CN108079172B

CN108079172B - Medicine for treating cough variant asthma

Info

Publication number: CN108079172B
Application number: CN201810056229.5A
Authority: CN
Inventors: 倪艳; 郝旭亮; 贺石麟; 刘聪; 李媛; 张雯霞; 赵怀舟; 岳永花
Original assignee: SHANXI TRADITIONAL CHINESE MEDICINE RESEARCH INSTITUTE
Current assignee: SHANXI TRADITIONAL CHINESE MEDICINE INSTITUTE (SHANXI PROVINCIAL HOSPITAL OF TRADITIONAL CHINESE MEDICINE)
Priority date: 2018-01-20
Filing date: 2018-01-20
Publication date: 2020-05-22
Anticipated expiration: 2038-01-20
Also published as: CN108079172A

Abstract

The invention discloses a medicine for treating cough variant asthma, which is prepared from 5-15 parts by weight of radix cynanchi wilfordii, 3-10 parts by weight of fritillaria ussuriensis, 5-12 parts by weight of bitter apricot seeds, 5-12 parts by weight of perilla seeds, 5-12 parts by weight of scutellaria baicalensis, 5-12 parts by weight of earthworms, 5-12 parts by weight of stiff silkworms, 3-12 parts by weight of peach kernels, 3-12 parts by weight of dark plums, 3-12 parts by weight of bitter oranges and 2-5 parts by weight of liquorice as raw medicines.

Description

Medicine for treating cough variant asthma

Technical Field

The invention relates to a medicament for treating asthma, in particular to a medicament for treating cough variant asthma based on Chinese herbal medicine raw materials.

Background

Cough Variant Asthma (CVA), also known as allergic asthma, is a specific type of asthma with chronic cough as the main or only clinical manifestation, and is also one of the common diseases of the respiratory system. Children have poor immunity because bronchi are still in a developmental stage, and are easy to cause cough variant asthma under the conditions of strenuous exercise, catching cold or contacting with allergens such as pollen and the like, and the morbidity is relatively high. If not treated in time, it is highly likely to develop into typical asthma.

Western medicine mainly aims at the characteristics of chronic inflammatory lesions of airways of cough variant asthma, adopts a mode of glucocorticoid inhalation for treatment, and does not adopt an individual treatment mode which is different from an adult according to the constitution of a child. The hormone medicine can generate certain adverse reactions to digestive tracts, growth and development and the like in the using process, and the compliance of the hormone medicine is poor.

Traditional Chinese medicine considers that chronic inflammation of an airway of cough variant asthma is related to wind and phlegm, and the main symptom is cough and is characterized by airway hyperresponsiveness. Therefore, cough variant asthma is often treated by using Chinese patent medicines or prescriptions such as compound fresh bamboo juice oral liquid, Maxingshigan decoction, modified Xiaoqinglong decoction and the like.

However, the internal organs of children are delicate and tender, and the spleen and the stomach are not healthy yet, so that the phenomena of frequent insufficiency of the spleen, invasion of exogenous pathogenic factors, stagnation and the like are easy to occur; the infant lung is delicate, tender and weak, but not strong, and different from adult lung in terms of tissue structure and physiological function. Although the traditional Chinese medicine preparation is more suitable for the infantile constitution than hormone medicines, the cough variant asthma treatment medicines specially aiming at the infantile constitution are not common at present.

Disclosure of Invention

The invention aims to provide a medicine for treating cough variant asthma based on the treatment principle of dispelling pathogenic qi and strengthening body resistance to consolidate constitution, so as to realize the phenotypic individualized treatment of the cough variant asthma in children.

The medicine for treating cough variant asthma is prepared from the following traditional Chinese medicines in parts by weight as raw material medicines:

5-15 of radix cynanchi wilfordii, 3-10 of fritillaria ussuriensis and 5-12 of bitter almonds

5-12 parts of perilla fruit, 5-12 parts of scutellaria root, 5-12 parts of earthworm

5-12 parts of stiff silkworm, 3-12 parts of peach kernel, 3-12 parts of dark plum and 3-12 parts of dark plum

3-12 parts of fructus aurantii and 2-5 parts of liquorice.

The selection and the dosage of the raw materials are obtained by the inventor through a large amount of grope and summary. Practice proves that the raw materials have good curative effect within the range of the weight portion.

Further, the medicine for treating cough variant asthma is prepared from the following raw material medicines in parts by weight:

caulis et folium Periplocae Forrestii 10-12 fritillaria ussuriensis 4-8 bitter almonds 6-10

6-10 parts of perilla fruit, 6-10 parts of scutellaria root, 6-10 parts of earthworm and 6-10 parts of earthworm

6-10 parts of stiff silkworm, 4-8 parts of peach kernel, 4-8 parts of dark plum and 4-8 parts of dark plum

4 to 8 portions of bitter orange and 3 to 5 portions of liquorice.

Furthermore, the most preferable raw material medicines of the medicine for treating cough variant asthma comprise, by weight:

caulis et folium Periplocae Forrestii, radix Rhodiolae Yunnanensis, Bulbus Fritillariae Ussuriensis, 5 semen Armeniacae amarum, 8

Fructus Perillae 8, Scutellariae radix 8, Lumbricus 8

Bombyx Batryticatus 8, semen Persicae, 6 mume fructus

Fructus Aurantii 6 and Glycyrrhrizae radix 3.

