CN113876844A

CN113876844A - Pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis and preparation method and application thereof

Info

Publication number: CN113876844A
Application number: CN202111180072.5A
Authority: CN
Inventors: 扎西次仁; 张水寒; 达瓦桑珠; 巴桑伦珠; 邵国强; 万丹; 李跃辉; 嘎尔琼; 巴桑
Original assignee: Shannan Tibetan Medicine Hospital
Current assignee: Shannan Tibetan Medicine Hospital
Priority date: 2021-10-11
Filing date: 2021-10-11
Publication date: 2022-01-04
Anticipated expiration: 2041-10-11
Also published as: CN113876844B

Abstract

The invention belongs to the technical field of traditional Chinese medicine Tibetan medicines, and discloses a pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis and a preparation method and application thereof. The Tibetan medicine is prepared from the following raw materials in parts by weight: 15-20 parts of sandalwood, 15-20 parts of Solms-Laubachia, 15-20 parts of Philippine violet herb, 10-15 parts of Aconitum tanguticum, 10-15 parts of Gentiana macrophylla flower and 10-15 parts of Qinghai gentiana scabra Bunge. The preparation method comprises the following operation steps: mixing sandalwood, Solms-Laubachia, purple chrysanthemum, Aconitum tanguticum, Gentiana macrophylla flower and Qinghai gentian, crushing into fine powder, and performing damp-heat sterilization at 115 ℃ for 30min to obtain the traditional Chinese medicine mixed powder, namely the pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis. The invention has reasonable formula, has the effects of clearing heat and removing toxicity, relieving sore throat and reducing swelling, and relieving cough and reducing sputum, and is used for treating symptoms such as sore throat, hoarseness, cough, expectoration, chest and back pain and the like caused by sphagitis and bronchitis.

Description

Pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis and preparation method and application thereof

Technical Field

The invention belongs to the technical field of traditional Chinese medicine Tibetan medicines, and particularly relates to a pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis and a preparation method and application thereof.

Background

Chronic bronchitis is a chronic nonspecific inflammation of the trachea, bronchial mucosa and surrounding tissues. Clinically, cough and expectoration are the main symptoms, particularly, the most obvious early morning phlegm is in the form of white mucus foam, the thick phlegm is not easy to be expectorated, and the symptoms are rapidly aggravated after infection or cold catching, the phlegm quantity is increased, the viscosity is increased or yellow purulent. Sometimes, there is blood in the cough-phlegm, and with the progress of the disease, there are cough-phlegm in the whole year, while in autumn and winter, even long-term attack may lead to emphysema.

The main cause of chronic bronchitis formation is the chronic nonspecific inflammation of the bronchi due to repeated infections with viruses or bacteria. When the temperature drops, the small vessels of the respiratory tract are spastic and ischemic, and the defense function is reduced, so that diseases are easy to cause; chronic irritation such as smoke dust, polluted atmosphere, etc. can also cause diseases; smoking causes bronchospasm, mucosal variation, decreased ciliary movement, and increased mucus secretion; allergic factors are also one of the causes of infection.

The western medicine for treating chronic tracheitis generally uses various antibiotic medicines, has high cost and poor treatment effect, and the taking of too much antibiotic is harmful to human body, which can cause the immunologic function of human body to decline, thus causing the disease condition to be difficult to cure repeatedly.

The traditional Chinese medicine has a long history in the aspect of treating chronic tracheitis and has a better treatment effect. There are many folk traditional Chinese medicine formulas for treating chronic tracheitis, but a traditional Chinese medicine formula with obvious effect and simple formula is lacked.

The Tibetan medicine is a unique national medicine in China, has the history of more than 2300 years, and is endowed with irreplaceable curative effect and medical value due to the special growth environment. The differences between the Tibetan herbs and the traditional Chinese herbs are expressed in the theoretical system, the collection of herbs, and the processing, besides the differences in the generation regions.

The theory of Tibetan medicine mainly focuses on the growth, nature, taste and effect of the drugs and five sources (water, soil, fire, wind and air), and in addition, the nature, taste and effect are the theoretical basis of clinical medication. The Tibetan medicine has six medicinal ingredients: sweet, sour, salty, bitter, pungent and astringent, etc., wherein the bitter taste is the best effect. In addition, as the Tibetan medicine generally grows on the snow peak with the altitude of 3800 meters, the medicine is not polluted, and the effective components of the medicine are not interfered and destroyed by other substances, so that the medicine effect of the Tibetan medicine is quicker than that of the traditional Chinese medicine. In addition, the biggest characteristic of Tibetan medicine is that it can directly enter into the focus of infection, directly inhibit virus and radically treat disease from the body.

The collection of Tibetan herbs is different from that of traditional Chinese herbs. The traditional Chinese medicine is collected along with seasons, while the Tibetan medicine is collected when the content of the active ingredients of the medicinal materials is the highest. The whole grass is generally collected when the plant grows vigorously, so that the drug effect is also highest when the plant is used as a medicine, and the whole grass has an excellent effect on treating diseases, is collected only once a year basically and is rare.

The processing terms of Tibetan herbs are different. The Tibetan medicine processing technology absorbs the advantages of the world medicine processing technologies such as ancient Rome, ancient Arabia, ancient India, Chinese traditional medicine Hanfang and the like, is effectively integrated with the plateau medicine processing technology, is very good at processing various mineral substances and heavy metals by hundreds of detoxification technologies to remove the toxicity and the survivability of the toxicity, and is scientifically compatible with animal and plant medicinal materials to form various compound compatible patent medicines.

In recent years, Tibetan medicines are increasingly researched for treating bronchitis, and reported medicaments such as lung rehabilitation powder, four-flavor rhodiola rosea decoction powder, ten-flavor gentian granules, sixteen-flavor rhododendron powder, seven-flavor grape powder, five-flavor sea buckthorn oral liquid, thirty-five-flavor agilawood powder and the like all achieve better curative effects on treating bronchitis. The formula of the invention is prepared by screening from the book of many Tibetan medicines and refining through clinical practice for many years.

Disclosure of Invention

In order to solve the defects in the prior art, the invention mainly aims to provide a pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis; the pure traditional Chinese medicine Tibetan medicine has no toxic or side effect, takes effect quickly, has obvious drug effect and is not easy to relapse after recovery.

The invention also aims to provide a preparation method of the pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis.

The invention also aims to provide application of the pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis.

The purpose of the invention is realized by the following technical scheme:

a pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis is prepared from the following raw materials: sandalwood, Solms-Laubachia, purple chrysanthemum, Aconitum tanguticum, Gentiana macrophylla flower and Qinghai gentian.

