Umbilical cord MSC exosome nanocarrier delivery system and application thereof in preparation of medicine for treating allergic rhinitis

Abstract

This invention belongs to the field of biomedical technology, specifically relating to an umbilical cord MSC exosome nanocarrier delivery system and its application in the preparation of drugs for treating allergic rhinitis. This invention prepares the umbilical cord MSC exosome nanocarrier delivery system by adding a specific concentration of diosgenin to the culture medium of umbilical cord mesenchymal stromal cells (MSCs), collecting the cell culture supernatant, and separating and extracting it. Animal experiments show that the delivery system prepared in this invention significantly improves the symptoms of nose scratching and sneezing in mice with allergic rhinitis. Its mechanism of action lies in effectively regulating the Th1 / Th2 immune balance: significantly inhibiting the overexpression of the pro-inflammatory factor IL-4, while simultaneously restoring the anti-inflammatory factor IL-10, thus avoiding immune dysregulation. This invention provides a safe and effective new treatment for allergic rhinitis and has good clinical application prospects.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to an umbilical cord MSC exosome nanocarrier delivery system and its application in the preparation of drugs for treating allergic rhinitis. Background Technology

[0002] Exosomes are vesicles secreted by cells with a lipid bilayer structure, ranging in diameter from 30 to 150 nm. This unique structure enables them to carry a variety of bioactive substances, including proteins, mRNA, microRNA, and lipids. Exosomes play a central role in immune responses, cell communication, tissue homeostasis, and tissue regeneration, serving as important mediators of intercellular communication and delivering bioactive substances to target cells.

[0003] Mesenchymal stromal cell (MSC) exosomes have attracted much attention due to their unique origin and biological characteristics. Umbilical cord MSC exosomes can precisely transport the active substances they carry into target cells through various mechanisms such as membrane fusion or endocytosis, thereby finely regulating the function of target cells.

[0004] Allergic rhinitis, a common chronic inflammatory disease of the upper respiratory tract, typically presents with symptoms including continuous sneezing, copious watery nasal discharge, nasal congestion, and nasal itching. Currently, medication is the primary treatment for allergic rhinitis. However, long-term medication use not only exposes patients to various adverse reactions, such as nasal dryness, bleeding, headache, and drowsiness, but also imposes a heavy financial burden. More challenging is the fact that no effective drug has yet been found to cure allergic rhinitis. Furthermore, with prolonged treatment, some patients develop drug resistance and tolerance, significantly reducing the effectiveness of medication and further complicating treatment. Therefore, actively exploring a safer and more effective treatment for allergic rhinitis is of crucial practical significance and clinical value for improving patients' quality of life, reducing their financial burden, and enhancing treatment outcomes. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention develops a nanocarrier delivery system for umbilical cord MSC exosomes containing diosgenin ketones based on the characteristics of exosomes. A mouse model of allergic rhinitis was established, and the efficacy of different umbilical cord MSC exosome nanocarrier delivery systems was evaluated. Animal experiment results show that the delivery system prepared in this invention significantly improves the symptoms of nose scratching and sneezing in mice with allergic rhinitis, significantly inhibits the overexpression of the pro-inflammatory factor IL-4, and simultaneously restores the anti-inflammatory factor IL-10, avoiding immune dysregulation. This provides a safe and effective new solution for the treatment of allergic rhinitis.

[0006] On one hand, the present invention provides a method for preparing an umbilical cord MSC exosome nanocarrier delivery system. The method includes the following steps: collecting umbilical cord tissue, cleaning and cutting it into tissue blocks, inoculating it in a culture medium containing diosgenin ketone for culture, collecting the cell culture supernatant, and separating and extracting the umbilical cord MSC exosome nanocarrier delivery system from the cell culture supernatant.

[0007] Furthermore, in the preparation method, the culture medium contains 0.01wt%~0.1wt% of diosgenin ketone.

[0008] Furthermore, in the preparation method, the culture medium contains 0.05 wt% diosgenin.

[0009] Furthermore, in the preparation method, the separation and extraction method is as follows: centrifuge the cell culture supernatant to remove cells and cell debris, and then use an ultrafiltration tube to concentrate the supernatant to obtain the final product.