The raw material medicaments can be prepared into any conventional oral dosage form in pharmaceutics by adopting a conventional method of Chinese medicinal preparations, but the dosage forms cannot be used for limiting the protection scope of the invention. It is emphasized that the active ingredients of the pharmaceutical compositions of the present invention need to be prepared prior to preparing any of the dosage forms.

The specific preparation method for preparing the active components of the medicine from the raw materials comprises the following steps: decocting the raw material medicines in parts by weight in water for 3 times, mixing decoctions, filtering, and concentrating to obtain the active pharmaceutical ingredients.

When the medicines are decocted by adding water, the water consumption is 8-12 times of the weight of the raw materials each time, and the decoction is carried out for 1-2 hours each time.

Generally, the invention concentrates the filtrate under reduced pressure into the active ingredients of the medicine with the relative density of 1.10-1.30 at room temperature.

The invention can further prepare the obtained active ingredients of the medicine into any common oral dosage form by a conventional Chinese medicine preparation method, such as tablets, capsules, granules, pills, mixtures, syrups, soft extracts and the like.

The medicine has the effects of clearing lung and relieving cough, and relieving spasm and asthma, and is mainly used for treating cough with lung heat, such as cough variant asthma, asthmatic bronchitis and the like, and the symptoms of cough lasting and paroxysmal cough or spasmodic cough which is not lasting day and night and aggravated after being contacted with pungent smell; red tongue with white coating, slippery pulse, purple-superficial stagnation of finger-prints.

In the formula of the medicine, the yang-invigorating ginseng is bitter in taste and slightly cold, enters the lung channel and the heart channel, has the effects of tonifying qi, relieving cough and asthma, clearing heat and reducing fire, and is used for treating diseases such as bronchitis, phthisis and the like; the fritillary bulb has the pharmacological actions of clearing lung, eliminating phlegm, relieving cough and asthma and the like. The combination of the fritillary bulb which has the functions of clearing lung-heat, relieving cough, tonifying qi and relieving asthma and participates in moistening lung, relieving cough, eliminating phlegm and descending qi, and the combination of clearing and moistening lung, relieving cough and relieving asthma and treating both symptoms and root causes is used as monarch drug together. The almond and the perilla seeds can disperse and descend, and regulate the vital organs of the pathogenic factors so as to disperse, descend and descend; bai Qin is indicated for damp-heat in lung and purging lung fire upwards, especially for intractable cough with syndrome of unremoved heat. The combination of the almond, the perilla fruit and the baical skullcap root is designed aiming at the pathogenesis of intractable cough, pertinence of excess heat and lung disorder and is a ministerial drug which can effectively strengthen the main drug for relieving cough and asthma. Cough and dyspnea, cough of lung of dark plum; in chronic diseases, the peach kernel promotes blood circulation and moistens lung; bombyx Batryticatus has effects of eliminating phlegm and resolving masses, calming endogenous wind and relieving spasm; earthworm clears heat and relieves asthma, clears the channels and activates the collaterals; fructus Aurantii moves qi to open chest, widens the middle energizer and removes distention. The five medicines of earthworm, stiff silkworm, dark plum, peach kernel and bitter orange are combined to be used as adjuvant medicines, which seems to be mainly used for treating the symptoms, but plays an important role in shortening the course of treatment and accelerating healing, and cannot be ignored. For chronic cough variant asthma, asthmatic bronchitis, etc., the combination of the medicines for relieving spasm and eliminating phlegm, promoting blood circulation and activating qi is essential. Clinical practice proves that the worm medicines such as earthworm and stiff silkworm have obvious effects of relieving spasm, calming panting and relieving cough, and the functions of dispelling wind, removing collaterals and expelling phlegm are far more obvious than those of the herbal medicines. For symptoms of cough, spasmodic cough, wheezy phlegm in throat and phlegm-qi deficiency, the stiff silkworm and the earthworm are combined with the products for promoting blood circulation and qi circulation, such as the bitter orange, the peach seed and the like, so that the effects of the herbal and the wood medicine and the insect medicine are enhanced, and the effects of mutual promotion and complement can be achieved. The lung is the delicate organ, the infant viscera are tender, and the compatibility of herbs is especially good. The common course of diseases such as cough variant asthma and asthmatic bronchitis in children is longer, and the requirements of eliminating pathogens without damaging healthy qi and relieving cough without converging pathogens are further considered. The use of dark plum in the adjuvant just reflects the compatibility spirit, the dark plum has the organic combination of astringing lung to stop chronic cough and opening the profit to relieve spasm of stiff dragon peach and trifoliate orange, the effect of relieving cough and eliminating phlegm is improved, and the disadvantages of astringing pathogen to stay in the nutmeg and attacking and hurting vital qi are avoided. Therefore, the five-flavor adjuvant drug is not only buckled with the pathological mechanism of the main disease, but also takes the constitutional features of the children into full consideration, and is a fine choice for the individual features of the children. Licorice root, radix Glycyrrhizae is an indispensable drug for blending various drugs, modifying the taste and nature of the drugs, and is used as a guiding drug. The medicines have the effects of clearing heat and dispersing lung and relieving cough and asthma, and have obvious clinical curative effect on intractable cough.