Preferably, the Tibetan medicine is prepared from the following raw materials in parts by weight: 15-20 parts of sandalwood, 15-20 parts of Solms-Laubachia, 15-20 parts of Philippine violet herb, 10-15 parts of Aconitum tanguticum, 10-15 parts of Gentiana macrophylla flower and 10-15 parts of Qinghai gentiana scabra Bunge.

More preferably, the Tibetan medicine is prepared from the following raw materials: 740g of sandalwood, 740g of Solms-Laubachia, 740g of Philippine violet herb, 600g of Aconitum tanguticum, 600g of Gentiana macrophylla flower and 600g of Qinghai-Tibet gentiana scabra Bunge.

The preparation method of the pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis comprises the following operation steps: mixing sandalwood, Solms-Laubachia, purple chrysanthemum, Aconitum tanguticum, Gentiana macrophylla flower and Qinghai gentian, crushing into fine powder, and performing damp-heat sterilization at 115 ℃ for 30min to obtain the traditional Chinese medicine mixed powder, namely the pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis.

The pure traditional Chinese medicine Tibetan medicine is applied to preparing the medicine for treating chronic tracheitis.

The medicine for treating chronic tracheitis is a tablet, and is prepared by the following steps: adding brown sugar and xylitol into the Tibetan medicine, uniformly mixing, preparing a soft material by using ethanol with the volume percentage concentration of 50%, granulating by using a 16-mesh sieve, drying at 50 ℃, finishing granules, adding menthol and magnesium stearate, uniformly mixing, and tabletting to obtain the medicinal tablet for treating chronic tracheitis.

The addition amount of the brown sugar is 33 percent of the mass of the pure traditional Chinese medicine Tibetan medicine; the addition amount of the xylitol is 33 percent of the mass of the pure traditional Chinese medicine Tibetan medicine; the addition amount of the menthol is 0.2 percent of the mass of the pure traditional Chinese medicine Tibetan medicine; the addition amount of the magnesium stearate is 0.5 percent of the mass of the pure traditional Chinese medicine Tibetan medicine.

Aconitum tanguticum, also known as Aconitum tanguticum Maxim, Capsicum annuum, Aconitum sinomontanum, Aconitum kusnezoffii, Aconitum calomel, Aconitum sudanense; echinacea purpurea (also known as Kanba Sec.); Qinghai-Tibet gentian, also known as Bangbu Zhengbao; Solms-Laubachia, Lasiosphaera angustifolia, Lasiosphaera indica and Lasiosphaera oleracea.

The purple-flower Asiatic chrysanthemum in the formula is bitter in taste and cold in nature, has the effects of clearing away heat and toxic materials, relieving sore throat and swelling, moistening lung and relieving cough, and is a monarch drug; the Qinghai-Tibet gentian has the functions of detoxifying, detumescence and throat benefiting, and the Gentiana macrophylla flower has the functions of clearing heat, detoxifying, relieving sore throat and producing voice and is used as a ministerial drug for assisting the monarch drug in detoxifying and relieving sore throat; the radix linderae tangutici has the effects of clearing heat and removing toxicity, the Solms-Laubachia herb has the effects of clearing heat, moistening lung, relieving cough and reducing phlegm, and the radix linderae, the bitter and cool, can clear lung, relieve cough and reduce phlegm together, and the sandalwood, which is pungent in flavor, warm in nature, can promote qi circulation and relieve pain to treat chest pain discomfort, are adjuvant drugs; the whole formula has the effects of clearing heat and removing toxicity, relieving sore throat and reducing swelling, and relieving cough and reducing sputum, and is used for treating symptoms such as sore throat, hoarseness, cough, expectoration, chest and back pain and the like caused by sphagitis and bronchitis.

Compared with the prior art, the invention has the following advantages and beneficial effects:

(1) the pure traditional Chinese medicine Tibetan medicine has the advantages of small number of medicinal ingredients, simple process and high dissolution rate, and is beneficial to medicine production and quality control.

(2) The pure traditional Chinese medicine Tibetan medicine has a reasonable formula, has a synergistic interaction effect among the raw materials, and has the effect of treating bronchitis.

(3) The pure traditional Chinese medicine Tibetan medicine has no toxic or side effect, good safety, is not easy to generate drug resistance, can be taken for a long time, and is beneficial to keeping the normal functions of organisms.

Drawings

FIG. 1 is a standard graph of phenol red.

FIG. 2 is a graph showing the effect (100X) of the pure traditional Chinese medicine Tibetan medicine on the pathological morphology of the bronchopulmonary tissues of the chronic bronchitis rat, wherein A is a normal group, B is a sham operation group, C is a model group, D is a positive medicine group, E is a high dose group, F is a medium dose group, and G is a low dose group.

FIG. 3 is a graph showing the effect of the pure traditional Chinese medicine Tibetan medicine on MyD 88/NF-kB/ICAM-1 protein bands of rat bronchopulmonary tissues with chronic bronchitis, wherein A is a normal group, B is a sham operation group, C is a model group, D is a positive medicine group, E is a high-dose group, F is a medium-dose group, and G is a low-dose group.

FIG. 4 is a graph showing immunohistochemical staining results (100X) for MyD88, NF-. kappa.B and ICAM-1 proteins in rat bronchopulmonary tissues of example 3, wherein A is a normal group, B is a sham group, C is a model group, D is a positive drug group, E is a high dose group, F is a medium dose group, and G is a low dose group.

Figure 5 is a gentiopicroside dissolution profile in different dosage forms.

Detailed description of the invention

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.

The sources of the medicinal raw materials used in the following examples are as follows:

sandalwood: guangxi, the origin, was identified as dry heartwood of Santalbum L. It should meet the relevant regulations of Sandalwood item on page 380 of pharmacopoeia of the people's republic of China 2015 edition.

Solms-Laubachia: is identified as dry root or whole plant of Solma-Laubachia eurycarpa (Maxim.) Blfsch belonging to Brassicaceae. Collected in full-bloom stage, cleaned and dried. Should conform to the WS₃The rules of BiC-0029 and 9529 Solms Laubachia.

Purple-flower chrysanthemum: dried aerial parts of Asteraceae plant Echinacea purpurea or Echinacea tenuifolia.

Monkshood tangut: identified as RanunculaceaeDried whole plants of the plants Aconitum tanguticum (Maxim.) Stapf and Aconitum boehmeri Aconitum naviculare (Bruhl.) Stapf. Digging root in late summer and early autumn, removing impurities, and drying in shade. Should conform to the WS₃-BC-0085-9585, under the Aconitum tanguticum.

