Compound essential oil for improving allergic rhinitis as well as preparation method and application of compound essential oil

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to compound essential oil for improving allergic rhinitis as well as a preparation method and application of the compound essential oil. The compound essential oil comprises the following components in parts by mass: 1-5 parts of peppermint essential oil and 1-5 parts of perilla leaf essential oil. According to the compound essential oil disclosed by the invention, the optimal production places of the mint and the perilla leaves are obtained through experiments, and the compound essential oil disclosed by the invention has an improvement effect on mice suffering from allergic rhinitis induced by ovalbumin, and can inhibit AR inflammatory response by regulating the balance of local Th1 / Th2, Th17 and Treg cells of nasal mucosa. According to the invention, nasal administration or sniffing is carried out in the form of mixed essential oil, the administration is convenient, the process is simple, the compliance of a patient is good, and a simpler, more convenient and more effective treatment method is provided for improving allergic rhinitis.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a compound essential oil for improving allergic rhinitis, a preparation method and an application thereof. Background Art

[0002] Allergic rhinitis (AR), also known as allergic rhinitis, is a common allergic disease worldwide. It is a chronic inflammatory disease of the nasal mucosa caused by IgE-mediated mediators produced after exposure to allergens in atopic individuals. Its pathogenesis is currently linked to a classic imbalance between Th1 / Th2 immune cells and the more recently discovered imbalance between Th17 / Treg immune cells. AR is characterized by sneezing, nasal itching, runny nose, and nasal congestion. While not life-threatening, it is prone to complications from other diseases or conditions that share the same pathogenic pathway. In severe cases, it can trigger bronchial asthma. Chronic swelling of the nasal mucosa can also cause sinusitis, otitis media, and even hypoxia, severely impacting patients' quality of life. Epidemiological surveys show an increasing incidence of the disease worldwide. Clinical treatments for AR primarily include avoidance of allergen exposure, medication, and immunotherapy. Western medications primarily include antihistamines, glucocorticoids, and anti-leukotrienes. Western medicine can quickly relieve symptoms, but long-term use can easily lead to drug resistance and dependence. Therefore, in recent years, more and more researchers have been committed to exploring safer and more effective treatments. Summary of the Invention

[0003] The purpose of the present invention is to provide a compound essential oil for improving allergic rhinitis, a preparation method and application thereof. The compound essential oil of the present invention has an improving effect on mice with allergic rhinitis induced by ovalbumin.

[0004] The invention provides a compound essential oil for improving allergic rhinitis. The compound essential oil comprises the following components in parts by mass: 1 to 5 parts of peppermint essential oil and 1 to 5 parts of perilla leaf essential oil.

[0005] As a preferred embodiment, the compound essential oil comprises the following components in parts by mass: 4 to 5 parts of peppermint essential oil and 1 part of perilla leaf essential oil.

[0006] As a preferred embodiment, the mint in the mint essential oil is produced in Ziyang, Sichuan; the perilla leaf in the perilla leaf essential oil is produced in Zhangjiajie, Hunan.

[0007] The present invention also provides a preparation method of the compound essential oil, comprising the following steps: respectively extracting peppermint essential oil and perilla leaf essential oil by a steam distillation method, and mixing the peppermint essential oil and perilla leaf essential oil to obtain the compound essential oil.

[0008] As a preferred solution, when preparing the peppermint essential oil, the mass volume ratio of the peppermint to water is 55-65g:550-650mL, and the steam distillation time is 2.5-3.5h.

[0009] As a preferred solution, when preparing the perilla leaf essential oil, the mass volume ratio of the perilla leaf to water is 45-55g:1100-1300mL, and the steam distillation time is 2-3h.

[0010] The present invention also provides the use of the compound essential oil in preparing a medicine for preventing and / or treating allergic rhinitis.

[0011] As a preferred embodiment, the drug for preventing and / or treating allergic rhinitis is used after diluting the compound essential oil with base oil;

[0012] The base oil includes olive oil, sweet almond oil, sesame seed oil, tea seed oil or jojoba oil.

[0013] As a preferred embodiment, the method of using the drug for preventing and / or treating allergic rhinitis includes nasal drops or sniffing.

[0014] As a preferred embodiment, the drug for preventing and / or treating allergic rhinitis is in the form of nasal drops, sprays or gels.

[0015] Beneficial Effects: The present invention provides a compound essential oil for improving allergic rhinitis, comprising the following components by weight: 1-5 parts peppermint essential oil and 1-5 parts perilla leaf essential oil. The present invention experimentally identified the optimal production areas of peppermint and perilla leaf. The compound essential oil improves ovalbumin-induced allergic rhinitis in mice and inhibits AR inflammatory responses by regulating the local Th1 / Th2 and Th17 / Treg cell balances in the nasal mucosa.

[0016] The compound essential oil of the present invention has a significant synergistic effect. In the examples, it was found that the compound essential oil is more effective than using peppermint essential oil or perilla leaf essential oil alone, mainly in reducing the release rate of β-hexosaminidase in P815 cells induced by C48 / 80 and the NO content in RAW264.7 cells induced by LPS, especially in inhibiting the release of NO.

[0017] The compound essential oil of the present invention has an improving effect on mice with ovalbumin-induced allergic rhinitis, and can inhibit AR inflammatory response by regulating the local Th1 / Th2 and Th17 / Treg cell balance in the nasal mucosa. The effect is comparable to that of budesonide nasal spray in terms of behavioral indicators and mRNA expression levels of IL-5, IL-13 and RORγt.

[0018] The compound essential oil of the present invention is composed of only two volatile oils of traditional Chinese medicines, has simple ingredients, and both are traditional Chinese medicines that are both medicinal and edible. It has both therapeutic efficacy and is easily available, safe, and inexpensive in daily life. It has a simple extraction method, is convenient for public preparation and use, and has broad application prospects and good economic value. The compound essential oil of the present invention can be administered by nasal drops or sniffing in the form of mixed essential oils, is convenient to administer, has a simple process, and has good patient compliance, providing a simpler, more convenient, and more effective treatment method for improving allergic rhinitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments are briefly introduced below.

