻塞性肺疾病急性加重期36例[J].四川中医,2008,26(11)：69.70.
[6]刘国华.中西医结合治疗慢性阻塞性肺疾病急性加重期38例[J].实用中医内科学,2007,21(6)：56.
[7]张顺利.中西医结合治疗慢性阻塞性肺疾病急性加重期痰热郁肺证30例临床观察[J].河北中医,2008,30(2)：164-165.
[8]张穗坚,方楚权.二陈汤合三子养亲汤治疗慢性阻塞性肺疾病48例疗效观察[J].新中医,2006,38(10)：24.25.
[9]赵菁华,杨广源,白燕,等.中西医结合治疗慢性阻塞性肺疾病 急性发作42例临床观察[J].内蒙古中医药,2007,10：19.20.
[10]彭波王桂珍李泽庚,等.化痰降气胶囊对慢性阻塞性肺疾病急性加重期患者肺功能的影响[J].北京中医,2006,25(11)：648.649.
[11]谢加富曹锐彬陈仕章,等.小青龙合剂配合基础治疗对慢性阻塞性肺疾病患者排痰效果的影响[J].实用医学杂志,2009,25(1)：144.146.
[12]郑柳鹏.中西医结合治疗老年人慢性阻塞性肺疾病30例疗效观察[J].实用心脑肺血管病杂志,2008,16(6)：73.74.
[13]周晓宏,谷雨.中西医结合治疗慢性阻塞性肺疾病急性发作期气道黏液高分泌临床观察[J].中华结核和呼吸杂志,1999,22(1)：30.32.
[14]杨兵文.中西医结合治疗慢性阻塞性肺疾病急性加重期48例疗效观察[J].新中医,2008,40(2)：56.57.
[15]许有志,王仕汉,张文东,等.中西医结合治疗慢性阻塞性肺疾病急性加重期100例[J].中医药临床杂志,2007,19(6)：579.580.
[16]汪丙柱,等.补肺通腑汤为主治疗慢性阻塞性肺疾病稳定期36例[J].浙江中医杂志,2009,44(1)：30.31.
[17]范发才,赵凤达,惠萍,等.苈黄导痰汤治疗慢性阻塞性肺疾病急性加重期临床观察[J].中医药学刊,2006,24(3)：566.568.
[18]李士瑾.通瘀泻脯汤治疗62例慢性阻塞性肺疾病临床观察[J].中医中医药现代远程教育,2008,6(4)：327.328.
[19]林女燕钊.通下法配合无创通气治疗慢性阻塞性肺疾病急性加重 期呼吸衰竭15例[J].新中医,2005,37(1)：41.43.
[20]郭文新,张凤宇.宣白承气汤加味联合沐舒坦对慢性阻塞性肺疾病急性加重期祛痰疗效观察[J].河北中医,2008,30(5)：502.503.
[21]肖波,刘晖,陈康桂,等.参麦注射液对慢性阻塞性肺疾病急性加重期的干预作用[J].新中医,2007,39(6)：11.12.
[22]陈红华.中西医结合治疗COPD急性加重期45例[J].浙江中西医结合杂志,2008,18(3)：171.172.
[23]谭光波,胡学军,卜献春.中西医结合治疗慢性阻塞性肺疾病42例临床观察[J].湖南中医杂志,2008,24(4)：12.13.
[24]茅万里.补肾法治疗慢性阻塞性肺疾病的近况[J].安徽中医临床杂志,2003,15(6)：544.545.
[25]许林生.中西医结合治疗对COPD稳定期中、重度患者肺功能的影响[J].中华中西医学杂志,2007,5(2)：55.56.
[26]柯新桥.固本防喘胶囊对慢性阻塞性肺疾病急性加重期患者治疗作用的研究[J].中国中西医结合急救杂志,2006,13(2)：76.79.
[27]张凤宇,郭文新,刘亚辉,等.中西医结合治疗慢性阻塞性肺疾病.缓解期50例临床观察[J].河北中医,2007,29(7)：615.616.
[28]瞿梅,许文清.中西医结合治疗慢性阻塞性肺疾病急性加重期46例总结[J].湖南中医杂志,2007,23(4)：6.8.
[29]刘小虹.活血化痰法对慢性阻塞性肺疾病稳定期患者肺功能和生存质量的影响[J].广州中医药大学学报,2006,23(4)：275.278.
[30]梁建萍,许萍,李平.通络活血和益气补肾法治疗慢性阻塞性 肺疾病的临床研究[J].中华中西医学杂志,2007,5(2)：55-56.
[31]孟强梅,李炜,李立民,等.宣肺祛瘀通络中药对慢性阻塞性肺疾病肺动脉高压伴高粘血症的影响[J].中国中医药信息杂志,2009,16(2)：70.71.