A traditional Chinese medicine composition for preventing and treating acute high altitude pulmonary edema and improving hypoxia tolerance, a pharmaceutical preparation, a preparation method and an application

Abstract

The application discloses a traditional Chinese medicine composition for preventing and treating acute high altitude pulmonary edema and improving hypoxia tolerance, a pharmaceutical preparation, a preparation method and application. The traditional Chinese medicine composition comprises, in parts by mass, 1-20 parts of ginseng, 1-20 parts of ophiopogon japonicus, 1-20 parts of schisandra chinensis and 1-20 parts of ziziphus jujuba. The traditional Chinese medicine composition can significantly improve the degree of pulmonary edema caused by low pressure hypoxia, reduce the lung dry / wet weight ratio, reduce the hypoxia factor content in alveolar lavage fluid caused by hypoxia, reduce the increase of pro-inflammatory factors caused by hypoxia, regulate the oxidation stress indexes GSH and SOD caused by hypoxia to approach normal levels, significantly relieve acute lung injury caused by high altitude hypoxia, effectively inhibit cell apoptosis caused by hypoxia through cell experiments, and thus improve the hypoxia tolerance of the organism. The four traditional Chinese medicine raw materials are food, traditional Chinese medicinal materials or health food raw materials, and can be prepared into the traditional Chinese medicine composition and the pharmaceutical preparation for preventing and treating acute high altitude pulmonary edema and improving hypoxia tolerance.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to a traditional Chinese medicine composition, pharmaceutical preparation, preparation method, and application for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance. Background Technology

[0002] People who rush to high altitudes or engage in intensive work at high altitudes are prone to serious harm due to the high altitude, cold, low oxygen, and low pressure environment, leading to acute mountain sickness (AMS). AMS is a common illness in high-altitude areas; if left uncontrolled, it can develop into high-altitude pulmonary edema, high-altitude cerebral edema, and high-altitude pulmonary hypertension, seriously endangering the health of those who rush to high altitudes and affecting their ability to work under hypoxic conditions. Currently, anti-altitude sickness drugs are mainly divided into traditional Chinese medicine, Tibetan medicine, Western medicine, and combined traditional Chinese and Western medicine, primarily working by reducing the body's oxygen consumption rate under hypoxic conditions, increasing the oxygen production rate of various tissues and organs, reducing hypoxic damage, and increasing the body's superoxide dismutase levels. High-altitude pulmonary edema involves functional changes in multiple organs and tissues, with a complex pathogenesis. Currently, there are no effective traditional Chinese medicine treatments for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance. Summary of the Invention

[0003] This application provides a traditional Chinese medicine composition, pharmaceutical preparation, preparation method, and application for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, and 1-20 parts Hovenia dulcis. This traditional Chinese medicine composition can significantly improve the degree of pulmonary edema caused by low-pressure hypoxia, thereby reducing the lung dry-wet weight ratio, reducing the content of hypoxia factors in bronchoalveolar lavage fluid (BALF) caused by hypoxia, alleviating the increase of pro-inflammatory factors caused by hypoxia, and regulating the oxidative stress indicators GSH and SOD caused by hypoxia to approach normal levels. It significantly alleviates acute lung injury and other symptoms caused by high-altitude hypoxia. Cell experiments have confirmed that this traditional Chinese medicine composition can effectively inhibit cell apoptosis caused by hypoxia, thereby improving the body's hypoxia tolerance, enhancing high-altitude adaptability, and improving work performance under hypoxic conditions at high altitudes. All four Chinese medicinal ingredients are food, Chinese medicinal materials, or health food ingredients. They are non-toxic, have no side effects, and are green and healthy. They can be prepared into Chinese medicinal compositions and drug preparations for the prevention and treatment of acute high-altitude pulmonary edema and to improve the ability to tolerate hypoxia.

[0004] In a first aspect, the present invention provides a traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, comprising the following raw materials in parts by mass: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, and 1-20 parts Hovenia dulcis.

[0005] In some embodiments, the ingredients include the following by weight: 1-5 parts ginseng, 1-5 parts Ophiopogon japonicus, 1-5 parts Schisandra chinensis, and 1-5 parts Hovenia dulcis.

[0006] In some embodiments, the ingredients include the following by weight: 1 part ginseng, 2 parts Ophiopogon japonicus, 1 part Schisandra chinensis, and 1 part Hovenia dulcis.

[0007] In some embodiments, the ingredients include the following by weight: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, 1-20 parts Hovenia dulcis, and 1-20 parts honey.

