CLKC1治疗小儿上呼吸道感染后咳嗽药效学研究及机理探讨
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摘要
小儿上呼吸道感染后咳嗽是指患儿感冒好转后遗留咳嗽久而不愈,又称感冒后咳嗽,简称CPI。上呼吸道感染是儿科常见病,多发于冬季,常常导致小儿上感后咳嗽。部分患儿在上呼吸道感染恢复后出现咳嗽的呼吸道症状,可持续数周至数月,经常被误诊为支气管炎、喘息性支气管炎或肺炎,反复使用抗生素、止咳药等无效,给患儿带来了一定的痛苦,也给父母造成了经济负担。由于小儿上呼吸道感染后咳嗽的发病率较高,近年来其药物治疗已经倍受关注。
目前,西医学治疗小儿上感后咳嗽的药物主要有组胺受体拮抗剂、中枢性镇咳药、麻醉剂、激素、肾上腺素能神经受体拮抗剂等。西药治疗小儿上感后咳嗽疗效欠佳。中医学治疗咳嗽历史悠久,经验丰富,与我国得天独厚的中药资源相结合,为中医药治疗小儿上呼吸道感染后咳嗽提供了广阔的空间。中成药携带方便,易于服用,因此将疗效显著的中药复方开发成治疗小儿上感后咳嗽的中成药,将具有广泛的经济效益和社会效益。
CLKC1是以临床疗效为基础,由中医经验方开发而成的颗粒剂。该方由黄芪、金荞麦等中药组成,具有清肺化痰,止咳利咽的功能。主治小儿痰热阻肺,肺失宣肃所致咳嗽不已,咳声重浊,痰稠色黄或黄白相兼,咽干不利,或大便秘结；小儿感冒或感冒后咳嗽见上述证候者。本校中药药剂教研室将其开发为颗粒剂,拟用于小儿上呼吸道感染后咳嗽的治疗。
目前研究认为上呼吸道感染后咳嗽的致病机理为气道高反应性。本课题以CLKC1的功能主治为依据,结合小儿上呼吸道感染后咳嗽的西医学发病机制,借助常规药理学动物模型对CLKC1的药效及其作用机制进行了研究。主要包括两部分研究内容：CLKC1治疗小儿上呼吸道感染后咳嗽的药效学研究；CLKC1治疗小儿上呼吸道感染后咳嗽的作用机制研究。在进行药效学研究的同时探讨CLKC1治疗小儿上呼吸道感染后咳嗽的作用机制,以期阐明其发挥药效的作用机理,为CLKC1的临床应用和新药的申报提供实验依据。
1CLKC1治疗小儿上呼吸道感染后咳嗽的药效学研究
1.1CLKC1镇咳作用的试验研究
目的：观察CLKC1对咳嗽动物模型咳嗽潜伏时和咳嗽次数的影响,以确定其是否具有镇咳作用。方法：分别采用二氧化硫引咳小鼠模型和枸橼酸引咳豚鼠模型,预防治疗给药后观察二氧化硫引咳小鼠和枸橼酸引咳豚鼠的咳嗽潜伏时和咳嗽次数。结果：CLKC1能够减少二氧化硫、枸橼酸引发的动物咳嗽的反应次数,并且大、中、小剂量组具有良好的量效关系(p<0.05)。但对咳嗽潜伏时的影响不明显。结论：CLKC1具有明显的镇咳作用。
1.2祛痰作用的试验研究
目的：观察CLKC1对小鼠气管段酚红排泌量的影响,确定其是否具有祛痰作用。方法：采用气管段酚红试验法,给药后观察气管段酚红的排泌量。结果：同对照组小鼠的气管段酚红排泌量相比较,CLKC1大、中、小三个剂量组小鼠的气管段酚红排泌量均增加,大剂量增加的更为明显(p<0.05)。结论：CLKC1可以促进气管酚红的排泌,具有较明显的祛痰作用。
1.3 CLKC1平喘作用的试验研究
目的：通过观察CLKC1对引喘动物喘息潜伏期和倒仆时间的影响评价其平喘作用。方法：采用Ach、组胺喷雾引喘法,观察给药后Ach、组胺引喘豚鼠的喘息潜伏期和倒仆时间的。结果：CLKC1各剂量组可明显减少豚鼠的咳嗽次数且具有良好的量效关系,但对咳嗽潜伏时无明显延长作用(p<0.05)。结论：CLKC1具有平喘作用。
1.4 CLKC1肠推进作用的试验研究
目的：通过观察CLKC1对小肠推进运动的影响,评价其通便的药理作用.方法：以伊文思蓝灌胃,分离从幽门端至回盲部的肠管,测量其在小鼠肠内的推进距离,计算伊文思蓝推进的百分率。结果：CLKC1各剂量组可明显促进小肠蠕动,提高小肠推进率(p<0.01)结论：CLKC1可促进肠推进运动,具有通便作用。
1.5 CLKC1抗炎作用的试验研究
目的：通过观察CLKC1对急慢性炎症动物模型的影响,确定其是否具有抗炎作用。方法：采用大鼠足跖皮下注射角叉菜胶造成的急性炎症模型,观察CLKC1对致炎后大鼠足肿胀的影响；采用大鼠腹股沟皮下埋置棉球造成的慢性炎症模型,观察CLKC1对致炎后大鼠棉球肉芽肿的影响。结果：CLKC1中、小剂量组在多个时间点可明显减轻大鼠的足肿胀度(p<0.01),但对大鼠棉球肉芽肿的干、湿重无明显影响。结论：CLKC1在炎症早期具有抗炎作用,而在炎症晚期无明显效果。
2 CLKC1治疗小儿上呼吸道感染后咳嗽的作用机制研究
2.1 CLKC1对哮喘大鼠气道高反应性的影响
目的：通过研究CLKC1对哮喘大鼠气道阻力,HMGB-1、IgE、IL-4、IL-13表达的影响,考察其对气道高反应性的抑制作用.方法：以卵蛋白和灭活结核杆菌菌苗激发大鼠哮喘模型,予CLKC1干预后测定其气道阻力、血清HMGB-1、肺匀浆中IgE、IL-4、IL-13的含量.结果：CLKC1可以抑制大鼠气道阻力的增加以及血清HMGB-1.肺匀浆中IgE、IL-4、IL-13含量的增高.结论：CLKC1可以通过抑制炎症介质、细胞因子含量的增高降低哮喘大鼠的气道高反应性。
2.2 CLKC1对哮喘小鼠气道高反应性的影响
目的：通过研究CLKC1对哮喘小鼠辣椒素咳嗽敏感性试验,NO、ET-1、IL-4、IL-13表达的影响,考察其对气道高反应性的抑制作用。方法：以卵蛋白激发Ba1b/c小鼠哮喘模型,予CLKC1干预后测定咳嗽敏感性试验辣椒素的阈值以及肺匀浆中NO、ET-1、IL-4、IL-13的含量。结果：CLKC1可以增加辣椒素咳嗽敏感性试验的阈值,抑制肺匀浆中NO、ET-1、IL-4、IL-13含量的增高。结论：CLKC1可能从减轻气道炎症、促进气道重建角度降低哮喘小鼠的气道高反应性。
2.3 CLKC1对晚期炎症介质HMGB-1含量的影响
目的：观察CLKC1对晚期炎症介质高迁移率族蛋白B-1(HMGB-1)含量的影响。方法：测定LPS尾静脉注射致急性肺损伤小鼠不同时段血清中HMGB-1的含量,取含量最高的时间点,CLKC1预防治疗给药,测定在含量最高时间点时急性肺损伤小鼠血清中HMGB-1的含量。结果：CLKC1能够抑制HMGB-1含量的增高(P<0.05)。结论：CLKC1对晚期炎症介质HMGB-1含量的增高具有一定的抑制作用。
结论
1.药效学研究表明CLKC1具有止咳、祛痰、平喘、通便、抗炎的药理作用。
2.作用机制研究表明CLKC1治疗小儿上呼吸道感染后咳嗽的机理可能为从抑制细胞因子及炎性介质含量的增高,促进气道重建等角度减小气道阻力,降低气道高反应性。
创新点
1.监测了急性肺损伤小鼠模型血清中HMGB-1的含量随时间的变化情况,研究了中药颗粒剂对晚期炎症介质HMGB-1含量的影响。
2.首次将辣椒素咳嗽敏感性试验与气道高反应动物模型相结合,以辣椒素的阈值检测气道高反应性,并研究了中药颗粒剂对气道高反应动物模型辣椒素咳嗽敏感性试验阈值的影响。
Pediatric coughing after upper respiratory tract infection refers to the children's coughing for a long time without healing while the cold restores, which is also called cough post infection, referred to as CPI. The upper respiratory tract infection is a common pediatric disease, frequently happens in winter, often leads to the post infection cough. For parts of the patients, respiratory symptoms of cough appear after the recovery of upper respiratory tract infection, lasting for several weeks to mohths, often misdiagnosed as bronchitis or pneumonia, and the repeated use of antibiotics, cough killer etc is of no effect. It not only brings the children of pain, but also causes economic burden for parents. Because of the high incidence of coughing after upper respiratory tract infection, the drug therapy has been more concerned in recent years.
