In vivo evaluation of the anti-asthmatic,antitussive and expectorant activities of extract and fractions from Elaeagnus pungens leaf

The leaf of Elaeagnus pungens thunb. (Family Elaeagnaceae) has been documented as an effective herb for the treatment of asthma and chronic bronchitis in traditional Chinese medicine. This study was aimed at evaluating the anti-asthmatic, antitussive and expectorant activities in vivo of the ethanolic extract and fractions from the leaf of Elaeagnus pungens. The results showed that the 70% ethanolic extract increased the preconvulsive time of asthma induced by the combination of histamine and acetylcholine chloride in guinea pigs at the medium dose of 1.379 g/kg. The water fraction significantly prolonged the preconvulsive time (P < 0.05) in guinea pigs, lengthened the latent period of cough (P < 0.05) as well as decreased the cough frequency caused by citric acid in guinea pigs (P < 0.01) and enhanced tracheal phenol red output in mice (P < 0.01). Moreover, the petroleum ether fraction significantly reduced the cough frequency induced in guinea pigs (P < 0.01) and improved tracheal phenol red output in mice (P < 0.01). It indicated that the petroleum ether fraction and the water fraction of Elaeagnus pungens leaf were effective on anti-asthmatic, antitussive and expectorant activities, which supplied for further research on chemical constituents and pharmacological mechanisms.     

葶苈子化学拆分组分止咳祛痰平喘作用研究

目的:探讨葶苈子的化学拆分组分的止咳祛痰及平喘作用。方法:分别采用氨水引咳法、气管酚红排泌法、组胺与乙酰胆碱致喘法、离体气管法观察葶苈子各化学拆分组分的止咳、祛痰、平喘作用。结果:葶苈子水煎液低、中、高剂量均能显著减少小鼠咳嗽次数、增加小鼠呼吸道的酚红排泌量,同时能延长豚鼠哮喘潜伏期,并且提高解痉率,与空白对照组比较,中剂量效果最好。与空白组相比,葶苈子20%乙醇组分、脂肪油组分、水部位组分均能显著减少浓氨水引起小鼠咳嗽的次数,并延长咳嗽潜伏期、增加酚红排泌量、延长豚鼠发生哮喘的潜伏期(P<0.01,P<0.05),此外,20%乙醇组分、水部位组分能使气管收缩幅度明显降低(P<0.01,P<0.05),具有一定的解痉挛作用。结论:葶苈子水煎液具有较好的止咳、祛痰、平喘作用,其有效部位为20%乙醇组分、脂肪油及水部位组分,其止咳祛痰和平喘作用最为显著,与水煎液中剂量作用相当。     

Pharmacological evaluation of Alstonia scholaris: Anti-tussive,anti-asthmatic and expectorant activities

Ethnopharmacological relevance

Alstonia scholaris (Apocynaceae) was documented as an effective herb for the treatment of chronic respiratory diseases in “dai” ethnopharmacy historically, and its leaf crude extract, used for releasing tracheitis and cold symptom, was approved to be a commercial formulation by State Food and Drugs Administration of China (SFDA).

Aim of the study

The investigation evaluates the anti-tussive and anti-asthmatic activities of the ethanolic extract, fractions and main alkaloids of Alstonia scholaris leaf to provide experimental evidence for its traditional and modern clinical use. For our most interesting, is to reveal the active components for further new drug development.

Materials and methods

The leaf of Alstonia scholaris was extracted with ethanol and then separated into different fractions. Furthermore, alkaloids were isolated by phytochemical method. The anti-tussive activity was evaluated using three different models including ammonia or sulfur dioxide induced mice coughing, and citric acid induced guinea pigs coughing. The anti-asthmatic activity was investigated on guinea pigs bronchoconstraction induced by histamine. The expectorant activity was evaluated by volume of phenol red in mice's tracheas.