[0008] Secondly, this application provides a method for preparing a traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, comprising the following steps: Step S1: Take the raw materials in the specified mass fractions; Step S2: After mixing the raw materials obtained in step S1, distilled water is added for decoction, and the mixture is filtered to obtain the first extract and the residue; wherein the mass ratio of the total mass of the raw materials in step S2 to the mass of the added distilled water is 1:8-16; the decoction time in step S2 is 1-4 hours. Step S3: Add distilled water to the dregs from step S2 and decoct at least once. After each decoction, filter and collect the filtrate and dregs. If multiple decoctions are performed, add distilled water to the dregs collected after the previous decoction and decoct again. Combine all the filtrates to obtain the second extract. The mass ratio of distilled water to dregs added in step S3 is 10-20:1. The decoction time in step S3 is 1-4 hours. Step S4: Mix the first extract obtained in step S2 and the second extract obtained in step S3 to obtain an aqueous extract of the traditional Chinese medicine composition; Step S5: Filter the aqueous extract of the traditional Chinese medicine composition obtained in step S4 to obtain the target filtrate, and concentrate the target filtrate under vacuum to obtain the traditional Chinese medicine composition.

[0009] In some embodiments, the decoction time in step S2 is 2 hours; the mass ratio of the total mass of each raw material to the mass of added distilled water in step S2 is 1:8; the total number of decoctions in step S3 is 1; the time of each decoction in step S3 is 1 hour; and the mass ratio of the mass of added distilled water to the mass of the dregs in each step S3 is 10:1.

[0010] In some embodiments, step S5 involves filtering the aqueous extract of the traditional Chinese medicine composition obtained in step S4 to obtain the target filtrate, and then concentrating the target filtrate under vacuum to 0.455 g crude drug / ml - 2.73 g crude drug / ml to obtain the traditional Chinese medicine composition.

[0011] Thirdly, this application provides a pharmaceutical preparation for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, the pharmaceutical preparation comprising the traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, as well as pharmaceutically acceptable excipients.

[0012] In some embodiments, the dosage form of the pharmaceutical preparation is tablets, capsules, granules, pills, powders, syrups, mixtures, or oral liquids; the pharmaceutically acceptable excipients are one or more of fillers, disintegrants, binders, lubricants, flavoring agents, preservatives, or suspending agents.

[0013] Fourthly, this application provides the use of a traditional Chinese medicine composition or pharmaceutical preparation in a drug for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance.

[0014] This invention provides a traditional Chinese medicine composition, pharmaceutical preparation, preparation method, and application for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, and 1-20 parts Hovenia dulcis. This traditional Chinese medicine composition can significantly improve the degree of pulmonary edema caused by low-pressure hypoxia, thereby reducing the lung dry-wet weight ratio, reducing the content of hypoxia factors in bronchoalveolar lavage fluid (BALF) caused by hypoxia, alleviating the increase of pro-inflammatory factors caused by hypoxia, and regulating the oxidative stress indicators GSH and SOD caused by hypoxia to approach normal levels. It significantly alleviates acute lung injury and other symptoms caused by high-altitude hypoxia. Cell experiments have confirmed that this traditional Chinese medicine composition can effectively inhibit cell apoptosis caused by hypoxia, thereby improving the body's hypoxia tolerance, enhancing high-altitude adaptability, and improving work performance under hypoxic conditions at high altitudes. All four Chinese medicinal ingredients are food, Chinese medicinal materials, or health food ingredients. They are non-toxic, have no side effects, and are green and healthy. They can be prepared into Chinese medicinal compositions and drug preparations for the prevention and treatment of acute high-altitude pulmonary edema and to improve the ability to tolerate hypoxia. Attached Figure Description

[0015] Figure 1 A flowchart is provided as an example of a method for preparing a traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, according to some embodiments. Figure 2 The graph shows the effects of a traditional Chinese medicine composition on the levels of inflammatory factors IL-1β, GSH, and ROS oxidative stress, as well as the content of the hypoxia factor HIF-1α in mouse BALF; where A represents the inflammatory factor IL-1β in BALF. β A is a schematic diagram of the content; B is a schematic diagram of the content of the hypoxia factor HIF-1α in BALF; C is a schematic diagram of the GSH oxidative stress level in BALF; D is a schematic diagram of the ROS oxidative stress level. Figure 3 This is a graph showing the effect of traditional Chinese medicine composition on the dry-wet weight ratio of the lungs in mice. Figure 4 A schematic diagram showing the pathological results of lung tissue in each group of mice; Figure 5 This is a schematic diagram showing the results of the anti-hypoxia pharmacological activity of HUVEC cells in each group. Detailed Implementation

[0016] To better understand the above technical solutions, the technical solutions of this application will be described in detail below through specific implementation methods.

