Composition for resisting cold and respiratory tract injury and preparation method thereof

Abstract

The invention belongs to the technical field of food and traditional Chinese medicine health care, and discloses an anti-cold and respiratory tract injury composition and a preparation method thereof, and the anti-cold and respiratory tract injury composition is prepared from the following medicinal materials in parts by mass: 2-10 parts of liquorice, 3-16 parts of sea-buckthorn and 3-9 parts of pericarpium citri reticulatae. According to the composition disclosed by the invention, by regulating and controlling a maternal expression gene 3 (MEG3) / micro RNA-223 (miR-223) / nucleotide to be combined with an oligomerization domain-like receptor protein 3 (NLRP3) axis, the effects of resisting cold and lung injury and treating symptoms such as cough and asthma can be achieved. The activity of the composition is improved, the composition has an innovative pharmacological mechanism, and the composition reduces the expression of MHC-II on the surface of pulmonary epithelial cells (AEC) so as to promote the amplification of CD4 + TRM cells, so that the lung immunity and young state are enhanced, the anti-cold activity is generated, and the composition does not have the activity similar to a compound. The composition and the preparation thereof have the effects of resisting cold and virus infection and improving the immune young state respiratory injury resisting activity, the curative effect is changed, and the mechanism is novel.

Description

Technical Field

[0001] The invention belongs to the fields of food, traditional Chinese medicine and health, and in particular relates to a composition for resisting colds and respiratory tract injuries and a preparation method thereof. Background Art

[0002] Colds, as defined by modern medicine, include the common cold and influenza. Influenza is an acute respiratory infectious disease caused by influenza, rhinovirus, coxsackievirus, and other viruses. It is highly contagious and can sometimes cause high fever, multiple body aches, significant fatigue, and mild respiratory symptoms. The disease is generally self-limiting. The early symptoms of the common cold include upper respiratory tract catarrhal symptoms: sneezing, runny nose, dry mouth, nose, and throat, sore throat, and a dry cough. After the virus infects the host, it replicates rapidly within eight hours, leading to the onset of catarrhal symptoms. Subsequently, the immune response is activated, resulting in aversion to cold. If a cold is also present, microcirculatory disturbances worsen, the condition worsens, and the prognosis is poor. The prominent symptoms of influenza are acute onset of chills, high fever, and generalized muscle aches. Headaches are also prominent, but upper respiratory tract catarrhal symptoms are relatively mild.

[0003] Currently, Western medicine often uses anti-inflammatory, anti-allergic, and anti-asthmatic therapies to treat various respiratory diseases, including the common cold, influenza, bronchitis, coughs and spitting, and asthma, as well as their derivatives. However, the results are often unsatisfactory, treating symptoms rather than the root cause, making it difficult to cure the disease and often accompanied by various side effects. Long-term use can easily lead to drug resistance, and in severe cases, surgery is required, which is expensive and causes great financial, physical, and mental pain to the patient. Traditional Chinese medicine treatment often requires a long course of treatment, and in the treatment of seriously ill patients, Traditional Chinese Medicine is often used only as an auxiliary treatment method to Western medicine, and cannot independently treat the condition. For patients with long-term and severe symptoms, the method of uniformly increasing the number of medications and the dosage is used to maintain the therapeutic effect, which places a great burden on organ metabolism.

[0004] Licorice, also known as guolao, micao, migan, and meicao, is the dried root and rhizome of the leguminous plant Glycyrrhiza uralensis, Glycyrrhiza glabra, or Glycyrrhiza glabra. It is sweet, mild, and non-toxic, and has the effects of tonifying the spleen and replenishing qi, clearing away heat and detoxifying, relieving pain, and harmonizing various medicinal herbs. my country was the first country in the world to recognize and study licorice, with its use documented in the Shennong Bencao Jing. Licorice plays a vital role in prescription combinations and is the most frequently used in traditional Chinese medicine (TCM) prescriptions, earning it the title of "King of Chinese Medicine." Many classic prescriptions feature licorice as a key ingredient, such as the famous Shaoyao Gancao Decoction from the Treatise on Febrile Diseases. Licorice is also frequently used in the preparation and combination of TCM herbs, such as when combined with aconite root. The diester alkaloids in aconite root are both its active and toxic components, which can cause severe cardiotoxicity if taken improperly. Licorice combined with licorice can mitigate toxicity and enhance efficacy by affecting the absorption, distribution, metabolism, and excretion of these toxic components. Licorice's chemical components primarily include triterpenoid saponins, flavonoids, and polysaccharides, exhibiting anti-inflammatory, anti-tumor, antibacterial, antiviral, antioxidant, immunomodulatory, anti-cardiovascular, anti-diabetic, and anti-liver damage effects. Licorice is sweet and mild in nature, possessing numerous health benefits, making it an economically valuable medicinal and edible plant. Rich in trace elements, it offers significant health benefits. However, the efficacy of licorice and its formulations remains suboptimal and in urgent need of improvement, failing to meet clinical needs. Through experimental discovery, new formulations, targets, mechanisms, and uses for licorice could ultimately benefit patients with various upper respiratory tract diseases, cough, wheezing, and other susceptible conditions. These innovations have significant scientific and practical value. There are reports of licorice treating colds and influenza, and also reports of its beneficial effects on lung injury recovery. However, we have discovered that fermentation of a compound extract of licorice, dried tangerine peel, and seabuckthorn exhibits novel anti-cold efficacy, resulting in a compositional shift.

[0005] Tangerine peel (Citrus tangerine) is the dried, mature peel of the citrus plant and its cultivated varieties, both of the Rutaceae family. It primarily contains flavonoids, volatile oils, alkaloids, and trace elements. Currently, extensive research has focused on its volatile oils, flavonoids, and trace elements. The volatile oils primarily contain D-limonene, β-myrcene, and α-pinene. Flavonoids include hesperidin, neohesperidin, tangeretin, dihydronobiletin, and 5-nordihydronobiletin. Trace elements include potassium, sodium, calcium, magnesium, copper, zinc, iron, and strontium. Modern pharmacological research has demonstrated that tangerine peel exhibits a variety of pharmacological effects, including antibacterial, anti-inflammatory, antioxidant, anti-tumor, digestive, expectorant, hepatoprotective, antihypertensive, and neuroprotective properties. In recent years, as one of the first ingredients to be used as both a medicine and a food, its medicinal value has become increasingly evident in clinical practice. However, the efficacy of tangerine peel and its prescriptions still needs to be improved. After the tangerine peel and licorice are combined through the best fermentation technology and extraction technology, a new product is obtained.

[0006] Seabuckthorn (Hippophae rhamnoides L.) is a shrub or small tree of the genus Acidosa, in the family Elaeagnaceae. It is also listed in the 2010 edition of the Chinese Pharmacopoeia (Volume 1). Its main functions and indications include relieving cough and expectoration, promoting digestion and relieving stagnation, and promoting blood circulation and dispersing blood stasis. It is used for cough with excessive sputum, indigestion, abdominal pain due to food stagnation, swelling caused by falls, fatigue, blood stasis, and amenorrhea. Seabuckthorn seeds, peels, fruits, and pomace all contain a certain amount of oil. Mature seabuckthorn seeds contain 8% to 20% oil, dried seabuckthorn fruits contain approximately 20% to 25% oil, and the pomace after extracting seabuckthorn juice still contains 15% to 20% oil. Research has found that seabuckthorn oil contains a variety of essential nutritional ingredients for the human body, including various unsaturated fatty acids, carotenoids and their derivatives, sterols, and natural vitamin E. Reported benefits include antibacterial, antiviral, anti-tumor, anti-ulcer, anti-aging, radiation protection, and immune-enhancing properties. However, the relevant activities of seabuckthorn still cannot meet the clinical needs of patients, and there is an urgent need to improve clinical efficacy through combination innovation. The combination of seabuckthorn, tangerine peel, licorice, and honeysuckle has a new therapeutic effect, especially for colds, with a new mechanism and significant innovation.

[0007] With the accelerated pace of life and environmental pollution leading to low immunity among more and more people, coupled with the accelerated rate of virus mutation, more and more people often suffer from various upper respiratory tract diseases, such as the common cold, lower respiratory tract pneumonia, respiratory tract damage and other diseases, and the difficulty of treatment has increased. The efficacy of previous drugs cannot meet the needs of patients. It is urgent to discover innovative drugs to improve lung immunity and drug efficacy, protect the health of the majority of patients, and at the same time reduce the economic burden on patients. Summary of the Invention

[0008] In response to the problems existing in the prior art, the present invention provides a composition for resisting colds and respiratory tract injuries and a preparation method thereof; provides a composition for preventing and treating colds, respiratory system injuries and improving immunity; and provides an application of the composition to prepare the composition into a preparation for treating respiratory diseases.

[0009] The present invention is realized in this way: a composition for resisting colds and respiratory tract damage is prepared according to the following medicinal material formula in parts by weight: 2-10 parts of licorice, 3-16 parts of seabuckthorn, and 3-9 parts of tangerine peel.

[0010] Preferably, the specific point values ​​of the proportions of licorice, seabuckthorn and tangerine peel can be selected as 5.1 parts, 5.2 parts and 8.1 parts respectively, and other specific point values ​​within the above numerical range of mass proportions can be selected.