Systematic pharmacodynamic tests prove that the medicine has the effects of relieving cough, eliminating phlegm, relieving asthma, resisting bacteria and treating cough variant asthma, and plays a role in treating cough variant asthma mainly through the effects of relieving inflammation, regulating the immune function of an organism and the like.

Drawings

FIG. 1 is the body temperature trend of the rats of the Streptococcus pneumoniae pulmonary heat model in different experimental groups.

FIG. 2 shows the histopathological effect of lung (HE X200) in different experimental groups of Streptococcus pneumoniae pulmonary heat model rats.

Detailed Description

The following examples are only preferred embodiments of the present invention and are not intended to limit the present invention in any way. Various modifications and alterations to this invention will become apparent to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Animal tests on the antitussive, expectorant, antiasthmatic and antibacterial antiallergic effects of the present invention are given in the following examples. All indexes in the test conclusion are expressed by mean and standard deviation (

) Shows that the whole of each group of data follows normal distribution and has uniform variance after normal test and variance homogeneity test (P>0.1), adopting one-factor variance analysis to each group of data, and adopting an LSD-t method to compare the multiple groups pairwise. The analysis of experimental data was done using the SPSS 22.0 software package,P<0.05 means statistically significant.

Example 1.

Decocting radix Rhodiolae 10g, Bulbus Fritillariae Ussuriensis 5g, semen Armeniacae amarum 8g, fructus Perillae 8g, Scutellariae radix 8g, Lumbricus 8g, Bombyx Batryticatus 8g, semen Persicae 6g, mume fructus 6g, fructus Aurantii 6g, and Glycyrrhrizae radix 3g in water for 3 times, each time with 760mL of water, and decocting for 1.5 h. Mixing the three decoctions, filtering, concentrating under reduced pressure to relative density of 1.2-1.3 (room temperature), spray drying, adding dextrin or sugar powder at a mass ratio of 1: 3, mixing, and making into granule.

Example 2.

Decocting radix Rhodiolae 12g, Bulbus Fritillariae Ussuriensis 6g, semen Armeniacae amarum 10g, fructus Perillae 10g, Scutellariae radix 10g, Lumbricus 10g, Bombyx Batryticatus 10g, semen Persicae 8g, mume fructus 8g, fructus Aurantii 8g, and Glycyrrhrizae radix 5g in water for 3 times, and decocting with 780mL water each time for 1.5 hr. And combining the three decoctions, filtering, concentrating under reduced pressure to obtain dry paste with the relative density of 1.1-1.3 (room temperature), adding sucrose accounting for 20-50% of the dry paste and a proper amount of preservative, adding water, boiling for dissolving, filtering, cooling, adding a proper amount of ethanol solution and water, stirring uniformly, and preparing syrup.

Example 3.

Decocting 8g of radix Rhodiolae Yunnanensis, 3g of Bulbus Fritillariae Cirrhosae, 6g of semen Armeniacae amarum, 6g of fructus Perillae, 6g of Scutellariae radix, 6g of Lumbricus, 6g of Bombyx Batryticatus, 4g of semen Persicae, 4g of mume fructus, 4g of fructus Aurantii, and 3g of Glycyrrhrizae radix with 620mL of water for 3 times, each time, and decocting for 1.5 h. And combining the three decoctions, filtering, and concentrating under reduced pressure to obtain dry paste, wherein 18-21 g of the active ingredients of the medicine are obtained. Adding sucrose accounting for about 5 percent of the mass of the active ingredients of the medicine, starch accounting for 5 to 10 percent of the mass of the active ingredients, dextrin accounting for about 20 percent of the mass of the active ingredients and a proper amount of magnesium stearate, uniformly mixing, preparing granules, and tabletting to prepare tablets.

Example 4.

Taking 9g of radix cynanchi wilfordii, 4g of fritillaria ussuriensis, 7g of bitter apricot seed, 7g of perilla seed, 7g of scutellaria baicalensis, 7g of earthworm, 7g of stiff silkworm, 5g of peach seed, 5g of dark plum, 5g of bitter orange and 3g of liquorice, adding water, carrying out reflux extraction for 3 times, using 730mL of water each time, and decocting for 1.5 h. And combining the three decoctions, filtering, and concentrating under reduced pressure to obtain dry paste, wherein 18-22 g of the active ingredients of the medicine are obtained. Adding adjuvants such as calcium carbonate and starch, mixing, drying, pulverizing into fine powder, sieving, and making into capsule.

Example 5.

Decocting 5g of radix Rhodiolae Yunnanensis, 3g of fritillaria ussuriensis, 6g of bitter apricot seed, 6g of perilla seed, 6g of scutellaria baicalensis, 6g of earthworm, 6g of stiff silkworm, 4g of peach kernel, 4g of dark plum, 4g of bitter orange and 2g of liquorice in water for 3 times, and decocting for 1.5 hours by using 580mL of water each time. Mixing the three decoctions, filtering, concentrating under reduced pressure to relative density of 1.1-1.3 (room temperature), adding ethanol, standing for 48h, filtering, recovering ethanol, concentrating to relative density of about 1.3, adding adjuvants including sodium benzoate (about 0.1%), sucrose and water, filtering, bottling, sterilizing, and making into mixture.