Large-leaved gentian flower: is identified as dried flower of Gentiana straminea Maxim. Collected in summer and autumn in flowering period, and dried in the shade. Should conform to the WS₃The prescription of Gentiana macrophylla flower at page-BC-0076-9576.

Qinghai-Tibet gentian: dried flowers of Gentiana Tinctoria Gentiana przewalski Maxim, Gentiana Tinctoria, Gentiana Przewalski Maxim, Gentiana, and Gentiana, Gentiana Przewalski Maxim. Collecting in full-bloom stage, removing impurities, cleaning, and drying in the sun. Should conform to the WS₃The rules under the section of-BC-0052-9552 page Qinghai-Tibet Gentiana.

Example 1:

a pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis is prepared from the following raw materials: 740g of sandalwood, 740g of Solms-Laubachia, 740g of Philippine violet herb, 600g of Aconitum tanguticum, 600g of Gentiana macrophylla flower and 600g of Qinghai-Tibet gentiana scabra Bunge. The preparation method of the Tibetan medicine comprises the following operation steps: mixing sandalwood, Solms-Laubachia, purple chrysanthemum, Aconitum tanguticum, Gentiana macrophylla flower and Qinghai gentian, crushing into fine powder, and performing damp-heat sterilization at 115 ℃ for 30min to obtain the traditional Chinese medicine mixed powder, namely the pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis.

Example 2: the pure traditional Chinese medicine Tibetan medicine has the effects of relieving cough and reducing sputum on mice

1. Materials used in this example:

1.1 animals:

100 ICR mice, each half of male and female, with body mass (20 +/-2) g; supplied by slaikatia laboratory animals ltd, Hunan province. Production license of experimental animal: SCXK (xiang) 2019-: SYXK (xiang) 2020-: 430727201101708058. feeding the seeds at the temperature of 23 +/-1 ℃ and the light and shade time table of 12: 12h alternating SPF barrier ambient conditions, free access to drinking water. The experimental procedures were carried out according to the SOP of the laboratory animal center of the institute of traditional Chinese medicine, Hunan province, all procedures were carried out according to the "nursing and use guide opinions of laboratory animals" made by the ministry of science and technology of China, and the serial numbers were approved: 2020-0092. Rats were allowed free access to water and food at constant temperature, humidity and light dark cycles (20-26 ℃; 40-70%; 07: 00-19: 00 light cycles).

1.2 drugs and reagents:

the test drugs are: the clinical application dose of the pure traditional Chinese medicine Tibetan medicine obtained in the embodiment 1 is 4 g/day, the mouse equivalent dose converted according to the body surface area is 0.52g/kg, and the high, medium and low doses of the pure traditional Chinese medicine Tibetan medicine are 4 times, 2 times and 1 time of the clinical equivalent dose; positive drugs: pentoxyverine citrate tablet (national pharmaceutical group, Capacity pharmaceutical Co., Ltd., batch No. 19022411, specification: 25mg, 9.75mg/kg), ambroxol hydrochloride sustained release tablet (cigarette Tai Dongcheng Dayang pharmaceutical Co., Ltd., batch No. 2024048, specification: 30mg, 0.024 g/kg); ammonia (Hunan Hui hong reagent, Inc.); sodium bicarbonate (mao chemical reagents, Tianjin); phenol red (shin & Fine chemical research institute, Tianjin).

1.3 Experimental instruments:

YLS-8A multifunctional cough-inducing asthma-inducing instrument (Jinan Yiyan science and technology development Co., Ltd.), and Envision2105 type enzyme-linked immunosorbent assay (PE Co., Ltd.).

2. The experimental methods and procedures used in this example:

2.1 mouse ammonia induced cough test:

after the mice are fed for 3 days adaptively, the mice are randomly divided into 5 groups, 10 groups are respectively a blank control group (normal saline), a positive control group (citric acid pentoxyverine tablets, 9.75mg/kg), a pure traditional Chinese medicine Tibetan medicine high, medium and low dose (2.08, 1.04 and 0.52g crude drug/kg) group, 7 days (dose 20mL/kg) are continuously administrated by gastric lavage, after l h is administrated for the last time, the mice are placed in a self-made closed glass bell jar, 0.2mL of concentrated ammonia water is sprayed on a cotton ball by an injector as uniformly as possible, the cotton ball is covered with a vessel for a certain time (about 10s) to induce cough, and the cough latency time and the cough frequency within 3min are recorded (note: the judgment of the cough of the mice is based on the fact that abdominal muscles are severely contracted and mouths are opened, and a slight cough sound is heard).

2.2 mouse phenol red excretion assay:

a certain amount of phenol red is accurately weighed and dissolved by adding a 5% sodium bicarbonate solution to prepare 1mL of solution containing 100 mu g, then the solution is sequentially diluted to 10.00, 5.00, 2.50, 1.25, 0.625 and 0 mu g.mL < -1 >, and the absorbance (A) is measured by a microplate reader at the position with the wavelength of 546 nm. A standard curve was prepared with phenol red dose (C) as abscissa and A as ordinate (as shown in FIG. 1). Taking 50 mice, randomly dividing into 5 groups, each group containing 10 mice, and respectively preparing blank control group (0.9% sodium chloride solution), ambroxol hydrochloride (0.024g/kg) positive control group, and high, medium and low dosage (2.08, 1.04, 0.52g crude drug/kg) groups of pure traditional Chinese medicine Tibetan medicine. Before administration, mice were fasted for 12h without water, and then administered by gavage at 20mL/kg body weight for 7 days continuously. 30min after the last administration, each mouse was intraperitoneally injected with 5% phenol red physiological saline solution at a dose of 0.01mL/g body weight, and 30min after injection, the mouse was sacrificed by dislocation of cervical vertebrae. Fixing the upper part of the trachea cannula on an operation plate, cutting the skin in front of the neck, separating the trachea, stripping the tissues around the trachea, cutting the lower edge of the thyroid cartilage to the trachea at the trachea bifurcation, separating a section of trachea with the same length from the lower part of the thyroid cartilage to the trachea bifurcation, putting each trachea section into a test tube which is previously filled with 2mL of 5 percent NaHCO3 solution, carrying out ultrasonic 35min, centrifuging at 3000r/min for 10min, taking the supernatant, measuring the absorbance A at the 546nm wavelength, and calculating the phenol red discharge amount by a standard curve method.

2.3 data processing:

SPSS17.0 is selected for statistical analysis, and the average + -standard deviation is adopted for measurement data

The method is characterized in that a t Test is adopted when the normality and the homogeneity of the variance are met by comparison between two groups, a Mann-Whitney Test is adopted when the normality or the homogeneity of the variance is not met by data, P < 0.05 shows that the statistical significance is achieved, and P < 0.01 shows that the tested difference has a very significant significance.