At present, the therapic drugs of CPI are mainly histamine receptor antagonist, central antitussive, anaesthetic, hormone, epinephrine never receptor antagonist, etc. Western medicine is not of great good effects in treating pediatric CPI. In treating cough, TCM has a long history and rich experience, our national unique traditional Chinese medicine resourse has provided a vast space for the application of TCM in treating CPI. Chinese patent medicines is convenient to carry and easy to use, so making Chinese prescription develope into Chinese patent medicines will have broad economic and social benefit based on the remarkable curative effect in treating CPI.
CLKC1 is developed from experienced prescription. The function is lung-clearing and expectorant, antitussive and throat-clearing. It has been made into granules by pharmaceutical department of Beijing University of Chinese Medicine, using for the treatment of CPI.
Generally speaking the main pathogenesis of CPI is AHR.With the function basis of CLKC1, combining the western medicine pathogenesis of CPI,we study pharmacological effect and function mechanism by conventional animal models. Two parts are mainly included:first a study on pharmacological effect; second the study on function mechanism. We try to clarify the function mechanism of CLKC1's pharmacological effect, in order to provide sufficient experimental data for the clinical using and new drug's application.
1 Study on pharmacodynamics of CLKC1 in CPI treatment
1.1 Study on antitussive effects of CLKC1
Objective:To observe the antitussive effects of CLKC1. Methods:Antitussive effects were observed through mice cough test induced by SO2steaming and through guinea pigs cough test induced by citric acid. Results:CLKC1 decreased the frequency of mice cough induced by ammonia and guinea pigs cough induced by citric acid, prolonged the cough latent period,dose groups have good concentration response relations (p<0.05). Conclusion: CLKC1 shows certain Antitussive effects.
1.2 Study on expectorant effects of CLKC1
Objective:To observe expectorant effect of CLKC1. Methods:Expectorant effectswere evaluated by measuring the excretion of phenol red in mice trachea. Results: Increased the excretion of phenol red from mice trachea, promoted the movement of prepared Chinese ink in the mice trachea (p<0.05). Conclusion:CLKC1 shows certain expectorant effects.
1.3 Study on Antiasthmatic effects of CLKC1
Objective:To observe antiasthmatic effect of CLKC1. Methods:Antiasthmatic actions in guinea pigs were observed by histamine and acetylcholine ultrasonic atomization test. Results:CLKC1 can inhibited the guinea pigs asthma introduced by histamine and acetylcholine solution (p<0.05). Conclusion:CLKC1 shows certain antiasthmatic effects. 1.4 Study on bowel propulsion effects of CLKC1
Objective:To observe the effect of CLKC1 on bowel propulsion in mice. Methods: Comput and compare the EB propulsion percentages in bowel in CLKC1 group, model group (treated with bowel propulsion supp ressants) and blank group, and the data were analyzed statistically 1. Results:CLKC1 dose groups could significantly increase EB propulsion percentages in mice(p<0.01).Conclusion:CLKC1 can promote intestine propulsive movement.
1.5 Study on anti-inflammatory effects of CLKC1
Objective:To observe anti-inflammatory effects of CLKC1. Methods:anti-inflammatory effects were observed by the swelling of hind paw in rats induced by carrageenin and granuloma induced by cotton pellets in rats. Results:CLKC1 medium and small doses groups can inhibite the paw edema induced by carrageenin (p<0.01), but not the granuloma induced by cotton pellet in SD rats. Conclusion:CLKC1 can inhibite early inflammation but without anti-inflammatory effects on later inflammation.
2 Study on Function mechanism of CLKC1 in CPI treatment
2.1 Study on effect of CLKC1 on AHR in allergic asthma rats
Objective:To investigate the effects of CLKC1 on the inhibition of airway hyperresponsiveness(AHR) in allergen induced asthma in rats. Methods:Model group and CLKC1 group were challenged with ovalbumin (OVA) to reproduce asthma., the blank group only receied an injection of same amount of Al(OH)3. On the 22nd day determine airway resistance, the content of HMGB-1、IgE、IL-4、IL-13 in lung homogenate. Results: CLKC1 can reduce the rat airway resistance, and inhibit the content's increase of HMGB-1、IgE、IL-13、IL-4 in lung homogenate(P<0.05). Conclusion:CLKC1 can reduce airway resistance and inhibit AHR.
2.2 Study on effect of CLKC1 on AHR of chronic asthma murine
Objective:To investigate the effects of CLKC1 on the inhibition of airway hyperresponsiveness(AHR) in allergen induced asthma in Balb/c rats. Methods:model group and CLKC1 group were challenged with ovalbumin (OVA) to reproduce asthma.,the blank group only receied an injection of same amount of Al(OH)3 On the 22nd day determine the threshold of capsaicin cough sensitivity test, the content of NO、ET-1、IL-4、IL-13 in lung homogenate and their physiological and pathological changes. Results:CLKC1 can increase the threshold of capsaicin cough sensitivity test, inhibit the content's increase of lung homogenate's NO、ET-1、IL-4、IL-13(P<0.05). Conclusion:CLKC1 can inhibit AHR.
2.3 Study on effect of CLKC1 on serum HMGB-1 level of sepsis mic
Objective:To observe the effects of CLKC1 on HMGB-1 protein expression in lung tissues of mice with endotoxin shock. Methods:Except the blank group, all other groups are injected LPS (5mg/kg)by the tails vein to establish the lung tissue injury models, The blank group only receied an injection of same amount of saline. Testing the serum HMGB-1 level at 24h,36h,48h,72h,96h. Study the effect of CLKC1 on serum HMGB-1 level of sepsis mic at the point of 72h. Results:72h after tail vail injection of LPS, serum HMGB-1 gets to the highest level; CLKC1 can inhibit the content's increase of HMGB-1 (P<0.05). Conclusion: CLKC1 can decrease serum HMGB-1 level.
Conclusion
1 The pharmacodynamics study shows that CLKC1 has antitussive, expectorant, antiasthmatic, bowel propulsion, anti-inflammatory effect pharmacological activity.
2 Study on function mechanism of CLKC1 in treating coughing after pediatric upper respiratory tract infection indicates that CLKC1 can reduce airway resistance, inhibit the content's increase of inflammatory mediators and cytokines, promote airway remodeling so as to decrease AHR.
Innovation
1 First studied the effect of Chinese medicine granule on the expression of. terminal inflammatory mediators HMGB-1.
2 Combine capsaicin cough sensitivity test with AHR animal models for the first time, detecting airway resistance by the threshold of capsaicin, and also study the effect of Chinese medicine granule on the threshold of capsaicin.
目前,西医学治疗小儿上感后咳嗽的药物主要有组胺受体拮抗剂、中枢性镇咳药、麻醉剂、激素、肾上腺素能神经受体拮抗剂等。西药治疗小儿上感后咳嗽疗效欠佳。中医学治疗咳嗽历史悠久,经验丰富,与我国得天独厚的中药资源相结合,为中医药治疗小儿上呼吸道感染后咳嗽提供了广阔的空间。中成药携带方便,易于服用,因此将疗效显著的中药复方开发成治疗小儿上感后咳嗽的中成药,将具有广泛的经济效益和社会效益。
CLKC1是以临床疗效为基础,由中医经验方开发而成的颗粒剂。该方由黄芪、金荞麦等中药组成,具有清肺化痰,止咳利咽的功能。主治小儿痰热阻肺,肺失宣肃所致咳嗽不已,咳声重浊,痰稠色黄或黄白相兼,咽干不利,或大便秘结；小儿感冒或感冒后咳嗽见上述证候者。本校中药药剂教研室将其开发为颗粒剂,拟用于小儿上呼吸道感染后咳嗽的治疗。
目前研究认为上呼吸道感染后咳嗽的致病机理为气道高反应性。本课题以CLKC1的功能主治为依据,结合小儿上呼吸道感染后咳嗽的西医学发病机制,借助常规药理学动物模型对CLKC1的药效及其作用机制进行了研究。主要包括两部分研究内容：CLKC1治疗小儿上呼吸道感染后咳嗽的药效学研究；CLKC1治疗小儿上呼吸道感染后咳嗽的作用机制研究。在进行药效学研究的同时探讨CLKC1治疗小儿上呼吸道感染后咳嗽的作用机制,以期阐明其发挥药效的作用机理,为CLKC1的临床应用和新药的申报提供实验依据。
1CLKC1治疗小儿上呼吸道感染后咳嗽的药效学研究
