Application of stone herba scrophulariae extract in preparation of medicine for treating allergic rhinitis

Abstract

The present application relates to the technical field of medicine, and particularly to application of an extract of Lithocarpus polystachyus Rehder in preparing a medicine for treating allergic rhinitis, wherein the extract of Lithocarpus polystachyus Rehder is found to take effect rapidly for the first time. The extract of Lithocarpus polystachyus Rehder is found to significantly relieve symptoms such as sneezing, rhinorrhea, nasal congestion, and nasal itching within 30 minutes after administration, which is a rapid-acting property that has not been reported in the field of traditional Chinese medicine for treating allergic rhinitis. The extract of Lithocarpus polystachyus Rehder discovered in the present application can treat the nose and lungs simultaneously. Through an allergic rhinitis animal model induced by OVA, it is confirmed that the extract of Lithocarpus polystachyus Rehder can not only reduce the nasal itching behavior of mice and other symptoms of rhinitis, but also significantly reduce the percentage of eosinophils in the bronchoalveolar lavage fluid, significantly reduce the levels of IgE, IL-5, and IL-13 in the serum, and improve the pathological damage of the lung tissue, thereby achieving a synergistic improvement on allergic rhinitis and related lower airway inflammation.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically the application of *Gynostemma pentaphyllum* extract in the preparation of drugs for treating allergic rhinitis. Background Technology

[0002] Allergic rhinitis is a chronic inflammatory disease of the nasal mucosa mediated by immunoglobulin E. Clinical manifestations include sneezing, runny nose, nasal congestion, nasal itching, and itchy eyes, severely impacting patients' quality of life. Epidemiological surveys show that the global prevalence of allergic rhinitis is as high as 10%-40%, and is showing an increasing trend year by year.

[0003] Currently, commonly used clinical treatments include nasal corticosteroids and oral antihistamines. However, existing drugs have limitations such as slow onset of action, easy relapse after discontinuation, and adverse reactions in some patients including drowsiness and nasal dryness. Traditional Chinese medicine has the advantages of multi-target and holistic regulation in the treatment of allergic diseases, but existing Chinese herbal formulas or extracts generally suffer from slow onset of action, unclear mechanisms of action, and a lack of modern pharmacological data to support their efficacy.

[0004] *Gynostemma pentaphyllum*, a plant belonging to the genus *Gynostemma* in the family Gesneriaceae, is commonly used in traditional Chinese medicine to treat respiratory ailments such as cough, excessive phlegm, and asthma. Existing literature reports confirm its antitussive, expectorant, and in vitro antioxidant effects. However, no research has been reported on its use in treating allergic rhinitis, particularly in rapidly relieving acute symptoms.

[0005] Therefore, developing a traditional Chinese medicine extract that is fast-acting, has a clear efficacy, and can simultaneously improve allergic inflammation in both the upper and lower airways has significant clinical importance and market value. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides the application of *Gynostemma pentaphyllum* extract in the preparation of drugs for treating allergic rhinitis, and its application in the preparation of drugs that simultaneously improve allergic rhinitis-related lung inflammation and oxidative stress, as detailed below: Application of *Gynostemma pentaphyllum* extract in the preparation of drugs for treating allergic rhinitis.

[0007] Furthermore, the aforementioned medication for treating allergic rhinitis is a drug that relieves one or more of the symptoms caused by allergic rhinitis, such as sneezing, runny nose, nasal congestion, nasal itching, and tearing.

[0008] Furthermore, the medication for treating allergic rhinitis is a drug that reduces lung inflammation caused by allergic rhinitis. This drug reduces the percentage of eosinophils in bronchoalveolar lavage fluid, reduces serum and / or bronchoalveolar lavage fluid levels of interleukin-5 and / or interleukin-13, and improves pathological damage to lung tissue.

[0009] Furthermore, the medication for treating allergic rhinitis is a drug that reduces the elevated level of oxidative stress in lung tissue caused by allergic rhinitis. This drug reduces the elevated level of oxidative stress in lung tissue caused by allergic rhinitis by increasing the activity of superoxide dismutase in lung tissue and / or reducing the malondialdehyde content in lung tissue.