Results

The alkaloids fraction significantly inhibited mice's frequency of cough induced by ammonia, increased mice's latent period of cough induced by sulfur dioxide, and increased guinea pigs’ latent period of cough and inhibited frequency of cough. Besides, the alkaloids fraction increased delitescence of convulsion, and tumble of guinea pigs in anti-asthmatic test, and enhanced tracheal phenol red output in expectorant evaluation. Moreover, the main alkaloid, picrinine exhibited anti-tussive and anti-asthmatic activities in vivo.

Conclusions

The alkaloids fraction was anti-tussive, anti-asthmatic and expectorant activities component of Alstonia scholaris leaf, and it may also be a valuable lead material for respiratory diseases drug development. Picrinine, the main anti-tussive and anti-asthmatic compound, could be applied in quality control of products from Alstonia scholaris leaf.     

桑白皮水煎液及化学拆分组分止咳祛痰平喘作用研究

目的：研究桑白皮水煎液及化学拆分组分的止咳祛痰平喘作用。方法：采用氨水诱咳法观察止咳作用;小鼠酚红排泌法观察祛痰作用;组胺与乙酰胆碱引喘法观察平喘作用;离体气管法观察解痉挛作用。结果：与空白组比较,桑白皮水煎液低、中、高剂量组均能显著减少小鼠咳嗽次数、增加小鼠呼吸道的酚红排泌量、延长豚鼠哮喘潜伏期、提高解痉率,以中剂量效果最好。各化学拆分组分与空白组相比,桑白皮30%、50%乙醇组分能明显减少浓氨水引起小鼠咳嗽的次数并延长咳嗽潜伏期、增加小鼠呼吸道的酚红排泌量（P<0.05或P<0.01）,30%乙醇组分、脂肪油组分能延长豚鼠哮喘潜伏期（P<0.05或P<0.01）,此外,30%乙醇组分能使气管收缩幅度明显降低,具有一定的解痉挛作用。结论：桑白皮水煎液具有较好的止咳、祛痰、平喘作用,其有效部位为30%乙醇组分,用量相当于中剂量的1/3,其止咳、祛痰、平喘作用最为显著,与水煎液中剂量作用相当。     

基于网络药理学的地龙平喘作用机制研究

目的 基于网络药理学方法研究地龙平喘的作用机制。方法 通过中国知网（CNKI）数据库、百链数据库、PubMed数据库文献检索和SwissADME平台筛选获得地龙活性化合物;通过SwissTargetPrediction预测活性成分的潜在靶点,并与OMIM、DrugBank数据库中哮喘的疾病靶点数据进行比对,获得地龙平喘的作用靶点;通过STRING 11.0平台分析靶点的蛋白质相互作用网络;通过Matascape数据库对地龙平喘靶点进行基因本体（GO）分析和京都基因与基因组百科全书（KEGG）通路富集分析;采用Cytoscape 3.7.1软件构建成分-靶点-通路网络;通过豚鼠离体气管平滑肌诱导收缩实验验证地龙提取物对胆碱能受体兴奋引起的气管平滑肌收缩的抑制作用。结果 共收集地龙平喘活性成分56个,包括氨基酸、脂肪酸、核苷酸等,预测其作用靶点包括前列腺素G/H合酶2、毒蕈碱型乙酰胆碱受体家族、β₂-肾上腺素受体等67个,潜在调控花生四烯酸代谢、胆碱能神经突触信号、色氨酸代谢等通路和生物过程。验证实验结果表明,地龙提取物对胆碱能受体兴奋引起的离体气管平滑肌收缩有明显的舒张作用。结论 地龙的平喘作用具有多成分、多靶点、多途径作用的特点,为地龙平喘作用机制的进一步研究提供了参考。     

咳喘清颗粒止咳平喘祛痰作用研究

目的通过相应的实验方法观察咳喘清颗粒的止咳、平喘、祛痰作用。方法通过小鼠二氧化硫引咳实验、小鼠氨水引咳实验观察其止咳作用;通过乙酰胆碱加组胺所致豚鼠哮喘及离体豚鼠气管平滑肌实验观察其平喘作用;通过大鼠毛细管排痰实验观察其排痰作用。结果咳喘清能显著减少小鼠由二氧化硫引起的咳嗽次数和显著延长其咳嗽潜伏期,能明显延长由氨水引起的咳嗽时间;咳喘清能延长由乙酰胆碱加组胺的引喘潜伏期,能明显抑制离体气管平滑肌的收缩;咳喘清能明显增加大鼠痰量的排出。结论咳喘清有明显的止咳、平喘、祛痰作用。     