Example 6.

Taking 15g of radix cynanchi wilfordii, 6g of bulbus fritillariae cirrhosae, 8g of bitter apricot seed, 8g of perilla seed, 8g of scutellaria baicalensis, 8g of earthworm, 8g of stiff silkworm, 7g of peach seed, 7g of dark plum fruit, 7g of fructus aurantii and 4g of liquorice, adding water, decocting for 3 times, and decocting for 1.5 hours by using 720mL of water each time. And combining the three decoctions, filtering, concentrating under reduced pressure to relative density of 1.1-1.3 (room temperature), adding about 20% of sucrose or simple syrup, heating to melt, mixing, concentrating to relative density of about 1.4, and cooling to obtain soft extract.

Example 7: cough-relieving experiments in mice.

120 Kunming young mice are taken and put into a 5L closed dryer one by one, and atomized 25% ammonia water is introduced into the dryer for 20s by an ultrasonic atomizer at the capacity of 4 mL/min. The mice were quickly transferred under an inverted funnel and the mice were listened for cough latency at the root of the funnel using a stethoscope with a bell-shaped connector. Sensitive mice with latency within 90s were selected as eligible animals for the experiment.

Taking 70 qualified mice, and randomly dividing the mice into a blank group, a model group, a positive western medicine group, a positive traditional Chinese medicine group, a low-dose group of the medicine, a medium-dose group of the medicine and a high-dose group of the medicine according to the weight, wherein each group comprises 10 mice and each half of the mice.

Mice in a blank group and a model group are drenched with equal volume of normal saline (0.2mL/10g) once a day, a positive traditional Chinese medicine group is administrated with 13.8mg/kg of particles for treating cough and asthma caused by lung heat of children, a positive western medicine group is administrated with 21.6mg/kg of bithionine, and the other test groups are drenched with 0.2mL/10g of low, medium and high dose medicines (equivalent to 11g/kg, 22g/kg and 44g/kg) of the invention respectively, and are continuously and prophylactically administrated for 7 d.

1h after the last administration, mice of each test group except the blank group were placed in a 5L closed desiccator, and atomized 25% ammonia water was introduced into the desiccator for 20s at a volume of 4mL/min using an ultrasonic atomizer. Mice were quickly transferred to the inverted funnel, and the mice in each test group were listened to and counted for cough latency and the number of coughs within 2min (no latency) using a stethoscope at the root of the funnel.

The period from the start of spraying to the appearance of cough was recorded as the cough latency of the mice. The cough frequency of the mouse within 2min is counted by calculating that the mouse has one cough when the abdominal muscle contracts or contracts the chest and the mouth is enlarged sometimes.

The statistical results of the effect of the drugs in each test group on the mice with the cough caused by ammonia are shown in table 1. Compared with the model group, the positive western medicine group, the positive Chinese medicine group and the low, medium and high dose groups of the medicine can obviously prolong the cough incubation period of the mice and reduce the cough frequency of the mice, and the difference is obvious (P<0.05). Compared with the positive western medicine group and the positive traditional Chinese medicine group, the low, medium and high dosage groups of the medicine of the invention can prolong the cough incubation period of mice without significant difference (P>0.05). The low, medium and high dose groups of the drug can reduce the cough frequency of mice, but the low dose group and the medium and high dose groups have statistical difference (P<0.05)。

Example 8: guinea pig cough test.

100 young guinea pigs are taken and placed in a 3L glass bell one by one day before the experiment, 17.5% citric acid solution is sprayed by an ultrasonic atomizer for 10s, the cough frequency within 5min from spraying is recorded, and the guinea pigs with the cough frequency more than 10 times are selected as qualified animals for the experiment.

70 qualified guinea pigs are randomly divided into a blank group, a model group, a positive western medicine group, a positive traditional Chinese medicine group, a low-dose group of the medicine, a medium-dose group of the medicine and a high-dose group of the medicine according to the weight, wherein each group comprises 10 male and female halves.

The guinea pigs in the blank group and the model group are irrigated with equal volume of normal saline (1mL/100g) once a day, the positive traditional Chinese medicine group is irrigated with lung heat cough and asthma granules for children at 6.5g/kg, the positive western medicine group is irrigated with expectorant at 10.1mg/kg, and the other test groups are irrigated with low, medium and high dose medicines (equivalent to 5g/kg, 10g/kg and 20g/kg)1mL/100g respectively, and are irrigated with continuous preventive administration for 7d 1 time a day.

1h after the last administration, the guinea pigs were placed in a 3L glass bell, stimulated with 17.5% citric acid solution as described above, and the cough latency (period from the start of spraying to the appearance of cough) and the number of coughs within 5min (no latency) were recorded in the guinea pigs of each test group. Guinea pigs cough loudly, and the number of coughs was calculated as the heard coughs.