[0010] Furthermore, the *Sedum spectabile* extract is a water or ethanol extract of *Sedum spectabile*.

[0011] Furthermore, the extract of *Sedum spectabile* is obtained by the following steps: take *Sedum spectabile* medicinal material, decoct with water or extract with ethanol, combine the extracts, filter, concentrate and dry the filtrate to obtain the extract.

[0012] Furthermore, the decoction or reflux, percolation, or cold soaking extraction is performed 1-5 times, with 4-20 times the amount of water or 10%~95% ethanol added each time, and the extraction time is 0.5-4 hours.

[0013] A medication for treating allergic rhinitis, the raw material of which is extract of *Gynostemma pentaphyllum*.

[0014] Compared with the prior art, the technical effects of this invention are reflected in: (1) This invention is the first to discover that the extract of Dendrobium nobile has a rapid onset of action. This invention is the first to discover that the extract of Dendrobium nobile can significantly relieve symptoms such as sneezing, runny nose, nasal congestion, and nasal itching within 30 minutes after administration. This rapid-acting characteristic has not been reported in the field of traditional Chinese medicine treatment of allergic rhinitis.

[0015] (2) The extract of *Sedum spectabile* discovered in this invention can treat both the nose and lungs. This invention has demonstrated through an OVA-induced animal model of allergic rhinitis that the extract of *Sedum spectabile* can not only alleviate behavioral symptoms of rhinitis such as nose scratching in mice, but also significantly reduce the percentage of eosinophils in bronchoalveolar lavage fluid, significantly reduce serum levels of IgE, IL-5, and IL-13, and improve pathological damage to lung tissue, thus achieving synergistic improvement of allergic rhinitis and related lower airway inflammation.

[0016] (3) This invention is the first to discover that the extract of *Sedum spectabile* also has antioxidant effects. Furthermore, this invention has found that the extract of *Sedum spectabile* can significantly increase the activity of superoxide dismutase in the lung tissue of mice with allergic rhinitis and reduce the content of malondialdehyde, providing new pharmacological evidence for the treatment of allergic rhinitis. Attached Figure Description

[0017] Figure 1 Timeline of establishment and treatment of a mouse model of allergic rhinitis.

[0018] Figure 2Bar chart showing the effect of *Gynostemma pentaphyllum* extract on mouse behavior. Note: NC is the normal control group; M is the model group; SDLL, SDLM, and SDLH are the low, medium, and high dose groups of *Gynostemma pentaphyllum* extract, respectively; DXM is the dexamethasone treatment group. ###P<0.001 vs. normal group; ***P<0.001 vs. model group.

[0019] Figure 3 Bar chart showing the effect of *Gynostemma pentaphyllum* extract on the percentage of eosinophils in bronchoalveolar lavage fluid of mice. Note: NC is the normal control group; M is the model group; SDLL, SDLM, and SDLH are the low, medium, and high dose groups of *Gynostemma pentaphyllum* extract, respectively; DXM is the dexamethasone treatment group. ###P<0.001 vs. normal group; *P<0.05, ***P<0.001 vs. model group.

[0020] Figure 4 Bar chart showing the effect of *Gynostemma pentaphyllum* extract on the levels of inflammatory factors and immunoglobulins in mouse serum and bronchoalveolar lavage fluid. Note: NC is the normal control group; M is the model group; SDLL, SDLM, and SDLH are the low, medium, and high dose groups of *Gynostemma pentaphyllum* extract, respectively; DXM is the dexamethasone treatment group. ##P<0.01, ###P<0.001 vs. normal group; *P<0.05, **P<0.01, ***P<0.001 vs. model group.

[0021] Figure 5 HE staining images of the effects of *Gynostemma pentaphyllum* extract on the pathological characteristics of lung tissue in mice with OVA-induced allergic rhinitis. Note: NC is the normal control group; M is the model group; SDLL, SDLM, and SDLH are the low, medium, and high dose groups of *Gynostemma pentaphyllum* extract, respectively; DXM is the dexamethasone treatment group.