1.1CLKC1镇咳作用的试验研究
目的：观察CLKC1对咳嗽动物模型咳嗽潜伏时和咳嗽次数的影响,以确定其是否具有镇咳作用。方法：分别采用二氧化硫引咳小鼠模型和枸橼酸引咳豚鼠模型,预防治疗给药后观察二氧化硫引咳小鼠和枸橼酸引咳豚鼠的咳嗽潜伏时和咳嗽次数。结果：CLKC1能够减少二氧化硫、枸橼酸引发的动物咳嗽的反应次数,并且大、中、小剂量组具有良好的量效关系(p<0.05)。但对咳嗽潜伏时的影响不明显。结论：CLKC1具有明显的镇咳作用。
1.2祛痰作用的试验研究
目的：观察CLKC1对小鼠气管段酚红排泌量的影响,确定其是否具有祛痰作用。方法：采用气管段酚红试验法,给药后观察气管段酚红的排泌量。结果：同对照组小鼠的气管段酚红排泌量相比较,CLKC1大、中、小三个剂量组小鼠的气管段酚红排泌量均增加,大剂量增加的更为明显(p<0.05)。结论：CLKC1可以促进气管酚红的排泌,具有较明显的祛痰作用。
1.3 CLKC1平喘作用的试验研究
目的：通过观察CLKC1对引喘动物喘息潜伏期和倒仆时间的影响评价其平喘作用。方法：采用Ach、组胺喷雾引喘法,观察给药后Ach、组胺引喘豚鼠的喘息潜伏期和倒仆时间的。结果：CLKC1各剂量组可明显减少豚鼠的咳嗽次数且具有良好的量效关系,但对咳嗽潜伏时无明显延长作用(p<0.05)。结论：CLKC1具有平喘作用。
1.4 CLKC1肠推进作用的试验研究
目的：通过观察CLKC1对小肠推进运动的影响,评价其通便的药理作用.方法：以伊文思蓝灌胃,分离从幽门端至回盲部的肠管,测量其在小鼠肠内的推进距离,计算伊文思蓝推进的百分率。结果：CLKC1各剂量组可明显促进小肠蠕动,提高小肠推进率(p<0.01)结论：CLKC1可促进肠推进运动,具有通便作用。
1.5 CLKC1抗炎作用的试验研究
目的：通过观察CLKC1对急慢性炎症动物模型的影响,确定其是否具有抗炎作用。方法：采用大鼠足跖皮下注射角叉菜胶造成的急性炎症模型,观察CLKC1对致炎后大鼠足肿胀的影响；采用大鼠腹股沟皮下埋置棉球造成的慢性炎症模型,观察CLKC1对致炎后大鼠棉球肉芽肿的影响。结果：CLKC1中、小剂量组在多个时间点可明显减轻大鼠的足肿胀度(p<0.01),但对大鼠棉球肉芽肿的干、湿重无明显影响。结论：CLKC1在炎症早期具有抗炎作用,而在炎症晚期无明显效果。
2 CLKC1治疗小儿上呼吸道感染后咳嗽的作用机制研究
2.1 CLKC1对哮喘大鼠气道高反应性的影响
目的：通过研究CLKC1对哮喘大鼠气道阻力,HMGB-1、IgE、IL-4、IL-13表达的影响,考察其对气道高反应性的抑制作用.方法：以卵蛋白和灭活结核杆菌菌苗激发大鼠哮喘模型,予CLKC1干预后测定其气道阻力、血清HMGB-1、肺匀浆中IgE、IL-4、IL-13的含量.结果：CLKC1可以抑制大鼠气道阻力的增加以及血清HMGB-1.肺匀浆中IgE、IL-4、IL-13含量的增高.结论：CLKC1可以通过抑制炎症介质、细胞因子含量的增高降低哮喘大鼠的气道高反应性。
2.2 CLKC1对哮喘小鼠气道高反应性的影响
目的：通过研究CLKC1对哮喘小鼠辣椒素咳嗽敏感性试验,NO、ET-1、IL-4、IL-13表达的影响,考察其对气道高反应性的抑制作用。方法：以卵蛋白激发Ba1b/c小鼠哮喘模型,予CLKC1干预后测定咳嗽敏感性试验辣椒素的阈值以及肺匀浆中NO、ET-1、IL-4、IL-13的含量。结果：CLKC1可以增加辣椒素咳嗽敏感性试验的阈值,抑制肺匀浆中NO、ET-1、IL-4、IL-13含量的增高。结论：CLKC1可能从减轻气道炎症、促进气道重建角度降低哮喘小鼠的气道高反应性。
2.3 CLKC1对晚期炎症介质HMGB-1含量的影响
目的：观察CLKC1对晚期炎症介质高迁移率族蛋白B-1(HMGB-1)含量的影响。方法：测定LPS尾静脉注射致急性肺损伤小鼠不同时段血清中HMGB-1的含量,取含量最高的时间点,CLKC1预防治疗给药,测定在含量最高时间点时急性肺损伤小鼠血清中HMGB-1的含量。结果：CLKC1能够抑制HMGB-1含量的增高(P<0.05)。结论：CLKC1对晚期炎症介质HMGB-1含量的增高具有一定的抑制作用。
结论
1.药效学研究表明CLKC1具有止咳、祛痰、平喘、通便、抗炎的药理作用。
2.作用机制研究表明CLKC1治疗小儿上呼吸道感染后咳嗽的机理可能为从抑制细胞因子及炎性介质含量的增高,促进气道重建等角度减小气道阻力,降低气道高反应性。
创新点
1.监测了急性肺损伤小鼠模型血清中HMGB-1的含量随时间的变化情况,研究了中药颗粒剂对晚期炎症介质HMGB-1含量的影响。
2.首次将辣椒素咳嗽敏感性试验与气道高反应动物模型相结合,以辣椒素的阈值检测气道高反应性,并研究了中药颗粒剂对气道高反应动物模型辣椒素咳嗽敏感性试验阈值的影响。
Pediatric coughing after upper respiratory tract infection refers to the children's coughing for a long time without healing while the cold restores, which is also called cough post infection, referred to as CPI. The upper respiratory tract infection is a common pediatric disease, frequently happens in winter, often leads to the post infection cough. For parts of the patients, respiratory symptoms of cough appear after the recovery of upper respiratory tract infection, lasting for several weeks to mohths, often misdiagnosed as bronchitis or pneumonia, and the repeated use of antibiotics, cough killer etc is of no effect. It not only brings the children of pain, but also causes economic burden for parents. Because of the high incidence of coughing after upper respiratory tract infection, the drug therapy has been more concerned in recent years.
At present, the therapic drugs of CPI are mainly histamine receptor antagonist, central antitussive, anaesthetic, hormone, epinephrine never receptor antagonist, etc. Western medicine is not of great good effects in treating pediatric CPI. In treating cough, TCM has a long history and rich experience, our national unique traditional Chinese medicine resourse has provided a vast space for the application of TCM in treating CPI. Chinese patent medicines is convenient to carry and easy to use, so making Chinese prescription develope into Chinese patent medicines will have broad economic and social benefit based on the remarkable curative effect in treating CPI.
CLKC1 is developed from experienced prescription. The function is lung-clearing and expectorant, antitussive and throat-clearing. It has been made into granules by pharmaceutical department of Beijing University of Chinese Medicine, using for the treatment of CPI.
Generally speaking the main pathogenesis of CPI is AHR.With the function basis of CLKC1, combining the western medicine pathogenesis of CPI,we study pharmacological effect and function mechanism by conventional animal models. Two parts are mainly included:first a study on pharmacological effect; second the study on function mechanism. We try to clarify the function mechanism of CLKC1's pharmacological effect, in order to provide sufficient experimental data for the clinical using and new drug's application.
1 Study on pharmacodynamics of CLKC1 in CPI treatment
1.1 Study on antitussive effects of CLKC1
Objective:To observe the antitussive effects of CLKC1. Methods:Antitussive effects were observed through mice cough test induced by SO2steaming and through guinea pigs cough test induced by citric acid. Results:CLKC1 decreased the frequency of mice cough induced by ammonia and guinea pigs cough induced by citric acid, prolonged the cough latent period,dose groups have good concentration response relations (p<0.05). Conclusion: CLKC1 shows certain Antitussive effects.
1.2 Study on expectorant effects of CLKC1
Objective:To observe expectorant effect of CLKC1. Methods:Expectorant effectswere evaluated by measuring the excretion of phenol red in mice trachea. Results: Increased the excretion of phenol red from mice trachea, promoted the movement of prepared Chinese ink in the mice trachea (p<0.05). Conclusion:CLKC1 shows certain expectorant effects.
1.3 Study on Antiasthmatic effects of CLKC1
Objective:To observe antiasthmatic effect of CLKC1. Methods:Antiasthmatic actions in guinea pigs were observed by histamine and acetylcholine ultrasonic atomization test. Results:CLKC1 can inhibited the guinea pigs asthma introduced by histamine and acetylcholine solution (p<0.05). Conclusion:CLKC1 shows certain antiasthmatic effects. 1.4 Study on bowel propulsion effects of CLKC1
Objective:To observe the effect of CLKC1 on bowel propulsion in mice. Methods: Comput and compare the EB propulsion percentages in bowel in CLKC1 group, model group (treated with bowel propulsion supp ressants) and blank group, and the data were analyzed statistically 1. Results:CLKC1 dose groups could significantly increase EB propulsion percentages in mice(p<0.01).Conclusion:CLKC1 can promote intestine propulsive movement.
1.5 Study on anti-inflammatory effects of CLKC1
Objective:To observe anti-inflammatory effects of CLKC1. Methods:anti-inflammatory effects were observed by the swelling of hind paw in rats induced by carrageenin and granuloma induced by cotton pellets in rats. Results:CLKC1 medium and small doses groups can inhibite the paw edema induced by carrageenin (p<0.01), but not the granuloma induced by cotton pellet in SD rats. Conclusion:CLKC1 can inhibite early inflammation but without anti-inflammatory effects on later inflammation.
2 Study on Function mechanism of CLKC1 in CPI treatment
2.1 Study on effect of CLKC1 on AHR in allergic asthma rats
Objective:To investigate the effects of CLKC1 on the inhibition of airway hyperresponsiveness(AHR) in allergen induced asthma in rats. Methods:Model group and CLKC1 group were challenged with ovalbumin (OVA) to reproduce asthma., the blank group only receied an injection of same amount of Al(OH)3. On the 22nd day determine airway resistance, the content of HMGB-1、IgE、IL-4、IL-13 in lung homogenate. Results: CLKC1 can reduce the rat airway resistance, and inhibit the content's increase of HMGB-1、IgE、IL-13、IL-4 in lung homogenate(P<0.05). Conclusion:CLKC1 can reduce airway resistance and inhibit AHR.