[0010] Furthermore, in the preparation method, the centrifugation conditions are 3000~4000g / min for 20~30min; the ultrafiltration conditions are 3000~3500g / min for 35~40min at room temperature.

[0011] In another aspect, the present invention also provides an umbilical cord MSC exosome nanocarrier delivery system prepared by the preparation method described herein.

[0012] Furthermore, the umbilical cord MSC exosome nanocarrier delivery system inhibits the expression of pro-inflammatory factor IL-4 and increases the expression of anti-inflammatory factor IL-10.

[0013] In another aspect, a pharmaceutical composition is also provided, comprising the umbilical cord MSC exosome nanocarrier delivery system described in this invention, and a pharmaceutically acceptable carrier or excipient.

[0014] Finally, the application of the umbilical cord MSC exosome nanocarrier delivery system described in this invention in the preparation of drugs for treating allergic rhinitis is also provided.

[0015] Furthermore, in the application, the drug alleviates allergic rhinitis symptoms by regulating the mRNA expression levels of IL-4 and IL-10 in nasal mucosal tissue.

[0016] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:

[0017] (1) This invention discloses for the first time a method for preparing an umbilical cord MSC exosome nanocarrier delivery system with therapeutic effects on allergic rhinitis by adding a specific concentration of diosgenin to the culture medium of umbilical cord mesenchymal stromal cells (MSCs). Animal experimental results show that the delivery system prepared in this invention significantly inhibits the overexpression of the pro-inflammatory factor IL-4, while moderately reducing the expression level of the anti-inflammatory factor IL-10, effectively avoiding immune disorders caused by excessive compensation of IL-10.

[0018] (2) This invention found that the exosomes prepared have the best therapeutic effect when the concentration of diosgenin in the culture medium is within a specific range (0.01wt%~0.1wt%), especially 0.05wt%. Furthermore, using diosgenin solution alone is almost ineffective. This indicates that the technical solution of this invention produces a synergistic effect.

[0019] (3) The delivery system prepared in this invention is derived from umbilical cord MSC exosomes, which has good biocompatibility and low immunogenicity, and high safety. Its excellent efficacy in animal models and its clear mechanism of action have laid a solid foundation for its clinical application, providing a safe and effective new treatment option for allergic rhinitis. Attached Figure Description

[0020] Figure 1 The relative expression levels of IL-4 mRNA in the nasal mucosa tissue of mice under different treatments. Figure 2 The relative expression levels of IL-10 mRNA in the nasal mucosa tissue of mice under different treatments. Detailed Implementation

[0021] The technical solution of the present invention will be described below with reference to the embodiments. However, the present invention is not limited to the following embodiments.

[0022] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings. However, the embodiments described are not intended to limit the present invention.

[0023] Unless otherwise specified, the experimental and detection methods described in the following embodiments are conventional methods; unless otherwise specified, the reagents and materials are commercially available.

[0024] Diosgenin, CAS No.: 6870-79-7, purity 98%.

[0025] Example 1 This embodiment describes the preparation of an umbilical cord MSC exosome nanocarrier delivery system.

[0026] Freshly collected umbilical cord tissue was washed with physiological saline containing penicillin-streptomycin solution and then cut into 1mm pieces using ophthalmic scissors. 2 Small pieces of the cells were transferred to cell culture flasks, and DMEM / F12 medium containing 0.005 wt% diosgenin and penicillin-streptomycin was added. The flasks were then incubated statically in a 5% CO2 incubator at 37°C. The medium was changed for the first time after one week, and then every 3-4 days thereafter. When the cells reached 80%-90% confluence, they were digested with trypsin / EDTA digestion solution and passaged, then transferred to new culture dishes for further culture. When the cell concentration reaches 80-90%, the medium is replaced with serum-free human mesenchymal stem cell culture medium and cultured for another 24 hours. The medium is then replaced and the cell culture supernatant is collected. This process is repeated three times for each batch of cells. The cell culture supernatant collected from the three times is centrifuged for 25 minutes (3500 g / min) to remove cells and cell debris. The cells are then centrifuged at 4000 g / min for 35 minutes at room temperature using a Centricon Plus-70 (100 kDa) ultrafiltration tube. This yields the umbilical cord MSC exosome nanocarrier delivery system, labeled as #1 umbilical cord MSC exosome nanocarrier delivery system.