[0022] Figure 6 Bar chart showing the effects of *Gynostemma pentaphyllum* extract on SOD activity and MDA content in lung tissue of mice with allergic rhinitis. Note: NC is the normal control group; M is the model group; SDLL, SDLM, and SDLH are the low, medium, and high dose groups of *Gynostemma pentaphyllum* extract, respectively; DXM is the dexamethasone treatment group. ##P<0.01, ###P<0.001 vs. normal group; *P<0.05, **P<0.01, ***P<0.001 vs. model group.

[0023] Figure 7 Results of the improvement of behavioral scores such as nose scratching in mice by ethanol extract of *Sedum spectabile* (Note: NC is the normal control group; M is the model group; SDL-YCL and SDL-YCH are the low- and high-dose groups of ethanol extract of *Sedum spectabile*, respectively; DXM is the dexamethasone treatment group. ##P<0.01 vs normal group; *P<0.05, **P<0.01 vs model group.)

[0024] Figure 8 Bar chart showing the effect of *Gynostemma pentaphyllum* ethanol extract on the percentage of eosinophils in bronchoalveolar lavage fluid of mice. Note: NC is the normal control group; M is the model group; SDL-YCL and SDL-YCH are the low- and high-dose groups of *Gynostemma pentaphyllum* ethanol extract, respectively; DXM is the dexamethasone treatment group. ##P<0.01 vs. normal group; **P<0.01 vs. model group. Detailed Implementation

[0025] The technical solution of the present invention will be further defined below with reference to specific embodiments, but the scope of protection is not limited to the description made.

[0026] A study on the anti-allergic rhinitis and antioxidant effects of *Gynostemma pentaphyllum* extract using an OVA-induced mouse model of allergic rhinitis. 1. Materials 1.1 Instruments YLS-8B Small Animal Nebulizer (Jinan Yiyan Technology Development Co., Ltd.); Allegra 64R Low-Temperature High-Speed ​​Centrifuge (Beckman Coulter, USA); IMS-20 Fully Automatic Snowflake Ice Maker (Changshu Xueke Electric Appliance Co., Ltd.); AE240 0.0001 ppm Electronic Balance (Mettler-Toledo Instruments [Shanghai] Co., Ltd.); Olympus CX41 Simple Polarizing Microscope and Mingmei MC-50 Microscopic Imaging System (Guangzhou Mingmei Optoelectronic Technology Co., Ltd.); EL240 Electronic Balance (Mettler-Toledo Instruments [Shanghai] Co., Ltd.); KQ-300DE CNC Ultrasonic Cleaner (Kunshan Ultrasonic Instruments Co., Ltd.); Micropipette (Eppendorf, Germany); 700 Series Ultra-Low Temperature Freezer (Thermo Fisher Scientific, USA); WP-UP-Ⅱ-20 Ultrapure Water System (Sichuan Wotel Technology Development Co., Ltd.); Model 680 Multifunctional Microplate Reader (Bio-Rad, USA).

[0027] 1.2 Test Drugs Ovalbumin (OVA, Sigma-Aldrich, batch No.: SLCH2414); aluminum hydroxide adjuvant (Thermo Fisher Scientific, USA, batch No.: WC320438); dexamethasone (Solarbio, batch No.: SD9530, specification: 100 mg); Wright staining solution (Solarbio, batch No.: 20200910); IL-5 (Cat. No.: ZC-37987), IL-13 (Cat. No.: ZC-37967) and IgE (Cat. No.: ZC-38496) kits were all purchased from Shanghai Zhuocai Biotechnology Co., Ltd.; malondialdehyde (Cat. No.: A003-1-2) and superoxide dismutase (Cat. No.: A001-3-2) assay kits were purchased from Nanjing Jiancheng Bioengineering Institute; phosphate buffered saline (PBS, pH 7.2~7.4); water was ultrapure water, and other reagents were all of analytical grade.