The results of the effect of each test group drug on the cough of guinea pig citric acid are shown in table 2. Compared with the model group, the positive western medicine group, the positive Chinese medicine group and the low, medium and high dose groups of the medicine can obviously prolong the cough latent period of the guinea pigs and reduce the cough frequency of the guinea pigs, and the difference is obvious (P<0.05). Compared with the positive traditional Chinese medicine, the low, medium and high dosage groups of the medicine can prolong the cough latent period of the guinea pig and reduce the cough frequency without obvious difference (P>0.05). Compared with the positive western medicine group, the low, medium and high dosage groups of the medicine can prolong the cough latency of guinea pigs and reduce the cough frequency, but the latency of each group has statistical difference (P<0.05). The low, medium and high dosage groups of the medicine have no significant difference (P>0.05)。

Example 9: mouse phenol red phlegm eliminating experiment.

Phenol red was dissolved in 5% sodium bicarbonate solution and prepared to have a concentration of 5 and 2, respectively.5. 1.25, 0.625, 0. mu.g/mL phenol red solution, and the OD at 546nm of each solution was measured with a spectrophotometer. Taking the concentration of the phenol red solution as a vertical coordinate and the OD value as a horizontal coordinate, calculating according to the concentration and the OD value of different phenol red standard solutions to obtain a linear regression equation of the concentration and the OD value of the phenol red asY=0.0956X+ 0.0452，r= 0.9997。

Taking 60 Kunming young mice, and randomly dividing the mice into a blank group, a positive western medicine group, a positive traditional Chinese medicine group, a low-dose group of the medicine, a medium-dose group of the medicine and a high-dose group of the medicine according to the weight, wherein each group comprises 10 mice and each half of the mice.

The mice in the blank group are drenched with equal volume of normal saline (0.2mL/10g) once a day, the positive traditional Chinese medicine group is administrated with 13.8g/kg of particles for treating the lung heat cough and asthma of the children, the positive western medicine group is administrated with 21.6mg/kg of Musultan, and the other test groups are drenched with 0.2mL/10g of low, medium and high dose medicines (equivalent to 11g/kg, 22g/kg and 44g/kg) respectively, and the continuous preventive administration is carried out for 7d 1 time a day.

1h after the last administration, carrying out intraperitoneal injection of 0.5mL of 2.5% phenol red physiological saline solution, 0.5h later, removing cervical vertebra of the mice to kill the mice, fixing the mice on an operation plate in an upward position, cutting off the skin of the neck, separating trachea, taking a trachea section from the thyroid cartilage to the trachea bifurcation, putting each trachea section into a container which is previously filled with 1.5mL of 5% sodium bicarbonate solution, fully shaking, standing at 4 ℃ for 24h, measuring the OD value of the solution, and calculating the content of phenol red according to a regression equation.

The results are shown in Table 3. Compared with the blank group, the positive western medicine group, the positive Chinese medicine group and the medicine high, medium and low dose group of the invention can increase the phenol red excretion amount of the trachea segment of the mouse to different degrees, and the difference is obvious (P<0.05). Compared with the positive western medicine group and the positive traditional Chinese medicine group, the high, medium and low dosage group of the medicine of the invention has no obvious difference (P>0.05). The high, medium and low dosage groups of the medicine have no significant difference (P>0.05)。

Example 10: rat capillary sputum excretion experiments.

60 young rats are randomly divided into a blank group, a positive western medicine group, a positive Chinese medicine group, a low-dose group of the medicine, a medium-dose group of the medicine and a high-dose group of the medicine according to the weight, wherein each group comprises 10 rats and each rat is male and female.

The rats in the blank group are drenched with isometric physiological saline (1mL/100g) once a day, the positive traditional Chinese medicine group is used for administering lung heat cough and asthma granules of the children by 7.4g/kg, the positive western medicine group is used for administering Musultan by 27.7mg/kg, and the other test groups are drenched with low, medium and high dose medicines (equivalent to 6g/kg, 12g/kg and 24g/kg) of the invention by 1mL/100g respectively, and the rats in the blank group are drenched with isometric physiological saline (1mL/100g) once a day, and the rats in the.

After the last administration for 1h, 20% urethane is injected for anesthesia, the back is fixed, the trachea is separated, a glass capillary tube with the length of about 3-5 cm and the diameter of 0.3mm is inserted at the position of the 3 rd cartilage ring under the thyroid cartilage ring, so that the glass capillary tube just contacts the bottom surface of the trachea, and the sputum is collected for 2 h. The length (cm) of the sputum in each group of glass capillaries within 2h is used as an index for analysis.

The analytical results are shown in Table 4. Compared with the blank group, the positive western medicine group, the positive Chinese medicine group and the low, medium and high dose group of the medicine can increase the sputum secretion of rats with obvious difference (P<0.05). Compared with the positive western medicine group and the positive traditional Chinese medicine group, the low, medium and high dosage group of the medicine of the invention has no significant difference (P>0.05). The low, medium and high dosage groups of the medicine have no significant difference (P>0.05)。

Example 11: guinea pig asthma test.