[0011] Preferably, 0.1-6 parts of honeysuckle are added to the licorice, sea buckthorn and tangerine peel in the composition, or 0.1-6 parts of other immune-boosting and anti-cold medicinal materials are added to the composition; the sea buckthorn in the composition can be replaced with 0.1-6 parts of honeysuckle.

[0012] Another object of the present invention is to provide a method for preparing a composition for resisting colds and respiratory tract injuries, comprising: taking the above raw materials, steam distilling licorice, and collecting the distillate for later use; combining the medicinal residue with tangerine peel and sea buckthorn medicinal materials, mixing them evenly, decocting them twice with water, each time for 0.1-1 hour, and collecting the decoction; combining the decoctions, filtering, and combining the filtrate with the licorice herbal extract; concentrating under reduced pressure to an extract with a relative density of 1.22g / ml (60°C), adding ethanol to make the alcohol content reach 60-75% (V / V), letting it stand for 24 hours, filtering, recovering ethanol from the supernatant under reduced pressure, concentrating, and adding the above licorice distillate; and finally adding pharmaceutically acceptable excipients and preparing various dosage forms through conventional processes.

[0013] Another object of the present invention is to provide a method for preparing a composition for resisting colds and respiratory tract damage, comprising: taking the above raw materials, soaking them in water for 1 hour, heating and boiling them 2-3 times, each time for 1-3 hours, filtering, combining the filtrates, and concentrating under reduced pressure to a relative density of 1.13-1.28 g / cm 3 The extract is heated to 60°C, ethanol is added to make the alcohol content reach 60-75% (V / V), the extract is allowed to stand for 20-24 hours, filtered, the supernatant is decompressed to recover the ethanol, and concentrated until there is no alcohol taste. Finally, acceptable excipients are added through conventional processes to prepare various commonly used dosage forms.

[0014] Another object of the present invention is to provide a method for preparing a composition for resisting colds and respiratory tract injuries, comprising: taking the above raw materials, soaking them in water for 1 hour, heating and decocting them twice, each time for 2 hours, filtering them, and finally adding acceptable excipients through conventional processes to prepare various usable dosage forms.

[0015] Another object of the present invention is to provide a method for preparing a composition for resisting colds and respiratory tract injuries, comprising: taking the above raw materials and crushing them into various commonly used preparations or foods such as tea bags, powders, ultrafine powders, candies, etc.

[0016] The composition of the present invention can be added with excipients required for product preparation, such as one or more of fillers, disintegrants, solubilizers, binders, flavoring agents, colorants, preservatives, lubricants, fragrances, and emulsifiers as needed, and then prepared into possible dosage forms: including but not limited to beverages, oral liquids, powders, tablets, granules, pills, capsules, ointments, pills, injections, nasal drops, aerosols, suppositories, or films.

[0017] Another object of the present invention is to provide a composition for resisting colds and respiratory tract damage for use in preparing medicines for resisting respiratory tract damage, sputum, sneezing, eye irritation, pharyngeal discomfort, and chest tightness.

[0018] Another object of the present application is to provide the use of the composition for resisting cold and respiratory tract injury in the preparation of a medicine for resisting inflammation, inhibiting inflammatory factors, resisting virus and cold.

[0019] Further, the composition plays a role through a new mechanism of MEG3 / miR-223 / NLRP3 axis, and exerts a curative effect by regulating a maternal expression gene 3 (MEG3) / microRNA-223 (miR-223) / nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) pathway.

[0020] The composition according to the present application is used in the preparation of a medicine for treating respiratory tract mucosa injury caused by chemical injury.

[0021] The composition according to the present application is used in the preparation of a medicine for treating respiratory tract mucosa injury caused by fire injury and the like.

[0022] The composition according to the present application is used in the preparation of a medicine for treating lung injury caused by various reasons.

[0023] In combination with the above technical solutions and solved technical problems, the technical solution to be protected by the present application has the following advantages and positive effects:

[0024] Firstly, the present application discloses a composition for relieving cold such as common cold, influenza and sequelae thereof, and respiratory tract injury, and preventing and treating lung injury and improving immunity, and the composition is a compound for treating respiratory tract injury, and the composition is widely used in the fields of medicine, health care product, food and beverage.

[0025] The composition according to the present application is prepared from traditional Chinese herbal medicine, and has refined prescription, definite curative effect and good clinical compliance.

[0026] The composition according to the present application is prepared from traditional Chinese herbal medicine, and has refined prescription, definite curative effect and good clinical compliance.

[0027] Thirdly, the technical solution of the present application solves the problems of the prior art in industrial application.

[0028] 1) Limitations of existing medicines for resisting cold and respiratory tract injury:

[0029] The most commonly used anti-cold and respiratory tract injury medications currently on the market include chemically synthesized drugs, which often come with side effects such as gastrointestinal discomfort and drowsiness. Long-term use can have negative effects on the human body. Traditional Chinese medicines are also commonly used, such as licorice, which is used for expectoration, cold relief, and antiviral effects, but their efficacy has not yet met patient needs. Respiratory diseases have developed new characteristics, and licorice cannot achieve its anti-cold and antiviral therapeutic goals, requiring new compound drugs to be effective.

[0030] Some Chinese medicines only relieve cold symptoms and cannot effectively solve symptoms such as razor throat, lung injury, and lung nodules. They have limited effects on respiratory tract repair and anti-inflammatory effects and fail to truly meet the clinical needs of patients.

[0031] 2) Insufficient effect on improving immune function:

[0032] In the existing technology, anti-cold drugs usually aim to relieve symptoms, but have weak effects on improving immune function and post-illness repair, making it difficult to prevent recurrence of colds.

[0033] Significant technical advancements of the present invention:

[0034] 1) Optimized medicinal formula and scientific ratio:

[0035] This invention utilizes a scientifically formulated combination of licorice, seabuckthorn, and dried tangerine peel, along with the optional addition of honeysuckle and other immune-boosting herbs, to create a synergistic composition. Licorice has anti-inflammatory, antitussive, and antiasthmatic properties, while seabuckthorn, rich in vitamin C and flavonoids, can enhance immunity and promote the repair of respiratory tract damage. Tangerine peel also has expectorant and cough-relieving properties. The combined formula alleviates cold symptoms while repairing respiratory tract damage and boosting immunity.

[0036] The introduction of optional ingredients (such as honeysuckle) gives the composition a certain degree of flexibility, which can be adjusted according to different diseases and individual differences, further improving the therapeutic effect.

[0037] 2) Efficient extraction and preparation methods:

[0038] The invention adopts a method combining steam distillation and multiple decoctions to extract the effective ingredients in the traditional Chinese medicine to the maximum extent, and further improves the purity and bioavailability of the medicine through reduced pressure concentration and alcohol precipitation processes.

[0039] The preparation method focuses on extracting the active ingredients of different medicinal materials and then mixing them, avoiding the problems of insufficient ingredient extraction or unbalanced compatibility in traditional Chinese medicine compositions, and significantly improving the efficacy.

[0040] 3) Multiple mechanisms of action:

[0041] The composition of the present invention can not only relieve cold symptoms (such as fever, cough, nasal congestion, etc.), but also has obvious anti-inflammatory and antioxidant effects, and has a significant effect on repairing damage to the respiratory tract.

[0042] By enhancing immune function (rich in polysaccharides and flavonoids in sea buckthorn and honeysuckle), the composition achieves effective control of cold prevention and recurrence.

[0043] 4) Wide applicability and safety:

[0044] The ingredients of the composition are all natural plant extracts with no obvious side effects, which is more in line with modern people's demand for drug safety and is especially suitable for children, the elderly and other people with weak constitutions.

[0045] It can be made into a variety of dosage forms (such as tablets, capsules, granules, oral liquids, etc.) to facilitate use by different patients.

[0046] 5) Industrialization advantages:

[0047] The invention adopts modern extraction technology and controllable production process in the preparation process, has high process repeatability, and is convenient for industrial production.

[0048] The efficient utilization of medicinal materials and multifunctional mechanism of action make the product more competitive in the market, which can not only meet consumer needs but also achieve improved economic benefits.

[0049] By optimizing the medicinal formula, incorporating a scientific preparation method, and utilizing multiple mechanisms of action, this invention addresses the significant side effects, limited functionality, and incomplete efficacy of traditional cold medications. It also avoids the compatibility and preparation deficiencies of existing Chinese medicine compositions. This unique technical solution significantly enhances the safety, efficacy, and applicability of cold and respiratory tract injury treatments, providing a novel approach and solution for the development and industrialization of green Chinese medicine compositions.