[0028] 1.3 Experimental Animals Healthy female Kunming mice (20±2 g) were purchased from Changsha Tianqin Biotechnology Co., Ltd., and were SPF grade animals with production license No. SCXK (Xiang) 2019-0014. Mice were housed in an environment with 12 h light / dark cycle, 18~25°C, and 50~70% relative humidity for one week before being used for experimental research, with free access to food and water during the period. All experimental protocols in this study were approved by the Animal Ethics Committee of Guizhou Medical University.

[0029] 2 Methods 2.1 Preparation of *Lysionotus pauciflorus* Extract *Lysionotus pauciflorus* was purchased from Bijie City, Guizhou Province, and was identified as *Lysionotus pauciflorus* Lysionotus pauciflorus Maxim. dried aerial parts by Researcher Liu Chunhua from Guizhou Provincial Key Laboratory of Pharmaceutics, Guizhou Medical University. Voucher specimens are stored in Guizhou Provincial Key Laboratory of Pharmaceutics, Guizhou Medical University. The medicinal material was crushed into coarse powder, 500 g was weighed, extracted by decoction with water for 3 times: 2 h for the first time (8 times volume (W / V) of ultrapure water), 1.5 h for the second time (6 times volume (W / V) of ultrapure water), and 0.5 h for the third time (6 times volume (W / V) of ultrapure water). The decoctions were combined, filtered, the filtrate was concentrated into a thick paste, and dried under reduced pressure at 60°C to obtain *Lysionotus pauciflorus* extract (yield: 13.71%).

[0030] 2.2 Grouping, Modeling and Administration of Animals Sixty mice were randomly divided into six groups: normal control group (NC), model group (M), low (5 g / kg crude drug, SDLL), medium (10 g / kg crude drug, SDLM), high (20 g / kg crude drug, SDLH) dose groups of *Sedum spectabile* extract, and dexamethasone (1 mg / kg, DXM) group, with 10 mice in each group. Except for the normal control group, mice in the other groups were sensitized by intraperitoneal injection of 0.1 mL of sensitization solution (containing 25 μg OVA and 2 mg aluminum hydroxide adjuvant) on days 0, 7, and 14 of the experiment. Mice in the normal control group were sensitized with an equal volume of physiological saline instead of sensitization solution, and the administration method was the same. From days 21 to 26, mice in the model group and each drug administration group were placed in a closed nebulizer and sensitized by nebulization with 2% OVA solution at a spray rate of 1 mL / min for 20 min. Mice in the normal control group were sensitized with physiological saline instead of sensitization solution. During mouse provocation, observe for positive reactions such as facial itching, sneezing or coughing, rapid breathing, restlessness, abdominal muscle twitching, and immobility. Starting on day 21, 30 minutes before nebulization, administer different doses of *Gynostemma pentaphyllum* and dexamethasone solutions in ultrapure water. Administer these solutions via gavage to mice in the *Gynostemma pentaphyllum* and dexamethasone groups, respectively, according to mouse body weight. The normal control and model groups are given ultrapure water. This process is repeated for 6 consecutive days. The establishment and treatment schedule for the allergic rhinitis model is as follows: Figure 1 As shown.

[0031] 2.3 Behavioral Indicators After the final nebulization, once the mice's fur was dried, behavioral indicators of allergic rhinitis, such as nose scratching, itching, and allergic rhinitis symptoms, were observed and scored within 10 minutes: 0 points for no nose scratching or itching, 1 point for 1-3 instances of nose scratching or itching, 2 points for 4-6 instances, and 3 points for 7 instances or more. The behavioral evaluation of the mice was conducted according to this standard.

[0032] 2.4 Collection of serum and bronchoalveolar lavage fluid (BALF) samples Twenty-four hours after the last challenge, blood was collected from the eyeballs of mice. After standing at room temperature, the blood was centrifuged at 3000 r / min for 10 min at 4°C to separate the serum. The serum was stored at -80°C for the detection of IgE, IL-5, and IL-13. Mice were euthanized by cervical dislocation after blood collection. Four mice from each group were randomly selected, their chest cavities were opened, and endotracheal tubes were inserted. The lungs were lavaged with pre-cooled PBS at a rate of 0.5 mL per lavage, twice in total. The lavage fluid was collected. The lavage fluid was centrifuged at 2500 r / min at 4°C for 5 min. The precipitate was resuspended in PBS, spread on a glass slide, and air-dried. After staining with Wright's stain, the total number of inflammatory cells and eosinophils were determined under a microscope. The supernatant was used to detect the IL-5 content.