3. As a result:

3.1 influence of pure traditional Chinese medicine Tibetan medicine on mouse cough caused by ammonia water:

compared with a blank control group, after the pure traditional Chinese medicine Tibetan medicine dry drug is used, the cough latency of mice is obviously prolonged, the cough frequency within 3min is obviously reduced, and the effect is the best (P is less than 0.05 and less than 0.01) with high dose and is consistent with the effect of positive citric acid pentoxyverine tablets. The pure traditional Chinese medicine Tibetan medicine has a certain cough relieving effect. The results are shown in Table 1.

TABLE 1 influence of Tibetan medicine on cough caused by ammonia water

n＝10)

Comparison with blank control:^*P＜0.05，^**P＜0.01

3.2 influence of pure traditional Chinese medicine Tibetan medicine on mouse tracheal phenol red secretion:

taking the concentration of phenol red as an abscissa and the OD value as an ordinate, and calculating a regression equation Y of 0.0776x +0.0245 according to the concentration and the OD value of different phenol red standard solutions²When the content is 0.9998, the mouse tracheal phenol red secretion is obtained, and compared with a blank control group, the concentration of the pure traditional Chinese medicine Tibetan medicine high-dose mouse tracheal phenol red secretion is obviously increased (P is less than 0.05, and P is less than 0.01). The pure traditional Chinese medicine Tibetan medicine has a certain phlegm reducing effect.

TABLE 2 influence of pure traditional Chinese medicine Tibetan medicine on mouse tracheal phenol red secretion amount (

n＝10)

Comparison with blank control:^*P＜0.05

4. and (4) conclusion:

the experiment adopts a chemical stimulation method, and ammonia water is used for inhalation to stimulate parasympathetic nervous system to cause cough. Test results show that the pure traditional Chinese medicine Tibetan medicine 2.08g/kg has obvious inhibition effect on the cough induced by ammonia water, can effectively inhibit the cough frequency and prolong the cough incubation period, and has obvious effect of relieving cough.

The sputum is an important factor for causing cough, and can dilute the sputum, improve the increase of respiratory secretion and play a role in eliminating phlegm. The experiment adopts a tracheal phenol red excretion method to observe the phlegm eliminating effect of the pure traditional Chinese medicine Tibetan medicine, and the experiment result shows that after the pure traditional Chinese medicine Tibetan medicine is administrated by gastric lavage for 7 days, the secretion of the phenol red in the trachea of a mouse can be obviously promoted, the secretion of the tracheal sputum of the mouse is increased, and the phlegm eliminating effect is obvious.

Example 3: the invention discloses the curative effect of pure traditional Chinese medicine Tibetan medicine on a chronic bronchitis rat model

1. Materials used in this example:

1.1 animals:

SD rat, 70, male, SPF grade, body mass 180-; supplied by slaikatia laboratory animals ltd, Hunan province. Production license of experimental animal: SCXK (xiang) 2019-: SYXK (xiang) 2020-: 430727211100275116. feeding the seeds at the temperature of 23 +/-1 ℃ and the light and shade time table of 12: 12h alternating SPF barrier ambient conditions, free access to drinking water. The experimental procedures were carried out according to the SOP of the laboratory animal center of the institute of traditional Chinese medicine, Hunan province, all procedures were carried out according to the "nursing and use guide opinions of laboratory animals" made by the ministry of science and technology of China, and the serial numbers were approved: 2021-0007. Rats were allowed free access to water and food at constant temperature, humidity and light dark cycles (20-26 ℃; 40-70%; 07: 00-19: 00 light cycles).

1.2 drugs and reagents:

the test drugs are: the clinical dosage of the pure traditional Chinese medicine Tibetan medicine obtained in the example 1 is 4 g/day, the rat equivalent dosage converted according to the body surface area is 0.36g crude drug/kg, and the high, medium and low dosages of the pure traditional Chinese medicine Tibetan medicine are 4 times, 2 times and 1 time (1.44, 0.72 and 0.36g crude drug/kg) of the clinical equivalent dosage; positive drugs: guilong Ruikuning tablet (Jiangxi Yaerren and pharmacy Co., Ltd., batch number: 200403, specification: 0.41g, 1.0 g/kg); lipopolysaccharide (LPS, Sigma Co., Lot: L2880, Specification: 100 mg); 10% of neutral formaldehyde; TNF-alpha and IL-10Elisa kits (Elapscience, Inc., batch: E-EL-R2856c, E-EL-R0016 c); MyD88 primary antibody (ABClona, lot: A0786), NF-. kappa.B primary antibody (Wuhan Proteintetech, lot: 66535-1-Ig), ICAM-1 primary antibody (Wuhan Proteintetech, lot: 10020-1-AP); SuperECL Plus hypersensitivity luminescent liquid (Advansta, USA, batch number: K-12045-D50).

1.3 Experimental instruments:

envision2105 microplate reader (PE); RM2245 rotary microtomes (Leica, germany); an Olympus BX53 type biomicroscope (Olympus corporation); 7DZ5-WS type low temperature low speed centrifuge (Hunan instrument laboratory instruments, Hunan); PANNORAMIC panorama section scanner (hungarian 3DHISTECH corporation); electrophoresis apparatus (six one in Beijing, China); horizontal agarose electrophoresis tank (six one in Beijing, China); ChemiDoc^TMXRS + -type gel imaging system (Bio-Rad, USA).

2. The experimental methods and procedures used in this example:

2.1 establishment of rat model of chronic bronchitis:

after the rats are adaptively fed for 5 days, the weight of the rats is randomly divided into: normal group (n-8), sham group (n-8) and model group (n-54), model group modeling method: the rats are placed in a smoking box for smoking twice every day, once in the morning and afternoon, each time is 1 hour, the smoking amount is 10 cigarettes each time, and the interval between the two smoking times is 4 hours (smoking is cancelled on the day of operation). 15d, model group rats were anesthetized with 1% sodium pentobarbital solution (50mg/kg) intraperitoneally, fixed on the plate, glottis was exposed, 200 μ g (2 μ g/μ l) of LPS solution was rapidly injected into the trachea, then the fixed plate was vertically rotated to uniformly distribute lipopolysaccharide to both lungs, and after the model was made, the rats were fasted for 6 hours without water restriction.

2.2 grouping and dosing:

after smoking for 15 days, the rats in the model group are randomly divided into a model group (n is 12), a positive drug (1.0 g/kg) group and a pure traditional Chinese medicine Tibetan drug high, medium and low dose (1.44, 0.72 and 0.36g/kg) group according to the weight, and 10 rats in each group are taken. The administration group started gavage administration intervention 15 days after the smoking treatment, on day 16 (D16), while continuing smoking treatment, and collected samples 30D after the administration.