[0027] Example 2 This embodiment describes the preparation of an umbilical cord MSC exosome nanocarrier delivery system.

[0028] Freshly collected umbilical cord tissue was washed with physiological saline containing penicillin-streptomycin solution and then cut into 1mm pieces using ophthalmic scissors. 2 Small pieces of the cells were transferred to cell culture flasks, and DMEM / F12 medium containing 0.01 wt% diosgenin and penicillin-streptomycin was added. The flasks were then incubated statically in a 5% CO2 incubator at 37°C. The medium was changed for the first time after one week, and then every 3-4 days thereafter. When the cells reached 80%-90% confluence, they were digested with trypsin / EDTA digestion solution and passaged, then transferred to new culture dishes for further culture. When the cell concentration reaches 80-90%, the medium is replaced with serum-free human mesenchymal stem cell culture medium and cultured for another 24 hours. The medium is then replaced and the cell culture supernatant is collected. This process is repeated three times for each batch of cells. The cell culture supernatant collected from the three times is centrifuged for 20 minutes (3000g / min) to remove cells and cell debris. The cells are then centrifuged at 3000g / min at room temperature for 40 minutes using a Centricon Plus-70 (100kDa) ultrafiltration tube. This yields the umbilical cord MSC exosome nanocarrier delivery system, labeled as #2 umbilical cord MSC exosome nanocarrier delivery system.

[0029] Example 3 This embodiment describes the preparation of an umbilical cord MSC exosome nanocarrier delivery system.

[0030] Freshly collected umbilical cord tissue was washed with physiological saline containing penicillin-streptomycin solution and then cut into 1mm pieces using ophthalmic scissors. 2 Small pieces of the cells were transferred to cell culture flasks, and DMEM / F12 medium containing 0.05 wt% diosgenin and penicillin-streptomycin was added. The flasks were then incubated statically in a 5% CO2 incubator at 37°C. The medium was changed for the first time after one week, and then every 3-4 days thereafter. When the cells reached 80%-90% confluence, they were digested with trypsin / EDTA digestion solution and passaged, then transferred to new culture dishes for further culture. When the cell concentration reaches 80-90%, the medium is replaced with serum-free human mesenchymal stem cell culture medium and cultured for another 24 hours. The medium is then replaced and the cell culture supernatant is collected. This process is repeated three times for each batch of cells. The cell culture supernatant collected from the three times is centrifuged for 28 minutes (4000 g / min) to remove cells and cell debris. The cells are then centrifuged at 3000 g / min for 40 minutes at room temperature using a Centricon Plus-70 (100 kDa) ultrafiltration tube. This yields the umbilical cord MSC exosome nanocarrier delivery system, labeled as the 3# umbilical cord MSC exosome nanocarrier delivery system.

[0031] Example 4 This embodiment describes the preparation of an umbilical cord MSC exosome nanocarrier delivery system.

[0032] Freshly collected umbilical cord tissue was washed with physiological saline containing penicillin-streptomycin solution and then cut into 1mm pieces using ophthalmic scissors. 2 Small pieces of the cells were transferred to cell culture flasks, and DMEM / F12 medium containing 0.1 wt% diosgenin and penicillin-streptomycin was added. The flasks were then incubated statically in a 5% CO2 incubator at 37°C. The medium was changed for the first time after one week, and then every 3-4 days thereafter. When the cells reached 80%-90% confluence, they were digested with trypsin / EDTA digestion solution and passaged, then transferred to new culture dishes for further culture. When the cell concentration reaches 80-90%, the medium is replaced with serum-free human mesenchymal stem cell culture medium and cultured for another 24 hours. The medium is then replaced and the cell culture supernatant is collected. This process is repeated three times for each batch of cells. The cell culture supernatant collected from the three times is centrifuged for 30 minutes (3500 g / min) to remove cells and cell debris. The cells are then centrifuged at 3500 g / min for 38 minutes at room temperature using a Centricon Plus-70 (100 kDa) ultrafiltration tube. This yields the umbilical cord MSC exosome nanocarrier delivery system, labeled as #4 umbilical cord MSC exosome nanocarrier delivery system.

[0033] Example 5 This embodiment describes the preparation of an umbilical cord MSC exosome nanocarrier delivery system.