[0033] 2.5 Detection of IgE, IL-5 and IL-13 The concentrations of IgE, IL-5, IL-13 in serum and IL-5 in BALF were determined using enzyme-linked immunosorbent assay (ELISA) according to the instructions of the corresponding kits.

[0034] 2.6 Pathological analysis of lung tissue Fresh mouse lungs were fixed in 10% neutral formaldehyde fixative and then dehydrated using a gradient of 70% to 100% ethanol. After trimming, embedding, sectioning, staining, and mounting, the lung tissue morphology and eosinophilic cell infiltration around the trachea and blood vessels were observed under an optical microscope. The main focus was on the degree of infiltration of inflammatory cells around the airway, especially eosinophils (EOS).

[0035] 2.7 Detection of SOD activity and MDA levels in lung tissue Accurately weigh the tissue and add 9 times the volume of physiological saline at a ratio of weight (g):volume (mL) = 1:9. Cut the tissue into small pieces, prepare a homogenate in an ice water bath, centrifuge at 2500~3000 r / min for 10 min, collect the supernatant, and determine the activity of SOD and the content of MDA according to the kit instructions.

[0036] 2.8 Statistical Methods All data were analyzed using GraphPad Prism 8.0.1 software. Experimental data are expressed as mean ± SD. One-way ANOVA was used to compare differences among multiple groups. Comparisons between two groups were performed using... t test, P A value <0.05 was considered statistically significant.

[0037] 3 Results 3.1 Behavioral changes in mice with allergic rhinitis After OVA modeling, the animals exhibited obvious symptoms of acute allergic rhinitis, such as facial itching, forelimb retraction and elevation, arched back, and abdominal breathing. Behavioral evaluation revealed that the model group mice had significantly higher comprehensive behavioral scores for allergic rhinitis than the control group. P <0.001); compared with the model group, the behavioral scores of allergic rhinitis symptoms were significantly reduced after treatment with SDL and DXM ( P <0.001), such as Figure 2 As shown.

[0038] 3.2 Effect of *Sedum spectabile* extract on the percentage of eosinophils in OVA-induced BALF in mice Eosinophils, as part of the immune system, are normally used to fight infection, but high levels can cause respiratory inflammation, exacerbating airway inflammation in patients with allergic rhinitis. To evaluate the therapeutic effect of *Gynostemma pentaphyllum* extract on OVA-induced allergic rhinitis in mice, the inhibitory effect of *Gynostemma pentaphyllum* extract on inflammatory cell infiltration was assessed by measuring the percentage of eosinophils in inflammatory cells. The results showed that the percentage of eosinophils (EOS) in the model group was significantly increased compared to the normal group. Compared to the model group, the percentage of eosinophils in the bronchoalveolar lavage fluid (BALF) of mice treated with SDL extract and DXM was significantly decreased. Figure 3 As shown.

[0039] 3.3 Effects of *Sedum spectabile* extract on serum and BALF levels of inflammatory factors and immunoglobulins Allergic rhinitis is significantly associated with elevated levels of specific IgE, and IgE is often used clinically as an important diagnostic indicator for allergic rhinitis. Figure 4 As shown in Figure A, compared with the normal group, the IgE level in the model group was significantly increased ( P <0.01); Compared with the model group, the serum IgE levels in mice treated with SDLM, SDLH, and DXM were significantly reduced ( P <0.05). Allergens can induce Th2 cells to secrete various cytokines, such as IL-4, IL-5, and IL-13. ELISA was used to further evaluate the expression of IL-5 and IL-13 in serum and BALF by *Gynostemma pentaphyllum* extract. Compared with the normal group, the levels of IL-5 and IL-13 in the model group were significantly increased (…). P <0.01), after treatment with SDL extract and DXM, the levels of IL-5 and IL-13 decreased significantly ( P <0.05) Figure 4 BD).