复方止咳颗粒药效学实验研究

目的:评价复方止咳颗粒的药效作用。方法:通过小鼠氨水引咳实验、豚鼠枸橼酸引咳实验观察其镇咳作用;通过大鼠毛细管祛痰实验观察其祛痰作用;通过磷酸组胺致豚鼠哮喘实验观察其平喘作用;通过二甲苯致小鼠耳肿胀实验、大鼠纽扣肉芽组织增生实验观察其抗炎作用。结果:复方止咳颗粒能显著抑制25%氨水诱发的小鼠咳嗽;能显著抑制枸橼酸诱发的豚鼠咳嗽;能有效抑制磷酸组胺诱发的豚鼠哮喘;能明显增加大鼠痰量的排出;对小鼠的急性炎症反应有抑制作用。结论:复方止咳颗粒具有止咳、祛痰、平喘、抗炎作用。     

药对麻黄地龙配比及平喘作用机制的研究

目的:考察药对麻黄、地龙用量配比及可能的平喘作用机制。方法:观察麻黄、地龙3种用量配比(1∶1,1∶3,1∶9)水煎液对10μmol.L-1卡巴胆碱(carbachol,CCh),10μmol.L^-1组胺(histamine,His)引起离体气管平滑肌张力增加的松弛作用,以及对10μmol.L^-1CCh引起的离体气管上皮短路电流增加的抑制作用。结果:对于10μmol.L^-1CCh引起的离体大鼠气管平滑肌收缩,麻黄地龙3种配比水提物的舒张作用随剂量增加而增强,1∶1,1∶3水提物的IC50分别为7.5,15 mg.mL^-1,1∶9水提物不能产生对CCh引发最大收缩抑制50%的效应;对于10μmol.L^-1His所引起的离体豚鼠气管平滑肌收缩,麻黄地龙3种配比水提物的舒张作用随剂量增加而增强,1∶9,1∶3,1∶1水提物的IC50分别为0.19,0.61,1.8 mg.mL-1;在抑制CCh引起的离体大鼠气管上皮短路电流增加方面,三者的作用强度顺序为1∶3>1∶1>1∶9,三者没有显著性差异。结论:麻黄、地龙药对具有舒张离体气管平滑肌和抑制气管上皮离子分泌的作用,其作用机制包括抗CCh和抗His作用。1∶3是麻黄、地龙药对中的最适配比。     

Anticough and antimicrobial activities of Psidium guajava Linn. leaf extract

The anticough activity of Psidium guajava Linn. (guava) leaf extract was evaluated in rats and guinea pigs. The results showed that water extract of the plant at doses of 2 and 5 g/kg, p.o. decreased the frequency of cough induced by capsaicin aerosol by 35 and 54%, respectively, as compared to the control, within 10 min after injection of the extract, (P < 0.01). However, the anticough activity is less potent than that of 3 mg/kg dextromethorphan which decreased frequency of cough by 78% (P < 0.01). An experiment on isolated rat tracheal muscle showed that the extract directly stimulated muscle contraction and also synergized with the stimulatory effect of pilocarpine. This effect was antagonized by an atropine. Moreover, growth of Staphylococcus aureus and beta-streptococcus group A, as determined by the disc diffusion method, was inhibited by water, methanol and chloroform extract of dry guava leaves (P < 0.001). The LD50 of guava leaf extract was more than 5 g/kg, p.o. These results suggest that guava leaf extract is recommended as a cough remedy.     

Antitussive,expectorant and bronchodilating effects of ethanol extract of Sorghum bicolor (L.) Moench roots

Ethnopharmacological relevance

Root of Sorghum bicolor (L.) Moench (RSB) is an herbal medicine in Traditional Chinese Medicine, still used in some rural areas in Central China as an alternative remedy to treat cough and asthma.