2.3 index detection:

2.3.1 general behavioral, signs and anatomical observations:

the animal body mass is weighed every week during the experiment process to adjust the dosage, and the conditions of the back hair, eyes, ears, tails, mental state, activity and the like of the animals are observed in detail.

2.3.2 sample Collection and preparation:

because each group of animals had a certain mortality rate after surgery, the next day after dosing, 7 rats were selected from each group, anesthetized with 1% pentobarbital sodium solution (50mg/kg), bled from the abdominal aorta, centrifuged (3500rpm, 10min), and the supernatant was taken and stored at-80 ℃ for future use. Then, opening the chest cavity of the rat, completely taking out all lung tissues, and freezing and storing the right lung by liquid nitrogen for later use; left lung tissue was fixed in 10% neutral formaldehyde solution for future use.

2.3.3 serum inflammatory factor assay:

after unfreezing the frozen serum sample at-20 ℃ and 4 ℃, balancing the kit to room temperature, preparing a washing solution, a standard working solution, a biotinylated antibody working solution, a medium and a substance working solution, loading according to the steps of an Elisa kit specification, incubating, washing a plate, and detecting the content change of TNF-alpha and IL-10 in the serum.

2.3.4 bronchopulmonary histopathological section HE staining:

1) preparing paraffin sections of the bronchus and the left lung tissue;

2) baking slices at 60 ℃ for 12 h;

3) slice dewaxing to water: the slices were first placed in xylene for 15min, 3 times. Then, the mixture was placed in 100%, 100%, 95%, 85% and 75% ethanol in turn for 5min per stage. Washing with distilled water for 5 min;

4) dyeing with hematoxylin for 1min, washing with distilled water, and bluing with PBS;

5) eosin staining for 0.5min, washing with distilled water;

6) gradient alcohol (95-100%) dehydration, 5min per stage (or baking the slices directly). Taking out, placing in xylene for 10min, sealing with neutral gum, and observing with microscope for 2 times;

7) the main manifestations of experimental modeling include inflammatory cell infiltration around the bronchus, whether exudate exists in the lumen, alveolar interstitial inflammation and alveolar emphysema change, whether the bronchus is dilated, etc.;

8) and (4) observing and analyzing by taking the lesion indexes of the chronic bronchitis as observation bases. Degree of bronchitis infiltration: and observing the cell infiltration condition of all bronchitis in the section, counting the number of inflammatory cell infiltration bronchi and non-infiltration bronchi, and performing statistical comparison.

2.3.5 Western Blot method for quantitatively detecting MyD 88/NF-kB/ICAM-1 protein expression:

1) protein extraction

Shearing 0.025g of tissue, washing the tissue with ice-precooled PBS, adding 300uL of RIPA lysate into a biological sample homogenizer, and repeatedly grinding the tissue until no tissue block can be seen; cracking for 10 minutes on ice; centrifuge at 12000rpm for 15min at 4 ℃. (centrifuge precooling in advance); the centrifuged supernatant was transferred to a 1.5mL centrifuge tube.

2) Glue making

Preparing 10% separation gel, adding TEMED, immediately shaking up, and filling gel. After pouring, sealing with isopropanol. Gelatin is said to have coagulated when the slightly inclined gel maker demarcation line no longer changes. Waiting for another 3min for the glue to solidify sufficiently, pouring off isopropanol on the upper layer of the glue and sucking it dry with filter paper. 4.8 percent of concentrated glue is prepared, and the concentrated glue is immediately shaken up after TEMED is added, so that glue can be filled. And (5) inserting the comb into the glass plate, filling the residual space with the concentrated glue, and waiting for the glue to solidify.

3) Sample preparation

And (3) taking 160uL of protein supernatant, adding 40uL of 5-loading buffer, uniformly mixing, boiling in boiling water for 5min, and placing in an ice box for quick cooling for later use.

4) Electrophoresis

The first well was spotted with 2uL of marker, and 10-20uL of denatured protein was loaded into each void. The electrophoresis was started, and the electrophoresis was performed at a constant voltage of 75V for 130 min. And stopping electrophoresis when the bromophenol blue electrophoresis reaches the bottom of the gel.

5) Rotary film

MyD88(37KD), NF-KB (65KD), ICAM-1(95-100KD) and actin (42KD) were cut separately. 6 pieces of filter paper having the same size as the glue and 1 piece of NC membrane were prepared, and the NC membrane was placed in the membrane transfer buffer together with the filter paper until the filter paper was completely soaked. 3 filter papers, NC membrane, glue and another 3 filter papers are put in sequence, and no air bubble is required in the middle. Covering the instrument, switching on the power supply, transferring the membrane to 300mA constant current, MyD88 for about 57min, NF-KB for about 85min, ICAM-1 for about 120min, actin for about 62 min. After the membrane transfer was completed, the membrane was taken out and washed in 1 × PBST 1 time.

6) Sealing of

5% skimmed milk powder was prepared from 1 × PBST, and the membrane was immersed and allowed to stand at room temperature for 90 min.

7) Primary antibody incubation

The primary antibody was diluted with 1 × PBST (specific ratios are shown in table below), and the membrane was incubated with the primary antibody for 60min, overnight at 4 ℃, and then left at room temperature for 30min the next day. At the end of incubation, 1 × PBST wash 3 times for 15min each.

8) Incubation with secondary antibody

HRP-labeled secondary antibodies (specific ratios shown in the table below) were diluted with 1 × PBST and the diluted secondary antibodies were incubated with the membrane for 90min at room temperature. At the end of incubation, 1 × PBST wash 3 times for 10min each.

9) Color development/Exposure

ECL color development exposure: incubating ECL chemiluminescence solution with membrane for 5min, sucking out liquid with filter paper, wrapping the membrane with plastic packaging membrane, and exposing with X-ray film in dark box for 1-30 min; and (5) developing and washing.