[0017] To address the aforementioned technical problems, this application provides a traditional Chinese medicine composition, preparation method, pharmaceutical formulation, and application for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance. The traditional Chinese medicine composition comprises the following raw materials by mass: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, and 1-20 parts Hovenia dulcis. This traditional Chinese medicine composition can significantly improve the degree of pulmonary edema caused by low-pressure hypoxia, thereby reducing the lung dry-wet weight ratio, reducing the content of hypoxia factors in bronchoalveolar lavage fluid (BALF) caused by hypoxia, alleviating the increase of pro-inflammatory factors caused by hypoxia, and regulating the oxidative stress indicators GSH and SOD caused by hypoxia to approach normal levels. It significantly alleviates acute lung injury and other symptoms caused by high-altitude hypoxia. Cell experiments have confirmed that this traditional Chinese medicine composition can effectively inhibit cell apoptosis caused by hypoxia, thereby improving the body's hypoxia tolerance, enhancing high-altitude adaptability, and improving work performance under hypoxic conditions at high altitudes. All four Chinese medicinal ingredients are food, Chinese medicinal materials, or health food ingredients. They are non-toxic, have no side effects, and are green and healthy. They can be prepared into Chinese medicinal compositions and drug preparations for the prevention and treatment of acute high-altitude pulmonary edema and to improve the ability to tolerate hypoxia.

[0018] This invention provides a traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, comprising the following raw materials by mass: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, and 1-20 parts Hovenia dulcis.

[0019] Ginsenosides in ginseng can improve hypoxic pulmonary vasoconstriction, while ophiopogonin in ophiopogon japonicus can protect the alveolar-capillary barrier. Schisandra chinensis lignans have antioxidant and anti-inflammatory effects. The synergistic effect of these three ingredients can address the core pathogenesis of high-altitude pulmonary edema, namely "deficiency of both qi and yin, and stagnation of fluids." In addition, Hovenia dulcis has anti-hypoxia and anti-fatigue effects. All four medicinal ingredients are food, medicinal materials, or health food ingredients, with no toxic side effects, and are green and healthy. They can be prepared into traditional Chinese medicine compositions and pharmaceutical preparations for the prevention and treatment of acute high-altitude pulmonary edema and the improvement of hypoxia tolerance.

[0020] In some embodiments, the ingredients include the following by weight: 1-5 parts ginseng, 1-5 parts Ophiopogon japonicus, 1-5 parts Schisandra chinensis, and 1-5 parts Hovenia dulcis.

[0021] In some embodiments, the ingredients include the following by weight: 1 part ginseng, 2 parts Ophiopogon japonicus, 1 part Schisandra chinensis, and 1 part Hovenia dulcis.

[0022] In some embodiments, the traditional Chinese medicine composition further includes honey. The traditional Chinese medicine composition comprises the following ingredients in parts by weight: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, 1-20 parts Hovenia dulcis, and 1-20 parts honey.

[0023] Common symptoms of altitude sickness include headache, insomnia, loss of appetite, and fatigue. Drinking a moderate amount of honey can alleviate these symptoms. Honey is rich in various nutrients such as sugars, amino acids, minerals, vitamins, and enzymes, which can replenish the body's nutrition. The glucose and fructose it contains can also provide energy, relieve fatigue caused by altitude sickness, and enhance the body's ability to adapt to high altitudes.

[0024] This application also provides a method for preparing a traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, such as... Figure 1 As shown, it includes the following steps S1-S5.

[0025] Step S1: Take the raw materials in the specified mass fractions.

[0026] Step S2: After mixing the raw materials obtained in step S1, add distilled water and decoct. After filtration, the first extract and the residue are obtained.

[0027] In step S2, the mass ratio of the total mass of the raw materials to the mass of the added distilled water is 1:8-16; the decocting time in step S2 is 1-4 hours.

[0028] In some embodiments, the decocting time in step S2 is 2 hours; the mass ratio of the total mass of each raw material to the mass of added distilled water in step S2 is 1:8.

[0029] Step S3: Add distilled water to the dregs from step S2 and decoct at least once. After each decoction, filter and collect the filtrate and dregs. If multiple decoctions are performed, add distilled water to the dregs collected after the previous decoction and decoct again. Combine all the filtrates to obtain the second extract.

[0030] In step S3, the mass ratio of distilled water to dregs added each time is 10-20:1; the decoction time in step S3 is 1-4 hours. In some embodiments, the total number of times of decoction in step S3 is 1; the time for each decoction in step S3 is 1 hour; and the mass ratio of the distilled water added each time in step S3 to the mass of the dregs is 10:1.

[0031] In some embodiments, the total number of decoctions in step S3 is 3. Step S3 specifically includes: adding distilled water to the dregs from step S2 for a first decoction; filtering and collecting the filtrate and dregs after decoction; then adding distilled water to the dregs for a second decoction; collecting the filtrate and dregs again after decoction; then adding distilled water to the dregs obtained after the second decoction for a third decoction; collecting the filtrate after decoction. The three collected filtrates are combined to obtain the second extract.

[0032] Step S4: Mix the first extract obtained in step S2 and the second extract obtained in step S3 to obtain an aqueous extract of the traditional Chinese medicine composition.

[0033] Step S5: Filter the aqueous extract of the traditional Chinese medicine composition obtained in step S4 to obtain the target filtrate, and concentrate the target filtrate under vacuum to obtain the traditional Chinese medicine composition.

[0034] In some embodiments, step S5 involves filtering the aqueous extract of the traditional Chinese medicine composition obtained in step S4 to obtain the target filtrate, and then concentrating the target filtrate under vacuum to 0.455 g crude drug / ml - 2.73 g crude drug / ml to obtain the traditional Chinese medicine composition.