2.2 Study on effect of CLKC1 on AHR of chronic asthma murine
Objective:To investigate the effects of CLKC1 on the inhibition of airway hyperresponsiveness(AHR) in allergen induced asthma in Balb/c rats. Methods:model group and CLKC1 group were challenged with ovalbumin (OVA) to reproduce asthma.,the blank group only receied an injection of same amount of Al(OH)3 On the 22nd day determine the threshold of capsaicin cough sensitivity test, the content of NO、ET-1、IL-4、IL-13 in lung homogenate and their physiological and pathological changes. Results:CLKC1 can increase the threshold of capsaicin cough sensitivity test, inhibit the content's increase of lung homogenate's NO、ET-1、IL-4、IL-13(P<0.05). Conclusion:CLKC1 can inhibit AHR.
2.3 Study on effect of CLKC1 on serum HMGB-1 level of sepsis mic
Objective:To observe the effects of CLKC1 on HMGB-1 protein expression in lung tissues of mice with endotoxin shock. Methods:Except the blank group, all other groups are injected LPS (5mg/kg)by the tails vein to establish the lung tissue injury models, The blank group only receied an injection of same amount of saline. Testing the serum HMGB-1 level at 24h,36h,48h,72h,96h. Study the effect of CLKC1 on serum HMGB-1 level of sepsis mic at the point of 72h. Results:72h after tail vail injection of LPS, serum HMGB-1 gets to the highest level; CLKC1 can inhibit the content's increase of HMGB-1 (P<0.05). Conclusion: CLKC1 can decrease serum HMGB-1 level.
Conclusion
1 The pharmacodynamics study shows that CLKC1 has antitussive, expectorant, antiasthmatic, bowel propulsion, anti-inflammatory effect pharmacological activity.
2 Study on function mechanism of CLKC1 in treating coughing after pediatric upper respiratory tract infection indicates that CLKC1 can reduce airway resistance, inhibit the content's increase of inflammatory mediators and cytokines, promote airway remodeling so as to decrease AHR.
Innovation
1 First studied the effect of Chinese medicine granule on the expression of. terminal inflammatory mediators HMGB-1.
2 Combine capsaicin cough sensitivity test with AHR animal models for the first time, detecting airway resistance by the threshold of capsaicin, and also study the effect of Chinese medicine granule on the threshold of capsaicin.
引文
[1]中华医学会呼吸病学分会哮喘学组.咳嗽的诊断与治疗指南(草案).中华结核和呼吸杂志,2005,28(11)：738-744.
[2]陆再英,终南山.内科学.7版[M].北京：人民卫生出版社,2004：70.
[3]陈庆,杨巍.奥亭加普米克令舒雾化吸入治疗上呼吸道感染后咳嗽的疗效观察[J].西南军医,2008,10(3)：9-10.
[4]王昀,程捷,祝振中.西替利嗪治疗感冒后咳嗽140例[J].现代中西医结合杂志,2009,18(5)：531.
[5]曹照龙.感冒后咳嗽[J].中国临床医生杂志,2007,35(1)：13-14.
[6]辛建保,向敏,陶晓南等.慢性干咳伴有气道高反应性即是咳嗽变异性哮喘吗?[J]中华结核和呼吸杂志,1998,21(3)：138-139.
[7]陆再英,终南山.内科学7版[M].北京：人民卫生出版社,2004：70.
[8]李清.阿斯美联合酮替酚改善上呼吸感染后咳嗽症状[N].中国医学论坛报,2006, 07-13(29).
[9]辛建保,向敏,陶晓南等.慢性干咳伴有气道高反应性即是咳嗽变异性哮喘吗?[J]中华结核和呼吸杂志,1998,21(3)：138-139.
[10]熊亮,陶晓南.气道高反应性发生机制及与之相关临床疾病[J].国际呼吸杂志,2006,26(3)：223-225.
[11]Braman SS. Postinfectious cough ACCP evidence-based clinical practice guidelines [J]. Chest,2006,129(1Suppl):138S-146S.
[12]黄秋芳.小儿感染后咳嗽的发病机理及治疗进展[J].中国药业,2009,18(15)：84-87.
[13]To Y, Dohi M, Tanaka Retal. Early interleuk in 4-dependent response can induce airway hyperreactivity before development of airway inflammation in a mouse model of asthma[J]. Lab Invest,2001,81:1385-1396.
[14]Monique S, Jean LB, Boris Vargaftig. Granulocyte depletion and dexamethasone differentially modulate airways hyperre activity, inflammation, mucus accumulation and sectetion induced by rm IL-13 or antigen[J]. Am J Respir Cell Mol Biol, 2002,26 (1):74-78.
[15]金伯泉.细胞和分子免疫学[M].北京：科学出版社,2001：141-142.
[16]Ming Simon PH, Peter JH, etal. Interleukin-13 mediates airway hyperre activity through the IL-4 receptor, alpha chain and STAT-6 independently of IL-5 and eotaxin[J]. Am JRespir Cell Mol Biol,2001; 2041-2044; 25:52-53.
[17]Finkelman FD, Yang M, Perkins C, etal [J]. J Immunol,2005,174:4630-4638.
[18]David M. Walter JJ, Mcintire GB, etal. Critical role for IL-13 in the development of allergen induced airway hyper reactivity [J]. J Immunol,2001; 167:4668-4675.
[19]MorcilloEJ, CortijoJ. Mucus and M U C in asthma[J]. Curr Opin Pulm Med,2006, 12(1):1-6.
[20]吴为群,江宏志,吴忠道等.血清白细胞介素IL-13、17与气道高反应性的关系[J].热带医学杂志,2004,4(5)：539-541,
[21]黄燕.Th1/Th2细胞因子网络失衡与毛细支气管炎[J].医学综述,2008,14(16)：2427—2430.
[22]Horwitz RJ, McGill KA, Buss WW.The role of leukotriene modifiers in the treatment of asthma[J]. Am J Respir Crit CareMed,1998,157(2):1363-71.
[23]马勇录,蒋昆.白三烯受体拮抗剂治疗哮喘的进展和临床评价[J].中国医院用药评价与分析,2004,4(5)：273-276.
[24]方超平,方凤.病毒感染与气道高反[J].应中华国际医学杂志,2002,2(2)：169-171.
[25]王欢,邹悦.细胞黏附分子在鼻息肉发病机制中的作用[J].中国耳鼻咽喉颅底外科杂志,2008,14(6)：478-480.
[26]何杰明,郝青林.细胞因子与急性肺损伤研究进展[J].医学综述,2009,15(3)： 385-389.
[27]Bernstein JM. The molecular biology of nasal polyposis [J]. Curr Allergy Asthma Rep, 2001,1 (3):262-267.
[28]任雁宏,秦晓群,管茶香.气道高反应中血管活性肠肽及受体的时空分布[J].中华结核和呼吸杂志,2004,24(4)：224-228.
[29]QINXiao-Qun, XIANGYang, LIUChi, etal. Therole of bronchiale Pithelial cells in airway hyperres Ponsiveness. Actaphysioloica Sinica,2007,59 (4):454-464.
[30]崔新乐,新元环.一氧化氮与哮喘[J].国外医学呼吸系统册,1996,16(1)：1.
[31]张慎峰.气道高反应性的研究进展[J].黑龙江医学,2003,27(10)：749-750.
[32]Ferrara N, Gerber HP, LeCouter J. The biology of VEGFand its receptors [J]. Nat Med, 2003,9 (6):669-676.
[33]Walters EH, Reid D, Soltani A, etal. Angiogenesis:apotentially critical part of remodelling in chronic airwaydiseases [J]. Pharmacol Ther,2008,118 (1):128-137.
[34]Hoshino M, Tanaka H, Abe S. Quantitative analysis ofbronchial wall vascularityin the medium and small airways of patients with asthma and COPD [J]. Chest,2005,127 (3):965-972.
[35]Hoshino M, Takahashi M, Aoike N, etal. Expression ofvascular endothelial growth factor, basic fibroblast growthfactor, and angiogenin immunoreactivity in asthmatic airways and its relationship to an giogenesis [J]. J Allergy Clin Immunol,2001,107 (2):295-301.
[36]Simcock DE, Kanabar V, Clarke GW, etal. Induction of angiogenesis by airway smooth muscle from patients with asthma [J]. Am J Respir Crit Care Med,2008,178 (5): 460-468.
[37]Heikkinen T, Jarvinen A. The common cold. Lancet,2003,361:51-59.
[38]王剑梅.酮替芬联合氨茶碱治疗感冒后咳嗽临床分析[J].医学理论与实践,2008,21(12)：1421-1422.
[39]王昀,程捷,祝振忠.西替利嗪治疗感冒后咳嗽140例[J].现代中西医结合杂志,2009,18(5)：531.
[40]王蓉,徐维国,张晓东.复方磷酸可待因溶液治疗感染后咳嗽的临床观察[J].华西医学,2008,23(5)：1128-1129.