[0034] Freshly collected umbilical cord tissue was washed with physiological saline containing penicillin-streptomycin solution and then cut into 1mm pieces using ophthalmic scissors. 2 Small pieces of the cells were transferred to cell culture flasks, and DMEM / F12 medium containing 0.5 wt% diosgenin and penicillin-streptomycin was added. The flasks were then incubated statically in a 5% CO2 incubator at 37°C. The medium was changed for the first time after one week, and then every 3-4 days thereafter. When the cells reached 80%-90% confluence, they were digested with trypsin / EDTA digestion solution and passaged, then transferred to new culture dishes for further culture. When the cell concentration reaches 80-90%, the medium is replaced with serum-free human mesenchymal stem cell culture medium and cultured for another 24 hours. The medium is then replaced and the cell culture supernatant is collected. This process is repeated three times per batch of cells. The cell culture supernatant collected from the three times is centrifuged for 25 minutes (4000g / min) to remove cells and cell debris. The cells are then centrifuged at 4000g / min for 35 minutes at room temperature using a Centricon Plus-70 (100kDa) ultrafiltration tube. This yields the umbilical cord MSC exosome nanocarrier delivery system, labeled as #5 Umbilical Cord MSC Exosome Nanocarrier Delivery System.

[0035] Comparative Example 1 The only difference between this comparative example and Example 2 is that diosgenin ketone is not added to the culture medium.

[0036] Freshly collected umbilical cord tissue was washed with physiological saline containing penicillin-streptomycin solution and then cut into 1mm pieces using ophthalmic scissors. 2 Small pieces of the cells were transferred to cell culture flasks, and DMEM / F12 medium containing 12% serum substitute and penicillin-streptomycin was added. The flasks were then incubated statically in a 5% CO2 incubator at 37°C. The medium was changed for the first time after one week, and then every 3-4 days thereafter. When the cells reached 80%-90% confluence, they were digested with trypsin / EDTA digestion solution and passaged, then transferred to new culture dishes for further culture. When the cell concentration reaches 80-90%, the medium is replaced with serum-free human mesenchymal stem cell culture medium and cultured for another 24 hours. The medium is then replaced and the cell culture supernatant is collected. This process is repeated three times per batch of cells. The cell culture supernatant collected from the three times is centrifuged for 20 minutes (3000 g / min) to remove cells and cell debris. The cells are then centrifuged at 3000 g / min for 40 minutes at room temperature using a Centricon Plus-70 (100 kDa) ultrafiltration tube to obtain the umbilical cord MSC exosome nanocarrier delivery system, which is labeled as the No. 1 umbilical cord MSC exosome nanocarrier delivery system contrast agent.

[0037] Comparative Example 2 This comparative example involves dissolving diosgenin in ethanol and then diluting it with physiological saline to prepare a 0.01 wt% diosgenin suspension. This suspension is labeled as contrast agent #2.

[0038] Example 6 This embodiment evaluates the therapeutic effects of different umbilical cord MSC exosome nanocarrier delivery systems on allergic rhinitis.

[0039] 6.1 Constructing an OVA-induced mouse model of allergic rhinitis Ninety Balb / c mice were acclimatized for one week at 20–24°C and 45%–65% relative humidity with alternating 12h / 12h light and dark conditions. Ten mice were randomly selected as a healthy control group, which were fed normally and did not participate in the modeling process. The remaining 80 mice underwent modeling. The modeling mice received intraperitoneal injections of ovalbumin-aluminum hydroxide adjuvant sensitizer for three weeks, with each mouse receiving 0.2 mL. The control group received saline. From the fourth week onwards, mice were challenged daily with 5% OVA nasal drops for one week. The control group received saline nasal drops. Behavioral observations were performed within ten minutes after the last three OVA challenges. The results showed that, compared with the control group, the model group scratched its nose 89.32 times / 10 min, while the control group scratched its nose 35.35 times / 10 min; the model group sneezed 76.81 times / 10 min, while the control group sneezed 6.29 times / 10 min. This indicates that the OVA-induced mouse allergic rhinitis model was successfully established.