2.3.6 immunohistochemistry method for detecting MyD 88/NF-kB/ICAM-1 protein expression:

1) taking fixed lung tissues for paraffin embedding, and cutting tissue sections with the thickness of 0.4 mu m by an automatic slicer; baking slices at 60 ℃ for 12 h;

2) slice dewaxing to water: the slices were first placed in xylene for 20min, 3 times. Then, the mixture was placed in 100%, 95%, 85% and 75% ethanol in turn for 5min per stage. Washing with distilled water for 5 min;

3) heat-repair antigen: soaking the slices in 0.01M citrate buffer solution (pH6.0), heating in electric furnace or microwave oven to boil, cutting off power, continuously decocting for 20min, cooling for 20min, taking out, and cooling to room temperature. After cooling, washing with 0.01M PBS (pH7.2-7.6) for 3min x 3 times;

4) 1% periodic acid was added at room temperature for 10 minutes to inactivate endogenous enzymes. Washing with PBS for 3min x 3 times; incubating the primary antibody: primary antibody (ICAM-1, MYD88, NF-KB) diluted appropriately was added dropwise over night at 4 ℃. Washing with PBS for 5min x 3 times;

5) incubation of secondary antibody: dripping 50-100 mu L of anti-rabbit, rabbit-IgG antibody-HRP polymer, incubating at 37 ℃ for 30min, and washing with PBS for 5min x 3 times;

6) DAB color development: dropwise adding 50-100 mu L of prepared color-developing agent DAB working solution, incubating at room temperature for 1-5 min, controlling the reaction time under a mirror, and washing with distilled water;

7) counterstaining with hematoxylin for 5-10 min, washing with distilled water, and returning blue with PBS;

8) dehydrating with 60-100% alcohol for 5min each stage. Taking out, placing in xylene for 10min, 2 times, sealing with neutral gum, and observing with microscope.

2.4 statistical treatment:

3. Experimental results for this example:

3.1 influence of pure traditional Chinese medicine Tibetan on the general condition of chronic bronchitis rats:

after the model is made, the fur gloss of the model group rat is gradually lost, the activity is reduced, the mood is tired, the water intake and the urine output are obviously increased, after three weeks, the airway wheezing sound and the phlegm wheezing sound can be heard, and the weight is obviously reduced; after the administration of pure traditional Chinese medicine Tibetan medicine, the wheeze sound of the air passage is obviously relieved. See table 3.

TABLE 3 influence of pure traditional Chinese medicine Tibetan medicine on body weight of chronic bronchitis rat (

n＝7)

Note: compared with the group of the pseudo-operation,^*P＜0.05，^**P＜0.01。

3.2 influence of pure traditional Chinese medicine Tibetan medicine on the contents of TNF-a and IL-10 in serum of rats with chronic bronchitis:

compared with the normal group and the false operation group, the content of TNF-a in the serum of the rat in the model group is obviously increased, and the content of IL-10 is obviously reduced; compared with the model group, the pure traditional Chinese medicine Tibetan medicine group with high and medium dosage can obviously reduce the content of serum TNF-a, but has no statistical difference (P is less than 0.05, P is less than 0.01) on the change of the content of IL-10. See table 4.

TABLE 4 influence of pure traditional Chinese medicine Tibetan medicine on TNF-a and IL-10 content in serum of rat with chronic bronchitis: (

n＝7)

Note: compared with the group of the pseudo-operation,^*P＜0.05，^**p is less than 0.01; in comparison with the set of models,^#P＜0.05，^##P＜0.01。

3.3 the effect of the pure traditional Chinese medicine Tibetan medicine on the pathological morphology of the bronchial lung tissue of the chronic bronchitis rat:

the normal group of rats has complete bronchial mucosa epithelial cells, no congestion and edema phenomenon, regular cell arrangement and no obvious pathological change of pulmonary tissue structure alveolus;

the model group rat bronchial mucosa epithelial cells are seriously damaged, a large amount of necrosis and desquamation exist, goblet cells are proliferated, a large amount of fibrous tissues on the tube wall are proliferated, the tube wall is obviously thickened, the tube cavity is narrow, the tube cavity is filled with secretion and exudates, the alveolar septa are widened, and inflammatory cell infiltration, capillary vessel expansion and congestion are realized to different degrees.

Compared with the model group, the positive medicine group rat has obvious reduction of pathological injury and inflammatory change of lung tissues, no serious damage of bronchial mucous epithelium, few necrotic cast-off cells and exudates in lumens and alveolar cavities, a small amount of fibrous tissues and inflammatory cell infiltration, and no obvious edema and congestion.

The rat bronchus mucosa epithelium of the high-dose group of the pure traditional Chinese medicine Tibetan medicine has a small degree of damage, a small amount of necrotic exfoliated cells and exudates are arranged in a lumen and an alveolar cavity, goblet cells are slightly proliferated, pathological damage and inflammatory change of lung tissues are reduced, the wall of the tube is thickened, alveolar septa are widened, and a small amount of fibrous tissues, inflammatory cell infiltration and edema are observed. See fig. 2 and table 5.

TABLE 5 influence of Tibetan medicine on chronic bronchitis in rat cell infiltration

n＝3)

3.4 Effect of pure traditional Chinese medicine Tibetan medicine on MyD 88/NF-kB/ICAM-1 protein expression of rat bronchopulmonary tissues of chronic bronchitis:

the Western Blot method quantitatively detects MyD 88/NF-kB/ICAM-1 protein gray values to find that compared with a sham operation group, the gray values of rat bronchopulmonary tissues MyD88, NF-kB and ICAM-1 protein of a model group are obviously increased (P is less than 0.05, and P is less than 0.01); compared with the model group, the gray values of the proteins MyD88, NF-kappa B and ICAM-1 of the rat bronchopulmonary tissues of the positive drug group and the pure traditional Chinese medicine Tibetan drug group are obviously reduced (P is less than 0.05, and P is less than 0.01). See fig. 3, table 6.

TABLE 6 influence of pure traditional Chinese medicine Tibetan medicine on MyD 88/NF-kB/ICAM-1 protein gray level value of rat bronchopulmonary tissue of chronic bronchitis ((

n＝3)

Note: compared with the group of the pseudo-operation,^*p is less than 0.05; in comparison with the set of models,^#P＜0.05，^##P＜0.01。

the immunohistochemical method detects the positive expression condition of MyD 88/NF-kB/ICAM-1 protein in rat bronchopulmonary tissues, the positive expression is carried out on yellow-brown particles in protein cytoplasm and nucleus, and the negative expression is carried out on blue-purple. Compared with a sham operation group, the positive rate of MyD88, NF-kappa B and ICAM-1 protein expression in rat bronchopulmonary tissues of the model group is obviously increased (P is less than 0.05, and P is less than 0.01); compared with the model group, the positive rate of the rat bronchopulmonary tissue MyD88, NF-kappa B and ICAM-1 protein expression of the positive drug group and the pure traditional Chinese medicine Tibetan drug group is obviously reduced, and the difference has statistical significance (P is less than 0.05 and P is less than 0.01). Consistent with the trend of results described above. See fig. 4, table 7.