[41]黄子庆.环甲膜注射地塞米松、利多卡因治疗慢性咳嗽[J].中国厂矿医学,2003,16(3)：224.
[42]蒋萍.爱全乐治疗感染后咳嗽40例治疗观察[J].临床肺科杂志,2008,13(4)：507-508.
[43]王小梅.复方甲氧那明和酮替芬治疗急性上呼吸道感染后咳嗽的疗效[J].基础医学·相关疾病与临床,2008,24(15)：1593-1595.
[44]唐红玉,叶巍岭,杨代秀等.普米克、万托林雾化吸入治疗上呼吸道感染后呼吸道症状的疗效观察[J].2007,23(23)：3747-3748.
[1]武维屏.中西医结合呼吸病学[M].北京：中国医药科技出版社,1998,4.
[2]周健.开肺行水法治疗外感咳嗽的理论分析与临床观察[J].现代医院,2006,6(7)：92.
[3]李际强,李振洁,罗翌.试述《金匮要略》中治肺止咳方法的特点[J].辽宁中医药大学学报,2007,9(4)：14.
[4]张秋霞.聂惠民教授治疗小儿外感咳嗽的经验[J].中华中医药杂志,2008,23(4)：335-337.
[5]来晓娟,司晓文.论温法在外感咳嗽中的应用[J].中国医药导报,2008,5(1)：76.
[6]吴琼.王素梅治疗小儿咳嗽经验[J].现代中西医结合杂志,2009,18(27：3362-3363.
[7]张颖.脏病治腑法在小儿外感咳嗽中的运用[J].山东中医杂志,2007,26(5)：343.
[8]叶贺平.小柴胡汤治疗郁火咳嗽60例疗效分析[J].实用中医内科杂,2006,20(1)：56.
[9]钟华,钟柏亭.王力宁因质辨治小儿外感咳嗽的经验[J].浙江中医杂志2006,41 (12)：688-689.
[10]王萍芬.中医儿科学[M].上海：上海科学技术出版社,1997,37.
[11]尹维东,任闪闪.小儿咳嗽的中医辨治[J].中医垂教.2006,22(316)：36-37.
[12]欧秀梅.止嗽散加减治疗小儿外感咳嗽40例[J].中医儿科杂志,2008,4(3)：20-21.
[13]王怡冰,张书亮.桑白皮汤加减治疗外感咳嗽疗效分析[J].医药论坛杂志,2006,27(13)：100-102.
[14]高季鸿,耿建国.麻杏甘石汤加味治疗外感后咳嗽的临床观察[J].四川中医,2007,25(8),66.
[15]肖湘.桑杏汤加减治疗燥咳54例[J].实用中医内科杂志,2007,21(2)：59.
[16]陈英妹,刘伟.咳嗽颗粒加味治疗小儿外感咳嗽72例[J].药物与临床,2009,3,103.
[17]孙学刚,马克林,崔燕.祛风止咳汤治疗外感咳178例[J].陕西中医学院学报,2005,28(5)：26-27.
[18]李桂君.湿热蕴肺致小儿咳嗽的辨治体会[J].新疆中医药,2009,27(4)：97-98.
[19]逵艳,张书俊,张莹.自拟止咳平喘汤治疗小儿外感咳嗽80例[J].国医论坛,2007,22(1)：27.
[20]虞金龙.中药贴敷治疗外感咳嗽与支气管炎疗效观察[J].浙江中西医结合杂志,2005,15(1)：41.
[21]乔林波.孔最穴封闭疗法临床应用体会[J].JCAM,2007,23(10)：28.
[22]郭素洁.推拿配合灸肺俞治疗小儿咳嗽67例[J].按摩与引导,2004,20,(5)：50.
[23]王玉.推拿加拔罐治疗小儿外感咳嗽90例[J].陕西中医,2008,29(11)：1518-1519.
[24]李炜奕.中药敷脐法治疗小儿外感咳嗽疗效观察[J].吉林医学,2008,29(3)：235.
[25]尹文仲.盐熨法治疗小儿外感咳嗽[J].湖北民族学院学报·医学版,2007,3,
[1]Anderssion U, Wang H, Palmblad K, etal. High mobility group 1 protein stimulates proinflammatory cytokine synthesis in humanmoco-cytes [J]. J.Exp. Med.2000,192: 5655-5570.
[2]姚咏明,盛志勇.高迁移率族蛋白-1在脓毒症发病中的作用和意义[J].解放军医学杂志,2002,27：752-756.
[3]Haniguchi N, Kawahara K, Yone K, etal. High mobility groupchrosomosomal protein 1 plays a role in the pathogenesis of rheumatoidarthritis as a novel cytokine [J]. Arthrtis Rheum,2003,48(4):971-981.
[4]Ullertis R, Jonsson IM, Verdrengh M, etal. High mobility groupchrosomosomal protein 1, a DNA binding cytokine, induces arthritis [J]. Arthritis Rheum,2003,48(6):1693-1 700.
[5]Wang H, Bloomo, Zhang M, etal. HMG-1 as a late mediatorof endotoxin lethality in mice[J]. Science 1999,285:248-251.
[6]Abraham E, Srcaroli J, Carmody A, et al. HMGB-1 as a media-tor of acute lung inflammation [J]. J. Immunol,2000,165:2950-2954.
[7]Ueno H, Matsuda T, Hashimoto S, etal. Contributions of highmobility group box protein in experimental and clincial acute lung in-jury [J]. Am J Respir,2004,17::185-189.
[8]孟志艳,李军,曹红.HMGB1在呼吸系统疾病中的作用[J].山东医药,2009,49(8)：17-108.
[9]Haniguchi N, Kawahara K, Yone K, etal. High mobility groupchrosomosomal protein 1 plays a role in the pathogenesis of rheumatoidarthritis as a novel cytokine [J]. Arthrtis Rheum,2003,48(4)::971-981.
[10]Ullertis R, Jonsson IM, Verdrengh M, etal. High mobility groupchrosomosomal protein 1, a DNA binding cytokine, induces arthritis [J]. Arthritis Rheum,2003,48(6):1693-1700.
[11]郭惠芳,刘淑霞,马丽艳等.类风湿关节炎患者血清中高迁移率族蛋白-1的作用及其与MMP2、 TIMP2的关系[J].中国现代医学杂志,2010,20(2)：192-195.
[12]Sass G, Heinlein S, Agli A, etal.Cytokine expression in threemouse models of experimental hepatitis [J]. Cytokine,2002,19(3):115-120.
[13]杨智勇,王春友,熊炯等.重症急性胰腺炎大鼠血清高迁移率族蛋白-1水平的时相变化及意义[J].华中科技大学学报(医学版),2004,33(4)：466-468.
[14]Sappington PL, Yang R, Yang H, etal. HMGB-1 B box increases the permeability of Caco-2 enteroytic monolayem and impairs intestinal barrier function in mice[J]. Gastroenterology,2002,123(3):790-802.
[15]栾正刚,何忠野,张成,等.丙酮酸乙酯对重症急性胰腺炎大鼠肠粘膜HMGB-1表达的影响[J].中国普通外科杂志,2008,17(3)：214-218.
[16]Kalinina N, Agrotis A, Antroprovaet etal. Increased expression ofthe DNA-binding cytokine HMGB-1 in human atherosclerotic le-sions.Arterioscler Thromb Vasc Biol, 2004,24(12):2320-2325.
[17]Ulfgren AK, Grundtman C, Borg k, et al. Dowin-regulation ofthe aberrant expression of the inflammation mediator high mobility groupchrosomosomal protein 1 in muscle tissure of patients with polymyositisand dermatomyositis treated with corticoste-roids.Arthritis Rheum,2004,50(5):1586-1594.
[18]Naghavi MH, Nowak P, Andersson J, et al. Intracelluar highmobility group protein represss HIV-1 LTR-directed transcription in a promotor and cell specific manner [J]. Virology,2003,314(1):179-189.
[19]朱海云,李银平.高迁移率族蛋白B-1的研究进展及其免疫作用[J].中国中西医结合急救杂志,2009,16(2)：124-125.
[1]Sant'Ambrogio G, Widdicombe J.Reflexes from airway rapidly adapting receptors [J]. Respir Physiol,2001,125(1-2):33-45.
[2]Karlsson JA, Fuller RW. Pharmacological regulation of the cough reflex from experimental models to antitussive effects in Man[J]. Pulm Pharmacol Ther,1999, 12(4):215-228.
[3]MyersA C, KajekarR, Undem B J. Allergic inflammation-inducedneuropeptide production in rapidly adapting afferent nerves in guinea pig airways [J]. Am J Physiol Lung Cell MolPhysio,12002,282:L775-L781.
[4]WinterC A, FlatakerL. The effects of drugs up on a graded cough response obtained in sensitized guinea pigs exposed to aerosol of specific antigen [J]. J Exp Med,1955, 101:17-24.
[5]Kamei J, MoritaK, KashiwazakiT, etal. Antitussive effect ofmoguisteine on allergic coughs in the guinea pig [J]. Eur J Pharmaco,11998,47:253-255.
[6]HouseA, CellyC, SkeansS, etal. Cough reflex in allergic dogs [J]. EurJ Pharmaco,2004, 492:251-258.
[7]Kamei J, Saitoh A, Morita K, Nagase H. Involvement of adenosinA1 receptors in antitussive effect in mice [J]. Life Science,1994; 5(20):383-8.