Aim of the study

The study was aimed at evaluating the antitussive, expectorant and bronchodilating effects of ethanol extract of RSB, support its folk use with scientific evidence, and lay a foundation for its further researches.

Materials and methods

RSB was extracted with 80% ethanol aqueous in reflux conditions, solutions were concentrated in reduced pressure, and lyophilized in vacuum to yield the RSB extract. Antitussive evaluations were carried out with three different models including ammonia liquor induced mice cough, capsaicin induced mice cough, and citric acid induced guinea pigs cough; phenol red secretion experiments in mice were performed to evaluate the expectorant ability; bronchodilating effects were evaluated with a bronchoconstrictive challenge induced by acetylcholine chloride and histamine in guinea pigs.

Results

In all the three antitussive tests, treatment of RSB significantly inhibited the frequency of cough, and prolonged the cough latent period in animals. And high dose of RSB (200 mg/kg in mice and 100 mg/kg in guinea pigs) created therapeutic activities as good as standard antitussive drug codeine phosphate (20 mg/kg). In the expectorant evaluation, 50, 100 and 200 mg/kg RSB treatment had significantly increased the amount of phenol red output for 0.39, 1.18, and 1.96 folds in mice tracheas. In the bronchodilating test, RSB treatment at 100 mg/kg extended the preconvulsive time for 44.84% compared with that of before treatment in guinea pigs.

Conclusions

RSB is an effective alternative medicine for the treatment of cough with potent antitussive, expectorant and bronchodilating activities.     

川贝雪梨膏主要药效学实验研究

目的：观察中药复方制剂“川贝雪梨膏”主要药理作用。方法：采用氨水和二氧化硫引咳法，整体平喘和离体试验，酚红祛痰和毛细管排痰法，干酵母和三联菌苗致热和二甲苯和角叉菜胶致炎等方法进行镇咳、平喘、祛痰、解热、抗炎的实验观察。结果：“川贝雪梨膏”能明显延长小鼠的咳嗽潜伏期；对豚鼠的引喘潜伏期有明显的延长，对豚鼠离体气管平滑肌有明显的松弛作用；对小鼠酚红的排出和大鼠的排痰量均有明显增加；对家兔和大鼠的体温升高模型有明显的解热作用；对由二甲苯和角叉菜胶引起的炎性肿胀均有明显的抑制作用。结论：川贝雪梨膏具有明显的镇咳、平喘、祛痰、解热及抗炎作用。     

甲基麻黄碱与伪麻黄碱镇咳平喘作用比较研究

目的:探讨甲基麻黄碱与伪麻黄碱在镇咳平喘方面的差异。方法:以枸橼酸引咳法、整体动物药物引喘法以及离体气管平滑肌法,研究二者镇咳平喘作用。结果:对枸橼酸引咳试验,甲基麻黄碱与伪麻黄碱均具有明显的镇咳作用;对整体动物哮喘试验,二者均具有明显的平喘作用,且二者的平喘作用存在显著差异(P0.05)。对离体气管平滑肌试验,二者均可舒张静息状态气管且可拮抗组织胺与乙酰胆碱对气管平滑肌的兴奋作用,对乙酰胆碱的拮抗作用二者存在显著差异(P0.05)。结论:二者均具有显著的镇咳平喘作用,且二者的平喘作用存在显著差异。     

The in vivo expectorant and antitussive activity of extract and fractions from Reineckia carnea

Aim of study

To evaluate the potential expectorant and antitussive activity of a traditional Chinese medicine.

Materials and methods

The water extract and four fractions of the aerial part of Reineckia carnea were orally administrated to coughing mice induced by ammonium hydroxide and mice injected with phenol red, respectively, to investigate their medification effect on coughing and mucus scretion.

