Traditional Chinese medicine preparation for treating allergic rhinitis and preparation method thereof

Abstract

The invention provides a traditional Chinese medicine preparation for treating allergic rhinitis and a preparation method thereof, and relates to the technical field of traditional Chinese medicine pharmacy, the traditional Chinese medicine preparation is prepared from radix asteris, schizonepeta spike, cynanchum paniculatum, perillaseed, spina gleditsiae, cottonrose hibiscus leaf, magnolia flower, semen brassicae, cortex lycii radicis, dendrobe, oroxylum indicum and rhodomyrtus tomentosa; an in-vitro experiment shows that the traditional Chinese medicine preparation can remarkably inhibit the activity of hyaluronidase, also can inhibit the release of beta-HEX in an RBL-2H3 cell degranulation model, reduces the HIS level in a cell supernatant, and effectively improves the morphological abnormality caused by degranulation of RBL-2H3 cells; in-vivo experiments show that the traditional Chinese medicine preparation can significantly reduce behavioral scores of mice suffering from allergic rhinitis, inhibit expression of inflammatory factors TNF-alpha, serum Ig E and OVE-sIg E levels and HA concentration, reduce histopathologic changes caused by allergic rhinitis, effectively repair immune injuries of nasal mucosa of the mice and improve the curative effect of the mice. And a safe and effective new strategy is provided for the treatment of allergic rhinitis.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine pharmaceutical technology, specifically to a traditional Chinese medicine preparation for treating allergic rhinitis and its preparation method. Background Technology

[0002] Allergic rhinitis (AR), also known as hay fever, is a chronic inflammatory disease of the nasal mucosa mediated by IgE. Clinical manifestations include paroxysmal sneezing, clear nasal discharge, nasal itching, and nasal congestion. Based on its characteristics, allergic rhinitis can be divided into seasonal (e.g., hay fever) and perennial (e.g., dust mite allergy). Epidemiological surveys show that the global prevalence of AR has reached 10%-40%, and the prevalence among adults in my country is approximately 17.6%, making it a significant public health concern. Currently, Western medicine treatments for allergic rhinitis mainly use antihistamines, nasal corticosteroids, and leukotriene receptor antagonists. While these drugs can relieve symptoms in the short term, long-term use may lead to adverse reactions such as nasal dryness, drug-induced rhinitis, and mucosal atrophy. Furthermore, they cannot fundamentally regulate immune imbalance, and relapse is common after discontinuation of medication.

[0003] In Traditional Chinese Medicine (TCM), allergic rhinitis falls under the category of "nasal congestion," and its pathogenesis is considered closely related to dysfunction of the lungs, spleen, and kidneys. TCM has accumulated over a thousand years of clinical experience in treating this condition, forming a unique system of syndrome differentiation and treatment. TCM clinical treatment emphasizes syndrome differentiation and treatment, harmonizing the Qi and blood of the internal organs, strengthening the body's resistance, and eliminating pathogenic factors, addressing the root cause of the condition to achieve the dual goals of controlling clinical symptoms and reducing recurrence rates. Existing classic formulas such as Yu Ping Feng San and Xin Yi Qing Fei Yin, while effective through multi-target immune regulation and improved nasal microcirculation, still face clinical bottlenecks in practical application, including a long onset of action (usually requiring 2-4 weeks of continuous use), the bitter taste of some formulas affecting medication adherence, and limited improvement in severe, persistent nasal congestion. Therefore, there is an urgent need to develop a TCM preparation with rapid onset of action, few side effects, and both symptom control and immune regulation functions. Summary of the Invention

[0004] (a) Technical problems to be solved

[0005] To address the shortcomings of existing technologies, this invention provides a traditional Chinese medicine preparation for treating allergic rhinitis and its preparation method.

[0006] (II) Technical Solution

[0007] To achieve the above objectives, the present invention provides the following technical solution:

[0008] This invention provides a traditional Chinese medicine preparation for treating allergic rhinitis, which is composed of the following raw materials in parts by weight: 5-25 parts of Aster tataricus, 3-23 parts of Schizonepeta tenuifolia spike, 3-22 parts of Cynanchum paniculatum, 3-22 parts of Perilla frutescens seed, 1-18 parts of Gleditsia sinensis thorn, 1-20 parts of Hibiscus mutabilis leaf, 1-18 parts of Magnolia biondii flower, 1-20 parts of Sinapis alba seed, 1-18 parts of Lycium chinense root bark, 1-18 parts of Dendrobium nobile, 1-20 parts of Oroxylum indicum, and 1-15 parts of Myrtle.

[0009] Furthermore, the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 10-20 parts of Aster tataricus, 8-18 parts of Schizonepeta tenuifolia spike, 8-18 parts of Cynanchum paniculatum, 8-18 parts of Perilla frutescens seed, 4-14 parts of Gleditsia sinensis thorn, 5-15 parts of Hibiscus mutabilis leaf, 2-14 parts of Magnolia biondii, 1-12 parts of Sinapis alba seed, 4-14 parts of Lycium chinense root bark, 4-14 parts of Dendrobium nobile, 3-15 parts of Oroxylum indicum, and 3-10 parts of Myrtle.

[0010] Furthermore, the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 15 parts of Aster tataricus, 13 parts of Schizonepeta tenuifolia spike, 13 parts of Cynanchum paniculatum, 13 parts of Perilla frutescens seed, 9 parts of Gleditsia sinensis thorn, 10 parts of Hibiscus mutabilis leaf, 7 parts of Magnolia biondii, 6 parts of Sinapis alba seed, 9 parts of Lycium chinense root bark, 9 parts of Dendrobium nobile, 8 parts of Oroxylum indicum, and 5 parts of Myrtle.