[8]Milos T, Giuseppe SA, Franca BSA. Laryngeal and tracheo bronchial cough in anesthetized dogs [J]. JAppl Physiol,1994; 76(6):2672-9.
[9]CaterinaMJ, LefflerA, MalmbergAB, etal. Impaired nociceptionand pain sensation in mice lacking the capsaicin receptor[J]. Science,2000,288:306-313.
[10]Ma QP. Expression of capsaicin receptor (VR1) by myelinated primary afferentneurons in rats [J]. NeurosciLett,2002,319:87-90.
[11]Takahama K, Araki T, Fuchikami J-I etal. Studies on the magnitude and the mechanism of cough potentiation by angiotensin converting enzyme inhibitors in guinea pigs involvement of bradykini in the potentiation[J]. JPharm Pharmacol,1996; 48:1027-33.
[12]陈如冲,赖克方,刘春丽等.辣椒素咳嗽激发试验方法的建立及其安全性评价[J].中华结核和呼吸杂志,2005,28(11)：751-754.
[13]Greene S A, Wolff R K, Hahn F F, etal. Sulfur dioxide-inducedchronic bronchitis in beagle dos[J]. JtoxicolEnvironHealth,1984,13:945-958.
[14]Takahama, Fuchikami J, KaiH, etal. Inhalation of phosphoradon, a neutral endopeptidase inhibitor, induces cough in awguinea-pigs[J]. Arch IntPharmacodyn Ther,1995,330: 241-250.
[15]Moreaux B, BeerensD, Gustin P. Development of a cough inductiontest in pigs:effects of SR 48968 and enalapri-1[J]. J Vet Pharmacol Ther,1999,22:387-389.
[16]Moreaux B, Advenier C, Gustin P. Role of bradykinin and tachyki-nins in the potentiation by enalaprilofcoughing induced by citric acid[J]. Fund Clin Pharmaco, 12001,15:23-29.
[1]徐叔云,卞如濂,陈修.药理实验方法学[M].北京：人民卫生出版社,1991,1：1821.
[2]周添浓,唐立海,黄诗冲等.五指毛桃镇咳及平喘作用研究[J].中药材,2009,32(4)：571-574.
[3]Sant'Ambrogio G, Widdicombe J. Reflexes from airway rapidly adapting receptors [J]. Respir Physiol,2001,125(1-2):33-45.
[4]Karlsson JA, Fuller RW. Pharmacological regulation of the cough reflex from experimental models to antitussive effects in Man[J]. Pulm Pharmacol Ther,1999, 12(4):215-228.
[5]Hunter DD, Undem BJ. Identification and substance P content of vagal afferent neurons innervating the epithelium of the guinea pig trachea [J]. Am J Respir Crit Care Med, 1999,159(6):1943-1948.
[1]陈奇.中药药理研究方法学[M].北京：人民卫生出版社,1994,80：140-148.
[2]魏东,姚斌,高兴亮等.少年红矾杏平喘糖浆镇咳、祛痰和平喘作用的实验研究[J].中医中药与微循环.2009,19(2)：29-32.
[1]Nakamura M,Yamaya M,Fukushima T,et al. Effect of mabuterol on t racheal muco-ciliary clearance of magnetized iron particles in anest hetized dogs[J]. Respiration, 1991,58(1):331.
[1]王科闯、杨宇.肺与大肠相表里的实验研究概况[J].《山东中医药大学学报》,2006,
[1]陈奇.中药药理研究方法学[M].北京：人民卫生出版社,1993,272-273.
[1]徐玉民,朱敏敏,戎海波,等.氯胺瞳雾化吸入对哮喘大鼠气道高反应性的影响[J].南京医科大学学报(自然科学版),2007,27(3)：235-238.
[2]李明华,殷凯生, 蔡映云.哮喘病学[M].北京：人民卫生出版社,2005,128.
[3]MorcilloEJ, CortijoJ. Mucus and M U C in asthma[J]. Curr Opin Pulm Med,2006, 12(1):1-6.
[4]吴为群,江宏志,吴忠道等.血清白细胞介素IL-13、17与气道高反应性的关系[J].热带医学杂志,2004,4(5)：539-541.
[1]Jung J, Hwang SW, Kwak J, et al. Capsaicin binds to the intracellular domain of the capsaicin-activated ion channel[J]. Jneurosci,1999,19(2):529-538.
[2]容朝晖,沈策.咳嗽反射机制的研究进展[J].国外医学呼吸系统分册,2005,25(3)：168-171.
[3]Forsberg K, Karlsso JA, The odorsson et al. Cough and bronchoconstriction mediated by capsaicin-sensitive sensory neurons in thguinea pig[J]. Pulm Pharmacol,1988,1:33-39.
[4]Takahama K, Araki T, Fuchikami J-I et al. Studies on the magnitude and the mechanism of cough potentiation by angiotensin converting enzyme inhibitors in guinea pigs
involvement of bradykini in the potentiation[J]. JPharm Pharmacol,1996,48:1027-1033.
[5]陈如冲,赖克方,刘春丽等.辣椒素咳嗽激发试验方法的建立及其安全性评价[J].中华结核和呼吸杂志,2005,28(11)：751-754.
[1]Anderssion, U, Wang H, Palmblad K, et al.High mobility group 1 protein stimulates proinflammatory cytokine synthesis in humanmoco-cytes. J.Exp. Med,2000,192:5 655-5570.
[2]Abraham E, Arcaroli J, Carmody A, et al. HMG-1 as a mediator of acute lung Inflam-mation[J]. The Journal of Immunology,2000,165(6):2950-2924.
[3]孟志艳,李军,曹红.HMGB-1在呼吸系统疾病中的作用[J].山东医药,2009,49(8)：107-108.
[4]Lin X, Yang H, Sakuragi T, et al. Alpha chemokine receptor blockade reduces high mobility group box-1 protein induced lung inflammarion and injury and improves survi-valin sepsis[J]. Am J PhysiolLung CellMol Physiol,2005,289(4):L583-590.
[5]姚咏明,盛志勇.高迁移率族蛋白-1在脓毒症发病中的作用和意义[J].解放军医学杂志,2002,27：752-756.
[2]陆再英,终南山.内科学.7版[M].北京：人民卫生出版社,2004：70.
[3]陈庆,杨巍.奥亭加普米克令舒雾化吸入治疗上呼吸道感染后咳嗽的疗效观察[J].西南军医,2008,10(3)：9-10.
[4]王昀,程捷,祝振中.西替利嗪治疗感冒后咳嗽140例[J].现代中西医结合杂志,2009,18(5)：531.
[5]曹照龙.感冒后咳嗽[J].中国临床医生杂志,2007,35(1)：13-14.
[6]辛建保,向敏,陶晓南等.慢性干咳伴有气道高反应性即是咳嗽变异性哮喘吗?[J]中华结核和呼吸杂志,1998,21(3)：138-139.
[7]陆再英,终南山.内科学7版[M].北京：人民卫生出版社,2004：70.
[8]李清.阿斯美联合酮替酚改善上呼吸感染后咳嗽症状[N].中国医学论坛报,2006, 07-13(29).
[9]辛建保,向敏,陶晓南等.慢性干咳伴有气道高反应性即是咳嗽变异性哮喘吗?[J]中华结核和呼吸杂志,1998,21(3)：138-139.
[10]熊亮,陶晓南.气道高反应性发生机制及与之相关临床疾病[J].国际呼吸杂志,2006,26(3)：223-225.
[11]Braman SS. Postinfectious cough ACCP evidence-based clinical practice guidelines [J]. Chest,2006,129(1Suppl):138S-146S.
[12]黄秋芳.小儿感染后咳嗽的发病机理及治疗进展[J].中国药业,2009,18(15)：84-87.
[13]To Y, Dohi M, Tanaka Retal. Early interleuk in 4-dependent response can induce airway hyperreactivity before development of airway inflammation in a mouse model of asthma[J]. Lab Invest,2001,81:1385-1396.
[14]Monique S, Jean LB, Boris Vargaftig. Granulocyte depletion and dexamethasone differentially modulate airways hyperre activity, inflammation, mucus accumulation and sectetion induced by rm IL-13 or antigen[J]. Am J Respir Cell Mol Biol, 2002,26 (1):74-78.
[15]金伯泉.细胞和分子免疫学[M].北京：科学出版社,2001：141-142.
[16]Ming Simon PH, Peter JH, etal. Interleukin-13 mediates airway hyperre activity through the IL-4 receptor, alpha chain and STAT-6 independently of IL-5 and eotaxin[J]. Am JRespir Cell Mol Biol,2001; 2041-2044; 25:52-53.
[17]Finkelman FD, Yang M, Perkins C, etal [J]. J Immunol,2005,174:4630-4638.
[18]David M. Walter JJ, Mcintire GB, etal. Critical role for IL-13 in the development of allergen induced airway hyper reactivity [J]. J Immunol,2001; 167:4668-4675.
[19]MorcilloEJ, CortijoJ. Mucus and M U C in asthma[J]. Curr Opin Pulm Med,2006, 12(1):1-6.
[20]吴为群,江宏志,吴忠道等.血清白细胞介素IL-13、17与气道高反应性的关系[J].热带医学杂志,2004,4(5)：539-541,
[21]黄燕.Th1/Th2细胞因子网络失衡与毛细支气管炎[J].医学综述,2008,14(16)：2427—2430.