[0035] In some embodiments, step S5 involves filtering the aqueous extract of the traditional Chinese medicine composition obtained in step S4 to obtain the target filtrate, and then concentrating the target filtrate under vacuum to 2.73 g crude drug / ml to obtain the traditional Chinese medicine composition.

[0036] In some embodiments, step S5 involves filtering the aqueous extract of the traditional Chinese medicine composition obtained in step S4 to obtain the target filtrate, and then concentrating the target filtrate under vacuum to 0.455 g crude drug / ml to obtain the traditional Chinese medicine composition.

[0037] In some embodiments, step S5 involves filtering the aqueous extract of the traditional Chinese medicine composition obtained in step S4 to obtain the target filtrate, and then concentrating the target filtrate under vacuum to 1.365 g crude drug / ml to obtain the traditional Chinese medicine composition.

[0038] This application also provides a pharmaceutical preparation for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, the pharmaceutical preparation comprising the traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, and pharmaceutically acceptable excipients.

[0039] In some embodiments, the dosage form of the pharmaceutical preparation is tablets, capsules, granules, pills, powders, syrups, mixtures, or oral liquids; the pharmaceutically acceptable excipients are one or more of fillers, disintegrants, binders, lubricants, flavoring agents, preservatives, or suspending agents.

[0040] This application also provides the use of a traditional Chinese medicine composition or pharmaceutical preparation in a drug for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance.

[0041] To illustrate the properties of the traditional Chinese medicine composition in the embodiments of this application, specific examples are provided below.

[0042] 1. Preparation of Traditional Chinese Medicine Compositions Experimental apparatus: 1 / 100,000 electronic balance (XSR205DU, Mettler Toledo International Ltd.) Experimental reagents: Ginseng (Beijing Tongrentang Technology Development Co., Ltd.), Ophiopogon japonicus (Beijing Tongrentang Technology Development Co., Ltd.), Schisandra chinensis (Beijing Tongrentang Technology Development Co., Ltd.), Hovenia dulcis (Beijing Tongrentang Technology Development Co., Ltd.) Experimental method: The traditional Chinese medicine composition comprises the following raw materials in parts by mass: 1 part ginseng, 2 parts Ophiopogon japonicus, 1 part Schisandra chinensis, and 1 part Hovenia dulcis. The preparation method of the traditional Chinese medicine composition includes the following steps: after mixing ginseng, Ophiopogon japonicus, Schisandra chinensis, and Hovenia dulcis, distilled water is added, and the mass ratio of the total mass of each raw material to the mass of the added distilled water is 1:8. The herbs were decocted for 2 hours, then separated by solid-liquid separation (filtration) to obtain the first aqueous extract and the residue. Distilled water was added to the residue again at a mass ratio of 10:1, and the decoction was continued for 1 hour. After solid-liquid separation, the second aqueous extract was obtained. The first and second aqueous extracts were mixed and then concentrated under vacuum to concentrations of 0.455 g crude drug / ml, 1.365 g crude drug / ml, and 2.73 g crude drug / ml, respectively, to obtain the corresponding low-dose traditional Chinese medicine composition (0.455 g crude drug / ml), medium-dose group (1.365 g crude drug / ml), and high-dose group (2.73 g crude drug / ml).

[0043] 2. Evaluation of the effects of traditional Chinese medicine compositions in preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance (in vivo animal experiments) Acute high-altitude pulmonary edema was induced in mice using a hypobaric hypoxia chamber. The levels of hypoxia factor HIF-1α and inflammatory factor IL-1 in mouse BALF were measured using ELISA. β The study evaluated the anti-hypoxia effect of the traditional Chinese medicine composition in three aspects: the content of oxidative stress indicators GSH and ROS; the determination of dry and wet weight of mouse lung tissue to assess lung injury caused by hypoxia; and the assessment of lung injury caused by hypoxia through pathological examination of mouse lung tissue.

[0044] Experimental instruments: IVC animal testing system (ZJ-4, Suzhou Fengshi Experimental Animal Equipment Co., Ltd.), low-pressure hypoxia animal testing chamber (FLYDWC50-IIC, Guizhou Fenglei Aviation Ordnance Co., Ltd.) Experimental animals: SPF-grade male Bagg ALBino (BALB / c) mice, 7 weeks old, weighing (20 ± 2) g (purchased from Beijing Sibeifu Biotechnology Co., Ltd.) Test sample: The low-dose traditional Chinese medicine composition of 0.455 g crude drug / ml, the medium-dose traditional Chinese medicine composition of 1.365 g crude drug / ml, and the high-dose traditional Chinese medicine composition of 2.73 g crude drug / ml prepared by the experimental method in "1. Preparation of Traditional Chinese Medicine Composition".