100 young guinea pigs with the weight of less than 200g are selected, the male and female guinea pigs are placed into a 3L closed dryer one by one, an ultrasonic atomizer is used for spraying a mixed solution of 0.1% histamine phosphate and 2% acetylcholine chloride with the same volume at 4mL/min, the mixed solution is sprayed for 10s, the incubation period of the asthma of the guinea pigs (the time from the beginning of spraying to the falling of the guinea pigs) is observed and recorded, and patients with the falling time of more than 180s are not selected.

70 qualified guinea pigs are selected and randomly divided into a blank group, a model group, a positive western medicine group, a positive traditional Chinese medicine group, a low-dose group of the medicine, a medium-dose group of the medicine and a high-dose group of the medicine, wherein each group comprises 10 male and female halves.

The guinea pigs in the blank group and the model group are drenched with the same volume of normal saline (1mL/100g) once a day, the particles for treating cough and asthma with lung heat in children are 6.5g/kg in the positive traditional Chinese medicine group, the aminophylline is 0.105g/kg in the positive western medicine group, and the low, medium and high dose drugs (5 g/kg, 10g/kg and 20g/kg) of the invention are drenched with 1mL/100g in the other test groups respectively, and are continuously and preventively administrated for 7 days 1 time a day.

1h after the last administration, an equal volume of mixed solution of 0.1% histamine phosphate and 2% acetylcholine chloride was sprayed with an ultrasonic nebulizer for 10s, and the asthma latency (i.e., the time from the start of spraying to the onset of asthma attack, dyspnea, until the twitch falls) was observed and recorded in guinea pigs of each test group.

The results of the observation are shown in Table 5. Compared with the model group, the positive western medicine group, the positive Chinese medicine group and the low, medium and high dose groups of the medicine can obviously prolong the asthma-inducing incubation period of the guinea pigs, and the difference is obvious (P<0.01). Compared with the positive western medicine group and the positive traditional Chinese medicine group, the low, medium and high dose groups of the medicine of the invention can prolong the incubation period of asthma induction of guinea pigs, but have statistical difference (P<0.05). The low, medium and high dosage groups of the medicine have no significant difference (P>0.05)。

Example 12: rat lung heat model test.

100 young rats with half each male and female are selected, and after the environment is adapted for one week, the body temperature is monitored in the morning and at the evening every day for 3 days continuously.

Rats with a body temperature excursion less than 0.5 ℃ were selected for a total of 70 rats. The medicine is randomly divided into a blank group, a model group, a positive western medicine group, a positive Chinese medicine group, a low-dose group of the medicine, a medium-dose group of the medicine and a high-dose group of the medicine. Except for the blank group, the other 6 groups of rats were administered with 1.0 concentration after ether inhalation for mild anesthesia10⁹The solution of streptococcus pneumoniae bacteria (0.5mL/kg) is slowly dropped into the nasal cavity of a rat by a syringe with a No. 4 half needle, the bacteria feeding speed is 0.05mL/min, and the bacteria are fed for 3 times every 12 hours for 1 time. The rats in the blank group were given an equal volume of saline to their nasal cavity and operated as above.

1d after modeling, 0.16g/kg of cefuroxime axetil tablets are taken by a positive western medicine group, 7.4g/kg of particles for treating the lung heat cough and asthma of the children are taken by a positive traditional Chinese medicine group, 1mL/100g of medicines with corresponding concentrations (equivalent to 6g/kg, 12g/kg and 24g/kg) are respectively filled into a low-dose group, a medium-dose group and a high-dose group of the medicines disclosed by the invention, the medicines are filled into the stomach 1 time every day, and purified water (1mL/100g) with the same volume is taken into a model group and a blank group, and the medicines are continuously taken.

The body temperatures of rats in each test group before molding, after molding, 3d for administration and 7d for administration are monitored and recorded.

The body temperature measurement results are shown in fig. 1. The body temperature of the rats before molding was relatively low. After streptococcus pneumoniae is dripped into the nasal cavity, the body temperature of the rat in the model group is obviously raised compared with that in the blank group, which indicates that the model building is successful; after the administration for 3d and 7d, the body temperatures of rats in the positive western medicine group and the positive traditional Chinese medicine group have obvious descending trends, the body temperatures of rats in the low, medium and high dose groups of the medicine also have descending trends, and the descending trends of the medium and high dose groups are more obvious.

After the last administration for 1h, the rats were injected with 10mL/kg of 20% urethane solution into the abdominal cavity, the exposed organs were cut at the center of the neck, a rounded 5mL syringe needle was inserted, 4mL of physiological saline was slowly injected into the trachea, aspiration was repeated several times, the wash solution was withdrawn, about 2mL of the wash solution was recovered, and the recovered wash solution was placed in a 5mL EP tube, left to stand for 1h, centrifuged at 3000r/min for 15min, and the supernatant was collected to obtain lung lavage fluid (BALF) for detection of inflammatory factors such as IL-10, IL-1 β, IL-6, IL-4, and Ig-E, TNF- α.