[0040] 3.4 Pathological changes in mouse lung tissue The area of ​​leukocyte infiltration in lung tissue is an important indicator for evaluating the severity of allergic rhinitis. HE staining was used to observe the morphological changes in lung tissue of mice with OVA-induced allergic rhinitis, and the bronchodilatory effect of *Gynostemma pentaphyllum* extract was evaluated using optical microscopy. In the normal group, the pleural structure of the lung tissue was relatively normal, with thin walls and no connective tissue hyperplasia or thickening; the bronchial structures at all levels were intact and clear, with normal epithelial cell morphology and no obvious degeneration, necrosis, or shedding. In contrast, the lung tissue of the model group showed pathological changes such as bronchial epithelial cell hyperplasia, alveolar epithelial cell degeneration and necrosis, alveolar septal thickening, and fibrous tissue hyperplasia. Compared with the model group, the degree of lung tissue lesions after intervention with *Gynostemma pentaphyllum* extract and DXM was relatively mild. Figure 5 As shown.

[0041] 3.5 Effects of *Sedum morganianum* on SOD and MDA in the lung tissue of mice with allergic rhinitis Recent studies have found that oxidative stress also participates in the pathogenesis of allergic rhinitis. Peroxides and their induced lipid peroxidation reactions participate in the inflammatory response of lung tissue in allergic rhinitis, exacerbating pathological changes in lung tissue. By detecting MDA levels and SOD activity in the lung tissue of mice with allergic rhinitis, it was found that compared with the normal group, the model group showed a significant increase in MDA content and a significant inhibition of SOD activity. This result further illustrates that oxidative damage occurred in mice with allergic rhinitis, and their antioxidant capacity was weakened. Compared with the model group, the MDA content in the medium- and high-dose SDL groups and the DXM group was significantly reduced. P <0.05), and SOD activity was significantly enhanced ( P <0.05), although there was no significant difference in the low-dose group of *Sedum spectabile*, there was a trend of regression, such as Figure 6 As shown in the figure. These results indicate that *Sedum morganianum* may enhance the antioxidant capacity of mice with allergic rhinitis and reduce the degree of oxidative damage by increasing SOD activity.

[0042] Ethanol extract case study: 1. Preparation of Stone Plant Extract The medicinal material was pulverized into a coarse powder, and 500 g was weighed out and extracted three times by refluxing with 50% ethanol. The first extraction was for 2 hours (8 times the amount (w / v) of 50% ethanol), the second for 1.5 hours (6 times the amount (w / v) of 50% ethanol), and the third for 0.5 hours (6 times the amount (w / v) of 50% ethanol). The extracts were combined, filtered, and the ethanol was recovered under reduced pressure. The extract was concentrated into a thick paste and dried under reduced pressure at 60°C to obtain the ethanol extract of *Sedum spectabile* (SDL-YC).

[0043] 2. Animal grouping, model establishment, and drug administration Fifty mice were randomly divided into five groups: normal control group (NC), model group (M), low-dose (5 g / kg crude drug, SDL-YCL), high-dose (20 g / kg crude drug, SDL-YCH) groups of *Sedum spectabile* extract, and dexamethasone (1 mg / kg, DXM) group, with 10 mice in each group. Except for the normal control group, mice in the other groups were sensitized on days 0, 7, and 14 by intraperitoneal injection of 0.1 mL of sensitization solution (containing 25 μg OVA and 2 mg aluminum hydroxide adjuvant). Mice in the normal control group received an equal volume of physiological saline instead of the sensitization solution, and the administration method was the same. From days 21 to 26, mice in the model group and each drug administration group were placed in a closed nebulizer and nebulized with 2% OVA solution to induce allergy. The nebulization rate was 1 mL / min, and the stimulation time was 20 min. Mice in the normal control group received physiological saline instead of the stimulation solution. During mouse provocation, observe for positive reactions such as facial itching, sneezing or coughing, rapid breathing, restlessness, abdominal muscle twitching, and immobility. Starting on day 21, 30 minutes before nebulization, administer different doses of *Gynostemma pentaphyllum* and dexamethasone solutions in ultrapure water. Administer these solutions via gavage to mice in the *Gynostemma pentaphyllum* and dexamethasone groups, respectively, according to mouse body weight. The normal control and model groups are given ultrapure water. This process is repeated for 6 consecutive days. The establishment and treatment schedule for the allergic rhinitis model is as follows: Figure 1 As shown.