[0011] This invention provides a method for preparing a traditional Chinese medicine preparation for treating allergic rhinitis. The medicine is prepared into any dosage form acceptable to the human body, including pills, powders, granules, decoctions, tablets, sprays, etc., by conventional traditional Chinese medicine manufacturing methods and with medically acceptable excipients.

[0012] Furthermore, the preparation method of the traditional Chinese medicine nasal spray includes the following steps:

[0013] (1) Weigh each Chinese herbal raw material according to the above weight proportions, wash and dry them, and then grind them into fine powder;

[0014] (2) Add the above fine powder to 70% ethanol at a material-to-liquid ratio of 1:8 (w / v), and reflux extract twice at 60℃ for 1.5h each time. Combine the extracts and concentrate under reduced pressure to obtain a clear extract with a relative density of 1.1-1.3 (measured at 60℃).

[0015] (3) Add an equal volume of distilled water to the clear paste to dilute it, use D101 macroporous adsorption resin, elute with 50% ethanol, and collect the eluent.

[0016] (4) Mix the above eluent with the pre-prepared excipients, adjust the pH to 5.0-6.0 with 0.1mol / L sodium hydroxide solution, filter through a 0.45μm microporous membrane, sterilize, and fill to obtain the nasal spray of the traditional Chinese medicine preparation of the present invention.

[0017] Further, in step (4), the amount of excipients added is 4-10 wt% of the eluent, which includes preservatives, humectants, and absorption promoters, mixed in a ratio of 1:3:1. The preservatives are one or more of benzalkonium chloride, potassium sorbate, and sodium benzoate; the humectants are one or more of glycerin, sorbitol, and mannitol; and the absorption promoters are one or more of polysorbate 80 and cyclodextrin.

[0018] This invention provides the application of the aforementioned traditional Chinese medicine preparation in the preparation of drugs for the prevention and / or treatment of allergic rhinitis.

[0019] The pharmacological effects of each Chinese herbal raw material in the Chinese herbal preparation of this invention are as follows:

[0020] Aster: It has a pungent and bitter taste, is warm in nature, and enters the lung meridian; it has the effects of moistening the lungs and lowering qi, eliminating phlegm and relieving cough; it is used for cough with excessive phlegm, cough that is new or chronic, and cough with blood due to overwork.

[0021] Schizonepeta tenuifolia spikes: pungent in taste, slightly warm in nature, and enters the lung and liver meridians; it has the effects of relieving exterior syndromes and dispelling wind, promoting rash eruption, and eliminating sores; it is used for colds, headaches, measles, rubella, and early-stage sores.

[0022] Xu Changqing: It has a pungent taste and warm nature, and enters the liver and stomach meridians; it has the effects of dispelling wind, resolving dampness, relieving pain, and relieving itching; it is used for rheumatic pain, stomach pain and bloating, toothache, back pain, traumatic injuries, urticaria, and eczema.

[0023] Perilla seed: It has a pungent taste and warm nature, and enters the lung meridian; it has the effects of lowering qi and resolving phlegm, relieving cough and asthma, and moistening the intestines and promoting bowel movement; it is used for phlegm accumulation and qi stagnation, cough and asthma, and constipation due to intestinal dryness.

[0024] Gleditsia sinensis thorns: pungent in taste and warm in nature, entering the liver and stomach meridians; has the effects of reducing swelling and promoting detoxification, draining pus and killing parasites; used for carbuncles in their early stages or when pus has formed but not yet ruptured; used externally for scabies and leprosy.

[0025] Hibiscus mutabilis leaves: pungent in taste, neutral in nature, and enters the lung and liver meridians; it has the effects of cooling blood, detoxifying, reducing swelling, and relieving pain; it is used for carbuncles, boils, shingles, red and swollen eyes, and traumatic injuries.

[0026] Magnolia flower bud: It has a pungent taste and warm properties, and enters the lung and stomach meridians; it has the effects of dispelling wind-cold and clearing nasal passages; it is used for wind-cold headache, nasal congestion and runny nose, nasal congestion, and sinusitis.

[0027] White mustard seed: It is pungent and warm in nature, and enters the lung meridian; it has the effects of warming the lungs, resolving phlegm and promoting qi circulation, dispersing nodules, unblocking the meridians and relieving pain; it is used for cough with cold phlegm, chest and rib pain, phlegm stagnation in the meridians, numbness and pain in the joints, phlegm and dampness, and carbuncles and boils.

[0028] Lycium bark: sweet in taste, cold in nature, enters the lung, liver and kidney meridians; has the effects of cooling blood and relieving steaming heat, clearing the lungs and reducing fire; used for yin deficiency with tidal fever, bone steaming and night sweats, lung heat cough, hemoptysis, epistaxis, and internal heat with thirst.

[0029] Dendrobium: It tastes sweet and is slightly cold in nature. It enters the stomach and kidney meridians. It has the effects of nourishing the stomach and promoting the production of body fluids, nourishing yin and clearing heat. It is used for febrile diseases with depletion of body fluids, dry mouth and thirst, insufficient stomach yin, poor appetite and dry retching, persistent low fever after illness, yin deficiency and fire excess, bone steaming fever, blurred vision, and weakness of muscles and bones.

[0030] Wood Oroxylum: It tastes bitter and sweet, and is cool in nature. It enters the lung, liver, and stomach meridians. It has the effects of clearing the lungs and relieving sore throat, soothing the liver and stomach. It is used for lung heat cough, sore throat, hoarseness, and liver and stomach pain.