TABLE 7 influence of pure traditional Chinese medicine Tibetan medicine on MyD 88/NF-kB/ICAM-1 protein expression positive rate of rat bronchopulmonary tissue of chronic bronchitis

(

n＝3)

4. conclusion

The pure traditional Chinese medicine Tibetan medicine can effectively improve the inflammatory infiltration of bronchopulmonary tissues, possibly reduce the release of inflammatory mediators TNF-a and the like, and possibly is related to the regulation and control of MyD 88/NF-kB/ICAM-1 signal channel expression.

Example 4: acute toxicity test of pure traditional Chinese medicine Tibetan medicine

1. Materials used in this example:

1.1 medicine:

the tested drugs are: the clinical dosage of the pure traditional Chinese medicine Tibetan medicine obtained in the example 1 is 4.0g crude drugs per day. During experiment, the medicine is prepared into the mouse with purified water, and the maximum concentration of the medicine for gastric lavage is 0.19 g/mL.

1.2 animals:

ICR mice, SPF grade, male and female half, weight 15 ~ 17g, 46. Supplied by the experimental animals of slagoka, Hunan province. Animals are placed in an SPF laboratory for breeding, the room temperature is 20-25 ℃, and the relative humidity is 50-70%; production license of experimental animal: SCXK (xiang) 2019-: SYXK (xiang) 2020-: 430727201101541286. feeding the seeds at the temperature of 23 +/-1 ℃ and the light and shade time table of 12: 12h alternating SPF barrier ambient conditions, free access to drinking water. The experimental procedures were carried out according to the SOP of the laboratory animal center of the institute of traditional Chinese medicine, Hunan province, all procedures were carried out according to the "nursing and use guide opinions of laboratory animals" made by the ministry of science and technology of China, and the serial numbers were approved: 2020-0082. Rats were allowed free access to water and food at constant temperature, humidity and light dark cycles (20-26 ℃; 40-70%; 07: 00-19: 00 light cycles).

2. Methods and steps used in the present example

2.1 preliminary experiments

6 animals to be tested are selected, half of the animals are selected, the animals are fasted for more than 12 hours before the experiment, the pure traditional Chinese medicine Tibetan medicine (0.19g/ml) suspension with the maximum concentration is given for intragastric administration (ig), the maximum volume is given for 40ml/kg, and the administration is carried out 2 times at intervals of 6 hours in one day.

2.2 official experimental dose setting

According to the results of acute toxicity preliminary experiments, since LD can not be obtained₅₀And limiting the drug concentration, the test adopts a maximum dose method, 40 ICR mice are selected and randomly divided into 2 groups, namely a control group and a pure traditional Chinese medicine Tibetan medicine group, each group is 20, fasting is carried out for more than 12 hours before the test, the mice of the administration group are subjected to intragastric perfusion (ig) by using pure traditional Chinese medicine Tibetan medicine suspension with the maximum concentration of 0.19g/ml, the maximum volume is 40ml/kg, purified water with the same volume as that of the intragastric perfusion of the control group is administered for 2 times at intervals of 6 hours in one day, and the cumulative administration dose is 15.2g/kg, which is detailed in a table 8.

TABLE 8 Experimental dose and group design

2.3 observation time and content:

animals were observed daily after dosing. On the day of administration, particularly, the poisoning performance and characteristics, the occurrence and recovery time of toxic reactions, the death condition and the like of each group of animals are closely observed and recorded within 0-4 hours after administration, and then the animals are observed for 2 times every day, 1 time in the morning and afternoon and continuously observed for 14 days.

The observation indexes include: animal appearance, behavioral activity, secretions, excretions, eating conditions, mortality (time to death, pre-mortem response), and the like. If the symptoms appear, the observation frequency is increased so as to observe the symptoms of the toxic reaction of the animals in detail, and the starting time, the severity, the duration and the reversibility of the symptoms. If the animal is dead or dying, the animal should be observed anatomically in time. At the end of the experiment, animals were weighed and dissected roughly, and the surface and sections of the major organs, such as liver, kidney, spleen, heart, brain, lung, etc., were observed visually, and when the general dissection was abnormal, the abnormal organs were recorded and histopathological examination was performed.

The body weight of the experimental animals was weighed before the administration on the day of administration and on the 4 th, 7 th, 10 th and 14 th days after the administration.

2.4 statistical analysis of data:

3. The experimental results are as follows:

3.1 Effect on weight gain in mice

Compared with a control group, the weight difference of the mice in the pure traditional Chinese medicine Tibetan medicine group has no statistical significance; the weight loss of the male mice at 7d after the administration is obviously different (P is less than 0.05); there was no significant difference after 10d and 14d (P > 0.05). See tables 9, 10, 11.

TABLE 9 Effect of pure Chinese medicinal Tibetan medicine on mouse body weight

g)

TABLE 10 Effect of pure Chinese medicinal Tibetan on female mouse weight

g)

Note: p < 0.05 compared to control group

TABLE 11 Effect of pure Chinese medicinal Tibetan medicine on body weight of Male mice: (

g)

Note: p < 0.05 compared to control group

3.2, effects on mouse major organs:

the results of animal dissection and visual observation show that no obvious abnormality is found in the tissues and organs of the heart, liver, spleen, lung, brain, kidney and the like of each group of animals.

4. And (4) conclusion:

after 14 days of observation after administration, no abnormal behavior of mice or animal death is observed in the pure traditional Chinese medicine Tibetan medicine and the control group. After the animals are sacrificed and dissected, no abnormality is found in heart, liver, spleen, lung, kidney, brain and the like through visual observation, which shows that the maximum tolerance of the pure traditional Chinese medicine Tibetan medicine can reach 15.2g/kg, which is 266 times of the clinical dosage of 70kg adults according to the conversion of kilogram body weight and 30.00 times according to the conversion of body surface area. See table 12.

TABLE 12 results of acute toxicity test of Tibetan medicine animals

5. Discussion of the related Art

The administration dosage is 0.19g crude drug/ml, when the dosage is taken twice a day, the behavior and the activity of mice are normal, the feces and the urine are normal, and the body weight of male animals is obviously reduced on the 7 th day, presumably because the content of fiber of the medicinal powder is high, the digestive function of the animals is influenced to a certain extent, but the difference of the total body weight has no statistical significance. The tissue and the organ of each group of animals are observed by naked eyes without obvious abnormality. The maximum tolerance of the mouse is measured to be 15.2g crude drug/kg, the clinical dosage of the mouse is 4g crude drug/day according to human, the maximum tolerance is 266 times of the clinical dosage of 70kg adult, and the maximum tolerance is 30.00 times of the clinical dosage according to the calculation of body surface area. No obvious toxic or side effect is seen, which shows that the pure traditional Chinese medicine Tibetan medicine is safe and reliable to take under the specified dosage.