[0044] 3. Behavioral indicators After the final nebulization, once the mice's fur was dried, behavioral indicators of allergic rhinitis, such as nose scratching, itching, and allergic rhinitis symptoms, were observed and scored within 10 minutes: 0 points for no nose scratching or itching, 1 point for 1-3 instances of nose scratching or itching, 2 points for 4-6 instances, and 3 points for 7 instances or more. The behavioral evaluation of the mice was conducted according to this standard.

[0045] 4. Collection of serum and bronchoalveolar lavage fluid (BALF) samples Twenty-four hours after the last challenge, blood was collected from the eyeballs of mice. After standing at room temperature, the blood was centrifuged at 3000 r / min for 10 min at 4°C to separate the serum. The serum was stored at -80°C for the detection of IgE, IL-5, and IL-13. Mice were euthanized by cervical dislocation after blood collection. Four mice from each group were randomly selected, their chest cavities were opened, and endotracheal tubes were inserted. The lungs were lavaged with pre-cooled PBS, 0.5 mL per lavage, for a total of 2 times. The lavage fluid was collected. The lavage fluid was centrifuged at 2500 r / min at 4°C for 5 min. The precipitate was resuspended in PBS, spread on a glass slide, and air-dried. After staining with Wright's stain, the total number of inflammatory cells and eosinophils were determined under a microscope.

[0046] 5. IgE and IL-13 detection The concentrations of IgE and IL-13 in serum were determined using enzyme-linked immunosorbent assay (ELISA) according to the instructions of the corresponding kits.

[0047] 6. Detection of MDA levels in lung tissue Accurately weigh the tissue and add 9 times the volume of physiological saline at a ratio of weight (g):volume (mL) = 1:9. Cut the tissue into small pieces, prepare a homogenate in an ice water bath, centrifuge at 2500~3000 r / min for 10 min, collect the supernatant, and determine the MDA content according to the kit instructions.

[0048] 7. Statistical methods All data were analyzed using GraphPad Prism 8.0.1 software. Experimental data are expressed as mean ± SD. One-way ANOVA was used to compare differences among multiple groups. Comparisons between two groups were performed using... t test, P A value <0.05 was considered statistically significant.

[0049] 8. Experimental Results Behavioral evaluation revealed that the comprehensive behavioral score of allergic rhinitis in the model group was significantly higher than that in the normal group. P <0.01); compared with the model group, the behavioral scores of allergic rhinitis symptoms were significantly reduced after treatment with SDL-YC and DXM ( P <0.05, P <0.01). See details. Figure 7 .

[0050] like Figure 8 As shown, compared with the normal group, the percentage of EOS in the model group was significantly higher ( P <0.01), compared with the model group, the percentage of EOS in BALF of mice treated with SDL-YC extract and DXM was significantly reduced ( P <0.01).

[0051] As shown in Table 1, compared with the normal group, the IgE level in the model group was significantly increased ( P <0.01); compared with the model group, the serum IgE levels in mice treated with SDL-YCL, SDL-YCH and DXM were significantly reduced ( P <0.05). Compared with the normal group, the Th2 cytokine IL-13 was significantly increased in the model group ( P <0.01), after treatment with SDL-YCL, SDL-YCH and DXM, IL-13 levels all decreased significantly ( P <0.05).

[0052] Simultaneously, the MDA level in lung tissue samples was measured. The results showed that, compared with the normal group, the MDA content in the model group was significantly increased; compared with the model group, the MDA content in the SDL-YCL, SDL-YCH, and DXM groups was significantly decreased. P <0.05), as shown in Table 1.