[0050] Although licorice and honeysuckle are also effective in treating colds in Western medicine, the present invention is formulated based on the detoxification theory of traditional Chinese medicine and the sticky-heat theory of ethnic medicine. The formulation and efficacy are different from various previous formulations and reports, the activity is significantly improved, and the new prescription is innovative. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments of the present invention. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0052] Figure 1This is a schematic diagram showing the effect of the composition provided in an embodiment of the present invention on the body weight of immunocompromised mice;

[0053] Figure 2 Schematic diagram of spleen index of an animal model prepared by adding attenuated virus to the composition provided in the embodiment of the present invention and the composition plus honeysuckle group;

[0054] Figure 3 Schematic diagram of the composition provided in the embodiments of the present invention and the composition adding honeysuckle to improve the thymus index of an animal model;

[0055] Figure 4 Schematic diagram of the effects of the composition and the honeysuckle group of the composition provided in the embodiments of the present invention on serum immunoglobulin IgM in animal models;

[0056] Figure 5 Schematic diagram of the effect of the composition provided in an embodiment of the present invention on immunoglobulin IgG in the serum of model mice;

[0057] Figure 6 Schematic diagram of the effect of the composition provided in an embodiment of the present invention on immunoglobulin IgA in the serum of model mice;

[0058] Figure 7 Schematic diagram of the effect of the composition provided in an embodiment of the present invention on the cytokine IFN-γ in the serum of model mice;

[0059] Figure 8 This is a schematic diagram showing the effect of the composition provided in an embodiment of the present invention on the cytokine IL-2 in the serum of model mice;

[0060] Figure 9 Schematic diagram of the effect of the composition provided in an embodiment of the present invention on the cytokine IL-6 in the serum of model mice;

[0061] Figure 10 This is a schematic diagram showing the effect of the composition provided in an embodiment of the present invention on the cytokine TNF-α in the serum of model mice;

[0062] Figure 11 Schematic diagram showing the effect of the composition provided in an embodiment of the present invention on lung injury and lung index in mice induced by attenuated virus;

[0063] Figure 12 Schematic diagram of the effect of the composition provided in an embodiment of the present invention on the lung pathological structure of a mouse model induced by an attenuated virus;

[0064] Figure 13 2 is a schematic diagram comparing IL-18 levels in lung tissues of various groups provided in an embodiment of the present invention;

[0065] Figure 142 is a schematic diagram comparing the IL-1β levels in lung tissues of various groups provided in an embodiment of the present invention;

[0066] Figure 15 2 is a schematic diagram comparing IL-6 levels in lung tissues of various groups provided in an embodiment of the present invention;

[0067] Figure 16 2 is a schematic diagram comparing the TNF-α levels in lung tissues of various groups provided in the embodiments of the present invention;

[0068] Figure 17 2. It is a comparative diagram of the antiviral factor IFN-β levels in lung tissues of various groups provided in an embodiment of the present invention;

[0069] Figure 18 Schematic diagram of the effect of the composition provided in the examples of the present invention on the level of miR-223 in mouse lung tissue;

[0070] Figure 19 Schematic diagram of the effect of the composition provided in an embodiment of the present invention on the level of MEG3 in mouse lung tissue;

[0071] Figure 20 Schematic diagram comparing NLRP3 levels in lung tissues of mice in various groups provided in the examples of the present invention;

[0072] Figure 21 Schematic diagram of the effects of the composition, MEG3 inhibitor and miR-223 inhibitor provided in the examples of the present invention on MEG3, miR-223 and NLRP3 in mouse lung tissue.

[0073] Figure 22 This is a graph showing the relative expression level of H2-Aa provided in an embodiment of the present invention.

[0074] Figure 23 This is a graph showing the relative expression level of H-2E provided in an embodiment of the present invention.

[0075] Figure 24 The CD4 provided by the embodiment of the present invention + T RM Relative expression detection results. DETAILED DESCRIPTION

[0076] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0077] An embodiment of the present invention provides a composition for resisting respiratory tract injury, which is prepared according to the following medicinal material formula in parts by weight: 2 to 10 parts of licorice, 3 to 16 parts of sea buckthorn, and 3 to 9 parts of tangerine peel.

[0078] Preferably, the specific point values ​​of the proportions of licorice, seabuckthorn and tangerine peel can be selected as 5.1 parts, 5.2 parts and 8.1 parts respectively, and other specific point values ​​within the above numerical range of mass proportions can be selected.

[0079] Preferably, 0.1-6 parts of honeysuckle are added to the licorice, sea buckthorn and tangerine peel in the composition, or 0.1-6 parts of other immune-boosting and anti-cold medicinal materials are added to the composition; the sea buckthorn in the composition can be replaced with 0.1-6 parts of honeysuckle.

[0080] The invention provides a preparation method of a composition for resisting respiratory tract injury, comprising: taking the above raw materials, steam distilling licorice, and collecting the distillate for later use; combining the medicinal residues with tangerine peel and seabuckthorn medicinal materials, mixing them evenly, decocting them twice with water, each time for 0.1-1 hour, and collecting the decoction; combining the decoctions, filtering, and combining the filtrate with the licorice herbal extract; concentrating under reduced pressure to obtain an extract with a relative density of 1.22 g / ml (60° C.), adding ethanol to adjust the alcohol content to 60-75% (V / V), allowing the extract to stand for 24 hours, filtering, recovering ethanol from the supernatant under reduced pressure, concentrating, and adding the above licorice distillate; and finally adding pharmaceutically acceptable excipients and preparing various dosage forms through conventional processes.

[0081] The present invention provides a method for preparing a composition for resisting respiratory tract injury, comprising: taking the above raw materials, soaking them in water for 1 hour, heating and boiling them 2-3 times, each time for 1-3 hours, filtering, combining the filtrate, and concentrating under reduced pressure to a relative density of 1.13-1.28 g / cm 3 The extract is heated to 60°C, ethanol is added to make the alcohol content reach 60-75% (V / V), the extract is allowed to stand for 20-24 hours, filtered, the supernatant is decompressed to recover the ethanol, and concentrated until there is no alcohol taste. Finally, acceptable excipients are added through conventional processes to prepare various commonly used dosage forms.

[0082] An embodiment of the present invention provides a method for preparing a composition for resisting respiratory tract injury, comprising: taking the above raw materials, soaking them in water for 1 hour, heating and decocting them twice, each time for 2 hours, filtering, and finally adding acceptable excipients through conventional processes to prepare various available dosage forms.

[0083] The embodiment of the present invention provides a method for preparing a composition for resisting respiratory tract injury, comprising: taking the above raw materials and crushing them into various commonly used preparations or foods such as tea bags, powders, ultrafine powders, candies, etc.

[0084] 2. Application Examples: In order to demonstrate the creativity and technical value of the technical solution of the present invention, this section provides application examples of the claimed technical solution on specific products or related technologies.

[0085] Example 1

[0086] Recipe: 3.3 parts licorice, 3.3 parts sea buckthorn, 3.3 parts tangerine peel

[0087] Preparation method:

[0088] (1) Weigh 15g of each of the above three herbs, crush and mix, soak in water for 1 hour, and heat and decoct twice, each time for 2 hours;

[0089] (2) Filter, combine the filtrates, and concentrate under reduced pressure to a relative density of 1.22 g / cm 3 (60℃) extract;

[0090] (3) Add ethanol to make the alcohol content reach 60% (V / V), let it stand for 24 hours, and filter;

[0091] (4) recovering ethanol from the supernatant under reduced pressure and concentrating until there is no alcohol smell;

[0092] (5) Oral solution 1000 ml is prepared directly or by adding auxiliary materials through conventional processes. Each 100 ml is equivalent to 4.5 grams. Other preparations can also be prepared.

[0093] Example 2

[0094] Recipe: 3 parts licorice, 6 parts sea buckthorn, 6 parts tangerine peel

[0095] Preparation method: Weigh 6g, 12g and 12g of licorice, seabuckthorn and tangerine peel respectively, take licorice and steam distill, collect the distillate and the liquid, set aside, combine the residue with seabuckthorn and tangerine peel, mix well, add water and boil twice, each time for 2 hours, combine the decoction, filter, combine the filtrate with the licorice liquid, and concentrate under reduced pressure to a relative density of 1.22g / cm 3 (60 ℃) extract, add ethanol to make the alcohol content reach 60% (V / V), let it stand for 24 hours, filter, recover ethanol from the supernatant under reduced pressure, concentrate, add the above distillate, stir well, refrigerate for 24 hours, filter, and freeze-dry the filtrate to obtain freeze-dried powder, which can also be used to prepare other preparations and solid foods.

[0096] Example 3

[0097] Recipe: 5 parts licorice, 5 parts sea buckthorn, 3 parts tangerine peel, 1 part honeysuckle

[0098] Preparation method: Weigh 10g, 10g, 6g and 1g of liquorice, seabuckthorn, tangerine peel and honeysuckle respectively, soak each in water for 1 hour and heat and decoct twice, each time for 2 hours, filter, take the filtrate of each medicinal material and mix them according to the weight ratio, and concentrate the mixture under reduced pressure to a relative density of 1.22g / cm 3The extract is heated to 60°C, ethanol is added to make the alcohol content reach 60% (V / V), the extract is allowed to stand for 24 hours, filtered, the supernatant is decompressed to recover the ethanol, and concentrated until there is no alcohol taste. Finally, it is prepared into capsules or oral solutions directly or by adding excipients acceptable in the pharmaceutical and food fields through conventional processes.

[0099] 3. Evidence of the effects of the embodiments: The embodiments of the present invention have achieved some positive effects during the development or use process, and indeed have great advantages over the existing technology. The following content describes them with reference to the data, charts, etc. of the experimental process.

[0100] The following pharmacological tests were performed on the above composition of the present invention.

[0101] Experiment 1: Safety test of the composition of Example 1

[0102] The maximum dose method was used to administer the extract suspension to mice via gavage to determine the acute toxicity of the drug. Results showed that no mice died after four gavages within 24 hours and two weeks of observation. No toxic reactions were observed in the mouse organs during autopsy. The maximum dose measured in mice was 352.4 g / kg, 230 times the normal dose, confirming the drug's safety.

[0103] Experiment 2: Protective effect of the composition of Example 1 on respiratory tract damage caused by chemical agents.