[0020] Figure 1 The effects of different essential oils in Example 1 on the cytotoxicity and β-hexosaminidase release of P815 cells, where A is the cell survival rate, B to G are the β-hexase release rates, B is magnolia and ligusticum, C is perilla leaf and mint, D is tangerine peel and wormwood leaf, E is patchouli and eucalyptus leaf, F is asarum and tea tree, and G is dexamethasone;

[0021] Figure 2 The effects of different essential oils in Example 2 on the cytotoxicity and NO production of RAW264.7 cells, where A is the cell survival rate and B is the NO release;

[0022] Figure 3 The effects of the compound essential oils in Example 3 on the cytotoxicity and β-hexosaminidase release of P815 cells are shown in Table 3, where A is the cell survival rate, B to E are the β-hexase release rates, F1 is Ligusticum chuanxiong: Magnolia bud: Mentha arvensis; F2 is Ligusticum chuanxiong: Magnolia bud: Perilla leaf; F3 is Ligusticum chuanxiong: Mentha arvensis: Perilla leaf; F4 is Ligusticum chuanxiong: Ligusticum chuanxiong; F5 is Ligusticum chuanxiong: Mentha arvensis; F6 is Ligusticum chuanxiong: Perilla leaf; F7 is Ligusticum chuanxiong: Mentha arvensis: Perilla leaf; F8 is Mentha arvensis: Perilla leaf; F9 is Ligusticum chuanxiong: Mentha arvensis; F10 is Ligusticum chuanxiong: Perilla leaf; F11 is Ligusticum chuanxiong: Magnolia bud: Mentha arvensis: Perilla leaf, and the proportions of the essential oils are all equal by mass.

[0023] Figure 4 The effects of the compound essential oils in Example 4 on the cytotoxicity and NO production of RAW264.7 cells, wherein A is the cell survival rate, B is the NO release, F1 is Ligusticum chuanxiong: Magnolia bud: Mentha arvensis; F2 is Ligusticum chuanxiong: Magnolia bud: Perilla leaf; F3 is Ligusticum chuanxiong: Mentha arvensis: Perilla leaf; F4 is Magnolia bud: Ligusticum chuanxiong; F5 is Magnolia bud: Mentha arvensis; F6 is Magnolia bud: Perilla leaf; F7 is Magnolia bud: Mentha arvensis: Perilla leaf; F8 is Mentha arvensis: Perilla leaf; F9 is Ligusticum chuanxiong: Mentha arvensis; F10 is Ligusticum chuanxiong: Perilla leaf; F11 is Ligusticum chuanxiong: Magnolia bud: Mentha arvensis: Perilla leaf, and the proportions of the essential oils are all equal by mass;

[0024] Figure 5 The effect of peppermint essential oils from different origins on the cytotoxicity and β-hexosaminidase release of P815 cells in Example 5, where A is the cell survival rate and B is the β-hexase release rate;

[0025] Figure 6 The effect of peppermint essential oils from different origins on the cytotoxicity and NO production of RAW264.7 cells in Example 6, where A is the cell survival rate and B is the NO release;

[0026] Figure 7 The effect of perilla leaf essential oils from different origins on the cytotoxicity and β-hexosaminidase release of P815 cells in Example 7, where A is the cell survival rate and B is the β-hexase release rate;

[0027] Figure 8 The effect of perilla leaf essential oils from different origins on the cytotoxicity and NO production of RAW264.7 cells in Example 8, where A is the cell survival rate and B is the NO release;

[0028] Figure 9 The effects of different ratios of peppermint and perilla leaf essential oils on P815 cell cytotoxicity and β-hexosaminidase release in Example 9, where A is the cell survival rate and B is the β-hexase release rate;

[0029] Figure 10 The effects of different ratios of mint and perilla leaf essential oils on the cytotoxicity and NO production of RAW264.7 cells in Example 10, where A is the cell survival rate and B is the NO release;

[0030] Figure 11 The effect of the compound essential oil in Example 12 on the number of sneezes and nose scratches within 10 minutes after the last nasal drop challenge in AR mice, where A is the number of sneezes within 10 minutes and B is the number of nose scratches within 10 minutes;

[0031] Figure 12 The effect of the compound essential oil in Example 12 on eosinophils in the blood of AR mice;

[0032] Figure 13 The effect of the compound essential oil in Example 12 on the degree of inflammatory infiltration in the nasal mucosa of AR mice is shown in FIG. 1 , where the black bar represents the scale, and the length represents 250 μm (upper figure) / 50 μm (lower figure);

[0033] Figure 14 The effect of the compound essential oil in Example 12 on goblet cells in the nasal mucosa of AR mice is shown in FIG. 1 , where the black bar represents the scale, and the length represents 250 μm (upper figure) / 50 μm (lower figure);

[0034] Figure 15The effect of the compound essential oil in Example 12 on the degree of inflammatory infiltration in the lung tissue of AR mice, where the black color represents the scale bar, and the length represents 250 μm (upper figure) / 50 μm (lower figure);

[0035] Figure 16 is the expression of Th1 / Th2 cytokine mRNA in the nasal mucosa of each group of mice in Example 12, wherein A is the relative expression of IFN-γ mRNA, B is the relative expression of IL-4 mRNA, C is the relative expression of IL-5 mRNA, and D is the relative expression of IL-13 mRNA. a is the blank group; b is the model group; c is the budesonide group; d is the high-dose compound essential oil group; and e is the low-dose compound essential oil group.

[0036] Figure 17 is the expression of Th1 / Th2 cell-specific transcription factor mRNA in the nasal mucosa of each group of mice in Example 12, wherein A is the relative expression level of T-bet mRNA, B is the relative expression level of GATA3 mRNA, a is the blank group; b is the model group; c is the budesonide group; d is the high-dose compound essential oil group; e is the low-dose compound essential oil group;

[0037] Figure 18 The expression of Th17 / Treg cell-specific transcription factor mRNA in the nasal mucosa of each group of mice in Example 12, wherein A is the relative expression level of RORγt mRNA, B is the relative expression level of Foxp3 mRNA, a is the blank group; b is the model group; c is the budesonide group; d is the high-dose compound essential oil group; and e is the low-dose compound essential oil group. DETAILED DESCRIPTION

[0038] The present invention provides a compound essential oil for improving allergic rhinitis, wherein the compound essential oil comprises the following components in parts by mass: 1 to 5 parts of peppermint essential oil and 1 to 5 parts of perilla leaf essential oil. As a specific embodiment, the mass parts of the compound essential oil can be 1 part of peppermint essential oil and 1 part of perilla leaf essential oil, 2 parts of peppermint essential oil and 1 part of perilla leaf essential oil, 3 parts of peppermint essential oil and 1 part of perilla leaf essential oil, 4 parts of peppermint essential oil and 1 part of perilla leaf essential oil, 5 parts of peppermint essential oil and 1 part of perilla leaf essential oil, 1 part of peppermint essential oil and 5 parts of perilla leaf essential oil, 1 part of peppermint essential oil and 4 parts of perilla leaf essential oil, 1 part of peppermint essential oil and 3 parts of perilla leaf essential oil or 1 part of peppermint essential oil and 2 parts of perilla leaf essential oil; as a specific embodiment, when the compound essential oil When the mass proportion of the essential oils is 4 parts of peppermint essential oil and 1 part of perilla leaf essential oil or 5 parts of peppermint essential oil and 1 part of perilla leaf essential oil, the β-hexosaminidase release rate of P815 cells induced by C48 / 80 and the NO content of RAW264.7 cells induced by LPS are reduced, and the compound essential oil has a synergistic effect, which is better than the effect of crude oil at the same dosage concentration. This proves that when the mass proportion of the essential oils is 4 parts of peppermint essential oil and 1 part of perilla leaf essential oil or 5 parts of peppermint essential oil and 1 part of perilla leaf essential oil, it is the optimal ratio of the compound essential oils.