[0045] Sodium chloride injection (0.9%) (Shijiazhuang No.4 Pharmaceutical Co., Ltd.) Dexamethasone (Chenxin Pharmaceutical Co., Ltd.), the administration dose is 1mg·kg -1 , which is equivalent to 1 times the clinically recommended dose.

[0046] Rhodiola Sachalinensis Capsules (Jiangsu Kanion Pharmaceutical Co., Ltd.), the preparation method of the Rhodiola Sachalinensis Capsule suspension is as follows: weigh a quantitative amount of Rhodiola Sachalinensis Capsules, use 0.9% normal saline as the dispersion medium for the powder in the capsules to prepare a concentration of 144mg·mL -1 suspension; the administration dose is 2.16g·kg -1 , which is equivalent to 6 times the clinically recommended dose.

[0047] Experimental method: 42 SPF-grade male Bagg ALBino (BALB / c) mice, 7 weeks old, weighing (20 ± 2) g, purchased from Beijing Sibeifu Biotechnology Co., Ltd. (License No.: SCXK (Beijing) 2024-0001). Animal feeding conditions: temperature 23 ± 2℃, 12-hour light / 12-hour dark alternating cycle, free access to food and water. The formal experiment was carried out after 7 days of adaptive feeding.

[0048] 42 mice were randomly divided into 7 groups: (1) Blank control group (Control); (2) Model control group (Model); (3) Positive drug dexamethasone group (DEX): administered with dexamethasone 1 mg·kg -1 ; (4) Positive drug Rhodiola Sachalinensis Capsule group (HJT): administered with Rhodiola Sachalinensis Capsules 2.16 g·kg -1 ; (5) Traditional Chinese medicine composition low-dose group (TPF L): administered with the traditional Chinese medicine composition 1.56g·kg -1 (calculated as crude drug); (6) Traditional Chinese medicine composition medium-dose group (TPF M): administered with the traditional Chinese medicine composition 4.68 g·kg -1 (calculated as crude drug); (7) Traditional Chinese medicine composition high-dose group (TPF H): administered with the traditional Chinese medicine composition 9.36 g·kg -1 (calculated as crude drug).

[0049] Administration method: The positive control group received 1 mg / kg via intraperitoneal injection. -1 All other groups were administered the medication by gavage. The blank control group and the model control group received an equal volume of physiological saline. All treatment groups were administered the medication once daily, with a gavage volume of 0.1 mL / 10 g. -1 weight.

[0050] Adaptive feeding: All 7 groups of mice underwent adaptive feeding for 7 days, with free access to food and water; Day 1-7: Pre-drug administration for 7 days. All 6 groups of mice (except for the positive drug dexamethasone group in group 3) were pre-drug administered for 7 days (the blank control group and the model control group were given the same volume of physiological saline). Days 8-10: Six groups of mice (except for the blank control group in group 1) were placed in the FLYDWC50-IIC low-pressure hypoxia animal experimental chamber (Guizhou Fenglei Aviation Ordnance Co., Ltd.) to induce an acute high-altitude hypoxia model. The conditions of the oxygen chamber were: ascent speed 10 m / s, descent speed 10 m / s, altitude 6000 m, and altitude maintained for 72 h. During this period, the rats had free access to food and water. Groups 3-7 were administered drugs for 3 consecutive days (the model control group inside the chamber and the blank control group outside the chamber were given the same volume of physiological saline).

[0051] (1) The levels of hypoxia factor HIF-1α and inflammatory factor IL-1 in mouse BALF were measured by ELISA. β In addition, the levels of oxidative stress indicators GSH and ROS were used to evaluate lung injury caused by hypoxia. Experimental instrument: Microplate reader (Multiskan GO, Thermotech Instruments, Finland) Experimental reagents: mouse IL-1 β ELISA kit from Wuhan Abbott Biotechnology Co., Ltd. (Catalog No.: RK04770); Mouse HIF-1α ELISA Kit, Wuhan Abbott Biotechnology Co., Ltd. (Catalog No.: RK02889); Mouse ROS ELISA Kit (Catalog No.: RK03201) - Wuhan Abbott Biotechnology Co., Ltd. Mouse GSH ELISA Kit (Catalog No.: RK09070) - Wuhan Abbott Biotechnology Co., Ltd.

[0052] Experimental methods: Preparation of bronchoalveolar lavage fluid (BALF): The left main bronchus of mice was ligated, and 1 mL of 4℃ pre-cooled PBS was drawn into the right lung with a syringe and slowly injected to lavage the lung. BALF was obtained and the process was repeated 3 times. After centrifugation (1000 rpm, 4℃, 15 min), the supernatant was collected and stored at -20℃ for later use.

[0053] The experiment was conducted according to the ELISA kit instructions from Beijing Abbott Biotechnology Co., Ltd., and the absorbance was measured at 450 nm using a microplate reader.

[0054] Experimental results are as follows Figure 2 As shown, A represents the inflammatory cytokine IL-1 in BALF. β A is a schematic diagram of the content; B is a schematic diagram of the content of the hypoxia factor HIF-1α in BALF; C is a schematic diagram of the GSH oxidative stress level in BALF; D is a schematic diagram of the ROS oxidative stress level.