The results are shown in Table 6 and Table 7, and the levels of IL-10, IL-1 β, IL-6, IL-4 and TNF- α in the model group BALF were increased in comparison with the blank group (see Table 7)P<0.05) indicating that the model is successfully made, compared with the model group, the low, medium and high dose groups of the drug of the invention have obviously reduced IL-1 β, IL-6, IL-4 and TNF- α levels, and the difference has statistical significance (P<0.05), while the IL-10 level in the high dose group was not significantly different from that in the model group (P>0.05), the water average of IL-1 β of the low, medium and high dose groups of the drug is far lower than that of a positive western medicine group, the water average of TNF- α is far lower than that of a positive traditional Chinese medicine group, but the difference has no statistical significance, the Ig-E content has no obvious difference among all test groups, and the result shows that the low and medium dose groups of the drug have obvious down-regulation trend on the water average of IL-10, IL-1 β, IL-6, IL-4 and TNF- α, and the down-regulation effect on the level of IL-1 β and TNF- α is better than that of a positive control drug.

The abdominal aorta of the rats of each test group is bled for detecting leucocytes and eosinophils, and the flow cytometry is used for detecting CD4/CD8T cells.

The results of the leukocyte and eosinophil measurements in each test group are shown in Table 8. Compared with the blank group, the number of the leucocytes and the eosinophils in the whole blood of the rats in the model group is obviously increased, and the difference has statistical significance (P<0.05), which indicates that the molding is successful; compared with the model group, the number of leucocytes and eosinophils in the whole blood of the rats in each administration group is obviously reduced, and the difference has statistical significance (1)P<0.05); there were no statistical differences between the groups administered.

CD3⁺CD4⁺、CD3⁺CD8⁺The results are shown in Table 9. Model set CD3 compared to blank set⁺CD4⁺、CD3⁺CD8⁺The percentage is remarkably reduced (P<0.01), and the Th/Ts ratio is significantly increased (P<0.01), indicating that the streptococcus pneumoniae-induced rat lung heat model destroys the relative balance of immune cells in rats; the Th/Ts ratios of all the groups were decreased compared to the model group, wherein the drug of the present invention was administered in the low, medium and high dose groups and positiveThe Th/Ts ratio of the western medicine group is closer to that of the blank group, and the adjustment effect of the low, medium and high dose group of the medicine on the immune imbalance caused by pneumonia is better than that of the positive traditional Chinese medicine and has equivalent curative effect with the positive western medicine.

Spleen and thymus of rats in each test group were isolated, and spleen index and thymus index were calculated.

The results of organ index measurements are shown in Table 10. The thymus and spleen indices of the model group were significantly reduced compared to the blank group (P<0.01); all the administered groups had significantly increased spleen index compared with the model group (P<0.05), the up-regulation level of the low, medium and high dosage groups of the medicine of the invention tends to blank group, which is superior to that of the positive control medicine; the thymus index of each administration group is increased compared with that of the model group, but the thymus index of each administration group is not statistically different (P>0.05)。

The other lung of the rat was removed, gross changes were observed under natural light, fixed with 10% formaldehyde solution, embedded in paraffin, sectioned, stained with hematoxylin-eosin (HE), and lung lesions were observed under light microscope.

The lung of the rat in the model group is observed by naked eyes, the lung is obviously scattered at a blood stasis point and has white needle-shaped or block-shaped abscess, a small number of the lung has larger area of consolidation and abscess, and the individual lung presents red or gray liver-like change.

The results of the optical microscopy are shown in FIG. 2. The lung tissue of the blank group is not abnormal; congestion and thickening of alveolar walls can be seen under a model group mirror, infiltration of red blood cells and neutrophils is formed in an alveolar cavity to form a small abscess, edema liquid can be seen in the alveolar cavity, and a plurality of small focitis cells are infiltrated under pleura. Compared with the model group, the positive western medicine group, the positive Chinese medicine group, the medicine middle-dosage group and the high-dosage group of the invention have no inflammatory cell infiltration and edema fluid, and pathological sections tend to a normal group; the low-dose group of the medicine can be seen in inflammatory cell infiltration mainly comprising neutrophils around local interstitium, blood vessels and bronchus. Therefore, the medium and high dose group of the medicine has a certain treatment effect on the lung heat rats infected by the streptococcus pneumoniae.

Example 13: guinea pig cough variant asthma model test.

70 young guinea pigs with the weight less than 200g are selected, and the weight is half female and half male. After 1 week of feeding, 60 guinea pigs with cough variant asthma were modeled, and 0.5mL of 4% OVA solution was injected into the No. 1d cavity and 2% Al (OH) was injected into the abdominal cavity₃0.2mL, one boost at 7 d; after 14d, the guinea pigs were challenged with 1% OVA solution by nebulization 1 time every other day for 120s for 7 times, to 29d to obtain molded guinea pigs.

The 60 molded guinea pigs were randomly divided into a model group, a positive western medicine group, a positive traditional Chinese medicine group, a low-dose group of the present invention, a medium-dose group of the present invention, and a high-dose group of the present invention, and 10 young guinea pigs that were not molded were used as blank groups.

The guinea pigs in the blank group and the model group are irrigated with equal volume of physiological saline (1mL/100g) once a day, the dexamethasone acetate suspension is given to the positive western medicine group at 1.2mg/kg, and the lung heat cough and asthma granules in the children are given to the positive traditional Chinese medicine group at 6.5 g/kg.