[0031] Myrtle: It tastes sweet and astringent, is neutral in nature, and enters the liver and spleen meridians; it has the effects of nourishing blood and stopping bleeding, astringing the intestines and consolidating essence; it is used for blood deficiency and weakness, hematemesis, epistaxis, coughing up blood due to overwork, hematochezia, metrorrhagia, seminal emission, leukorrhea, dysentery, rectal prolapse, burns, and external bleeding.

[0032] (III) Beneficial Effects

[0033] This invention provides a traditional Chinese medicine preparation for treating allergic rhinitis and its preparation method. The preparation consists of twelve herbs: Aster tataricus, Schizonepeta tenuifolia, Cynanchum paniculatum, Perilla frutescens, Gleditsia sinensis, Hibiscus mutabilis leaf, Magnolia biondii, Sinapis alba seed, Lycium chinense root bark, Dendrobium nobile, Oroxylum indicum, and Myrtle. This formula addresses the root cause of lung congestion, focusing on "opening the lungs and clearing the nasal passages." It combines the medicinal properties of each herb to clear the nasal passages, dispel wind and resolve phlegm to alleviate the symptoms, while also clearing heat, nourishing yin, and harmonizing and protecting the body's foundation. Aster tataricus and Schizonepeta tenuifolia are the principal herbs. Aster tataricus moistens the lungs, lowers qi, resolves phlegm, and stops coughing, thus relieving cough and reducing phlegm secretion. Schizonepeta tenuifolia is light and dispersing, effectively dispelling wind and pathogens, relieving symptoms such as nasal congestion, sneezing, and runny nose. Together, they work synergistically to open the lungs, dispel pathogens, resolve phlegm, and stop coughing, targeting the main pathogenesis of allergic rhinitis: wind-evil attacking the lungs and internal accumulation of phlegm. The formula uses Cynanchum paniculatum, Perilla frutescens, Gleditsia sinensis thorns, and Hibiscus mutabilis leaves as assistant herbs. Cynanchum paniculatum dispels wind, unblocks the meridians, and relieves pain, assisting the principal herb in dispelling wind and releasing the exterior. Perilla frutescens lowers qi, resolves phlegm, stops cough, and relieves asthma. When used in conjunction with Aster tataricus, it can enhance the effect of resolving phlegm and stopping cough. Gleditsia sinensis thorns reduce swelling, drain pus, and clear the nasal passages. Hibiscus mutabilis leaves clear heat, detoxify, cool the blood, stop bleeding, reduce swelling, and relieve pain, playing an auxiliary role in reducing swelling (for nasal turbinate swelling) or inhibiting heat toxicity. The four herbs work together to assist the principal herb from different angles, jointly targeting the concurrent symptoms of allergic rhinitis and resolving the core symptom cluster. Magnolia flower, white mustard seed, lycium bark, dendrobium, and oroxylum indicum are used as adjuvant herbs. Magnolia flower disperses wind-cold and clears nasal passages; white mustard seed warms the lungs, resolves phlegm, promotes qi circulation, disperses stagnation, and can warm and transform cold phlegm (nasal discharge), helping to clear deep nasal congestion; lycium bark cools the blood, eliminates steaming heat, clears lung heat, and prevents latent heat from damaging yin; dendrobium benefits the stomach, generates fluids, nourishes yin, clears heat, moistens dryness, and soothes the throat, while also working with lycium bark to counteract the drying properties of magnolia flower, schizonepeta spike, and perilla seed; oroxylum indicum clears the lungs, soothes the throat, calms the liver and stomach, moistens the lungs, and opens the voice, thus clearing the throat. Each herb performs its specific function, enhancing the therapeutic effect on the main and secondary symptoms while also harmonizing the properties of the herbs. Myrtle is used as the guiding herb, with its astringent properties, possessing the dual effects of "guiding the medicine to its proper channels" and "harmonizing the properties of the herbs."

[0034] In vitro experiments showed that the traditional Chinese medicine preparation of the present invention has significant anti-allergic activity. Pharmacodynamic evaluation showed that the traditional Chinese medicine preparation of the present invention has a significant inhibitory effect on hyaluronidase activity. In the RBL-2H3 cell degranulation model, the traditional Chinese medicine preparation of the present invention can significantly inhibit β-HEX release and reduce the HIS level in the cell supernatant. In addition, the traditional Chinese medicine preparation of the present invention can also effectively improve the morphological abnormalities of RBL-2H3 cells caused by degranulation.

[0035] In vivo experiments showed that the traditional Chinese medicine preparation of this invention has a significant therapeutic effect on mice with allergic rhinitis, significantly reducing the behavioral scores of mice with allergic rhinitis and alleviating symptoms such as sneezing and runny nose; it can also inhibit the expression of pro-inflammatory cytokine TNF-α and the levels of serum IgE, OVE-sIgE and HA concentration, effectively improving the body's inflammatory response; in addition, the traditional Chinese medicine preparation of this invention can reduce the infiltration of inflammatory cells in nasal mucosa tissue, reduce goblet cell proliferation, effectively repair immune damage to the nasal mucosa in mice, and reduce the histopathological changes of allergic rhinitis, providing a safe and effective new strategy for the treatment of allergic rhinitis. Attached Figure Description

[0036] Figure 1 This section compares the hyaluronidase inhibition rates of each drug group. Note: Compared with the chlorpheniramine group, * P<0.05, ** P<0.01.

[0037] Figure 2 The results of toluidine blue staining of cells in each group.