[0053] Table 1. Effects of ethanol extract of *Sedum spectabile* on plasma IL-13, IgE, and MDA levels in mouse lung tissue.

[0054] Note: NC represents the normal control group; M represents the model group; SDL-YCL and SDL-YCH represent the low- and high-dose groups of *Gynostemma pentaphyllum* ethanol extract, respectively; DXM represents the dexamethasone treatment group. P <0.01 vs. normal group;** P <0.01 vs model group.

[0055] These results indicate that the extract of *Gynostemma pentaphyllum* using ethanol as a solvent also has a good anti-allergic rhinitis effect.

[0056] Typical human data: 1. Case Information The patient is a 37-year-old female with a 9-year history of seasonal allergic rhinitis (spring). Main symptoms include: sneezing (more than 5 times consecutively), runny nose with clear discharge, nasal congestion, nasal itching, and itchy eyes. These symptoms last approximately 6-8 weeks each spring.

[0057] 2. Administration method During the spring flare-up period, administer 30g of *Gynostemma pentaphyllum* orally when symptoms appear. Assess symptom changes before administration, 30 minutes after administration, 1 hour after administration, and 2 hours after administration.

[0058] 3. Observation Results

[0059] 4. Conclusion This patient used *Sedum spectabile* during the spring flare-ups for two consecutive years, and similar rapid relief was observed in both cases, with no significant adverse reactions. The results of this case are corroborated by the results of an animal model (OVA-induced reduction of nose scratching symptoms in mice), supporting the potential value of *Sedum spectabile* in treating allergic rhinitis.

[0060] Finally, it should be noted that the above embodiments are merely representative examples of the present invention. Obviously, the technical solution of the present invention is not limited to the above embodiments, and many variations are possible. All variations that can be directly derived or conceived by those skilled in the art from the content disclosed in this invention should be considered within the scope of protection of this invention.

Claims

1. Application of *Gynostemma pentaphyllum* extract in the preparation of drugs for treating allergic rhinitis.

2. The application according to claim 1, characterized in that, The medication for treating allergic rhinitis is a drug that relieves one or more of the symptoms caused by allergic rhinitis, such as sneezing, runny nose, nasal congestion, nasal itching, and tearing.

3. The application according to claim 1, characterized in that, The medication mentioned is for treating allergic rhinitis and is designed to reduce lung inflammation caused by allergic rhinitis.

4. The application according to claim 3, characterized in that, The drug that reduces lung inflammation caused by allergic rhinitis is a drug that reduces the percentage of eosinophils in bronchoalveolar lavage fluid, reduces serum and / or bronchoalveolar lavage fluid levels of interleukin-5 and / or interleukin-13, and improves pathological damage to lung tissue.

5. The application according to claim 1, characterized in that, The medication mentioned is for treating allergic rhinitis and is designed to reduce the increased oxidative stress levels in lung tissue caused by allergic rhinitis.

6. The application according to claim 5, characterized in that, The drug that reduces the increased oxidative stress level in lung tissue caused by allergic rhinitis is a drug that increases the activity of superoxide dismutase in lung tissue and / or reduces the malondialdehyde content in lung tissue.

7. The application according to claim 1, characterized in that, The extract of *Sedum spectabile* is a water or ethanol extract of *Sedum spectabile*.

8. The application according to claim 7, characterized in that, The extract of *Sedum spectabile* is obtained by the following steps: take *Sedum spectabile* medicinal material, decoct with water or extract with ethanol, combine the extracts, filter, concentrate and dry the filtrate to obtain the extract.

9. The application according to claim 8, characterized in that, The extraction is carried out by decocting or reflux, percolation, or cold soaking 1-5 times, with 4-20 times the amount of water or 10%~95% ethanol added each time, and the extraction time is 0.5-4 hours.

10. A medication for treating allergic rhinitis, characterized in that, Its raw material is extract from *Gynostemma pentaphyllum*.

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