[0055] The core pathological mechanism of acute high-altitude pulmonary edema (HAPE) involves hypoxia-induced pulmonary vasoconstriction, an inflammatory cascade, and oxidative stress damage, ultimately leading to disruption of the alveolar-capillary barrier. Detecting these indicators in bronchoalveolar lavage fluid (BALF) can accurately reflect the real-time dynamics of the local pulmonary microenvironment. HIF-1α, as a core transcription factor in the hypoxia response, shows a decrease in its level, directly indicating improved oxygen supply to lung tissue; IL-1... β These are key pro-inflammatory factors that trigger the inflammatory cascade; their decreased levels indicate suppressed macrophage activation and reduced vascular leakage. The reduction in ROS and the rise in GSH together confirm enhanced free radical scavenging capacity and the restoration of redox homeostasis. When treatment is effective, these four indicators simultaneously tend towards normal values, indicating that the vicious cycle of hypoxia, inflammation, and oxidative stress has been broken, alveolar permeability has been restored, and the effective reversal of HAPE has been confirmed from a pathophysiological perspective.

[0056] Compared with the normal group, the level of hypoxia factor HIF-1α in the BALF of the model group was significantly increased ( P <0.001), indicating that low-pressure hypoxia exposure successfully induced a hypoxic response in lung tissue. Pro-inflammatory cytokine IL-1 β ( P The significantly increased levels of <0.001) indicate a marked inflammatory response in the lung tissue. Among the oxidative stress markers, the level of reactive oxygen species (ROS) was significantly elevated (…). P <0.001), antioxidant reduced glutathione (GSH) P The level of <0.01 was significantly reduced ( P <0.001) indicates severe oxidative stress damage to lung tissue and weakened antioxidant defense capabilities.

[0057] Compared with the model group, the positive drug dexamethasone ( P <0.001), Rhodiola rosea capsules ( P <0.001) and the dosage groups of the traditional Chinese medicine composition ( P The levels of HIF-1α in BALF (<0.001) were significantly reduced, suggesting that the drug could effectively alleviate pulmonary hypoxia. Regarding inflammatory factors, the positive control drug dexamethasone (… P <0.001), dosage in traditional Chinese medicine composition ( P <0.001) IL-1 β Inflammatory factors were significantly reduced in the positive drug Rhodiola rosea capsule group ( P <0.05) IL-1 β The levels of inflammatory factors were significantly reduced, and all dosage groups of the traditional Chinese medicine composition were significantly better than the positive control drug, Rhodiola rosea capsules. Regarding oxidative stress, the ROS levels in all dosage groups of the traditional Chinese medicine composition were significantly reduced (…). P <0.001), GSH levels were significantly elevated ( P <0.05, P <0.001), and the above indicators also showed similar improvement trends in the positive control drug dexamethasone group and the Rhodiola rosea capsule group. In terms of GSH level, the high-dose group of traditional Chinese medicine composition was superior to Rhodiola rosea capsule, and in terms of ROS level, all dose groups of traditional Chinese medicine composition were superior to the positive control drug Rhodiola rosea capsule.

[0058] The above results indicate that the traditional Chinese medicine composition can significantly improve the hypoxic state of lung tissue induced by low pressure and hypoxia, inhibit inflammatory response, reduce oxidative stress damage, and has a good protective effect on the prevention and treatment of acute high-altitude pulmonary edema and the improvement of hypoxia tolerance.

[0059] (2) The determination of dry and wet weight of mouse lung tissue was used to evaluate lung injury caused by hypoxia: Experimental method: The surface moisture of the left lung tissue was absorbed with filter paper and the wet weight was immediately recorded. After baking in an oven at 60℃ for 72 hours, the dry weight was recorded.

[0060] Mouse lung dry-wet weight ratio % = lung dry weight (g) / lung wet weight (g) × 100%.

[0061] Experimental results are as follows Figure 3 As shown, hypoxia damages pulmonary capillaries, leading to fluid extravasation and accumulation, increasing lung wet weight, and significantly raising the wet-to-dry weight ratio (W / D). This ratio directly reflects the severity of pulmonary edema; a decrease in this ratio indicates a reduction in pulmonary fluid and edema resolution. Compared with the normal group, ### P <0.001; compared with the model group, P <0.05, P <0.001. Experimental results showed that, compared with the normal group, the wet-to-dry weight ratio of lung tissue in the model group mice was significantly increased ( P <0.001 indicates that low-pressure hypoxia exposure successfully induced pulmonary edema. The positive result was achieved with dexamethasone (…). P <0.001), Rhodiola rosea capsules ( P <0.05) and various dosage groups of traditional Chinese medicine compositions ( P <0.01, P The concentration of the herbal composition (<0.001) significantly reduced the increase in lung tissue wet and dry weight. Compared with the model group, the low-dose group of the herbal composition showed a significant decrease in both lung wet and dry weight ratios, and was superior to the positive control drug, Rhodiola rosea capsules. These results indicate that the medium and high-dose groups of the herbal composition can significantly reduce the degree of pulmonary edema induced by low pressure and hypoxia, improve lung tissue fluid metabolism, and play a role in preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance.