[0038] Figure 3 HE staining results of nasal mucosa tissue from each group of mice. Note: A is the control group; B is the model group; C is the loratadine group; D is the low-dose group of the traditional Chinese medicine preparation of this invention; E is the medium-dose group of the traditional Chinese medicine preparation of this invention; F is the high-dose group of the traditional Chinese medicine preparation of this invention. Detailed Implementation

[0039] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0040] Example 1

[0041] A traditional Chinese medicine preparation for treating allergic rhinitis, the preparation comprising the following raw materials in parts by weight: 15 parts Aster tataricus, 13 parts Schizonepeta tenuifolia, 13 parts Cynanchum paniculatum, 13 parts Perilla frutescens, 9 parts Gleditsia sinensis, 10 parts Hibiscus mutabilis leaves, 7 parts Magnolia biondii, 6 parts Sinapis alba, 9 parts Lycium chinense root bark, 9 parts Dendrobium nobile, 8 parts Oroxylum indicum, and 5 parts Myrtle.

[0042] Example 2

[0043] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 5 parts of Aster tataricus, 3 parts of Schizonepeta tenuifolia spike, 3 parts of Cynanchum paniculatum, 3 parts of Perilla frutescens seed, 1 part of Gleditsia sinensis thorn, 1 part of Hibiscus mutabilis leaf, 1 part of Magnolia biondii, 1 part of Sinapis alba seed, 1 part of Lycium chinense root bark, 1 part of Dendrobium nobile, 1 part of Oroxylum indicum, and 1 part of Myrtle.

[0044] Example 3

[0045] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 25 parts of Aster tataricus, 23 parts of Schizonepeta tenuifolia spike, 22 parts of Cynanchum paniculatum, 22 parts of Perilla frutescens seed, 18 parts of Gleditsia sinensis thorn, 20 parts of Hibiscus mutabilis leaf, 18 parts of Magnolia biondii, 20 parts of Sinapis alba seed, 18 parts of Lycium chinense root bark, 18 parts of Dendrobium nobile, 20 parts of Oroxylum indicum, and 15 parts of Myrtle.

[0046] Example 4

[0047] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 12 parts of Aster tataricus, 10 parts of Schizonepeta tenuifolia spike, 10 parts of Cynanchum paniculatum, 9 parts of Perilla frutescens seed, 9 parts of Gleditsia sinensis thorn, 10 parts of Hibiscus mutabilis leaf, 9 parts of Magnolia biondii, 8 parts of Sinapis alba seed, 8 parts of Lycium chinense root bark, 9 parts of Dendrobium nobile, 7 parts of Oroxylum indicum, and 5 parts of Myrtle.

[0048] Example 5

[0049] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 9 parts of Aster tataricus, 8 parts of Schizonepeta tenuifolia spike, 7 parts of Cynanchum paniculatum, 7 parts of Perilla frutescens seed, 5 parts of Gleditsia sinensis thorn, 3 parts of Hibiscus mutabilis leaf, 3 parts of Magnolia biondii, 5 parts of Sinapis alba seed, 3 parts of Lycium chinense root bark, 3 parts of Dendrobium nobile, 5 parts of Oroxylum indicum, and 3 parts of Myrtle.

[0050] Example 6

[0051] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 22 parts of Aster tataricus, 21 parts of Schizonepeta tenuifolia spike, 20 parts of Cynanchum paniculatum, 20 parts of Perilla frutescens seed, 16 parts of Gleditsia sinensis thorn, 18 parts of Hibiscus mutabilis leaf, 16 parts of Magnolia biondii, 18 parts of Sinapis alba seed, 16 parts of Lycium chinense root bark, 16 parts of Dendrobium nobile, 18 parts of Oroxylum indicum, and 13 parts of Myrtle.

[0052] Example 7

[0053] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 7 parts Aster tataricus, 5 parts Schizonepeta tenuifolia, 4 parts Cynanchum paniculatum, 4 parts Perilla frutescens, 2 parts Gleditsia sinensis, 3 parts Hibiscus mutabilis leaves, 2 parts Magnolia biondii, 3 parts Sinapis alba, 2 parts Lycium chinense root bark, 2 parts Dendrobium nobile, 3 parts Oroxylum indicum, and 2 parts Myrtle.

[0054] Example 8

[0055] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 20 parts of Aster tataricus, 17 parts of Schizonepeta tenuifolia spike, 18 parts of Cynanchum paniculatum, 18 parts of Perilla frutescens seed, 15 parts of Gleditsia sinensis thorn, 16 parts of Hibiscus mutabilis leaf, 13 parts of Magnolia biondii, 15 parts of Sinapis alba seed, 14 parts of Lycium chinense root bark, 15 parts of Dendrobium nobile, 16 parts of Oroxylum indicum, and 9 parts of Myrtle.

[0056] Example 9

[0057] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 18 parts of Aster tataricus, 15 parts of Schizonepeta tenuifolia spike, 15 parts of Cynanchum paniculatum, 15 parts of Perilla frutescens seed, 12 parts of Gleditsia sinensis thorn, 13 parts of Hibiscus mutabilis leaf, 9 parts of Magnolia biondii, 10 parts of Sinapis alba seed, 12 parts of Lycium chinense root bark, 13 parts of Dendrobium nobile, 12 parts of Oroxylum indicum, and 7 parts of Myrtle.

[0058] Experimental Example 1

[0059] 1. Experimental Materials

[0060] 1.1 Cell lines

[0061] The RBL-2H3 cell line was purchased from Peking Union Medical College Cell Resource Center.

[0062] 1.2 Test Drugs

[0063] Drug group 1: 13 parts of Schizonepeta tenuifolia spike, 13 parts of Cynanchum paniculatum, 13 parts of Perilla frutescens seed, 9 parts of Gleditsia sinensis thorn, 10 parts of Hibiscus mutabilis leaf, 9 parts of Lycium chinense root bark, 9 parts of Dendrobium nobile, 8 parts of Oroxylum indicum, and 5 parts of Myrtle.