[0040] 6.2 Evaluation of the therapeutic effects of different umbilical cord MSC exosome nanocarrier delivery systems on allergic rhinitis The allergic rhinitis mouse model constructed in 6.1 was randomly divided into 8 groups, with 10 mice in each group, labeled as test group 1#, test group 2#, test group 3#, test group 4#, test group 5#, control group 1#, control group 2#, and model group. Healthy control mice raised in 6.1 served as the healthy control group, and were fed routinely. Mice in test group 1# received 10 μL of the umbilical cord MSC exosome nanocarrier delivery system prepared in Example 1 via intranasal drops once daily. Mice in test group 2# received 10 μL of the umbilical cord MSC exosome nanocarrier delivery system prepared in Example 2 via intranasal drops once daily. Mice in test group 3# received 10 μL of the umbilical cord MSC exosome nanocarrier delivery system prepared in Example 3 via intranasal drops once daily. Mice in test group 4# received 10 μL of the umbilical cord MSC exosome nanocarrier delivery system prepared in Example 4 via intranasal drops once daily. Mice in the standard control group received 10 μL of the No. 5 umbilical cord MSC exosome nanocarrier delivery system prepared in Example 5 via intranasal instillation once daily. Mice in the control group received 10 μL of the No. 5 umbilical cord MSC exosome nanocarrier delivery system prepared in Comparative Example 1 via intranasal instillation once daily. Mice in the model group received 10 μL of physiological saline via intranasal instillation once daily. Healthy controls received 10 μL of physiological saline via intranasal instillation once daily. After 7 days of continuous nasal instillation treatment, and then for 3 consecutive days following day 14, the room was kept in absolute quiet. Simultaneously, for 3 consecutive days, 10 μL of 5% OVA was slowly instilled into each nostril using a pipette, taking care to avoid choking. Mice were gently placed in their cages, and the duration was 10 minutes. The number of sneezes was recorded during this time, and videos were also taken. The number of sneezes and nose scratchings was verified by reviewing the video footage. The behavior of each group of mice was statistically analyzed, and the results are shown in Table 1.

[0041] Table 1. Behavioral statistics of mice in different treatment groups

[0042] Table 1 shows that the model group mice scratched their noses 91.35 times / 10 min and sneezed 34.82 times / 10 min, which were significantly higher than the healthy control group (18.29 times / 10 min and 6.15 times / 10 min), further verifying the successful establishment of the allergic rhinitis model. Experimental group #3 showed the best therapeutic effect with 20.35 times / 10 min of nose scratching and 8.31 times / 10 min of sneezing, approaching the level of the healthy control group. Experimental group #2 (29.41 times / 10 min of nose scratching and 13.49 times / 10 min) and experimental group #4 (35.85 times / 10 min of nose scratching and 15.76 times / 10 min of sneezing) showed moderate therapeutic effects. Experimental groups #1 and #5 were comparable to control groups #1 and #2, showing some therapeutic effect, but their therapeutic effect was far lower than that of experimental groups #2, #3, and #4. Control group #2 was comparable to the model group, with no significant therapeutic effect.

[0043] After the final treatment, mice were sacrificed 21 days later. Head tissue was harvested, skin was removed, mandibles were excised, and the brain was removed along the suture line connecting the top of the head and the back of the eye sockets, preserving the nasal cavity. Connective tissue and muscle on the surface of the skull were removed, and intact nasal mucosa tissue was obtained under a dissecting microscope. The relative expression levels of IL-4 and IL-10 mRNA in the nasal mucosa tissue were detected by Q-PCR. The results are shown below. Figure 1 and Figure 2 As shown.

[0044] Depend on Figure 1 The relative expression level of IL-4 mRNA in the model group reached 14.38, exhibiting typical Th2-type inflammatory response characteristics, confirming the successful construction of the allergic rhinitis model. Compared with the model group, experimental group #3 significantly reduced IL-4 mRNA expression, decreasing it by 83.5%, closest to the level of the healthy control group; indicating its optimal effect. Experimental group #2 significantly reduced IL-4 mRNA expression compared with the model group, decreasing it by 59.5%; experimental group #4 reduced IL-4 expression by 54.7%. Experimental groups #1, #5, and control group #1 only reduced IL-4 expression by 35-38%, while control group #2 had a similar IL-4 expression level to the model group.