[0039] As a specific embodiment, the compound essential oil of the present invention can be treated by nasal administration, which is simple to operate, efficient, safe, and has good patient compliance. Through respiratory administration, targeted therapy can be performed and the toxic and side effects of systemic medication can be reduced. The compound essential oil is blended from two single essential oils, and may smell better, have a longer-lasting and more comprehensive therapeutic effect, and can be directly applied to the skin. Due to differences in species and ingredients, plant essential oils may exhibit different activities. Through deployment and combination, single essential oils can exhibit synergistic activity when used in combination.

[0040] As a specific embodiment, the mint in the mint essential oil is produced from Jiangsu, Anhui, Jiangxi, Hubei, Sichuan and Henan, among which the mint essential oil produced from mint in Ziyang, Sichuan has the best effect; the perilla leaf essential oil is produced from perilla leaves in Guangxi, Anhui, Hunan and Yunnan, among which the perilla leaf essential oil produced from perilla leaves in Zhangjiajie, Hunan has the best effect.

[0041] The present invention also provides a preparation method of the compound essential oil, comprising the following steps: respectively extracting peppermint essential oil and perilla leaf essential oil by a steam distillation method, and mixing the peppermint essential oil and perilla leaf essential oil to obtain the compound essential oil.

[0042] When preparing the peppermint essential oil of the present invention, the mass volume ratio of the peppermint to water is 55-65g:550-650mL, and the steam distillation time is 2.5-3.5h; as a specific embodiment, the mass volume ratio of the peppermint to water can be 55g:550mL, 57g:550mL, 59g:550mL, 61g:550mL, 63g:550mL, 65g:550mL, 55g:560mL, 57g:560mL, 59g:560mL, 61g:560mL, 63g:560mL, 65g:560mL, 55g:570mL, 57g:570mL, 59g:570mL, 61g:570mL, 63g:570mL, 65g:570mL, 55g:580mL, 57g:580mL, 59g:580mL, 61g:580mL, 63g:580mL, 65g:580mL, 60g:600mL, 55g:640mL, 57g:640mL, 59g:640mL, 61g:640mL, 63g:640mL, 65g:640mL, 55g:650mL, 57g:650mL, 59g:650mL, 61g:650mL, 63g:650mL or 65g:650mL; as a specific embodiment, the steam distillation time can be 2.5h, 2.6h, 2.7h, 2.8h, 2.9h, 3.0h, 3.1h, 3.2h, 3.3h, 3.4h or 3.5h.

[0043] When preparing the perilla leaf essential oil of the present invention, the mass volume ratio of the perilla leaf to water is 45-55g:1100-1300mL, and the steam distillation time is 2-3h; as a specific embodiment, the mass volume ratio of the perilla leaf to water can be 45g:1100mL, 46g:1100mL, 47g:1100mL, 48g:1100mL, 49g:1100mL, 50g:1100mL, 51g:1100mL, 52g:1100mL, 53g:1100mL, 54g:1100mL, 55g:1100mL, 45g:1200mL, 46g:1200mL, 47g:1200mL, 48g:1200mL, 49g:1200mL, : 50g: 1200mL, 51g: 1200mL, 52g: 1200mL, 53g: 1200mL, 54g: 1200mL, 55g: 1200mL, 45g: 1300mL, 46g: 1300mL, 47g: 1300mL, 48g: 1300mL, 49g: 1300mL, 50g: 1300mL, 51g: 1300mL, 52g: 1300mL, 53g: 1300mL, 54g: 1300mL or 55g: 1300mL; as a specific embodiment, the steam distillation time can be 2.0h, 2.1h, 2.2h, 2.3h, 2.4h, 2.5h, 2.6h, 2.7h, 2.8h, 2.9h or 3.0h.

[0044] The present invention also provides the use of the compound essential oil in preparing a drug for preventing and / or treating allergic rhinitis. In one embodiment, the drug for preventing and / or treating allergic rhinitis is prepared by diluting the compound essential oil with a base oil; the base oil may include olive oil, sweet almond oil, sesame seed oil, tea seed oil, or jojoba oil. In one embodiment, the drug for preventing and / or treating allergic rhinitis may be administered by nasal drops or sniffing. In one embodiment, the drug for preventing and / or treating allergic rhinitis may be administered in the form of a nasal drop, spray, or gel.

[0045] As a specific embodiment, the compound essential oil of the present invention has an improving effect on mice with allergic rhinitis induced by ovalbumin, and can inhibit AR inflammatory response by regulating the local Th1 / Th2 and Th17 / Treg cell balance in the nasal mucosa. It is comparable to the effect of budesonide nasal spray in terms of behavioral indicators, IL-5, IL-13 and RORγt mRNA expression levels. The composition of the compound essential oil of the present invention is composed of only two volatile oils of traditional Chinese medicine, with simple ingredients. Both are traditional Chinese medicines with medicinal and edible properties. They have both therapeutic efficacy and are easily available, safe, inexpensive, and simple extraction methods in daily life. They are easy to prepare and use by the public, and have broad application prospects and good economic value.

[0046] To further illustrate the present invention, the following detailed description of a compound essential oil for improving allergic rhinitis provided by the present invention, its preparation method and application is provided in conjunction with examples, but they should not be construed as limiting the scope of protection of the present invention.

[0047] Unless otherwise specified, the present invention has no special requirements for the raw materials, and commercially available products known to those skilled in the art can be used.

[0048] The experimental materials and reagents used in the present invention are shown in Table 1.