[0064] Drug group 2: 15 parts of Aster tataricus, 13 parts of Cynanchum paniculatum, 13 parts of Perilla frutescens, 9 parts of Gleditsia sinensis thorns, 10 parts of Hibiscus mutabilis leaves, 7 parts of Magnolia biondii, 6 parts of Sinapis alba, 8 parts of Oroxylum indicum, and 5 parts of Myrtle.

[0065] Drug group 3: 15 parts of Aster tataricus, 13 parts of Schizonepeta tenuifolia spike, 9 parts of Gleditsia sinensis thorn, 10 parts of Hibiscus mutabilis leaf, 7 parts of Magnolia biondii, 6 parts of Sinapis alba seed, 9 parts of Lycium chinense root bark, 9 parts of Dendrobium nobile, and 5 parts of Myrtle.

[0066] Drug group 4: Aster tataricus 15 parts, Schizonepeta tenuifolia 13 parts, Cynanchum paniculatum 13 parts, Perilla frutescens 13 parts, Magnolia biondii 7 parts, Sinapis alba 6 parts, Lycium chinense root bark 9 parts, Dendrobium nobile 9 parts, Oroxylum indicum 8 parts.

[0067] Drug group 5: Aster tataricus 15 parts, Schizonepeta tenuifolia spike 13 parts, Cynanchum paniculatum 13 parts, Perilla frutescens seed 13 parts, Gleditsia sinensis thorn 9 parts, Hibiscus mutabilis leaf 10 parts, Magnolia biondii 7 parts, Sinapis alba seed 6 parts, Lycium chinense root bark 9 parts, Dendrobium nobile 9 parts, Oroxylum indicum 8 parts, Myrtle 5 parts.

[0068] Weigh each raw material according to the above weight proportions, wash, dry, grind into powder, add 10 times (m / V) of distilled water, soak for 30 minutes, decoct for 1 hour, obtain filtrate, add 8 times of distilled water again and decoct for 1 hour, combine the filtrates, concentrate to a drug mass concentration of 1g / mL to obtain the original Chinese medicine solution of drug group 1-5, and store at 4℃ for later use.

[0069] 2 Experimental Methods

[0070] 2.1 Hyaluronidase Inhibition Test

[0071] The Elson-Morgan hyaluronidase inhibition assay was performed, following the procedures outlined in Table 1 below, with 0.03 mg / mL chlorpheniramine as the positive control. The obtained data were recorded, and the anti-allergic activity was calculated using the following formula.

[0072] Hyaluronidase inhibition rate (%) = [A 对照组 -(A 样品组 -A 样品对照组 )] / A 对照组 ×100%

[0073] In the formula: A is the absorbance value of each group.

[0074] Table 1 Hyaluronidase Inhibition Experiment

[0075]

[0076]

[0077] Note: Both hyaluronidase and sodium hyaluronate were prepared using acetate buffer solution; the acetylacetone solution was prepared by adding 3.5 mL of acetylacetone to 50 mL of Na₂CO₃ solution (1.0 mol·L⁻¹). -1 The P-DAB colorimetric reagent is a mixture of p-dimethylaminobenzaldehyde (0.8g), concentrated hydrochloric acid (15mL), and an equal amount of glacial acetic acid.

[0078] 2.2 The intervention effects of each group of drugs on the RBL-2H3 cell degranulation model

[0079] Logarithmic growth phase RBL-2H3 cells were digested with trypsin to adjust the cell concentration to 4.0 × 10⁻⁶. 5 ·mL -1Cells were seeded in 24-well plates at 500 μL per well and incubated for 24 h. After complete cell adhesion, the cells were randomly divided into a control group, a model group, and experimental groups 1-5. Except for the control group, each well contained an appropriate concentration of 0.125 μg / mL. -1 Stimulation with anti-DNF-IgE monoclonal antibody for 24 h. Then, wash three times with benchtop buffer. Afterwards, except for the control group, add 200 μL of a solution containing 62.5 ng / mL to each well. -1 The cells were incubated with DNP-HAS buffer at 37°C for 1 hour to elicit an allergic reaction. The control group was treated with the same amount of DNP-HAS buffer. The reaction was then terminated by incubating on ice for 10 minutes. Control group cells were added to 1% Triton X-100 cell lysis buffer. After complete cell lysis, the lysis buffer was collected and centrifuged (4°C, 1000 rpm). -1 After 10 min, collect 50 μL of the supernatant and transfer it into a 96-well plate for the determination of total enzyme β-hexosaminease (β-HEX).

[0080] Collect 50 μL of cell supernatant from each group and transfer it to a 96-well plate. Add 50 μL of chromogenic reagent (0.1 mol·L⁻¹) to each well. -1 The 4-nitrobenzene-N-acetyl-β-D-glucosinolate solution (solute: sodium citrate buffer) was incubated at 37°C for 45 min. The reaction was terminated by adding 200 μL of stop solution, and the OD value at 405 nm was measured using a microplate reader to calculate the β-HEX release rate. Histamine (HIS) levels in the supernatant of each group were detected according to the ELISA kit instructions.

[0081] β-HEX release rate (%) = 100% × (OD value of sample group - OD value of blank well) / (OD value of total enzyme well - OD value of blank well)

[0082] After the cells had fully adhered to the culture vessel, add anti-DNP-IgE monoclonal antibody at a concentration of 125.0 ng / mL. -1 Cells were cultured in MEM basal medium for 24 hours. The medium was discarded, and the cells were incubated with basal medium for 30 minutes. The medium was then discarded again, and the cells were washed three times with TerraMaster buffer. DNP-HAS at a concentration of 62.5 ng / mL was added to the model group. -1 200 μL of Benchtop buffer was used for the control group, and the cells were incubated in an incubator for 1 h to stimulate RBL-2H3 cell activation and degranulation. The reaction was terminated by ice bath for 10 min, the supernatant was taken out, fixed with methanol for 5 min and discarded, then stained with toluidine blue solution for 10 min, washed 3 times with purified water, and the cell morphology was observed under a microscope and images were collected.