[0062] (3) Evaluation of hypoxia-induced lung injury through histopathological examination: Experimental methods: After drug administration, lung tissue from mice in each group was collected, soaked in 4% paraformaldehyde for 48 h, and dehydrated with xylene and ethanol. The tissues were then embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE) to observe pathological changes.

[0063] Experimental results are as follows Figure 4 As shown, Figure 4 This is a schematic diagram of the pathological results of lung tissue in mice of each group, analyzed by HE staining, 200×.

[0064] HE staining of lung tissue is a classic method for assessing the degree of pathological damage to lung tissue. HE staining was used to observe the pathological morphological changes in the lung tissue of mice in different groups to evaluate the ameliorative effect of a traditional Chinese medicine composition on lung tissue damage caused by high-altitude hypoxia.

[0065] The experimental results showed that the lung tissue of the normal group mice was clear and intact, with no significant thickening of the alveolar septa, no obvious exudate in the alveolar cavities, no obvious inflammatory cell infiltration, and normal lung tissue morphology. In the model group mice, after exposure to low pressure and hypoxia, the lung tissue exhibited severe pathological damage, characterized by significant destruction of the alveolar structure, significant thickening of the alveolar septa, and extensive infiltration of inflammatory cells (as shown by the blue arrows). In contrast, intervention with the positive control drug dexamethasone, Rhodiola rosea capsules, and different doses of traditional Chinese medicine combinations significantly improved the pathological changes in the mouse lung tissue, showing a reduction in inflammatory cell infiltration and effective alleviation of alveolar wall thickening. This suggests that the above drugs can effectively inhibit low pressure and hypoxia-induced pulmonary inflammatory response and tissue damage, prevent acute high-altitude pulmonary edema, and improve hypoxia tolerance.

[0066] 3. Evaluation of the effect of traditional Chinese medicine composition on improving hypoxia tolerance (in vitro cell experiments): Hypoxia-induced mitochondrial-mediated apoptosis was evaluated using Annexin V-FITC / PI double staining combined with flow cytometry.

[0067] Experimental methods: Preparation of Traditional Chinese Medicine Composition: A traditional Chinese medicine composition with a concentration of 0.455 g crude drug / ml was prepared according to the experimental method in "1. Preparation of Traditional Chinese Medicine Composition".

[0068] Cell culture: Collect HUVEC cells in the logarithmic growth phase. Use a pipette to transfer 10 μL of cell dilution buffer into a hemocytometer, and count the cells using a microscope. Adjust the cell suspension concentration based on the calculated results, and seed 1.5 × 10⁴ cells per well in a 96-well plate, adding 100 μL of cell suspension to each well. Incubate the culture plate at 37°C with 5% CO₂ for 24 h before proceeding with subsequent experiments.

[0069] Administration method: After cells adhered for 24 h, the control group was given complete culture medium without CoCl2 induction; the model group was induced with CoCl2 only; and the drug-treated group was given 50 μL of drug-containing culture medium per well to make the final drug concentration in each well 750 μg / mL. After incubation in an incubator for 1 h, 50 μL of CoCl2-containing culture medium was added to each well to make the final CoCl2 concentration in the well 300 μM / mL. The cells were then incubated in a 37℃, 5% CO2 incubator for 24 h.

[0070] Annexin V-FITC / PI double staining flow cytometry for apoptosis detection: The above-mentioned herbal composition with a concentration of 0.455 g crude drug / ml was further filtered to obtain a filtrate. The filtrate was centrifuged at 3000 rpm for 10 min, and the supernatant was collected, concentrated under reduced pressure, and then freeze-dried to obtain the freeze-dried powder of the herbal composition. The freeze-dried powder of the herbal composition was weighed and dissolved in DMSO to prepare a solution of 100 mg·mL. -1 Reserve the stock solution. Collect HUVEC cells from each group after treatment, wash twice with pre-chilled PBS, centrifuge at 1000 rpm for 5 min, and discard the supernatant. Resuspend the cells in each tube with 500 μL of Annexin V binding buffer and adjust the cell density to 1×10⁻⁶. 6Cells / mL. Transfer 100 μL of cell suspension to a flow cytometry tube, add 5 μL of Annexin V-FITC and 5 μL of PI staining solution, mix gently, and incubate at room temperature in the dark for 15 min. After incubation, add 400 μL of Annexin V binding buffer to each tube, mix gently, and immediately analyze using flow cytometry. Annexin V-FITC positive cells show green fluorescence, and PI positive cells show red fluorescence. Collect 10,000-20,000 cells per tube and analyze using FlowJo software. Early apoptotic cells are Annexin V positive / PI negative, late apoptotic cells are Annexin V positive / PI positive, and necrotic cells are Annexin V negative / PI positive.