[0045] Depend on Figure 2 It was found that IL-10 expression in the model group increased compensatorily to 6.31-fold, indicating a self-protective response to inflammation in the mice. However, excessively high IL-10 levels suggest immune dysregulation. In experimental group #3, treated with the #3 umbilical cord MSC exosome nanocarrier delivery system, IL-10 levels were closest to normal, maintaining appropriate anti-inflammatory capacity while avoiding overcompensation. Experimental groups #2 and #4 showed some improvement, but levels remained high. In experimental groups #1, #5, and control group #1, IL-10 levels remained high. Control group #2 was almost ineffective, similar to the model group.

[0046] In summary, the results show that in experimental group #3, the addition of 0.05 wt% diosgenin to the culture medium was the most effective way to restore the Th1 / Th2 immune balance, significantly inhibiting the overexpression of the pro-inflammatory factor IL-4 and regulating the anti-inflammatory factor IL-10 to near-normal levels, thus avoiding compensatory increases. Experimental groups #2 (0.05 wt% diosgenin added to the culture medium, 0.01 wt%) and #4 (0.05 wt% diosgenin added to the culture medium, 0.1 wt%) effectively suppressed inflammation, but the degree of immune balance restoration was slightly less than that of experimental group #3. Experimental groups #1, #5, and control group #1 exhibited significant Th2-type inflammation, showing some effect in reducing the inflammatory response, but failing to effectively correct the immune imbalance. In control group #2, the use of diosgenin suspension alone had almost no therapeutic effect, remaining at the same level as the model group.

[0047] The umbilical cord MSC exosome nanocarrier delivery system prepared by adding 0.05 wt% diosgenin to the culture medium was able to most effectively regulate the Th1 / Th2 immune balance in allergic rhinitis mice: it significantly inhibited the expression of the pro-inflammatory factor IL-4 (from 14.38-fold to 2.37-fold), while moderately restoring the anti-inflammatory factor IL-10 to near-normal levels (from 6.31-fold to 2.43-fold), and restored the inflammation balance index (IL-4 / IL-10 ratio) from 2.28 to 0.98, close to that of the healthy control group, which is the best example.

[0048] As described above, the basic principles, main features, and advantages of the present invention have been well described. The above embodiments and specifications are merely descriptions of preferred embodiments of the present invention, and the present invention is not limited to the above embodiments. Various changes and improvements made to the technical solutions of the present invention by those skilled in the art without departing from the spirit and scope of the present invention should fall within the protection scope defined by the present invention.

Claims

1. A method for preparing an umbilical cord MSC exosome nanocarrier delivery system, characterized in that, Includes the following steps: Umbilical cord tissue was collected, cleaned, cut into tissue blocks, and inoculated into a culture medium containing diosgenin ketone for culture. The cell culture supernatant was collected, and the umbilical cord MSC exosome nanocarrier delivery system was isolated and extracted from the cell culture supernatant.

2. The preparation method according to claim 1, characterized in that, The culture medium contains 0.01wt% to 0.1wt% diosgenin.

3. The preparation method according to claim 2, characterized in that, The culture medium contains 0.05 wt% diosgenin.

4. The preparation method according to claim 1, characterized in that, The separation and extraction method is as follows: centrifuge the cell culture supernatant to remove cells and cell debris, and then use an ultrafiltration tube to concentrate the supernatant to obtain the final product.

5. The preparation method according to claim 4, characterized in that, The centrifugation conditions are 3000~4000g / min for 20~30min; the ultrafiltration conditions are 3000~3500g / min for 35~40min at room temperature.

6. The umbilical cord MSC exosome nanocarrier delivery system prepared by the preparation method according to any one of claims 1 to 5.

7. The umbilical cord MSC exosome nanocarrier delivery system according to claim 6, characterized in that, The umbilical cord MSC exosome nanocarrier delivery system inhibits the expression of pro-inflammatory factor IL-4 and increases the expression of anti-inflammatory factor IL-10.

8. A pharmaceutical composition, characterized in that, It includes the umbilical cord MSC exosome nanocarrier delivery system of claim 6, and a pharmaceutically acceptable carrier or excipient.

9. The use of the umbilical cord MSC exosome nanocarrier delivery system according to claim 6 in the preparation of a drug for treating allergic rhinitis.

10. The application according to claim 9, characterized in that, The drug alleviates allergic rhinitis symptoms by regulating the mRNA expression levels of IL-4 and IL-10 in nasal mucosa tissue.

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