[0083] 2.3 Statistical Methods

[0084] Data analysis was performed using SPSS 25 software. Following the procedure for analysis of variance, a homogeneity of variance test was first conducted, followed by pairwise comparisons of the means among multiple experimental groups and a control group. If variances were unequal, a rank-sum test was used.

[0085] 3 Results

[0086] 3.1 Effects of each drug group on hyaluronidase inhibitory activity

[0087] Depend on Figure 1 It can be seen that each group of drugs has a certain inhibitory ability on hyaluronidase. Among them, drug group 5 has the highest hyaluronidase inhibitory activity, with an inhibition rate of up to (68.21±3.23)% and no significant difference from the positive drug chlorpheniramine group (P>0.05). This indicates that the traditional Chinese medicine preparation of the present invention has high activity in inhibiting hyaluronidase and exhibits significant anti-allergic effects.

[0088] 3.2 The intervention effects of each drug group on the RBL-2H3 cell degranulation model

[0089] As shown in Table 2, compared with the control group, the β-HEX release rate and HIS content in the supernatant of RBL-2H3 cells in the model group were significantly higher than those in the control group (P<0.01), indicating that the model was successfully established. Compared with the model group, drug groups 1-5 could inhibit the release of β-HEX and HIS from cells to varying degrees. Among them, drug group 5 had the most significant inhibitory effect on β-HEX release (P<0.01), and its HIS content was also significantly reduced (P<0.01), indicating that the traditional Chinese medicine preparation of the present invention may exert its anti-allergic effect by stabilizing the mast cell membrane and inhibiting the degranulation process.

[0090] Figure 2 The results of toluidine blue staining are shown. Under the microscope, the cells in the blank group have clear outlines, smooth and intact cell membranes, and are stained dark blue by toluidine blue. There are purplish-red metachromatic granules in the cytoplasm. In the model group, the degranulated cells exhibit diverse morphologies, with uneven and indistinct cell membranes, granule extravasation, and scattered granule traces around the cells, which are stained light blue by toluidine blue. The cells in each drug-treated group have clearer outlines, are stained dark blue by toluidine blue, and the purplish-red metachromatic granules in the cytoplasm are less obvious. Granule extravasation is significantly reduced compared to the model control. The drug group 5 showed the most significant improvement in cell morphology.

[0091] Table 2 Comparison of β-HEX release rate and HIS level in each group

[0092] Group β-HEX release rate (%) HIS level (ng / mL) control group 36.12±0.36 3.93±0.59 Model group <![CDATA[72.54±2.89 ## ]]> <![CDATA[7.21±0.18 ## ]]> Drug group 1 65.03±2.69 <![CDATA[6.20±0.48 * ]]> Drug group 2 <![CDATA[57.29±10.06 * ]]> <![CDATA[5.27±0.32 ** ]]> Drug group 3 <![CDATA[56.13±9.17 * ]]> <![CDATA[4.79±0.34 ** ]]> Drug group 4 <![CDATA[41.22±4.60 ** ]]> <![CDATA[5.02±0.41 ** ]]> Drug group 5 <![CDATA[37.40±1.73 ** ]]> <![CDATA[3.62±0.12 ** ]]>

[0093] Note: Compared with the control group, # P<0.05, ## P<0.01; compared with the model group, *P < 0.05, ** P < 0.01.

[0094] 4 Conclusions

[0095] The traditional Chinese medicine preparation of the present invention has significant anti-allergic activity. In vitro pharmacodynamic evaluation shows that the traditional Chinese medicine preparation of the present invention has a significant inhibitory effect on the activity of hyaluronidase; in the RBL-2H3 cell degranulation model, the traditional Chinese medicine preparation of the present invention can significantly inhibit the release of β-HEX and simultaneously reduce the HIS level in the cell supernatant; in addition, the traditional Chinese medicine preparation of the present invention can also effectively improve the morphological abnormalities of RBL-2H3 cells caused by degranulation. The above experimental results reveal the mechanism of action of the traditional Chinese medicine preparation of the present invention in regulating allergic reactions through multiple targets, providing a basis for its subsequent treatment of allergic rhinitis.

[0096] Experimental Example 2

[0097] 1 Experimental Materials

[0098] 1.1 Experimental Animals

[0099] SPF-grade BALB / c mice, male, 72 in number, 6 weeks old, 17 - 21 g, bred by Beijing Speveto Biotechnology Co., Ltd. The production license number of experimental animals: SCXK (Beijing) 2012-0001, and the quality certificate number of experimental animals: 110324221101200036. After the animals were purchased, they were adaptively raised in the barrier environment of this animal room and then the experiment began. The license number for using this experimental animal room: SYXK (Beijing) 2018-0008. The temperature of the animal room: 22℃ - 25℃, relative humidity: 55% - 70%.