[0049] Table 1 Experimental materials and reagents information

[0050]

[0051]

[0052] Extraction of essential oils from each Chinese medicinal material of the present invention: Refer to the 2020 edition of the "Chinese Pharmacopoeia" Part IV 2204 General Rules for the Determination of Volatile Oils Method A to extract essential oils from mint, ligusticum chuanxiong, magnolia, perilla leaves, patchouli, tangerine peel, asarum and artemisia argyi.

[0053] The present invention conducts β-hexosaminidase release test and NO content determination experiment because: β-hex has a long half-life and its detection method is simple, so it can be regarded as a preferred indicator for detecting mast cell activation; regulating the polarization of macrophages and inhibiting their release of NO and inflammatory cytokines has the potential to improve allergic diseases such as AR.

[0054] Example 1 Effects of different single essential oils on the toxicity and β-hexosaminidase release of C48 / 80-induced P815 cells

[0055] 1. Experimental Methods

[0056] (1) Cell viability assay

[0057] P815 cells in the logarithmic growth phase were gently pipetted down with complete culture medium and cultured at a rate of 5×10 5Cells were seeded in 96-well plates at 100 μL per well and placed in a 37°C, 5% CO2 incubator. The culture medium was discarded after 12 hours. Dosing was performed according to the following groupings, and a zero-adjustment group was set up without inoculation: blank and zero-adjustment groups received 100 μL of DMEM high-glucose medium; the dosing groups received 100 μL of different concentrations of volatile oil (peppermint essential oil, Ligusticum chuanxiong essential oil, Magnolia flos floridana essential oil, Perilla frutescens leaf essential oil, Patchouli chinensis essential oil, Tangerine peel essential oil, Asarum essential oil, Eucalyptus leaf essential oil, Tea Tree essential oil, or Artemisia argyi essential oil) or dexamethasone (the stock solution was prepared in DMSO and diluted to 80 μg / mL, 40 μg / mL, 20 μg / mL, and 10 μg / mL in DMEM high-glucose medium immediately before use). Incubate in a 37°C, 5% CO2 incubator for 4 hours, then discard the supernatant. Add 100 μL of modified benchtop buffer to the blank and zero-adjustment groups, and 100 μL of modified benchtop buffer containing 10 μg / mL C48 / 80 to the drug-treated group. Incubate in the incubator for 30 minutes. Add 10 μL / well of MTT solution (250 mg of MTT powder in 50 mL of PBS to a concentration of 5 mg / mL). Incubate for 4 hours, discard the supernatant, and add 150 μL of DMSO to each well. Shake until the formazan crystals are completely dissolved. Measure the OD value at 490 nm to calculate cell viability.

[0058] Cell survival rate (%) = [OD (drug group) - OD (zero adjustment group)] / [OD (blank group) - OD (zero adjustment group)] × 100%

[0059] (2) Single essential oil β-hexosaminidase release test

[0060] P815 cells in the logarithmic growth phase were gently pipetted down with complete culture medium and cultured at a rate of 5×10 5 Cells were inoculated into 96-well plates at 100 μL per well in a 37°C, 5% CO2 incubator for 12 hours. The culture medium was discarded. The cells were administered according to the following groups, and a zero-treatment group was set up without inoculation:

[0061] 100 μL of DMEM high-glucose medium was added to each of the blank, model, lysis, and zero-adjustment groups. 100 μL of different concentrations of volatile oil / dexamethasone (the stock solution was prepared with DMSO and diluted with DMEM high-glucose medium to 40 μg / mL, 20 μg / mL, and 10 μg / mL immediately before use) was added to the drug-treated group. The cells were cultured in a 37°C, 5% CO2 incubator for 4 hours. The supernatant was discarded.

[0062] The blank group and the zero-adjustment group were each added with 100 μL of modified benchtop solution, the model group and the drug-treated group were each added with 100 μL of modeling agent C48 / 80 (10 μg / mL), and the lysis group was added with 100 μL of 0.1% Triton X-100. The cells were incubated at 37°C and 5% CO2 for 30 min. 50 μL of the supernatant was added to a new 96-well plate, and 50 μL of the color developing solution (13.692 mg of 4-nitrophenyl N-acetyl-β-D-glucosamine was accurately weighed and dissolved in 40 mL of 0.1 mol / L sodium citrate buffer by ultrasonication, and filtered through a 0.22 μm filter) was added to each well. The plates were incubated at 37°C for 1.5 h, and 200 μL of the stop solution (2.12 g of Na2CO3 was weighed, diluted to 200 mL with pure water, and completely dissolved by ultrasonication to obtain 0.1 mol / mL Na2CO3; 0.42 g of NaHCO was weighed, diluted to 50 mL with pure water, and completely dissolved by ultrasonication to obtain 0.1 mol / mL NaHCO3; 0.1 mol / mL Na2CO3 and 0.1 mol / mL NaHCO3 were mixed at a ratio of 9:1 before use) was added to each well. The OD value was measured at 405 nm, and the β-Hex release rate was calculated.

[0063] β-Hex release rate = [(OD sample supernatant - OD zero adjustment) / (OD lysis - OD zero adjustment)] × 100%

[0064] β-Hex inhibition rate = [(release rate of model group - release rate of drug-treated group) / (release rate of model group - release rate of blank group)] × 100%

[0065] 2. Experimental Results

[0066] Figure 1 Figure A shows that when the 10 single essential oils and dexamethasone were administered at a concentration of ≤40 μg / mL, the survival rate of P815 cells was ≥90%, with no obvious toxic side effects. Therefore, three final concentrations of 40 μg / mL, 20 μg / mL, and 10 μg / mL were selected as the administration concentrations for P815 cells. Figure 1 Figures B through G show that wormwood, patchouli, eucalyptus, and tea tree essential oils had no effect on the release rate of β-hexosaminidase. Tangerine peel essential oil showed a significant effect starting at a medium concentration (P < 0.05), and Asarum showed a significant effect at a high concentration (P < 0.01). However, magnolia, ligusticum, perilla, and peppermint essential oils, as well as the positive drug dexamethasone, showed significant effects at all three concentrations (P < 0.01), with inhibition rates of 42.35 ± 3.83%, 27.06 ± 5.50%, 38.03 ± 8.62%, 37.32 ± 2.95%, and 29.89 ± 5.60%, respectively. This indicates that the top three essential oils with a stronger effect than the positive drug are magnolia, perilla, and peppermint essential oils.