[0104] Clinical cases: 24 patients with respiratory tract injury caused by chlorine gas leak were randomly selected from the respiratory department. Among them, 16 were male and 8 were female, aged 15 to 53 years (average 28.6 years); 1 patient had a history of chronic bronchitis and 2 had a history of hypertension. The degree of respiratory tract chlorine injury: 6 cases of irritation, 13 cases of mild, and 5 cases of moderate. The scoring standard for patients' respiratory tract injury is 1 point for mild, 2 points for moderate, and 3 points for severe. 10 patients who refused treatment were selected for follow-up, and 10 patients each who were treated with licorice, sea buckthorn, and tangerine peel decoctions were selected for observation. The symptoms and scores of these 40 patients were not statistically different from those of the combination decoction treatment group using one-way analysis of variance, P>0.05.

[0105] Clinical manifestations: 20 patients experienced eye irritation and tearing, 21 experienced sneezing, 24 experienced pharyngeal discomfort, and 16 experienced chest tightness. Cough and expectoration were reported in 24 patients. Twenty patients presented with white sputum, four with yellow sputum, and one with blood in the sputum. Sixteen patients experienced tachypnea, 14 with increased heart rate, six with dry rales, three with moist rales, nine with elevated white blood cell counts, and six with increased neutrophils. X-rays revealed thickened lung markings in 10 patients. Liver and kidney function tests were normal. Eight patients underwent fiberoptic bronchoscopy, which revealed varying degrees of tracheal and bronchial mucosal congestion and edema. Two patients presented with petechiae, five with sticky sputum, and three with yellowish and white sputum.

[0106] Treatment Method: Patients willing to undergo treatment were observed in the outpatient clinic with self-oxygen therapy. Mild and moderate patients were hospitalized for bed rest and gavage with 1.5 g / kg of the crude drug prepared from the composition in Example 1. A 150 ml decoction was decocted and administered orally, once in the morning and evening. Patients who declined treatment returned home to recover and were followed up.

[0107] Results: Among the patients who actively accepted treatment, 7 patients with irritation reactions were observed in the oxygen outpatient clinic. All respiratory tract injury symptoms, including eye, pharyngeal, expectorant, and chest tightness, essentially disappeared after 3 days, and treatment was completed. All 17 patients with mild to moderate symptoms were discharged after 3-4 days without any complications or sequelae. The symptoms of patients who refused treatment remained unresolved after 4 days, with statistical significance shown in one-way analysis of variance (P < 0.05).

[0108] Conclusion: Chlorine gas injury mainly causes upper respiratory tract damage, including mucosal damage, and further damages the small airways and alveoli of the lungs when inhaled at high concentrations. The 24 patients who actively received treatment in this study all had varying degrees of eye and respiratory irritation symptoms. The symptoms of chest tightness and decreased respiratory function caused by small airway and alveolar damage were obvious. All of them were cured within 4 days of treatment, and there was no sign of lung damage. However, patients who refused treatment still had symptoms after 4 days of follow-up, and there was a statistical difference, P < 0.01. Compared with the combination, the efficacy scores of patients in the other single medicinal extract treatment groups were all lower than those in the combination treatment group, with a statistical difference, P < 0.05

[0109] Chlorine is a highly irritating gas with active chemical properties. Upon contact with water, chlorine partially decomposes into hydrochloric acid and nascent oxygen. Respiratory tract damage occurs when chlorine reacts with water to form hydrochloric acid, which irritates and burns the tracheal and bronchial mucosa, causing edema, congestion, and even mucosal necrosis. The Chinese medicine composition of the present invention is highly effective in treating respiratory tract damage and is therefore clinically effective in treating chlorine-induced respiratory tract damage.

[0110] Experiment 3: Effect of the composition of Example 2 on the immunity of the immunodeficiency model

[0111] 1. Materials and Methods

[0112] 1.1 Experimental Animals: SPF-grade male BALB / c mice, weighing 20-25 g and aged 6-8 weeks, were housed in our animal facility for 5 days prior to the experiment. The temperature was maintained at 20-25°C, the relative humidity was 40%-70%, and the mice were housed on a 12-hour light-dark cycle. The mice had free access to food and water. Samples used in the composition group were prepared according to the method described in Example 2.

[0113] 1.2 Experimental methods

[0114] 1.2.1 Preparation and grouping of immunocompromised animal models

[0115] Eighty-eight healthy male BALB / c mice were randomly divided into 11 groups, each consisting of eight mice, each. These groups included a normal control group (Nor), a model group (Mod, for establishing an immunosuppressive model), a licorice root group (administered with licorice root extract, serving as both a positive control and a single licorice root group; Con), a combination group, a tangerine peel group, a seabuckthorn group, a licorice root + tangerine peel group (referred to as the "Ganchen group"), a licorice root + seabuckthorn group (referred to as the "Gansha group"), a tangerine peel + seabuckthorn group (referred to as the "Chensha group"), a combination group + honeysuckle group (referred to as the "combination honeysuckle group" to investigate the efficacy of the combination in enhancing the activity of the medicinal materials), and a licorice root, tangerine peel, and honeysuckle group (referred to as the "Ganchen honeysuckle group" to investigate the activity of seabuckthorn in place of honeysuckle in the combination). Except for the normal control group, which received an intraperitoneal injection of 10 ml / kg / day of 0.9% sodium chloride injection, mice in all other groups were injected with 80 mg / kg / day of cyclophosphamide for three consecutive days to establish an immunosuppressive model. The drug was administered orally starting on the fourth day for 14 consecutive days. The normal control group and the model control group were gavaged with 1 ml / (kg·d) of purified water, and the positive control group was gavaged with 100 mg / (kg·d) of licorice extract, 100 mg / (kg·d) of the combination group, 100 mg / (kg·d) of the combination + honeysuckle extract intervention group, 100 mg / (kg·d) of licorice, tangerine peel and honeysuckle intervention group, 100 mg / (kg·d) of tangerine peel intervention group, 100 mg / (kg·d) of sea buckthorn intervention group, 100 mg / (kg·d) of licorice + tangerine peel intervention group, 100 mg / (kg·d) of tangerine peel + sea buckthorn intervention group, and 100 mg / (kg·d) of licorice + sea buckthorn intervention group.

[0116] 1.2.2 Determination of organ index to study the immune-boosting effect of the composition

[0117] After the last administration, the mice were fasted for 12 hours but not water. The body weight of each group of mice was measured, and blood was collected from the eyes of the mice. The mice were killed, and the spleen and thymus were removed. The surrounding fat or other tissues were removed, and the residual blood and water on the surface were wiped with filter paper. The organs were weighed and the organ index was calculated as follows: spleen index (mg / g) = spleen weight (mg) / body weight (g); thymus index (mg / g) = thymus weight (mg) / body weight (g)

[0118] 1.2.3 Determination of peripheral blood cells to study immune capacity

[0119] Blood was collected from the medial canthal vein of mice and placed in anticoagulant tubes. The concentrations of white blood cells, red blood cells, platelets, lymphocytes, and mean corpuscular hemoglobin were determined using an automatic blood cell analyzer.

[0120] 1.2.4 Determination of cytokines and immunoglobulins to study anti-inflammatory and antiviral effects

[0121] Orbital blood was taken, and left at room temperature for 30 min, centrifuged (4000 r / min, 10 min, 4℃), and the supernatant was taken. The concentration changes of immunoglobulin IgA, IgG, IgM and the content level changes of cytokines IL-2, IFN-γ, TNF-α, IL-6 were detected by enzyme-linked immunoassay according to the operation steps of the ELISA kit.

[0122] 1.2.5 Statistical method

[0123] IBM Statistics SPSS23.0 software was used, and the data were expressed by mean ± standard deviation , the data were subjected to homogeneity of variance test and normal distribution test, and single factor variance analysis method, data transformation, LSD and the like were used to study the differences between groups according to the situation, and P<0.05 was the standard of statistical significance.

[0124] Experimental results

[0125] 2.1 Effect of the composition on the immune function of the immunocompromised mice

[0126] Compared with the blank group, the body weight, spleen index and thymus index of the model group were significantly reduced (#P<0.05, ##P<0.01), indicating that the immunocompromised mouse model was successfully prepared. Compared with the model group, the positive drug group, the composition group, the pericarpium citri reticulatae group, the hippophae rhamnoides group, the composition honeysuckle group, the licorice pericarpium citri reticulatae group, the licorice hippophae rhamnoides group and the pericarpium citri reticulatae hippophae rhamnoides group were significantly increased (*P<0.05, P<0.01), indicating that the above administration groups could promote the development of the immune organs of the immunocompromised mice and had the activity of improving the immunity (see Figure 1 , 2 , 3). Compared with the composition group, the positive drug group, the pericarpium citri reticulatae group, the hippophae rhamnoides group, the licorice pericarpium citri reticulatae group, the licorice hippophae rhamnoides group and the pericarpium citri reticulatae hippophae rhamnoides group were significantly reduced (&P<0.05 or &P<0.01), indicating that the composition group and the administration groups of the application could improve the immunity, and the efficacy of the composition group of the application was obviously better than those of the other groups, and could promote the growth of the immunocompromised mice. At the same time, it was found for the first time that the composition obtained by adding honeysuckle to the composition was also better than the other administration groups and the positive control group (licorice group) except the composition group of the application, which belonged to the first discovery.