Example 5: medicine tablet for treating chronic tracheitis

740g of sandalwood, 740g of Solms-Laubachia, 740g of Philippine violet herb, 600g of aconitum tanguticum, 600g of large-leaved gentian flower and 600g of Qinghai-Tibetan gentian are mixed, crushed into fine powder and sterilized by moist heat at 115 ℃ for 30min to obtain traditional Chinese medicine mixed powder; adding brown sugar which accounts for 33 percent of the mass of the traditional Chinese medicine mixed powder and xylitol which accounts for 33 percent of the mass of the traditional Chinese medicine mixed powder into the traditional Chinese medicine mixed powder, uniformly mixing, preparing a soft material by 50 percent of ethanol, granulating by a 16-mesh sieve, drying at 50 ℃, finishing granules, adding menthol which accounts for 0.2 percent of the mass of the traditional Chinese medicine mixed powder and magnesium stearate which accounts for 0.5 percent of the mass of the traditional Chinese medicine mixed powder, uniformly mixing, and tabletting to obtain 1000 tablets of the medicine for treating chronic tracheitis.

Example 6:

sample 1 buccal tablets: 740g of sandalwood, 740g of Solms-Laubachia, 740g of Philippine violet herb, 600g of aconitum tanguticum, 600g of large-leaved gentian flower and 600g of Qinghai-Tibetan gentian are mixed, crushed into fine powder and sterilized by moist heat at 115 ℃ for 30min to obtain traditional Chinese medicine mixed powder; adding brown sugar accounting for 33 percent of the mass of the traditional Chinese medicine mixed powder and xylitol accounting for 33 percent of the mass of the traditional Chinese medicine mixed powder into the traditional Chinese medicine mixed powder, uniformly mixing, preparing a soft material by 50 percent of ethanol, granulating by a 16-mesh sieve, drying at 50 ℃, finishing granules, adding menthol accounting for 0.2 percent of the mass of the traditional Chinese medicine mixed powder and magnesium stearate accounting for 0.5 percent of the mass of the traditional Chinese medicine mixed powder, uniformly mixing and tabletting.

Sample 2 water-honeyed pill: 740g of sandalwood, 740g of Solms-Laubachia, 740g of Philippine violet herb, 600g of aconitum tanguticum, 600g of large-leaved gentian flower and 600g of Qinghai-Tibetan gentian are mixed, crushed into fine powder and sterilized by moist heat at 115 ℃ for 30min to obtain traditional Chinese medicine mixed powder; adding 50g of Mel and appropriate amount of water into 100g of powder, making into pill, and drying at 60 deg.C.

Chromatographic conditions are as follows: a chromatographic column: sepax Bio-C18 (4.6X 250mm, 5 μm) SN: 9F18808, PN: 106185-4625; mobile phase methanol (a): 0.1% phosphoric acid water (B) 20: 80; flow rate: 1 ml/min; column temperature: 30 ℃; the detection wavelength is 270 nm; sample introduction amount: 5 μ l.

Preparation of control solutions

Taking appropriate amount of gentiopicroside reference substance, precisely weighing, and adding methanol to obtain solution containing 0.50mg per 1ml, to obtain reference substance solution.

And (3) determination of dissolution rate: the pulp method is adopted, the rotating speed is 100r/min, and the temperature is (37.0 +/-2) DEG C. Taking 900ml of purified water subjected to ultrasonic treatment, 6 parts of each purified water; samples 1 are taken, proper amount of buccal tablets and proper amount of water-honeyed pills of the samples 2 are taken, 3 parts of each group are respectively placed in dissolution media for testing. Sampling 5ml at fixed point for 5, 10, 15, 20, 30, 45, 60, 90min, 2h, immediately supplementing 5ml of fresh pure water solution at the same temperature, filtering the filtrate with 0.45 μm microporous membrane, collecting the filtrate, measuring according to the above chromatographic conditions, sampling 5 μ l, and calculating cumulative dissolution percentage. The dissolution profile is shown in FIG. 5.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. A pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis is characterized in that: the Tibetan medicine is prepared from the following raw materials: sandalwood, Solms-Laubachia, purple chrysanthemum, Aconitum tanguticum, Gentiana macrophylla flower and Qinghai gentian.

2. The Tibetan medicine for treating chronic tracheitis of claim 1, wherein the Tibetan medicine comprises the following components: the Tibetan medicine is prepared from the following raw materials in parts by weight: 15-20 parts of sandalwood, 15-20 parts of Solms-Laubachia, 15-20 parts of Philippine violet herb, 10-15 parts of Aconitum tanguticum, 10-15 parts of Gentiana macrophylla flower and 10-15 parts of Qinghai gentiana scabra Bunge.

3. The Tibetan medicine for treating chronic tracheitis of claim 1, wherein the Tibetan medicine comprises the following components: the Tibetan medicine is prepared from the following raw materials: 740g of sandalwood, 740g of Solms-Laubachia, 740g of Philippine violet herb, 600g of Aconitum tanguticum, 600g of Gentiana macrophylla flower and 600g of Qinghai-Tibet gentiana scabra Bunge.

4. The preparation method of the pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis according to any one of claims 1 to 3, which is characterized by comprising the following operation steps: mixing sandalwood, Solms-Laubachia, purple chrysanthemum, Aconitum tanguticum, Gentiana macrophylla flower and Qinghai gentian, crushing into fine powder, and performing damp-heat sterilization at 115 ℃ for 30min to obtain the traditional Chinese medicine mixed powder, namely the pure traditional Chinese medicine Tibetan medicine for treating chronic tracheitis.

5. The use of a pure traditional Chinese medicine Tibetan medicine according to any one of claims 1 to 3 in the preparation of a medicament for treating chronic tracheitis.

6. Use according to claim 5, characterized in that: the medicine for treating chronic tracheitis is a tablet, and is prepared by the following steps: adding brown sugar and xylitol into the Tibetan medicine, uniformly mixing, preparing a soft material by using ethanol with the volume percentage concentration of 50%, granulating by using a 16-mesh sieve, drying at 50 ℃, finishing granules, adding menthol and magnesium stearate, uniformly mixing, and tabletting to obtain the medicinal tablet for treating chronic tracheitis.

7. Use according to claim 6, characterized in that: the addition amount of the brown sugar is 33 percent of the mass of the pure traditional Chinese medicine Tibetan medicine; the addition amount of the xylitol is 33 percent of the mass of the pure traditional Chinese medicine Tibetan medicine; the addition amount of the menthol is 0.2 percent of the mass of the pure traditional Chinese medicine Tibetan medicine; the addition amount of the magnesium stearate is 0.5 percent of the mass of the pure traditional Chinese medicine Tibetan medicine.