[0067] Example 2 Effects of different single essential oils on the NO content of LPS-induced RAW264.7 cells

[0068] 1. Experimental Methods

[0069] (1) Cell viability assay

[0070] RAW264.7 cells in the logarithmic growth phase were gently pipetted down with complete culture medium and plated at 8×10 5 Cells were inoculated into 96-well plates at 100 μL per well and placed in a 37°C, 5% CO2 incubator. After 24 hours, the culture medium was discarded. The cells were then administered according to the following groups, with a separate zero-treatment group not inoculated with cells:

[0071] The blank and zero-adjustment groups were treated with 200 μL of DMEM high-glucose medium. The drug-treated groups were treated with 100 μL of different concentrations of essential oil / dexamethasone (the stock solution was prepared with DMSO and diluted with DMEM high-glucose medium to 320 μg / mL, 160 μg / mL, 80 μg / mL, and 40 μg / mL immediately before use) and 100 μL of LPS (2 μg / mL). The final concentrations in each well after dilution were 160 μg / mL, 80 μg / mL, 40 μg / mL, and 20 μg / mL. The cells were cultured in a 37°C, 5% CO2 incubator for 24 hours. MTT solution (10 μL / well) was added and cultured for another 4 hours. The supernatant was discarded, and 150 μL of DMSO was added to each well. The cells were shaken until the formazan crystals were completely dissolved. The OD value was measured at a wavelength of 490 nm, and the cell viability was calculated.

[0072] Cell survival rate (%) = [OD (drug group) - OD (zero adjustment group)] / [OD (blank group) - OD (zero adjustment group)] × 100%

[0073] (2) NO content determination

[0074] RAW264.7 cells in the logarithmic growth phase were gently pipetted down with complete culture medium and plated at 8×10 5The cells were inoculated with 100 μL of each well in a 96-well plate and placed in an incubator at 37°C and 5% CO2. The culture medium was discarded after 24 hours. The mice were divided into the following groups for drug administration: the blank group received 200 μL of DMEM high-glucose medium; the model group received 100 μL of LPS (2 μg / mL) and 100 μL of DMEM high-glucose medium; the drug administration group received 100 μL of different concentrations of volatile oil / dexamethasone (the stock solution was prepared with DMSO and diluted with DMEM high-glucose medium to 160 μg / mL, 80 μg / mL, and 40 μg / mL before use) and 100 μL of LPS (2 μg / mL). The final concentrations after dilution were 80 μg / mL, 40 μg / mL, and 20 μg / mL. After culturing in an incubator at 37°C and 5% CO2 for 24 hours, 50 μL of the supernatant was taken and first added with 50 μL of solution A of the NO test kit, then with 50 μL of solution B. After shaking on an oscillator for 5 minutes, the OD value was measured at a wavelength of 540 nm and substituted into the NO content standard curve to obtain the NO release amount.

[0075] NO release amount = OD value × 82.03-4.3004

[0076] 2. Experimental Results

[0077] Figure 2 Figure A shows that the survival rate of RAW264.7 cells was ≥90% when the 10 single essential oils and dexamethasone were ≤80 μg / mL, with no obvious toxic side effects. Therefore, three final concentrations of 80 μg / mL, 40 μg / mL, and 20 μg / mL were selected as the dosing concentrations for RAW264.7 cells. Figure 2 Figure B shows that except for Asarum, the other nine essential oils and positive drugs had significant effects compared with the model group (P < 0.01), among which mint, Ligusticum chuanxiong and Perilla leaf essential oils had the best effect in inhibiting NO production, ranking in the top three.

[0078] Example 3 Effects of different compound essential oils on the toxicity and β-hexosaminidase release of C48 / 80-induced P815 cells

[0079] Based on the results of Examples 1 and 2, essential oils of magnolia bud, perilla leaf, peppermint, and ligusticum were selected and mixed in equal proportions to form a crude oil formula. Diluted according to the method of Experimental Example 1, the final concentrations of the compound oils were 40 μg / mL, 20 μg / mL, and 10 μg / mL, for a total of 11 formulas. The methods for cell viability assay and β-hexosaminidase release assay were similar to those of Example 1.

[0080] Depend on Figure 3 As shown in Figure A, the survival rates of the compound essential oils in different groups on P815 cells at three dosages were all ≥90%, indicating that they were non-toxic to P815 cells. Figure 3BE analysis revealed that the compound essential oils in groups F9, F10, and F11 had no effect on the release rate of β-hexosaminidase, while the other groups showed significant effects at high concentrations. F7 (magnolia: mint: perilla leaf) and F8 (mint: perilla leaf) showed significant inhibition at all three concentrations, with inhibition rates of 41.76±2.46% and 39.41±5.50%, respectively. Compared with the inhibition rates of the individual essential oils (magnolia: 42.35±3.83%; perilla leaf: 38.03±8.62%; mint: 37.32±2.95%), F8 (mint: perilla leaf) exhibited a synergistic effect.

[0081] Example 4 Effects of different compound essential oils on NO content in LPS-induced RAW264.7 cells

[0082] Based on the results of Experimental Examples 1 and 2, essential oils of magnolia bud, perilla leaf, peppermint, and ligusticum were selected and mixed in equal proportions to form a crude oil formulation. Diluted according to the method of Experimental Example 2, the final concentrations of the administered essential oils were 80 μg / mL, 40 μg / mL, and 20 μg / mL, for a total of 11 formulations. The methods for cell viability and NO content determination were similar to those in Example 2.

[0083] Depend on Figure 4 As shown in Figure A, the survival rates of different groups of compound essential oils on RAW264.7 cells at doses of 80 μg / mL, 40 μg / mL, and 20 μg / mL were all ≥90%, indicating that they had no toxicity to RAW264.7 cells. Figure 4 As shown in Figure B, all compound essential oils showed significant differences at high concentrations compared to the LPS model group (P < 0.01), demonstrating that different compound essential oils can inhibit NO production to a certain extent; the effects are: F3 > F9 > F8 > F10 > F7 > F1 > F2 > F4 > F6 > F5 > F11; compared with the effects of individual essential oils, some groups showed synergistic effects. Since the F8 group in Experimental Example 3 showed a synergistic effect on inhibiting the β-hexosaminidase release rate of C48 / 80-induced P815 cells and ranked third in this experimental example, F8: Peppermint: Perilla Leaf Essential Oil was selected for further investigation of its origin and in vivo efficacy in improving allergic rhinitis.