[0127] 2.2 Effect of the composition on the number of peripheral blood leukocytes of the mice

[0128] Compared with the blank control group, the model group showed significantly decreased white blood cell (WBC), red blood cell (RBC), platelet (PLT), and lymphocyte (LYM) counts (P < 0.01 or P < 0.0001), and significantly increased mean corpuscular hemoglobin (MCHC) concentration. Compared with the model group, the white blood cell, red blood cell, and platelet counts in the positive control group and the combination group were significantly increased (P < 0.01, P < 0.05, or P < 0.001), indicating recovery. Compared with the model group, the composition of the present invention had the best activity, and the addition of honeysuckle still had high activity, indicating that the composition of the present invention also has innovation and applicability when adding a single medicinal material. The composition of the present invention maintained its activity even after replacing seabuckthorn with honeysuckle, indicating that the composition also has innovation and applicability after replacing seabuckthorn. Compared with the model group, the number of lymphocytes in the combination group showed an increasing trend (see Table 1), suggesting that the combination may upregulate peripheral blood immune cells to enhance immunity.

[0129] Table 1 Comparison of blood routine indicators of mice in each group (Mean±SD, n=8)

[0130]

[0131] Note: Compared with the Nor group, #P<0.05, ##P<0.01, ###P<0.001; compared with the Mod group, *P<0.05, P<0.01, *P<0.001

[0132] 2.3 Effects of the composition on the levels of immunoglobulins IgM, IgG, and IgA and immune capacity

[0133] Depend on Figure 4 、 5As shown in Figure 6, compared with the blank group, the serum immunoglobulin IgA, IgG, and IgM levels of the model group mice were significantly decreased (#P<0.05 or ##P<0.01), indicating that the cyclophosphamide immunosuppression mouse model was successfully established. Compared with the model group, the serum immunoglobulin IgA, IgG, and IgM levels of the positive drug group, the combination group, the tangerine peel group, the sea buckthorn group, the combination honeysuckle group, the Ganchen honeysuckle group, the licorice and tangerine peel group, the licorice and sea buckthorn group, and the tangerine peel and sea buckthorn group were significantly increased (*P<0.05 or P<0.01); compared with the combination group, the serum immunoglobulin IgA, IgG, and IgM levels of the positive drug group, the tangerine peel group, the sea buckthorn group, the Ganchen honeysuckle group, the licorice and tangerine peel group, the licorice and sea buckthorn group, and the tangerine peel and sea buckthorn group were significantly decreased (&P<0.05 or &P<0.01), indicating that the combination is innovative and practical. The composition still has high activity after adding honeysuckle, indicating that the composition of the present invention is innovative and applicable even if a single herbal ingredient is added. The composition of the present invention still maintains activity even if seabuckthorn is replaced with honeysuckle, indicating that the composition is innovative and applicable even after the seabuckthorn is replaced.

[0134] 2.4 Effects of the composition on cytokines IL-2, IFN-γ, TNF-α, IL-6 and anti-inflammatory ability in mouse serum

[0135] Depend on Figure 7 、 8 , 9, 10 showed that compared with the blank control group, the levels of IL-2, IFN-γ, TNF-α, and IL-6 in the serum of the mice in the model control group were significantly decreased (#P<0.001); compared with the model group, the positive drug group, the combination group, the tangerine peel group, the sea buckthorn group, the combination honeysuckle group, the ganchen honeysuckle group, the liquorice tangerine peel group, the liquorice sea buckthorn group, and the tangerine peel sea buckthorn group all significantly increased the levels of IL-2, IFN-γ, TNF-α, and IL-6 (*P<0.05 or P<0.01), which suggests that the drug-treated group may alleviate the immunosuppression caused by cyclophosphamide by upregulating the content of cytokines in the serum of immunocompromised mice; compared with the combination group, the activities of the positive drug group, tangerine peel group, sea buckthorn group, licorice and honeysuckle group, licorice and tangerine peel group, licorice and sea buckthorn group, and tangerine peel and sea buckthorn group were significantly reduced (&P<0.05 or &P<0.01), indicating that the combination group upregulates the activity of immune cytokines and is better than other groups in regulating immune function. The combination still has high activity after adding honeysuckle, indicating that the addition of a medicinal material to the composition of the present invention is also innovative and applicable. The composition of the present invention maintains its activity even when sea buckthorn is replaced with honeysuckle, indicating that the composition is also innovative and applicable after the sea buckthorn is replaced.

[0136] in conclusion

[0137] The immune system includes immune organs, immune cells, and immune molecules, which work together to protect the body from pathogens. The thymus and spleen are the main immune organs and play an important role in the production and maintenance of immune cells. As the center of humoral immunity and cellular immunity, the spleen is the main place where T and B lymphocytes settle, so the spleen index can reflect the level of the body's immunity. Cyclophosphamide is a type of alkylating agent and is widely used in cancer treatment. However, it not only has various toxicities to cancer cells, but also to rapidly dividing cells such as immune cells and epithelial cells. It is often used to establish immunosuppressive models. In this study, after intraperitoneal injection of cyclophosphamide into mice, the body weight, spleen index, thymus index, and immunoglobulin concentration levels of the mice were significantly lower than those of the blank control group (##P<0.01), indicating that the immunosuppressive model mice were successfully prepared. After two weeks of continuous treatment with the composition of the present invention, compared with the model control group, all treatment groups showed improved immune capacity. The efficacy of the composition group was significantly superior to that of the other groups, demonstrating a significant improvement in efficacy, a finding not previously reported. Furthermore, the new combination obtained by adding honeysuckle to the composition was also superior to the other treatment groups and the positive control group (licorice group), demonstrating that the composition still exhibits high activity even with the addition of honeysuckle, demonstrating the innovative and applicable nature of the composition. Even when seabuckthorn was replaced with honeysuckle in the composition of the present invention, the activity of the composition was maintained, demonstrating the innovative and applicable nature of the composition after the addition of a single herbal ingredient. The composition also demonstrated its innovative nature. The peripheral blood leukocyte, lymphocyte, and platelet counts, as well as spleen and thymus indexes, of mice in the composition groups were significantly increased (P < 0.05 or P < 0.01), indicating that the composition can promote the development of immune organs in immunocompromised mice. ELISA testing revealed significant increases in the concentrations of immunoglobulins IgA, IgG, and IgM in the composition group and the positive control group (P < 0.01), demonstrating that the composition helps enhance immune function. In this study, both the composition and the positive drug group significantly increased the levels of IL-2, IFN-γ, TNF-α, and IL-6 in the serum of immunocompromised mice (P<0.05 or P<0.01), indicating that the composition can regulate immune function and alleviate cyclophosphamide-induced immunosuppression by increasing the levels of cytokines in the serum of immunocompromised mice.

[0138] In summary, the composition can increase the immune organ index and the number of peripheral blood leukocytes, lymphocytes, and platelets, improve pathological changes in immune organs, upregulate the expression levels of immune-related proteins and cytokines, alleviate the immunodeficiency caused by CTX, and exhibit certain immunoregulatory activity.

[0139] Experiment 4: Using an influenza mouse model to study the maternally expressed gene 3 (MEG3) / microRNA-223 (miR-223) / nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) axis mechanism of the composition sample against lung injury

[0140] 1. Materials and Methods

[0141] 1.1 Materials

[0142] 1.1.1 Experimental animals: Male C57BL / 6 mice weighing (20.5 ± 0.5) g, aged 6–8 weeks, fed with standard animal feed and free access to water, were placed on a 12-h light-dark cycle.

[0143] 1.1.2 Virus: H7N9 low pathogenicity virus strain, the virus was inoculated into 10-day-old chicken embryos and incubated in a 37°C incubator for 48 h. The chicken embryo allantoic fluid was collected and centrifuged to prepare a 5×107 half-egg infectious dose (egg 50% infectious dose, EID50) / ml virus suspension for later use.

[0144] 1.2 Methods

[0145] 1.2.1 Animal grouping, modeling, and drug administration

[0146] Eighty-eight mice were randomly divided into 11 groups, each consisting of eight mice. The control group consisted of a normal control (Nor), a model group (Mod), a licorice treatment group (i.e., a positive control group, Con), a combination group (composition treatment group of the present invention), a tangerine peel treatment group, a seabuckthorn treatment group, a licorice + tangerine peel treatment group (referred to as the "Ganchen group"), a licorice + seabuckthorn treatment group (referred to as the "Gansha group"), a tangerine peel + seabuckthorn treatment group (referred to as the "Chensha group"), a combination group + honeysuckle treatment group (referred to as the "composition honeysuckle group"), and a licorice, tangerine peel, and honeysuckle treatment group (referred to as the "Ganchen honeysuckle group"). All groups, except the blank control group, were intranasally administered with a 5×105 EID50 dose of H7N9 virus suspension. The blank group was intranasally administered with an equal volume of phosphate buffer. On the day of modeling, the drugs were administered orally. The blank control group and the model control group were gavaged with 1 mL / (kg·d) of purified water. The positive control group was gavaged with 50 mg / (kg·d) of licorice tablets, 150 mg / (kg·d) of the combination group, 200 mg / (kg·d) of the combination + honeysuckle intervention group, 150 mg / (kg·d) of the licorice, tangerine peel and honeysuckle intervention group, 50 mg / (kg·d) of the tangerine peel intervention group, 50 mg / (kg·d) of the seabuckthorn intervention group, 100 mg / (kg·d) of the licorice + tangerine peel intervention group, 100 mg / (kg·d) of the tangerine peel + seabuckthorn intervention group, and 100 mg / (kg·d) of the licorice + seabuckthorn intervention group. The gavage volume was 10 mL / (kg·d). Mice were killed 24 hours after administration.