[0100] 1.2 Experimental Drugs

[0101] 15 parts of Aster tataricus L.f., 13 parts of Schizonepeta tenuifolia Briq., 13 parts of Cynanchum paniculatum (Bunge) Kitagawa, 13 parts of Perilla frutescens (L.) Britt., 9 parts of Gleditsia sinensis Lam., 10 parts of Hibiscus mutabilis L., 7 parts of Magnolia liliflora Desr., 6 parts of Brassica alba (L.) Boiss., 9 parts of Lycium chinense Mill., 9 parts of Dendrobium officinale Kimura et Migo, 8 parts of Oroxylum indicum (L.) Kurz., 5 parts of Myrtus communis L. Weigh each raw material according to the above weight parts, wash, dry, powder, add 10 times the amount (m / V) of distilled water, soak for 30 min, then decoct for 1 h to obtain a filtrate, add 8 times the amount of distilled water again and decoct for 1 h, combine the filtrates, and concentrate to a drug mass concentration of 1 g / mL to obtain the traditional Chinese medicine stock solution of the present invention, which is stored in a 4℃ refrigerator for later use.

[0102] 2 Experimental Methods

[0103] 2.1 Animal Modeling, Grouping and Administration

[0104] After one week of acclimatization feeding, mice were randomly divided into four groups according to body weight: a control group, a model group, a loratadine (P) group, a low-dose (TL) group, a medium-dose (TM) group, and a high-dose (TH) group of the traditional Chinese medicine preparation of this invention, with 12 mice in each group. The first two weeks were the basal sensitization phase. 3.000g of aluminum hydroxide was added to 100mL of physiological saline to prepare a suspension. 100mg of ovalbumin (OVA) was added to 10mL of the aluminum hydroxide suspension, and 0.1mL was injected intraperitoneally into each mouse. The control group received an equal volume of physiological saline instead. Injections were given every other day for a total of 7 days. Starting on day 15 of modeling, the challenge phase began. 150mg of OVA was dissolved in 3mL of physiological saline, and 10μL was instilled into each nostril of the mice daily. The control group received an equal volume of physiological saline instead. Challenge was continued for 7 days, and behavioral observations evaluated the success of modeling. Starting from the day of model establishment, the loratadine group was administered a clinically equivalent dose of loratadine by gavage. The low-dose group, medium-dose group, and high-dose group of the traditional Chinese medicine preparation of this invention were administered 1000 mg / kg / d, 2000 mg / kg / d, and 4000 mg / kg / d of the original traditional Chinese medicine solution of this invention by gavage, respectively. The control group and the model group were administered an equal volume of ultrapure water by gavage. All treatments were administered once a day for 7 consecutive days.

[0105] 2.2 Detection Indicators

[0106] 2.2.1 Behavioral Evaluation

[0107] After the stimulation ended and the last administration, the rats were observed and recorded for nose scratching, sneezing, and runny nose within 30 minutes, and scored according to the severity. The scoring criteria are shown in Table 3.

[0108] Table 3 Behavioral Scoring Criteria

[0109]

[0110]

[0111] 2.2.2 ELISA method for detecting serum-related indicators

[0112] After the final behavioral observation, blood was collected by enucleation, allowed to stand for two hours, centrifuged to remove serum, and stored at -80℃. The levels of immunoglobulin E (IgE), ovalbumin-specific immunoglobulin E (OVA-sIgE), histamine (HA), and tumor necrosis factor-α (TNF-α) were measured according to the ELISA kit instructions.

[0113] 2.2.3 HE staining of nasal mucosa tissue

[0114] Mouse nasal mucosa tissue was isolated, fixed in 4% paraformaldehyde for 48 h, and decalcified. After decalcification, the tissue was trimmed into blocks and dehydrated in gradients of ethanol at different concentrations. The dehydrated tissue was then embedded in paraffin and sectioned at a thickness of 4 μm. After HE staining, the pathological morphological changes of the nasal mucosa were observed under a microscope.

[0115] 2.2.4 Statistical Methods

[0116] All data were processed using Graphpad Prism 8.0 software, and the data are expressed as mean ± standard deviation. This means that when all data series are normally distributed, ANOVA is used for analysis of variance; when variances are homogeneous, LSD-test is used for comparison between groups; when variances are unequal, Dunnett's test is used for comparison between groups; when one or more data series are not normally distributed, nonparametric tests are used.

[0117] 3 Results

[0118] 3.1 Effects of each drug administration group on the behavioral scores of mice

[0119] As shown in Table 4, compared with the control group, the model group mice exhibited nose scratching, sneezing, and runny nose behaviors, and their behavioral scores were significantly increased (P<0.01). Compared with the model group, the behavioral scores of mice in each treatment group were reduced, and the symptoms of nose scratching, sneezing, and runny nose were alleviated to varying degrees. This indicates that the traditional Chinese medicine preparation of the present invention has a significant therapeutic effect in relieving allergic symptoms in mice, with the high-dose group showing the most significant effect.

[0120] Table 4 Comparison of behavioral scores of mice in each group

[0121]

[0122]

[0123] Note: Compared with the control group, # P<0.05, ## P<0.01; compared with the model group, * P<0.05, ** P<0.01.

[0124] 3.2 Effects of each drug administration group on relevant serum indicators in mice

[0125] As shown in Table 5, compared with the control group, the serum levels of IgE, OVA-sIgE, HA, and TNF-α in the model group mice were significantly increased (P<0.01); compared with the model group, the serum levels of IgE, OVA-sIgE, HA, and TNF-α in each treatment group mice were decreased to varying degrees, indicating that the traditional Chinese medicine preparation of the present invention can reduce the body's inflammatory response and alleviate allergic reactions.

[0126] Table 5 Comparison of serum-related indicators in mice of different groups

[0127]

[0128] Note: Compared to the Control group, ## P<0.01, # P<0.05; compared with the Model group, ** P<0.01, * P<0.05.