[0084] Example 5 Effects of peppermint essential oils from different origins on the toxicity and β-hexosaminidase release of C48 / 80-induced P815 cells

[0085] 14 batches of mint (see Table 2) were extracted into volatile oils and diluted according to the method of Experimental Example 1. The final dosage concentration was 40 μg / mL. The methods of cell viability determination and β-hexosaminidase release test were referred to Example 1. Figure 5 As shown in Figure A, at this dose, the cell survival rates of the 14 peppermint essential oils were all ≥90%, indicating that they had no toxicity to P815 cells. Figure 5As shown in Figure B, the effects of 14 peppermint essential oils on reducing the release of β-hexosaminidase are arranged from strong to weak as follows: B12 > B1 > B8 > B13 > B3 > B5 > dexamethasone > B14 > B7 > B2 > B4 > B6 > B11 > B9 > B10.

[0086] Table 2 Origin of mint from different origins

[0087] serial number Origin serial number Origin B1 Nantong, Jiangsu B8 Jiangxi B2 Jiangsu B9 Hubei B3 Jiangsu B10 Shennongjia, Hubei B4 Anhui B11 Sichuan B5 Fuyang, Anhui B12 Ziyang, Sichuan B6 Bozhou, Anhui B13 Henan B7 Jiangxi B14 Henan

[0088] Example 6 Effects of peppermint essential oils from different origins on the NO content in LPS-induced RAW264.7 cells

[0089] 14 batches of mint (see Table 2) were extracted into volatile oils and diluted according to the method of Example 2. The final dosage concentration was 80 μg / mL. The methods for cell viability determination and NO content determination were referred to Example 2. Figure 6 As shown in Figure A, at this dose, the cell viability of the 14 peppermint essential oils was ≥90%, indicating that they had no toxicity to RAW264.7 cells. Figure 6 As shown in Figure B, the effects of 14 peppermint essential oils in reducing NO content are arranged from strong to weak as follows: B4>B13>B12>B1>B10>B9>B7>B2>B14>B5>B8>B11>B6>B3>dexamethasone, and all peppermint essential oils have significant inhibitory effects.

[0090] Example 7 Effects of Perilla Leaf Essential Oils from Different Origins on Toxicity and β-Hexosaminidase Release in C48 / 80-Induced P815 Cells

[0091] Ten batches of perilla leaves (see Table 3) were extracted into volatile oils and diluted according to the method of Example 1. The final dosage concentration was 40 μg / mL. The methods of cell viability determination and β-hexosaminidase release test were referred to Example 1. Figure 7 As shown in Figure A, when the dosage was 40 μg / mL, the cell survival rates of the 10 kinds of perilla leaf essential oils were all ≥90%, indicating that they had no toxicity to P815 cells. Figure 7 As shown in Figure B, the effects of the 10 perilla leaf essential oils in reducing the release of β-hexosaminidase are arranged from strong to weak as follows: Z5=Z6>Z4>Z1>dexamethasone>Z2>Z9>Z3>Z7>Z10>Z8, among which Z5, Z6, Z4 and Z1 showed significant inhibition compared with the model group.

[0092] Table 3 Sources of Perilla leaves from different origins

[0093] serial number Origin serial number Origin Z1 Yulin, Guangxi Z6 Shaoyang, Hunan Z2 Yulin, Guangxi Z7 Yunnan Z3 Yulin, Guangxi Z8 Dali, Yunnan Z4 Changsha, Hunan Z9 Bozhou, Anhui Z5 Zhangjiajie, Hunan Z10 Lu'an, Anhui

[0094] Example 8 Effects of Perilla Leaf Essential Oils from Different Origins on NO Content in LPS-Induced RAW264.7 Cells

[0095] Ten batches of perilla leaves (see Table 3) were extracted into volatile oils and diluted according to the method of Example 2. The final dosage concentration was 80 μg / mL. The methods for cell viability determination and NO content determination were referred to Example 2. Figure 8 As shown in Figure A, the cell viability of the 10 perilla leaf essential oils at this dose was ≥90%, indicating that they had no toxicity to RAW264.7 cells.

[0096] Depend on Figure 8 As shown in Figure B, the effects of the ten perilla leaf essential oils on reducing NO content are arranged from strong to weak as follows: Z3>Z2>Z1>Z5>Z4>Z6>Z7>dexamethasone>Z9>Z8>Z10, and all ten perilla leaf essential oils have significant inhibitory effects.

[0097] Example 9 Effects of different ratios of peppermint and perilla leaf essential oils on the toxicity and β-hexosaminidase release of C48 / 80-induced P815 cells

[0098] According to Examples 5, 6, 7 and 8, the crude oils of Z5 perilla leaves and B12 mint were mixed in different proportions, diluted according to the method of Example 1, and administered at a final concentration of 40 μg / mL for cell experiments. The methods for cell viability determination and β-hexosaminidase release test were referred to Example 1. Figure 9 As shown in Figure A, this dose is a safe dose. The effects of different proportions of essential oils on reducing the release of β-hexosaminidase are arranged from strong to weak as follows: 4B1Z>5B1Z>3B1Z=mint (B12)>1B1Z=1B3Z>2B1Z>1B4Z=1B2Z=perilla leaf (Z5)>1B5Z (B represents mint, Z represents perilla leaf, such as 4B1Z refers to a mixture of 4 parts mint and 1 part perilla leaf). Compared with the model group, there are significant differences, among which the combination of 4B1Z and 5B1Z essential oils is more effective than the single formula: B12 mint essential oil / Z5 perilla leaf essential oil.

[0099] Example 10 Effects of different ratios of peppermint and perilla leaf essential oils on the NO content in LPS-induced RAW264.7 cells

[0100] The essential oils in different proportions in Example 9 were combined and diluted according to the method of Example 2, and the final concentration was 80 μg / mL for cell experiments. The methods for cell viability determination and NO content determination were referred to Example 2. Figure 10 As shown in Figure A, when the dosage was 80 μg / mL, except for 5B1Z with a survival rate of 86.17%, the cell survival rates of the other essential oil combinations at different ratios were all ≥90%, indicating that they had no toxicity to RAW264.7 cells. Figure 10As shown in Figure B, the effects of different proportions of essential oil combinations on reducing NO content are arranged from strong to weak as follows: 5B1Z>4B1Z>3B1Z>2B1Z>1B3Z>1B1Z>1B2Z>1B4Z>mint (B12)>1B5Z>perilla leaf (Z5). The essential oil combinations in different proportions all have significant inhibitory effects, and most essential oil combinations are better than the single formula: B12 mint essential oil / Z5 perilla leaf essential oil.