[0147] 1.2.2 Lung injury and lung index detection

[0148] Mice were immediately sacrificed, lung tissues were dissected, rinsed with saline, and surface moisture was removed. Lung index was calculated (lung index = mouse lung mass / body mass).

[0149] 1.2.3 HE staining to observe lung tissue morphology

[0150] The left lungs of each mouse group were fixed in 10% formaldehyde, rinsed with pre-cooled phosphate buffer, dehydrated with graded ethanol solutions, cleared with xylene, immersed in paraffin, and sectioned at 5 μm thickness. Sections were dewaxed in xylene, treated with graded ethanol solutions, stained with hematoxylin, counterstained with eosin, dehydrated with graded ethanol solutions, cleared with xylene, and mounted for observation of lung tissue morphology under a microscope.

[0151] 1.2.4ELISA detection of IL-1β, IL-18, IL-6, TNF-α, and IFN-β levels in lung tissue

[0152] Part of the right lung tissue of each group of mice was taken, and the lung tissue was homogenized by adding phosphate buffer containing protease inhibitors. The lung tissue was centrifuged at 3000 r / min and 4°C for 15 min. The supernatant was the lung tissue homogenate. The levels of IL-1β, IL-18, IL-6, TNF-α, and IFN-β in the lung tissue were detected strictly according to the operating instructions of the mouse test kit.

[0153] 1.2.5 RT-qPCR detection of MEG3 and miR-223 levels in lung tissue

[0154] Right lung tissue was collected from each group of mice and minced. Total RNA was extracted using the Trizol method, and cDNA was synthesized using a first-strand cDNA synthesis kit. The levels of MEG3 and miR-223 in lung tissue were measured by RT-qPCR. The MEG3 forward primer was 5'-GTGAAGGTCGGAGTGAACG-3', and the reverse primer was 5'-CTCGCTCCTGGAAGATGGTG-3'; the β-actin forward primer was 5'-TGGAATCCTGTGGCATCCATGAAAC-3', and the reverse primer was 5'-TAAAACGCAGCTCAGTAACAGTCCG-3'; the miR-223 forward primer was 5'-TGGCTGTCAGTTTGTCAAAT-3', and the reverse primer was 5'-CTGCAGGGTCCGAGGT-3'; and the U6 forward primer was 5'-CTCGCTTCGGCAGCACA-3', and the reverse primer was 5'-AACGCTTCACGAATTTGCGT-3'. Sample loading system: 2× Mix 10 μl, cDNA (200 ng / μl) 1 μl, forward primer / reverse primer (10 μmol / L) 0.5 μl each, ddH2O 8 μl. Reaction conditions: 95°C 120 s; 94°C 5 s, 60°C 30 s, 40 cycles. -ΔΔCT The relative expression levels of MEG3 and miR-223 were calculated by the method.

[0155] 1.2.6 Detection of NLRP3, ASC, caspase-1, and pro-caspase-1 protein levels in lung tissue by western blotting

[0156] Part of the lung tissue from the right lung of each group of mice was minced, added to a protein lysis buffer containing protease inhibitors, ground on ice, and centrifuged at 12,000 rpm at 4°C for 20 minutes. The supernatant was the total protein. 20 ng of protein was subjected to gel electrophoresis, PVDF transfer, and blocked with 5% skim milk powder at room temperature for 2 hours. After rinsing with TBST, the corresponding primary antibodies NLRP3, ASC, caspase-1, pro-caspase-1, and β-actin were added, respectively, and incubated at 4°C overnight; rinsed with TBST, and secondary antibodies were added and incubated at room temperature for 2 hours. DAB color development solution was used in the dark, and the protein gel imaging system was used to photograph and quantitatively analyze.

[0157] 1.3 Statistical analysis

[0158] GraphPad Prism 8.0 software was used to perform statistical analysis on all data. P<0.05 indicates statistically significant differences.

[0159] 2. Results

[0160] 2.1 Effects of the composition on lung injury and lung index in influenza mice

[0161] Depend on Figure 11 It can be seen that compared with the blank group, the lung index of the model group was significantly increased (#P<0.05 or ##P<0.01), indicating that the lung injury model was successfully established; compared with the model group, the lung index of the positive drug group, the combination group, the tangerine peel group, the sea buckthorn group, the combination honeysuckle group, the Ganchen honeysuckle group, the licorice tangerine peel group, the licorice sea buckthorn group, and the tangerine peel sea buckthorn group were all significantly reduced (*P<0.05 or P<0.01); compared with the combination group, the lung index of the positive drug group, the tangerine peel group, the sea buckthorn group, the Ganchen honeysuckle group, the licorice tangerine peel group, the licorice sea buckthorn group, and the tangerine peel sea buckthorn group were all significantly higher than that of the combination group of the present invention (&P<0.05 or &P<0.01), indicating that the combination group and the combination plus honeysuckle group were better than the other groups in reducing the lung index, proving that the efficacy of the combination against lung injury was significantly improved. The addition of honeysuckle to the combination still had high activity, indicating that the addition of a single medicinal material to the composition of the present invention also has innovation and applicability. The activity of the composition of the present invention is maintained even when the seabuckthorn is replaced with honeysuckle, which indicates that the composition is innovative and applicable even after the seabuckthorn is replaced.

[0162] 2.2 Effects of the composition on the pathological structure of mouse lung tissue

[0163] The alveolar and bronchial morphology and structure of the lung tissue of the mice in the control group were intact, and there was no exudation in the cavity; the alveolar wall congestion of the lung tissue of the mice in the model group was obvious, and obvious inflammatory exudation and inflammatory cell infiltration were found in the cavity; the congestion of the alveolar wall of the lung tissue of the mice in the combination group and the combination honeysuckle group, as shown in FIG Figure 12 .

[0164] 2.3 Effects of the composition on inflammatory factors such as IL-1β, IL-18, IL-6, and TNF-α in lung tissue

[0165] Compared with the blank group, the levels of IL-1β, IL-18, IL-6 and TNF-α in the lung tissue of the model group were significantly increased (#P<0.05 or ##P<0.01); compared with the model group, the levels of IL-1β, IL-18, IL-6 and TNF-α in the lung tissue of the positive drug group, combination group, tangerine peel group, sea buckthorn group, combination honeysuckle group, Ganchen honeysuckle group, licorice and tangerine peel group, licorice and sea buckthorn group, and tangerine peel and sea buckthorn group were decreased (*P<0.05 or P<0.01); compared with the combination group, the levels of IL-1β, IL-18, IL-6 and TNF-α in the lung tissue of the positive drug group, tangerine peel group, sea buckthorn group, Ganchen honeysuckle group, licorice and tangerine peel group, licorice and sea buckthorn group, and tangerine peel and sea buckthorn group were all increased (&P<0.05 or &P<0.01), indicating that the anti-inflammatory activity of the combination was significantly higher than that of the single medicinal material and the pairwise combination of medicinal materials. The composition is added with a medicinal material honeysuckle which has strong anti-inflammatory activity and is beneficial to resist lung damage, such as Figure 13 、 14 , 15, and 16. The composition still has high activity after adding honeysuckle, indicating that the composition of the present invention is innovative and applicable even with the addition of a single herbal ingredient. The composition of the present invention maintains its activity even after replacing seabuckthorn with honeysuckle, indicating that the composition is innovative and applicable even after replacing seabuckthorn.

[0166] 2.4 Effect of the composition on the antiviral ability of the antiviral factor IFN-β

[0167] Compared with the blank group, the IFN-β levels in the lung tissue of the model group were significantly increased (#P<0.05 or ##P<0.01); compared with the model group, the IFN-β levels in the lung tissue of the positive drug group, the combination group, the tangerine peel group, the sea buckthorn group, the combination honeysuckle group, the Ganchen honeysuckle group, the licorice and tangerine peel group, the licorice and sea buckthorn group, and the tangerine peel and sea buckthorn group were increased (*P<0.05 or P<0.01); compared with the combination group, the IFN-β levels in the lung tissue of the positive drug group, the tangerine peel group, the sea buckthorn group, the Ganchen honeysuckle group, the licorice and tangerine peel group, the licorice and sea buckthorn group, and the tangerine peel and sea buckthorn group were decreased (&P<0.05 or &P<0.01), indicating that the ability of the combination to enhance antiviral activity is significantly superior to that of a single herb or a combination of two herbs. The addition of honeysuckle to the combination still maintained high activity, indicating that the addition of a single herb to the composition of the present invention is also innovative and applicable. The composition of the present invention can still maintain its activity even when the seabuckthorn is replaced with honeysuckle, which shows that the composition is innovative and applicable even after the seabuckthorn is replaced. Figure 17 shown.

[0168] 2.5 Effects of the composition on MEG3 and miR-223 levels in lung tissue

[0169] Compared with the control group, the MEG3 level in the lung tissue of the model group was increased (##P<0.001), and the miR-223 level was decreased (##P<0.001); compared with the model group, the MEG3 level in the lung tissue of the positive drug group, the combination group, the tangerine peel group, the sea buckthorn group, the combination honeysuckle group, the ganchen honeysuckle group, the liquorice and tangerine peel group, the liquorice and sea buckthorn group, and the tangerine peel and sea buckthorn group were decreased (*P<0.05 or P<0.01), and the miR-223 level was increased. Compared with the combination group, the MEG3 levels in the positive drug group, tangerine peel group, seabuckthorn group, honeysuckle group, liquorice tangerine peel group, liquorice seabuckthorn group, and tangerine peel seabuckthorn group were increased (&P<0.05 or &P<0.01), and the miR-223 levels were decreased (&P<0.05 or &P<0.01), indicating that the combination exerts its therapeutic effect through the MEG3 / miR-223 / NLRP3 inflammasome pathway. Figure 18 and Figure 19 .