[0129] 3.3 Effects of each drug administration group on histopathological changes of mouse nasal mucosa

[0130] Figure 3 The image shows the pathological examination results of the nasal mucosa of AR mice, with the control group ( Figure 3 A) The nasal mucosal epithelial cells of mice showed intact morphology and structure, with no ciliary loss, no inflammatory cell infiltration, and no glandular hyperplasia in the underlying connective tissue; the model group ( Figure 3 B) The nasal mucosa of mice was incomplete, with disordered arrangement of mucosal cilia, irregular cell arrangement, and visible inflammatory cell infiltration. Submucosal connective tissue showed glandular hyperplasia and goblet cell hyperplasia. Compared with the model group, the integrity of the nasal mucosa structure in each treatment group was improved to varying degrees, with reduced inflammatory cell infiltration, lessened submucosal connective tissue glandular hyperplasia, and reduced goblet cell hyperplasia, especially in the high-dose group. Figure 3 The improvement effect of F) is the most significant.

[0131] 4. Conclusion

[0132] The above experimental results show that the traditional Chinese medicine preparation of this invention exerts anti-allergic, anti-inflammatory, and immunomodulatory effects through multiple targets and pathways, and has a significant therapeutic effect on mice with allergic rhinitis. It can significantly reduce the behavioral scores of mice with allergic rhinitis and alleviate symptoms such as sneezing and runny nose. It can also inhibit the expression of pro-inflammatory cytokine TNF-α and the levels of serum IgE, OVE-sIgE and HA concentration, effectively improving the inflammatory response. In addition, the traditional Chinese medicine preparation of this invention can reduce inflammatory cell infiltration, alleviate goblet cell proliferation, effectively repair immune damage to the nasal mucosa of mice, and reduce the histopathological changes of allergic rhinitis, providing a safe and effective new strategy for the treatment of allergic rhinitis.

[0133] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine preparation for treating allergic rhinitis, characterized in that, The traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 5-25 parts of Aster tataricus, 3-23 parts of Schizonepeta tenuifolia spike, 3-22 parts of Cynanchum paniculatum, 3-22 parts of Perilla frutescens seed, 1-18 parts of Gleditsia sinensis thorn, 1-20 parts of Hibiscus mutabilis leaf, 1-18 parts of Magnolia biondii flower, 1-20 parts of Sinapis alba seed, 1-18 parts of Lycium chinense root bark, 1-18 parts of Dendrobium nobile, 1-20 parts of Oroxylum indicum, and 1-15 parts of Myrtle.

2. A traditional Chinese medicine preparation for treating allergic rhinitis according to claim 1, characterized in that, The traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 10-20 parts of Aster tataricus, 8-18 parts of Schizonepeta tenuifolia spike, 8-18 parts of Cynanchum paniculatum, 8-18 parts of Perilla frutescens seed, 4-14 parts of Gleditsia sinensis thorn, 5-15 parts of Hibiscus mutabilis leaf, 2-14 parts of Magnolia biondii flower, 1-12 parts of Sinapis alba seed, 4-14 parts of Lycium chinense root bark, 4-14 parts of Dendrobium nobile, 3-15 parts of Oroxylum indicum, and 3-10 parts of Myrtle.

3. A traditional Chinese medicine preparation for treating allergic rhinitis according to claim 1, characterized in that, The traditional Chinese medicine preparation is composed of the following raw materials in parts by weight: 15 parts of Aster tataricus, 13 parts of Schizonepeta tenuifolia spike, 13 parts of Cynanchum paniculatum, 13 parts of Perilla frutescens seed, 9 parts of Gleditsia sinensis thorn, 10 parts of Hibiscus mutabilis leaf, 7 parts of Magnolia biondii, 6 parts of Sinapis alba seed, 9 parts of Lycium chinense root bark, 9 parts of Dendrobium nobile, 8 parts of Oroxylum indicum, and 5 parts of Myrtle.

4. The method for preparing a traditional Chinese medicine preparation according to any one of claims 1 to 3, characterized in that, The drug is prepared into any dosage form acceptable to the human body, including pills, powders, granules, decoctions, tablets, sprays, etc., using conventional Chinese medicine manufacturing methods and with medically acceptable excipients.

5. The preparation method according to claim 4, characterized in that, The preparation method of the traditional Chinese medicine nasal spray includes the following steps: (1) Weigh each Chinese herbal raw material according to the above weight proportions, wash and dry them, and then grind them into fine powder; (2) Add the above fine powder to 70% ethanol at a material-to-liquid ratio of 1:8 (w / v), and reflux extract twice at 60℃ for 1.5h each time. Combine the extracts and concentrate under reduced pressure to obtain a clear extract with a relative density of 1.1-1.3 (measured at 60℃). (3) Add an equal volume of distilled water to the clear paste to dilute it, use D101 macroporous adsorption resin, elute with 50% ethanol, and collect the eluent. (4) Mix the above eluent with the pre-prepared excipients, adjust the pH to 5.0-6.0 with 0.1mol / L sodium hydroxide solution, filter through a 0.45μm microporous membrane, sterilize, and fill to obtain the nasal spray of the traditional Chinese medicine preparation of the present invention.

6. The method for preparing the nasal spray of traditional Chinese medicine preparation according to claim 5, characterized in that, In step (4), the amount of excipients added is 4-10 wt% of the eluent, which includes preservatives, humectants, and absorption promoters, mixed in a ratio of 1:3:

1. The preservatives are one or more of benzalkonium chloride, potassium sorbate, and sodium benzoate; the humectants are one or more of glycerin, sorbitol, and mannitol; and the absorption promoters are one or more of polysorbate 80 and cyclodextrin.

7. The use of the traditional Chinese medicine preparation according to any one of claims 1 to 3 in the preparation of a medicament for the prevention and / or treatment of allergic rhinitis.

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