[22]Horwitz RJ, McGill KA, Buss WW.The role of leukotriene modifiers in the treatment of asthma[J]. Am J Respir Crit CareMed,1998,157(2):1363-71.
[23]马勇录,蒋昆.白三烯受体拮抗剂治疗哮喘的进展和临床评价[J].中国医院用药评价与分析,2004,4(5)：273-276.
[24]方超平,方凤.病毒感染与气道高反[J].应中华国际医学杂志,2002,2(2)：169-171.
[25]王欢,邹悦.细胞黏附分子在鼻息肉发病机制中的作用[J].中国耳鼻咽喉颅底外科杂志,2008,14(6)：478-480.
[26]何杰明,郝青林.细胞因子与急性肺损伤研究进展[J].医学综述,2009,15(3)： 385-389.
[27]Bernstein JM. The molecular biology of nasal polyposis [J]. Curr Allergy Asthma Rep, 2001,1 (3):262-267.
[28]任雁宏,秦晓群,管茶香.气道高反应中血管活性肠肽及受体的时空分布[J].中华结核和呼吸杂志,2004,24(4)：224-228.
[29]QINXiao-Qun, XIANGYang, LIUChi, etal. Therole of bronchiale Pithelial cells in airway hyperres Ponsiveness. Actaphysioloica Sinica,2007,59 (4):454-464.
[30]崔新乐,新元环.一氧化氮与哮喘[J].国外医学呼吸系统册,1996,16(1)：1.
[31]张慎峰.气道高反应性的研究进展[J].黑龙江医学,2003,27(10)：749-750.
[32]Ferrara N, Gerber HP, LeCouter J. The biology of VEGFand its receptors [J]. Nat Med, 2003,9 (6):669-676.
[33]Walters EH, Reid D, Soltani A, etal. Angiogenesis:apotentially critical part of remodelling in chronic airwaydiseases [J]. Pharmacol Ther,2008,118 (1):128-137.
[34]Hoshino M, Tanaka H, Abe S. Quantitative analysis ofbronchial wall vascularityin the medium and small airways of patients with asthma and COPD [J]. Chest,2005,127 (3):965-972.
[35]Hoshino M, Takahashi M, Aoike N, etal. Expression ofvascular endothelial growth factor, basic fibroblast growthfactor, and angiogenin immunoreactivity in asthmatic airways and its relationship to an giogenesis [J]. J Allergy Clin Immunol,2001,107 (2):295-301.
[36]Simcock DE, Kanabar V, Clarke GW, etal. Induction of angiogenesis by airway smooth muscle from patients with asthma [J]. Am J Respir Crit Care Med,2008,178 (5): 460-468.
[37]Heikkinen T, Jarvinen A. The common cold. Lancet,2003,361:51-59.
[38]王剑梅.酮替芬联合氨茶碱治疗感冒后咳嗽临床分析[J].医学理论与实践,2008,21(12)：1421-1422.
[39]王昀,程捷,祝振忠.西替利嗪治疗感冒后咳嗽140例[J].现代中西医结合杂志,2009,18(5)：531.
[40]王蓉,徐维国,张晓东.复方磷酸可待因溶液治疗感染后咳嗽的临床观察[J].华西医学,2008,23(5)：1128-1129.
[41]黄子庆.环甲膜注射地塞米松、利多卡因治疗慢性咳嗽[J].中国厂矿医学,2003,16(3)：224.
[42]蒋萍.爱全乐治疗感染后咳嗽40例治疗观察[J].临床肺科杂志,2008,13(4)：507-508.
[43]王小梅.复方甲氧那明和酮替芬治疗急性上呼吸道感染后咳嗽的疗效[J].基础医学·相关疾病与临床,2008,24(15)：1593-1595.
[44]唐红玉,叶巍岭,杨代秀等.普米克、万托林雾化吸入治疗上呼吸道感染后呼吸道症状的疗效观察[J].2007,23(23)：3747-3748.
[1]武维屏.中西医结合呼吸病学[M].北京：中国医药科技出版社,1998,4.
[2]周健.开肺行水法治疗外感咳嗽的理论分析与临床观察[J].现代医院,2006,6(7)：92.
[3]李际强,李振洁,罗翌.试述《金匮要略》中治肺止咳方法的特点[J].辽宁中医药大学学报,2007,9(4)：14.
[4]张秋霞.聂惠民教授治疗小儿外感咳嗽的经验[J].中华中医药杂志,2008,23(4)：335-337.
[5]来晓娟,司晓文.论温法在外感咳嗽中的应用[J].中国医药导报,2008,5(1)：76.
[6]吴琼.王素梅治疗小儿咳嗽经验[J].现代中西医结合杂志,2009,18(27：3362-3363.
[7]张颖.脏病治腑法在小儿外感咳嗽中的运用[J].山东中医杂志,2007,26(5)：343.
[8]叶贺平.小柴胡汤治疗郁火咳嗽60例疗效分析[J].实用中医内科杂,2006,20(1)：56.
[9]钟华,钟柏亭.王力宁因质辨治小儿外感咳嗽的经验[J].浙江中医杂志2006,41 (12)：688-689.
[10]王萍芬.中医儿科学[M].上海：上海科学技术出版社,1997,37.
[11]尹维东,任闪闪.小儿咳嗽的中医辨治[J].中医垂教.2006,22(316)：36-37.
[12]欧秀梅.止嗽散加减治疗小儿外感咳嗽40例[J].中医儿科杂志,2008,4(3)：20-21.
[13]王怡冰,张书亮.桑白皮汤加减治疗外感咳嗽疗效分析[J].医药论坛杂志,2006,27(13)：100-102.
[14]高季鸿,耿建国.麻杏甘石汤加味治疗外感后咳嗽的临床观察[J].四川中医,2007,25(8),66.
[15]肖湘.桑杏汤加减治疗燥咳54例[J].实用中医内科杂志,2007,21(2)：59.
[16]陈英妹,刘伟.咳嗽颗粒加味治疗小儿外感咳嗽72例[J].药物与临床,2009,3,103.
[17]孙学刚,马克林,崔燕.祛风止咳汤治疗外感咳178例[J].陕西中医学院学报,2005,28(5)：26-27.
[18]李桂君.湿热蕴肺致小儿咳嗽的辨治体会[J].新疆中医药,2009,27(4)：97-98.
[19]逵艳,张书俊,张莹.自拟止咳平喘汤治疗小儿外感咳嗽80例[J].国医论坛,2007,22(1)：27.
[20]虞金龙.中药贴敷治疗外感咳嗽与支气管炎疗效观察[J].浙江中西医结合杂志,2005,15(1)：41.
[21]乔林波.孔最穴封闭疗法临床应用体会[J].JCAM,2007,23(10)：28.
[22]郭素洁.推拿配合灸肺俞治疗小儿咳嗽67例[J].按摩与引导,2004,20,(5)：50.
[23]王玉.推拿加拔罐治疗小儿外感咳嗽90例[J].陕西中医,2008,29(11)：1518-1519.
[24]李炜奕.中药敷脐法治疗小儿外感咳嗽疗效观察[J].吉林医学,2008,29(3)：235.
[25]尹文仲.盐熨法治疗小儿外感咳嗽[J].湖北民族学院学报·医学版,2007,3,
[1]Anderssion U, Wang H, Palmblad K, etal. High mobility group 1 protein stimulates proinflammatory cytokine synthesis in humanmoco-cytes [J]. J.Exp. Med.2000,192: 5655-5570.
[2]姚咏明,盛志勇.高迁移率族蛋白-1在脓毒症发病中的作用和意义[J].解放军医学杂志,2002,27：752-756.
[3]Haniguchi N, Kawahara K, Yone K, etal. High mobility groupchrosomosomal protein 1 plays a role in the pathogenesis of rheumatoidarthritis as a novel cytokine [J]. Arthrtis Rheum,2003,48(4):971-981.
[4]Ullertis R, Jonsson IM, Verdrengh M, etal. High mobility groupchrosomosomal protein 1, a DNA binding cytokine, induces arthritis [J]. Arthritis Rheum,2003,48(6):1693-1 700.
[5]Wang H, Bloomo, Zhang M, etal. HMG-1 as a late mediatorof endotoxin lethality in mice[J]. Science 1999,285:248-251.
[6]Abraham E, Srcaroli J, Carmody A, et al. HMGB-1 as a media-tor of acute lung inflammation [J]. J. Immunol,2000,165:2950-2954.
[7]Ueno H, Matsuda T, Hashimoto S, etal. Contributions of highmobility group box protein in experimental and clincial acute lung in-jury [J]. Am J Respir,2004,17::185-189.
[8]孟志艳,李军,曹红.HMGB1在呼吸系统疾病中的作用[J].山东医药,2009,49(8)：17-108.
[9]Haniguchi N, Kawahara K, Yone K, etal. High mobility groupchrosomosomal protein 1 plays a role in the pathogenesis of rheumatoidarthritis as a novel cytokine [J]. Arthrtis Rheum,2003,48(4)::971-981.
[10]Ullertis R, Jonsson IM, Verdrengh M, etal. High mobility groupchrosomosomal protein 1, a DNA binding cytokine, induces arthritis [J]. Arthritis Rheum,2003,48(6):1693-1700.