[0101] Example 11 Compound essential oil for improving allergic rhinitis

[0102] Steam distillation was performed on 60g of mint in 600mL of water and 50g of perilla leaf in 1200mL of water, respectively. The mint was kept at a slight boil for 3 hours, and the perilla leaf was kept at a slight boil for 2.5 hours to obtain mint essential oil or perilla leaf essential oil. The mint essential oil and perilla leaf essential oil were prepared in different ratios (1:1, 2:1, 3:1, 4:1, 5:1, 1:5, 1:4, 1:3, 1:2) to obtain compound essential oils with different ratios.

[0103] Example 12 Therapeutic Effect Test on Allergic Rhinitis Mice

[0104] 1. Purpose of the experiment

[0105] This study used mice with an allergic rhinitis model as the research subjects. The number of sneezes and nose scratches within 10 minutes after the last nasal drop challenge, blood eosinophils, nasal mucosa and lung tissue pathological observation, and mRNA expression levels of Th1 / Th2 and Th17 / Treg cell-related factors were used as observation indicators. Budesonide nasal spray was used as a positive control drug to analyze the pharmacodynamics of mint and perilla leaf compound essential oil in the treatment of allergic rhinitis.

[0106] 2. Experimental Grouping

[0107] Fifty SPF female Balb / c mice, 6-8 weeks old, were randomly divided into five groups: a blank group, an allergic rhinitis model group, a budesonide group (33.28 μg / kg), a high-dose compound essential oil group (prepared according to the method of Example 11 and then added with olive oil) (25 mg / kg), and a low-dose compound essential oil group (6.25 mg / kg), with 10 mice in each group.

[0108] 3. Experimental Methods

[0109] Sensitization was performed on days 0, 3, 6, 9, 12, and 15: each mouse in the blank group was intraperitoneally injected with 200 μl of normal saline, and each mouse in the other groups was intraperitoneally injected with 200 μl of normal saline containing 2 mg of aluminum hydroxide and 100 μg of ovalbumin. On the 21st day, nasal challenge and drug administration began. After the mice were anesthetized, 20 μl of olive oil was administered to each mouse in the blank group and model group. 20 μl of olive oil containing essential oils of different doses (i.e., high-dose group and low-dose group) was administered to each mouse in the compound essential oil high-dose group and low-dose group. 20 μl of a suspension containing budesonide (diluted with normal saline) was administered to each mouse in the budesonide group. One hour after drug administration, nasal challenge was performed. 20 μl of normal saline was administered to the blank group, and 20 μl of normal saline containing 200 μg of ovalbumin was administered to the mice in the other groups. After 10 days of continuous drug administration and nasal challenge, within 10 minutes after the last nasal challenge, personnel who were blinded to the experiment recorded the number of sneezes and nose scratches for each mouse. 24 hours after the last intranasal challenge, samples were collected, including blood, nasal mucosa, and lungs (i.e., starting from the 21st day, the drug was administered every day, and the intranasal challenge was performed 1 hour after the drug administration. On the last day of intranasal challenge, that is, the 30th day, the intranasal challenge was performed 1 hour after the drug administration. The number of sneezes and nose scratches of each mouse was counted within 10 minutes; samples were collected on the 31st day).

[0110] 4. Experimental indicators

[0111] After the last nasal drop challenge, blinded subjects were asked to count the number of sneezes and nose scratches; HE staining (Yuan Yuze. Research on the pathogenesis of allergic airway inflammation based on microbiome and metabolomics [D]. Jilin University, 2022. DOI: 10.27162 / d.cnki.gjlin.2022.007527) was used to observe the pathological changes of nasal mucosa and lungs; PAS staining (Yuan Yuze. Research on the pathogenesis of allergic airway inflammation based on microbiome and metabolomics [D]. Jilin University, 2022. DOI: 10.27162 / d.cnki.gjlin.2022.007527.) was used to observe the changes of goblet cells in the nasal mucosa; Giemsa staining (Qian Xingyi. Based on NF -κB signaling pathway to explore the mechanism of action of Chimonanthus chinensis leaf granules and volatile oil in the treatment of allergic rhinitis [D]. Jiangxi University of Traditional Chinese Medicine, 2023. DOI: 10.27180 / d.cnki.gjxzc.2023.000607.) The eosinophils in the blood of mice were stained; the mRNA expressions of Th1 cytokines: IFN-γ, Th2 cytokines: IL-4, IL-5, IL-13, Th1 cell transcription factor: T-bet, Th2 cell transcription factor: GATA3, Th17 cell transcription factor: RORγt and Treg cell transcription factor: Foxp3 in the nasal mucosa were observed by real-time fluorescence quantitative polymerase chain reaction. The primer sequences are shown in Table 4.

[0112] Table 4 Real-time quantitative PCR primer sequences

[0113]

[0114] 5. Statistical methods

[0115] All experimental data were expressed as mean ± standard deviation and statistically processed using GraphPad Prism 8. One-way ANOVA was used to compare multiple groups, and P < 0.05 was considered statistically significant.

[0116] 6. Experimental Results

[0117] (1) Mouse behavioral performance

[0118] Within 10 minutes after the last stimulation of the mouse nasal cavity, the number of sneezing and nose scratching in the model group mice was higher than that in the blank group, and the differences were significant. The number of sneezing and nose scratching in the positive drug group and the different doses of compound essential oil groups were significantly lower than those in the model group, and in terms of nose scratching, the different doses of compound essential oil groups were better than the positive group. Figure 11 .

[0119] (2) Staining results of eosinophils in mouse blood

[0120] Depend on Figure 12 It can be seen that compared with the blank group, the eosinophils in the blood of the model group increased significantly. After the administration of the high and low doses of compound essential oil and the positive drug, the eosinophils in the blood of mice decreased significantly.

[0121] (3) Effects of compound essential oils on inflammatory response of nasal mucosal tissue in mice

[0122] Depend on Figure 13 It can be seen that the nasal mucosal tissue of the blank group mice is intact, the cells are arranged neatly, the ciliated columnar epithelium is covered continuously, and no inflammatory cell infiltration is seen. In the model group nasal mucosa, the visible mucosal epithelium is partially damaged and broken, the cells are arranged relatively irregularly, and a large amount of obvious inflammatory cell infiltration is seen, and the mucosal layer and submucosal layer are obviously swollen. After the intervention of the compound essential oil and budesonide, compared with the model group, the inflammatory cell infiltration is reduced, the mucosal tissue is relatively arranged neatly, and the degree of mucosal swelling is reduced. In the low and high dose groups of the compound essential oil, the above-mentioned effect gradually becomes obvious as the drug dosage increases. It can be obtained from this that the present invention can significantly reduce the degree of nasal mucosal inflammation infiltration caused by allergic rhinitis caused by ovalbumin.