1h after the last dose, the guinea pigs were placed in a 5L desiccator using 10^-4The mol/L capsaicin solution was atomized for 2min, and cough latency (period from the start of atomization to the appearance of cough) and the number of coughs within 5min (no latency) of the guinea pigs were observed and recorded.

The statistical results are shown in Table 11. The cough latency in model guinea pigs was significantly reduced compared to the blank group (P<0.05), significant increase in cough frequency within 5 min: (P<0.01), indicating that model group guinea pigs are sensitive to capsaicin challenge. The incubation period of guinea pigs in each administration group was significantly prolonged as compared with that in the model group (P<0.05), the number of coughs was significantly reduced within 5min, and the differences were statistically significant (P<0.01)。

The guinea pig is injected with 10mL/kg 20% urethane solution into abdominal cavity, the right middle lobe lung is ligated, the exposed organ is cut at the center of the neck, a rounded 5mL syringe needle is inserted, 4mL physiological saline is slowly injected into the trachea, the aspiration is repeated several times, the wash solution is extracted, about 2mL of the wash solution is recovered, the recovered wash solution is placed in a 5mL EP tube, the recovered wash solution is left to stand for 1h, centrifuged at 3000r/min for 15min, and the supernatant is collected to obtain lung lavage fluid (BALF) for testing inflammatory factors such as IL-5, IL-10, LTE4, NGF, TNF- α and inflammatory cells such as leucocyte, neutrophil, lymphocyte, monocyte, eosinophil, basophil and the like.

The results of the inflammatory factor detection are shown in Table 12, compared with the blank group, the IL-5, IL-10, LTE4, NGF and TNF- α contents in the model group BALF are obviously increased (P<0.05) compared with the model group, each administration group has down-regulation effect on IL-5, IL-10, NGF and TNF- α and has up-regulation effect on LTE4P<0.05) the callback effect of the low, medium and high dose groups of the medicine of the invention to IL-5, IL-10, LTE4 and TNF- α is equivalent to the effect of a positive western medicine group, the down regulation effect of the medicine to NGF is better than that of a positive Chinese medicine group, but the difference is not statistically different (P>0.05)。

The results of inflammatory cell detection are shown in tables 13 and 14. Compared with the blank group, the numbers of neutrophils, lymphocytes, monocytes, eosinophils and basophils in the BALF of the model group are all obviously increased, and the difference has statistical significance (theP<0.01), white blood cell count is reduced (P<0.05); compared with the model group, the positive western medicine group has obvious down-regulation effect on the numbers of neutrophils, lymphocytes, eosinophils and basophils (P<0.05), the number of lymphocytes, monocytes and eosinophils is obviously reduced by the positive traditional Chinese medicine group (P<0.05); the low, medium and high dosage groups of the medicine have the callback function to the numbers of leucocytes, lymphocytes and eosinophilsTendency (1)P<0.05)。

Spleen and thymus of guinea pigs were isolated and spleen and thymus indices were calculated.

The results are shown in Table 15. The spleen index of the model group was significantly reduced compared to that of the blank group (P<0.01), the thymus index decreased but the difference was not statistically significant (P>0.05); spleen indices were significantly increased in the administered group compared with those in the model group (P<0.05), the up-regulation level of the low, medium and high dosage groups of the medicine of the invention tends to blank group, which is superior to that of the positive traditional Chinese medicine; the thymus index of each administration group is increased compared with that of the model group, but the thymus index of each administration group is not statistically different (P>0.05)。

The thymus index and the spleen index can reflect the immune function state of the organism. The above studies showed that the administration group guinea pigs had enhanced immune function to various degrees.

Claims

1. A medicine for treating cough variant asthma is prepared from the following raw material medicines in parts by weight:

3-12 parts of fructus aurantii and 2-5 parts of liquorice.

2. The drug for treating cough variant asthma according to claim 1, which is prepared from the following raw material drugs in parts by weight:

4 to 8 portions of bitter orange and 3 to 5 portions of liquorice.

3. The drug for treating cough variant asthma according to claim 1, which is prepared from the following raw material drugs in parts by weight:

Fructus Perillae 8, Scutellariae radix 8, Lumbricus 8

Bombyx Batryticatus 8, semen Persicae, 6 mume fructus

Fructus Aurantii 6 and Glycyrrhrizae radix 3.

4. The preparation method of the medicine for treating cough variant asthma as claimed in claim 1, comprises decocting the raw materials in water for 3 times, mixing decoctions, filtering, and concentrating to obtain the active pharmaceutical ingredient.

5. The method for preparing a medicament for treating cough variant asthma as claimed in claim 4, wherein the amount of water used for each time is 8-12 times of the weight of the raw materials, and the decoction is carried out for 1-2 hours each time.

6. The method of claim 4, wherein the filtrate is concentrated under reduced pressure to a pharmaceutically active component having a relative density of 1.10 to 1.30 at room temperature.

7. The method of claim 4, wherein the active pharmaceutical ingredient is formulated into any one of the commonly used oral dosage forms.

8. The method of claim 7, wherein the oral dosage form comprises tablets, capsules, granules, pills, mixtures, syrups, and electuaries.