[11]郭惠芳,刘淑霞,马丽艳等.类风湿关节炎患者血清中高迁移率族蛋白-1的作用及其与MMP2、 TIMP2的关系[J].中国现代医学杂志,2010,20(2)：192-195.
[12]Sass G, Heinlein S, Agli A, etal.Cytokine expression in threemouse models of experimental hepatitis [J]. Cytokine,2002,19(3):115-120.
[13]杨智勇,王春友,熊炯等.重症急性胰腺炎大鼠血清高迁移率族蛋白-1水平的时相变化及意义[J].华中科技大学学报(医学版),2004,33(4)：466-468.
[14]Sappington PL, Yang R, Yang H, etal. HMGB-1 B box increases the permeability of Caco-2 enteroytic monolayem and impairs intestinal barrier function in mice[J]. Gastroenterology,2002,123(3):790-802.
[15]栾正刚,何忠野,张成,等.丙酮酸乙酯对重症急性胰腺炎大鼠肠粘膜HMGB-1表达的影响[J].中国普通外科杂志,2008,17(3)：214-218.
[16]Kalinina N, Agrotis A, Antroprovaet etal. Increased expression ofthe DNA-binding cytokine HMGB-1 in human atherosclerotic le-sions.Arterioscler Thromb Vasc Biol, 2004,24(12):2320-2325.
[17]Ulfgren AK, Grundtman C, Borg k, et al. Dowin-regulation ofthe aberrant expression of the inflammation mediator high mobility groupchrosomosomal protein 1 in muscle tissure of patients with polymyositisand dermatomyositis treated with corticoste-roids.Arthritis Rheum,2004,50(5):1586-1594.
[18]Naghavi MH, Nowak P, Andersson J, et al. Intracelluar highmobility group protein represss HIV-1 LTR-directed transcription in a promotor and cell specific manner [J]. Virology,2003,314(1):179-189.
[19]朱海云,李银平.高迁移率族蛋白B-1的研究进展及其免疫作用[J].中国中西医结合急救杂志,2009,16(2)：124-125.
[1]Sant'Ambrogio G, Widdicombe J.Reflexes from airway rapidly adapting receptors [J]. Respir Physiol,2001,125(1-2):33-45.
[2]Karlsson JA, Fuller RW. Pharmacological regulation of the cough reflex from experimental models to antitussive effects in Man[J]. Pulm Pharmacol Ther,1999, 12(4):215-228.
[3]MyersA C, KajekarR, Undem B J. Allergic inflammation-inducedneuropeptide production in rapidly adapting afferent nerves in guinea pig airways [J]. Am J Physiol Lung Cell MolPhysio,12002,282:L775-L781.
[4]WinterC A, FlatakerL. The effects of drugs up on a graded cough response obtained in sensitized guinea pigs exposed to aerosol of specific antigen [J]. J Exp Med,1955, 101:17-24.
[5]Kamei J, MoritaK, KashiwazakiT, etal. Antitussive effect ofmoguisteine on allergic coughs in the guinea pig [J]. Eur J Pharmaco,11998,47:253-255.
[6]HouseA, CellyC, SkeansS, etal. Cough reflex in allergic dogs [J]. EurJ Pharmaco,2004, 492:251-258.
[7]Kamei J, Saitoh A, Morita K, Nagase H. Involvement of adenosinA1 receptors in antitussive effect in mice [J]. Life Science,1994; 5(20):383-8.
[8]Milos T, Giuseppe SA, Franca BSA. Laryngeal and tracheo bronchial cough in anesthetized dogs [J]. JAppl Physiol,1994; 76(6):2672-9.
[9]CaterinaMJ, LefflerA, MalmbergAB, etal. Impaired nociceptionand pain sensation in mice lacking the capsaicin receptor[J]. Science,2000,288:306-313.
[10]Ma QP. Expression of capsaicin receptor (VR1) by myelinated primary afferentneurons in rats [J]. NeurosciLett,2002,319:87-90.
[11]Takahama K, Araki T, Fuchikami J-I etal. Studies on the magnitude and the mechanism of cough potentiation by angiotensin converting enzyme inhibitors in guinea pigs involvement of bradykini in the potentiation[J]. JPharm Pharmacol,1996; 48:1027-33.
[12]陈如冲,赖克方,刘春丽等.辣椒素咳嗽激发试验方法的建立及其安全性评价[J].中华结核和呼吸杂志,2005,28(11)：751-754.
[13]Greene S A, Wolff R K, Hahn F F, etal. Sulfur dioxide-inducedchronic bronchitis in beagle dos[J]. JtoxicolEnvironHealth,1984,13:945-958.
[14]Takahama, Fuchikami J, KaiH, etal. Inhalation of phosphoradon, a neutral endopeptidase inhibitor, induces cough in awguinea-pigs[J]. Arch IntPharmacodyn Ther,1995,330: 241-250.
[15]Moreaux B, BeerensD, Gustin P. Development of a cough inductiontest in pigs:effects of SR 48968 and enalapri-1[J]. J Vet Pharmacol Ther,1999,22:387-389.
[16]Moreaux B, Advenier C, Gustin P. Role of bradykinin and tachyki-nins in the potentiation by enalaprilofcoughing induced by citric acid[J]. Fund Clin Pharmaco, 12001,15:23-29.
[1]徐叔云,卞如濂,陈修.药理实验方法学[M].北京：人民卫生出版社,1991,1：1821.
[2]周添浓,唐立海,黄诗冲等.五指毛桃镇咳及平喘作用研究[J].中药材,2009,32(4)：571-574.
[3]Sant'Ambrogio G, Widdicombe J. Reflexes from airway rapidly adapting receptors [J]. Respir Physiol,2001,125(1-2):33-45.
[4]Karlsson JA, Fuller RW. Pharmacological regulation of the cough reflex from experimental models to antitussive effects in Man[J]. Pulm Pharmacol Ther,1999, 12(4):215-228.
[5]Hunter DD, Undem BJ. Identification and substance P content of vagal afferent neurons innervating the epithelium of the guinea pig trachea [J]. Am J Respir Crit Care Med, 1999,159(6):1943-1948.
[1]陈奇.中药药理研究方法学[M].北京：人民卫生出版社,1994,80：140-148.
[2]魏东,姚斌,高兴亮等.少年红矾杏平喘糖浆镇咳、祛痰和平喘作用的实验研究[J].中医中药与微循环.2009,19(2)：29-32.
[1]Nakamura M,Yamaya M,Fukushima T,et al. Effect of mabuterol on t racheal muco-ciliary clearance of magnetized iron particles in anest hetized dogs[J]. Respiration, 1991,58(1):331.
[1]王科闯、杨宇.肺与大肠相表里的实验研究概况[J].《山东中医药大学学报》,2006,
[1]陈奇.中药药理研究方法学[M].北京：人民卫生出版社,1993,272-273.
[1]徐玉民,朱敏敏,戎海波,等.氯胺瞳雾化吸入对哮喘大鼠气道高反应性的影响[J].南京医科大学学报(自然科学版),2007,27(3)：235-238.
[2]李明华,殷凯生, 蔡映云.哮喘病学[M].北京：人民卫生出版社,2005,128.
[3]MorcilloEJ, CortijoJ. Mucus and M U C in asthma[J]. Curr Opin Pulm Med,2006, 12(1):1-6.
[4]吴为群,江宏志,吴忠道等.血清白细胞介素IL-13、17与气道高反应性的关系[J].热带医学杂志,2004,4(5)：539-541.
[1]Jung J, Hwang SW, Kwak J, et al. Capsaicin binds to the intracellular domain of the capsaicin-activated ion channel[J]. Jneurosci,1999,19(2):529-538.
[2]容朝晖,沈策.咳嗽反射机制的研究进展[J].国外医学呼吸系统分册,2005,25(3)：168-171.
[3]Forsberg K, Karlsso JA, The odorsson et al. Cough and bronchoconstriction mediated by capsaicin-sensitive sensory neurons in thguinea pig[J]. Pulm Pharmacol,1988,1:33-39.
[4]Takahama K, Araki T, Fuchikami J-I et al. Studies on the magnitude and the mechanism of cough potentiation by angiotensin converting enzyme inhibitors in guinea pigs
involvement of bradykini in the potentiation[J]. JPharm Pharmacol,1996,48:1027-1033.
[5]陈如冲,赖克方,刘春丽等.辣椒素咳嗽激发试验方法的建立及其安全性评价[J].中华结核和呼吸杂志,2005,28(11)：751-754.
[1]Anderssion, U, Wang H, Palmblad K, et al.High mobility group 1 protein stimulates proinflammatory cytokine synthesis in humanmoco-cytes. J.Exp. Med,2000,192:5 655-5570.
[2]Abraham E, Arcaroli J, Carmody A, et al. HMG-1 as a mediator of acute lung Inflam-mation[J]. The Journal of Immunology,2000,165(6):2950-2924.
[3]孟志艳,李军,曹红.HMGB-1在呼吸系统疾病中的作用[J].山东医药,2009,49(8)：107-108.
[4]Lin X, Yang H, Sakuragi T, et al. Alpha chemokine receptor blockade reduces high mobility group box-1 protein induced lung inflammarion and injury and improves survi-valin sepsis[J]. Am J PhysiolLung CellMol Physiol,2005,289(4):L583-590.
[5]姚咏明,盛志勇.高迁移率族蛋白-1在脓毒症发病中的作用和意义[J].解放军医学杂志,2002,27：752-756.