Results and discussion

90% Ethanol fraction significantly lengthened the latent period of cough and decreased cough frequency caused by ammonium hydroxide at the dose of 0.372 g/kg (p < 0.05). Sixty percent ethanol fraction reduced the cough frequency as well as the mucus secretion from mouse tracheas obviously at the dose of 0.570 g/kg (p < 0.05) by measuring the tracheal output of phenol red in mice. The medication effects in multiple doses of the active fractions were then performed and it has been proved that the 60% ethanol and 90% ethanol fraction were curatively effective on expectoration and coughing, respectively both at the high and middle dose, which supplied proofs for the further research on chemical constituents in both of two effective fractions.     

Bronchospasm potentiating effect of methanolic extract of Ficus religiosa fruits in guinea pigs

Ethnopharmacological relevance

The sacred tree Peepal (Ficus religiosa family: Moraceae) has numerous therapeutic utility in folk medicine.

Aim of the study

It has been reported to be used in ethno medical treatment of asthma and also in epilepsy due to its high serotonin content, which has been implicated in pathophysiology of asthma, this led us to carry out the present study.

Materials and methods

The in vivo studies of histamine induced bronchospasm in guinea pigs and in vitro isolated guinea pig tracheal chain and ileum preparation.

Results

Pre-treatment of guinea pigs with ketotifen (1 mg/kg, p.o.) has significantly delayed the onset of histamine aerosol induced pre-convulsive dyspnea, compared with vehicle control (281.8^a ± 11.7 vs. 112.2 ± 9.8). The administration of methanolic extract (125, 250 and 500 mg/kg, p.o.) did not produced any significant effect on latency to develop histamine induced pre-convulsive dyspnea. On the other hand, methanolic extract of the fruits at the doses employed (i.e., 0.5, 1 and 2 mg/ml) has significantly potentiate the EC₅₀ doses of both histamine and acetylcholine in isolated guinea pig tracheal chain and ileum preparation. In addition, HPLC analysis of the methanolic extract showed the presence of high amounts of serotonin (2.89%, w/w).

Conclusions

On the basis of data, it may be concluded that Ficus religiosa fruits have been found to be ineffective against histamine induced bronchospasm in guinea pigs. In addition, methanolic extract of the fruits have shown to potentiate the bronchoconstriction induced by both histamine and acetylcholine on guinea pig tracheal chain preparation.     

银杏露镇咳、祛痰及平喘的药效学研究

目的：观察银杏露的镇咳、祛痰及平喘作用。方法：采用小鼠氨水引咳法及气管酚红排泌法观察银杏露的镇咳及祛痰作用,采用哮喘发作潜伏期及离体气管平滑肌松弛试验观察银杏露的平喘作用。结果：银杏露能抑制氨水引起的小鼠咳嗽,促进小鼠气管酚红排泌,对磷酸组胺引起的豚鼠哮喘具有保护作用,能明显延长哮喘潜伏期;对磷酸组胺引起的豚鼠气管收缩具有抑制作用。结论：银杏露具有镇咳、祛痰及平喘作用。     

罗汉果皂苷Ⅴ的镇咳、祛痰及解痉作用研究

 目的筛选罗汉果镇咳祛痰的有效部位,并进一步研究罗汉果皂苷Ⅴ的镇咳、祛痰及解痉作用。方法通过观察罗汉果水提部位、体积分数95%醇提部位及体积分数50%醇提部位对浓氨水致小鼠咳嗽次数及咳嗽潜伏期的影响,筛选出罗汉果镇咳的有效部位为体积分数50%醇提部分。进一步观察了50%醇提部分中的罗汉果皂苷Ⅴ对小鼠酚红排泄量及组胺致豚鼠离体回肠及离体气管痉挛的影响。结果罗汉果水煎剂50g生药·kg^-1,罗汉果体积分数50%醇提部位50g生药·kg^-1及罗汉果皂苷Ⅴ300,150,75mg·kg^-1能明显减少小鼠的咳嗽次数,罗汉果体积分数50%醇提部位50g生药·kg^-1及罗汉果皂苷Ⅴ300mg·kg^-1还能明显延长小鼠咳嗽潜伏期;罗汉果皂苷Ⅴ150mg·kg-1能明显增加小鼠气管酚红排泄量,提示有一定的祛痰作用;5g·L-1的罗汉果皂苷Ⅴ能明显拮抗组胺引起的回肠收缩;2.5及1.25g·L^-1剂量对组胺引起的气管痉挛有明显的拮抗作用。结论罗汉果皂苷Ⅴ显示出一定的镇咳、祛痰、解痉活性,推测为罗汉果镇咳的主要活性成分。     