[0170] 2.6 Effects of the Composition on Inflammasome NLRP3 in Lung Tissue

[0171] Compared with the control group, the level of NLRP3 in the lung tissue of the model group increased (#P<0.01); compared with the model group, the level of NLRP3 in the positive drug group, the composition group, the pericarpium citri reticulatae group, the hippophae rhamnoides group, the composition honeysuckle group, the licorice pericarpium citri reticulatae group, the licorice hippophae rhamnoides group, the pericarpium citri reticulatae hippophae rhamnoides group was reduced (*P<0.05 or P<0.01); compared with the composition group, the level of NLRP3 in the positive drug group, the pericarpium citri reticulatae group, the hippophae rhamnoides group, the licorice pericarpium citri reticulatae group, the licorice hippophae rhamnoides group, the pericarpium citri reticulatae hippophae rhamnoides group was increased (&P<0.05 or &P<0.01), indicating that the composition exerts activity through NLRP3 inflammasome, and the previous results prove that the composition exerts activity through the MEG3 / miR-223 / NLRP3 inflammasome mechanism, and the mechanism is innovative. See Figure 20 .

[0172] 2.7 Targeting relationship between MEG3 and miR-223, and miR-223 and NLRP3

[0173] Compared with the model group, the content of MEG3 in the composition group was reduced (#P<0.01), the content of miR-223 was increased (#P<0.01), and the content of NLRP3 was reduced (#P<0.01); compared with the composition group, the content of MEG3 was increased *P<0.05, the content of miR-223 was reduced, and the content of NLRP3 was increased *P<0.05 after adding the composition+MEG3 inhibitor; compared with the composition group, the content of MEG3 was unchanged, the content of miR-223 was increased (&P<0.05), and the content of NLRP3 was reduced (&P<0.05) after adding the composition+miR-223 inhibitor, indicating that there is a targeting relationship between MEG3 and miR-223, and miR-223 and NLRP3, and the composition exerts an impact on influenza infection lung injury by regulating the MEG3 / miR-223 axis, see Figure 21 .

[0174] 3. Conclusion

[0175] In this study, the combination reduced lung pathological damage and elevated lung index in a mouse model of influenza virus infection, suggesting that the combination significantly alleviated inflammatory exudates and inflammatory cell infiltration in the lungs. In this study, elevated expression of IL-1β, IL-18, IL-6, TNF-α, and IFN-β was observed in the lungs of mice infected with H7N9 avian influenza. This suggests that H7N9 infection leads to elevated expression of chemokines, proinflammatory factors, and antiviral factors, ultimately leading to increased inflammation and tissue damage. Administration of the combination reduced levels of chemokines and proinflammatory factors in lung tissue, while levels of the antiviral factor IFN-β continued to increase, thereby inhibiting viral infection, reducing inflammation, and alleviating damage. MEG3 was highly expressed in the model group in this study, suggesting that elevated MEG3 expression may play a role in lung damage caused by H7N9 avian influenza virus infection. This study identified a targeted regulatory relationship between miR-223 and MEG3. Further research has revealed that miR-223 primarily functions to suppress inflammatory responses, preventing infection and complications, and playing a role in innate immunity to viral infections. Promoting miR-223 expression inactivates the NLRP3 inflammasome, thereby alleviating influenza A virus-induced inflammatory damage in lung epithelial cells. This study further revealed a targeted regulatory relationship between miR-223 and NLRP3, with miR-223 regulating NLRP3 expression. The NLRP3 inflammasome contributes to multiple signaling pathways, including environmental stimuli, endogenous signals, pathogens, and mitochondrial damage. It promotes the activation and secretion of IL-1β and IL-18, leading to inflammation-related damage. ASC and caspase-1 are both components of the NLRP3 inflammasome. In this study, miR-223 expression was low in the model group, potentially impairing its inhibitory effect on inflammation. High expression of NLRP3 inflammasome proteins resulted in significant lung inflammation in H7N9 avian influenza virus-infected lungs. After adding the composition, the level of miR-223 increased, and the levels of NLRP3, ASC, and caspase-1 / pro-caspase-1 proteins decreased, suggesting that the composition can promote the expression of miR-223 and inhibit the NLRP3 inflammasome, thereby alleviating lung damage caused by H7N9 avian influenza virus infection.

[0176] In summary, the composition can alleviate lung damage caused by H7N9 avian influenza virus infection in mice, which is closely related to the MEG3 / miR-223 / NLRP3 axis.

[0177] Experiment 5. Using a mouse model infected with SARS-CoV-2 virus to study the effect of the composition sample on enhancing lung immune function and its mechanism

[0178] 1. Materials and Methods

[0179] 1.1 Materials

[0180] 1.1.1 Experimental animals: 56 SPF-grade BALB / c mice, half male and half female, weighing (12 ± 2) g, 5 weeks old, fed with standard animal feed, with free access to water and a 12-h light-dark cycle.

[0181] 1.1.2 Virus: Vero E6 cells, an adrenal cell line derived from female African green monkeys, were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% anti-antibody at 37°C in 5% CO2. Virus titration was performed in Vero E6 cells at 10-fold dilutions. CPE was scored 3 days after inoculation, and the TCID was calculated using the Reed-Muench formula. 50 .

[0182] 1.2 Methods

[0183] 1.2.1 Animal grouping, modeling, and drug administration

[0184] Fifty-six BALB / c mice, half male and half female, were randomly divided into 7 groups (8 mice each), including a normal control group (Nor), a model group (Mod), a positive control group (Lianhua Qingwen group, Con), a combination group, a tangerine peel group, a licorice group, and a seabuckthorn group. The mice were anesthetized with tribromoethanol (250 mg / kg) and, except for the blank control group, were infected intranasally with 50 μl of DMEM medium per mouse for 7 × 10 5 TCID 50 The blank control group was given an equal volume of phosphate buffer nasal drops. On the day of model establishment, the blank control group and the model control group were given purified water 10 mL / (kg·d), the positive control group was given Lianhua Qingwen Capsules 560 mg / (kg·d), the combination group was given the formula 150 mg / (kg·d), the licorice group was given licorice tablets 50 mg / (kg·d), the tangerine peel group was given tangerine peel 50 mg / (kg·d), and the sea buckthorn group was given sea buckthorn 50 mg / (kg·d). The gavage volume was 10 mL / (kg·d) and the administration was continued for 7 days.

[0185] 1.2.2 Effect of the Combination on the Expression of MCH-II Subclasses in Lung Epithelial Cells

[0186] 1.2.2.1 Digestion of Mouse Lungs for FACS Sorting

[0187] The lungs of euthanized mice were perfused with PBS, intubated, and then infused with 1 ml of preheated Disase solution. The trachea was ligated, the lungs were removed from the chest cavity, and then minced with scissors. Two ml of Dispase and DNase I (20 U / ml) were added to the resulting slurry, and the sample was digested on a shaker at 37°C for 20-30 minutes. The resulting cell suspension was gently mixed and filtered through 100 μm and 40 μm filters, respectively, and the filter was then rinsed with excess DMEM. The single cell suspension was centrifuged (5 min, 300 g) and the pellet was resuspended in 1 mL of pre-chilled FACS buffer. The cell suspension was resuspended in MACS buffer, mixed with anti-CD45, anti-CD31, and anti-Ter119 microbeads, and incubated at 4°C for 15 minutes. After washing, the cells were loaded onto a pretreated MS column placed in the magnetic field of a MACS separator, and the effluent containing unlabeled cells was collected. These enriched LECs were then blocked, stained, and washed using CD16 / CD32 Fc-Block before analysis by flow cytometry on an LSRFortessa flow cytometer. Fluorescence minus one controls were used for sorting.

[0188] 1.2.2.2 FACS sorting of lung epithelial cells

[0189] Epithelial cell subpopulations from stained single-cell suspensions isolated from elastase-digested lungs were sorted on ice into RPMI 1640 containing 20% ​​FBS using FACS-Aria II SORP.

[0190] 1.2.2.3 Real-time fluorescence quantitative PCR to measure the expression of MCH-II subclasses in lung epithelial cells

[0191] Total RNA was extracted from mouse hippocampal tissue using the Trizol method. 50 mg of tissue was added to 1 mL of Trizol solution for homogenization and lysed on ice for 15 minutes. Subsequently, 200 μl of chloroform was added, mixed by inversion, and allowed to stand on ice for 15 minutes. Centrifuged at 4°C 14,000 rpm for 15 minutes, then 200 μl of supernatant was transferred to an RNase-free centrifuge tube, an equal volume of isopropanol was added, and the tube was placed on ice for 5 minutes. Centrifuged at 4°C 14,000 rpm for 10 minutes, the supernatant was discarded, and the tube was washed with 75% ethanol, centrifuged at 4°C 14,000 rpm for 5 minutes, the ethanol was discarded, and the tube was dried. Then, 50 μl of DEPC water was added to dissolve the extracted RNA. The concentration and purity of RNA were detected by Quickdrop. Using 1 μg of RNA as a template, a reverse transcription kit (StarScript II two-step method

[0192] RT-PCR kit) was reversed into cDNA according to the instructions. Then cDNA was pre-mixed with 2X probe method real-time fluorescence quantitative PCR reaction system kit, with β-actin as internal reference and 2 -△△CT The expression levels of the detected genes were calculated.