[0123] (4) Effects of compound essential oils on goblet cells in the nasal mucosa of mice

[0124] In the context of allergic rhinitis, goblet cells are overexpressed, resulting in excessive mucus in the nasal cavity. Goblet cells are characterized by a narrow base and a bulged top, and appear purple after PAS staining. Figure 14 It can be seen that compared with the blank group mice, a large number of proliferating purple goblet cells were seen in the nasal mucosa of the model group; compared with the model group, the goblet cells in the nasal mucosa of the low-dose and high-dose compound essential oil groups and the positive group were significantly reduced, and there was no significant difference in the effect between the high-dose compound essential oil and the positive group.

[0125] (5) Effect of compound essential oil composition on the degree of inflammatory infiltration in mouse lung tissue

[0126] Depend on Figure 15 It can be seen that compared with the blank group mice, the lungs of the model group mice showed obvious thickening of the tracheal wall, irregular lumen, and obvious inflammatory cell infiltration in the connective tissue around the bronchi and blood vessels; after intervention with compound essential oil and budesonide, compared with the model group, the infiltration of inflammatory cells in the lungs was reduced, the tracheal wall became thinner, and the effect became better with the increase of essential oil dosage. The effects of the high-dose compound essential oil group and the budesonide group were comparable.

[0127] (6) Expression of Th1 / Th2 cell-related cytokine mRNA in the nasal mucosa of mice in each group

[0128] Depend on Figure 16 It can be seen that the mRNA levels of Th2 cell-related cytokines IL-4, IL-5, and IL-13 in the nasal mucosa of mice in the model group were significantly higher than those in the blank group, while the mRNA level of Th1 cell-related cytokine IFN-γ was significantly lower than that in the blank group; compared with the model group, the high-dose compound essential oil group and the budesonide group significantly reduced the mRNA levels of IL-4 and IL-13, and the low-dose compound essential oil group showed a downward trend; compared with the model, the mRNA levels of IL-5 in each drug-treated group were significantly reduced, and the high-dose compound essential oil group was better than the positive drug group; compared with the model, the mRNA levels of Th1 cell-related cytokine IFN-γ in the three drug-treated groups were increased, but not significantly.

[0129] (7) Expression of Th1 / Th2 cell-specific transcription factor mRNA in the nasal mucosa of mice in each group

[0130] Depend on Figure 17 It can be seen that the mRNA level of GATA3, a Th2 cell-specific transcription factor, in the nasal mucosa of mice in the model group was significantly higher than that in the blank group, while the mRNA level of T-bet, a Th1 cell-specific transcription factor, was significantly lower than that in the blank group; after administration, the mRNA level of T-bet in the high-dose compound essential oil group and the budesonide group showed a trend of recovery, and the mRNA level of GATA3 in the three drug-administered groups showed a trend of decrease.

[0131] (8) Expression of Th17 / Treg cell-specific transcription factor mRNA in the nasal mucosa of mice in each group

[0132] Depend on Figure 18 It can be seen that the mRNA level of Th17 cell-specific transcription factor RORγt in the nasal mucosa of mice in the model group was significantly higher than that in the blank group, while the mRNA level of Treg cell-specific transcription factor Foxp3 was significantly lower than that in the blank group; after administration, the mRNA level of RORγt in the high-dose group of compound essential oil showed a significant downward trend, and the mRNA levels of Foxp3 in the three drug-administered groups showed a significant upward trend.

[0133] 7. Summary

[0134] Animal experiments showed that the mint and perilla leaf compound essential oil of the present invention has a certain improvement effect on the ovalbumin-induced allergic rhinitis mouse model, which may be achieved by improving the inflammatory infiltration of the nasal mucosa and lungs, reducing goblet cell proliferation, and regulating the local Th1 / Th2 and Th17 / Treg cell balance in the nasal mucosa to inhibit AR inflammatory response. The high-dose group of the compound essential oil had an effect comparable to that of budesonide nasal spray in behavioral indicators, nasal mucosa and lung pathology, and IL-5, IL-13, and RORγt mRNA expression levels.

[0135] Thus, the compound essential oil of the present invention has a better effect in treating rhinitis than the individual essential oils in the compound composition at the same dosage, and has a synergistic effect. The compound essential oil of the present invention involves essential oils of two medicinal plants that are both medicinal and edible, and is used to prevent and alleviate the symptoms of rhinitis. It has significant and stable efficacy, is safe and reliable, and is low in cost, and has practical application value.

[0136] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A compound essential oil for improving allergic rhinitis, characterized in that: The compound essential oil comprises the following components in parts by mass: 1 to 5 parts of peppermint essential oil and 1 to 5 parts of perilla leaf essential oil.

2. The compound essential oil according to claim 1, wherein The compound essential oil comprises the following components in parts by mass: 4 to 5 parts of peppermint essential oil and 1 part of perilla leaf essential oil.

3. The compound essential oil according to claim 1 or 2, wherein The mint in the mint essential oil is produced in Ziyang, Sichuan; The production area of the perilla leaves in the perilla leaf essential oil includes Zhangjiajie, Hunan.

4. The method for preparing the compound essential oil according to any one of claims 1 to 3, characterized in that: The method comprises the following steps: respectively extracting peppermint essential oil and perilla leaf essential oil by a steam distillation method, and mixing the peppermint essential oil and the perilla leaf essential oil to obtain the compound essential oil.

5. The preparation method according to claim 4, characterized in that When preparing the peppermint essential oil, the mass volume ratio of the peppermint to water is 55-65 g:550-650 mL, and the steam distillation time is 2.5-3.5 h.

6. The preparation method according to claim 4, characterized in that When preparing the perilla leaf essential oil, the mass volume ratio of the perilla leaf to water is 45-55g:1100-1300mL, and the steam distillation time is 2-3h.

7. Use of the compound essential oil according to any one of claims 1 to 3 or the compound essential oil prepared by the preparation method according to any one of claims 4 to 6 in the preparation of a medicament for preventing and / or treating allergic rhinitis.

8. The use according to claim 7, characterized in that The drug for preventing and / or treating allergic rhinitis is used after diluting the compound essential oil with base oil; The base oil includes olive oil, sweet almond oil, sesame seed oil, tea seed oil or jojoba oil.

9. The use according to claim 7, characterized in that The method of using the drug for preventing and / or treating allergic rhinitis includes nasal drops or sniffing.

10. The use according to claim 7, characterized in that The preparation forms of the drug for preventing and / or treating allergic rhinitis include nasal drops, sprays or gels.

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