复方半夏水提取物的镇咳、祛痰、平喘作用研究

[目的]研究复方半夏水提取物的镇咳、祛痰、平喘作用.[方法]用乙酰胆碱和组胺诱发豚鼠哮喘,研究药物对引喘潜伏期的影响;用人工气胸后组胺致痉实验研究药物对肺溢流量的影响;用氨水引咳实验研究药物对咳嗽潜伏期及咳嗽次数的影响;用酚红分泌实验研究复方半夏水提取物的祛痰作用.[结果]复方半夏水提取物可剂量依赖性地延长引喘潜伏期,降低肺溢流量,显著延长咳嗽潜伏期并减少咳嗽次数,1.2g/kg复方半夏水提取物可显著增加小鼠呼吸道分泌的酚红量.[结论]复方半夏水提取物具有平喘、镇咳、祛痰作用.     

宣肺化痰方标准煎剂与煮散剂药理作用对比

目的: 对比观察宣肺化痰方标准煎剂及煮散剂的药理作用。 方法: 大鼠分为8组,分别灌入等体积蒸馏水、急支糖浆、宣肺化痰方标准煎剂高、中、低剂量(含生药46.80,23.40,11.70 g·kg^-1)、宣肺化痰方煮散剂高、中、低剂量(含生药23.40, 11.70,5.85 g·kg^-1),观察对小鼠毛细血管通透性的影响、对氨水致小鼠咳嗽的影响、对磷酸组胺诱发豚鼠哮喘的影响、对小鼠气管酚红排泌的影响、对慢性支气管炎大鼠超氧化物歧化酶(SOD)、丙二醛(MDA)的影响。 结果: 宣肺化痰方标准煎剂和煮散剂均能显著降低小鼠毛细血管通透性;延长小鼠咳嗽潜伏期和减少咳嗽次数;延长豚鼠引喘潜伏期;增加小鼠气管酚红排泌量;升高慢性支气管炎大鼠血清SOD活性,降低MDA含量。宣肺化痰方标准煎剂和煮散剂比较,两者在同剂量组级别(高、中、低)之间作用无显著性差异。 结论: 宣肺化痰方标准煎剂和煮散剂均有显著抗炎、止咳、平喘和化痰作用,两者作用无显著性差异。     

还阳参止咳平喘活性部位的研究

目的比较还阳参不同提取物止咳平喘作用,为寻找确定其止咳平喘有效部位提供实验基础。方法制备还阳参不同提取物,观察其对浓氨引起的小鼠咳嗽以及组胺乙酰胆碱引起的豚鼠哮喘的影响。结果还阳参石油醚提取物的H2-1部位对氨水刺激引起的小鼠咳嗽有明显的止咳作用;正丁醇提取物的HD-2部位对组胺和乙酰胆碱引起的豚哮喘有抑制作用。结论明确了还阳参止咳平喘的物质基础,为还阳参止咳平喘的临床功效提供了科学依据。     

咳喘口服液的药理作用研究

目的：研究咳喘口服液祛痰、镇咳、平喘和抗炎的药理作用。方法：采用小鼠酚红祛痰法、小鼠氨水引咳法、豚鼠肺支气管灌流法、豚鼠离体气管法、大鼠足跖肿胀法模型，观察咳喘口服液的祛痰、镇咳、平喘和抗炎作用。结果：咳喘口服液能明显增加小鼠气管纤毛对酚红的分泌排出量；延长小鼠咳嗽潜伏期和减少小鼠咳嗽次数；对组胺致痉的豚鼠离体气管平滑肌有明显的解痉作用，对卵蛋白致敏豚鼠肺支气管的灌流量有显著增加作用；对大鼠足肿有明显的抑制作用。结论：咳喘口服液具有明显的止咳、祛痰、平喘和抗炎的作用。