[0193] β-actin forward primer: 5'-TGGAATCCTGTGGCATCCATGAAAC-3', reverse primer: 5'-TAAAACGCAGCTCAGTAACAGTCCG-3',

[0194] H2-Aa forward primer: 5'-GACCTCCCAGAGACCAGGAT-3', reverse primer: 5'-GGAACACAGTCGCTTGAGGA-3',

[0195] H-2E forward primer: 5′-GATGCTGGAGACGGTTCCTC-3′, reverse primer: 5′-AGGCCTGGGTCAGGGATAAA-3′.

[0196] 1.2.3 Flow cytometry to count CD4 + T RM Cell number

[0197] To count lung CD4 + For the extravascular and intravascular fractions of T cells, anesthetized mice were injected intraperitoneally with 2 μg of anti-CD45.2 antibody 3 minutes before euthanasia. Lungs were collected in RPMI1640 containing 10% FBS before processing for flow cytometry. Lung tissue was digested with type 2 collagenase and DNase I to prepare a single-cell suspension. Cells were blocked with TruStain αCD16 / CD32 Fc-Block. Cells were stained with the fluorescent antibodies and isotype controls listed in the table below. Flow cytometry was performed using an LSR II flow cytometer. High-dimensional multiparameter spectral flow cytometry was performed on a Cytek SpectraFlo (Cytek) software using the ordinary least squares algorithm for spectral unmixing of the data, and the data were analyzed using FlowJo software.

[0198] Table 2 Antibodies required for flow cytometry

[0199]

[0200] 1.2.4 Statistical analysis

[0201] GraphPad Prism 8.0 software was used to perform statistical analysis on all data. P<0.05 indicates statistically significant differences.

[0202] 2. Results

[0203] Compared with the blank group, the relative expression of AEC H2-Aa and H-2E mRNA in the model group was significantly decreased (P<0.0001); compared with the model group, the relative expression of AEC H2-Aa and H-2E mRNA in the combination group was significantly increased (P<0.0001), while there was no significant difference in the relative expression of AEC H2-Aa and H-2E mRNA in the tangerine peel group, seabuckthorn group, and licorice group. + T RM The number of cells was significantly decreased (*P<0.001); compared with the model group, the CD4 + T RM The number of CD4 cells increased significantly (P<0.01), while the CD4 + T RM There was no significant difference in cell number. Figure 22-24 .

[0204] 3. Conclusion

[0205] The composition increases the expression of MCH-Ⅱ class molecules in lung epithelial cells, thereby regulating CD4 + T RM The increase in cell number plays a role in improving lung immunity.

[0206] Application of the composition in treating respiratory mucosal damage caused by chemical injury: The anti-cold and respiratory injury composition of the present invention can be used in the preparation of a remedy for respiratory mucosal damage caused by chemical injury (such as chemical gases, smoke, chemical solvents, etc.). By regulating inflammatory pathways, the composition can repair damaged mucosa, alleviate local inflammatory reactions, and improve the patient's respiratory function, demonstrating a significant protective effect.

[0207] Use of the composition in treating respiratory mucosal injuries caused by burns and other causes: The composition provided by this invention has unique therapeutic efficacy in treating respiratory mucosal injuries caused by fire, hot gas inhalation, or burns. By reducing the release of inflammatory factors, repairing damaged cell structures, and restoring mucosal barrier function by targeting specific molecular mechanisms, the composition significantly improves patient healing outcomes.

[0208] Application of the composition in the preparation of drugs for lung injury: The composition of the present invention exhibits multi-target therapeutic advantages in the treatment of lung injury caused by various causes, such as viral infection, physical injury, or chemical exposure. By regulating inflammatory factors and immune responses, the composition can alleviate pathological lung damage, promote lung tissue repair, and enhance lung function.

[0209] The composition works through the novel MEG3 / miR-223 / NLRP3 axis: The anti-cold and respiratory injury composition of the present invention works through the novel MEG3 / miR-223 / NLRP3 signaling axis. By regulating the maternally expressed gene 3 (MEG3), microRNA-223 (miR-223), and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) pathways, the composition exerts anti-inflammatory and repair effects, inhibiting damage expansion and accelerating the repair and functional recovery of damaged tissues.

[0210] The composition inhibits MHC-II expression on lung epithelial cells and promotes TRM cell proliferation: A mouse model demonstrated that the composition can inhibit abnormal MHC-II expression on the surface of lung epithelial cells (AECs) while significantly promoting the proliferation and activation of CD4+ TRM cells. This effect effectively enhances the body's ability to resist viral infection, thereby forming a powerful protective mechanism against respiratory viral invasion.

[0211] A novel composition combats colds and respiratory tract damage through a novel mechanism: The composition of this invention exhibits a novel mechanism of action in treating colds and respiratory tract damage, combining antiviral, anti-inflammatory, and immunomodulatory functions. By jointly regulating multiple molecular targets and pathways, the composition not only prevents and treats respiratory tract damage, but also accelerates tissue repair and significantly reduces the risk of secondary infection, providing a safe and efficient drug preparation solution.

[0212] The above description is only a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent substitutions and improvements made by any technician familiar with this technical field within the technical scope disclosed by the present invention and within the spirit and principles of the present invention should be covered by the scope of protection of the present invention.

Claims

1. A composition for resisting colds and respiratory tract damage, characterized in that: The composition is prepared according to the following medicinal material formula in parts by weight: 2 to 10 parts of licorice, 3 to 16 parts of sea buckthorn, and 3 to 9 parts of tangerine peel.

2. The anti-cold and anti-respiratory tract damage composition according to claim 1, characterized in that: The specific amounts of licorice, seabuckthorn and tangerine peel are 5.1 parts, 5.2 parts and 8.1 parts respectively.

3. The composition for resisting colds and respiratory tract damage according to claim 1, characterized in that: The raw materials of the composition are: 0.1-6 parts of honeysuckle are added to licorice, sea buckthorn and tangerine peel, or 0.1-6 parts of other immune-boosting and anti-cold medicinal materials are added to the composition; the sea buckthorn in the composition can be replaced by 0.1-6 parts of honeysuckle.

4. The method for preparing the anti-cold and anti-respiratory tract injury composition according to any one of claims 1 to 3, characterized in that: include: The method comprises the following steps: steam distilling liquorice, collecting the distillate for later use; combining the liquorice residue with dried tangerine peel and seabuckthorn medicinal materials, mixing them evenly, decocting them twice with water, each time for 0.1-1 hour, and collecting the decoction; combining the decoctions, filtering, and combining the filtrate with liquorice extract; concentrating under reduced pressure to obtain an extract with a relative density of 1.22 g / ml (60° C.), adding ethanol to adjust the alcohol content to 60-75% (V / V), allowing the extract to stand for 24 hours, filtering, recovering the ethanol from the supernatant under reduced pressure, concentrating, and adding the liquorice distillate; and finally adding pharmaceutically acceptable excipients and preparing various dosage forms through conventional processes.

5. The method for preparing the anti-cold and anti-respiratory tract damage composition according to any one of claims 1 to 3, characterized in that: include: Soak in water for 1 hour, heat and boil 2-3 times, 1-3 hours each time, filter, combine the filtrate, and concentrate under reduced pressure to a relative density of 1.13-1.28g / cm 3 The extract is heated to 60°C, ethanol is added to make the alcohol content reach 60-75% (V / V), the extract is allowed to stand for 20-24 hours, filtered, the supernatant is decompressed to recover the ethanol, and concentrated until there is no alcohol taste. Finally, acceptable excipients are added through conventional processes to prepare various commonly used dosage forms.

6. The method for preparing the anti-cold and anti-respiratory tract damage composition according to any one of claims 1 to 3, characterized in that: include: Soak in water for 1 hour, heat and boil twice, each time for 2 hours, filter, and finally add acceptable excipients through conventional procedures to prepare various available dosage forms.

7. The method for preparing the anti-cold and anti-respiratory tract damage composition according to any one of claims 1 to 3, characterized in that: include: The raw materials are crushed and prepared into various commonly used preparations or foods such as tea bags, powders, ultrafine powders, candies, etc.

8. The anti-cold and anti-respiratory tract damage composition according to any one of claims 1 to 3, characterized in that: Excipients required for product preparation may be added as needed, including one or more of fillers, disintegrants, solubilizers, binders, flavoring agents, colorants, preservatives, lubricants, fragrances, and emulsifiers, and the dosage forms prepared include but are not limited to beverages, oral liquids, powders, tablets, granules, pills, capsules, ointments, pills, injections, nasal drops, aerosols, suppositories, or films.

9. Use of the anti-cold and anti-respiratory tract injury composition according to any one of claims 1 to 3 in the preparation of a medicament for treating symptoms of respiratory tract injury, expectoration, sneezing, eye irritation, pharyngeal discomfort, and chest tightness.

10. Use of the anti-cold and anti-respiratory tract damage composition according to any one of claims 1 to 3 in the preparation of anti-inflammatory, anti-inflammatory, anti-viral and anti-cold drugs.

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