[0071] The apoptosis rate of HUVEC cells in each group was detected by Annexin V-FITC / PI double staining combined with flow cytometry. The results are as follows: Figure 5 .

[0072] Experimental results showed that the apoptosis rate in the control group was lower, and early apoptosis (Annexin V) was also lower. + / PI - ) and late apoptosis (Annexin V) + / PI - The proportion of cells undergoing apoptosis was relatively low, with a total apoptosis rate of approximately 7.32%. Compared to the control group, the proportion of cells undergoing early and late apoptosis in the CoCl2-induced hypoxia model group was significantly increased, and the total apoptosis rate was significantly higher than that in the control group. P <0.001). This result indicates that hypoxia induced apoptosis in HUVEC cells. Compared with the model group, the proportions of early and late apoptotic cells in the drug-treated group were significantly reduced, and the total apoptosis rate was significantly lower than that in the model group. P The result (<0.001) indicates that the traditional Chinese medicine composition can effectively inhibit hypoxia-induced apoptosis, thus confirming that the traditional Chinese medicine composition has a good effect on improving hypoxia tolerance.

[0073] This experiment used a hypobaric chamber to simulate a 6000 m high-altitude environment to establish an acute high-altitude hypoxia model in mice. The effects of a traditional Chinese medicine (TCM) composition on the prevention and treatment of high-altitude pulmonary edema were evaluated by measuring the dry and wet weight of the lungs in each group of mice, the levels of pro-inflammatory factors, hypoxia factors, and oxidative stress in bronchoalveolar lavage fluid, and by combining these with pathological morphological changes in lung tissue. Furthermore, the effects of CoCl2-induced hypoxia in HUVEC cells on hypoxia-induced apoptosis were evaluated, confirming at the cellular level that the TCM composition can effectively inhibit hypoxia-induced apoptosis, thereby improving the body's tolerance to hypoxia. These results indicate that the TCM composition of this application has good pharmacological activity against high-altitude pulmonary edema, significantly improving hypoxia, oxidative stress, and inflammatory responses induced by hypobaric hypoxia, reducing hypoxia-induced tissue pathological damage, and effectively inhibiting hypoxia-induced apoptosis. Therefore, it can significantly alleviate acute lung injury and other symptoms caused by high-altitude hypoxia, demonstrating significant preventive and therapeutic effects against acute high-altitude pulmonary edema and improved hypoxia tolerance.

[0074] This application provides a traditional Chinese medicine composition, pharmaceutical preparation, preparation method, and application for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, and 1-20 parts Hovenia dulcis. This traditional Chinese medicine composition can significantly improve the degree of pulmonary edema caused by low-pressure hypoxia, thereby reducing the lung dry-wet weight ratio, reducing the content of hypoxia factors in bronchoalveolar lavage fluid (BALF) caused by hypoxia, alleviating the increase of pro-inflammatory factors caused by hypoxia, and regulating the oxidative stress indicators GSH and SOD caused by hypoxia to approach normal levels. It significantly alleviates acute lung injury and other symptoms caused by high-altitude hypoxia. Cell experiments have confirmed that this traditional Chinese medicine composition can effectively inhibit cell apoptosis caused by hypoxia, thereby improving the body's hypoxia tolerance, enhancing high-altitude adaptability, and improving work performance under hypoxic conditions at high altitudes. All four Chinese medicinal ingredients are food, Chinese medicinal materials, or health food ingredients. They are non-toxic, have no side effects, and are green and healthy. They can be prepared into Chinese medicinal compositions and drug preparations for the prevention and treatment of acute high-altitude pulmonary edema and to improve the ability to tolerate hypoxia.

[0075] It will be readily understood by those skilled in the art that the above-described advantageous methods can be freely combined and superimposed without conflict. The above are merely preferred embodiments of this application and are not intended to limit this application. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of this application should be included within the protection scope of this application. The above are merely preferred embodiments of this application. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of this application, and these improvements and modifications should also be considered within the protection scope of this application.

Claims

1. A traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, characterized in that, The ingredients include the following ingredients by weight: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, and 1-20 parts Hovenia dulcis.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The ingredients include the following ingredients by weight: 1-5 parts ginseng, 1-5 parts Ophiopogon japonicus, 1-5 parts Schisandra chinensis, and 1-5 parts Hovenia dulcis.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The ingredients, by weight, include the following: 1 part ginseng, 2 parts ophiopogon japonicus, 1 part schisandra chinensis, and 1 part jujube fruit.

4. The traditional Chinese medicine composition according to claim 1, characterized in that, The ingredients, by weight, include the following: 1-20 parts ginseng, 1-20 parts Ophiopogon japonicus, 1-20 parts Schisandra chinensis, 1-20 parts Hovenia dulcis, and 1-20 parts honey.

5. A method for preparing a traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance as described in any one of claims 1-4, characterized in that, Includes the following steps: Step S1: Take the raw materials in the specified mass fractions; Step S2: After mixing the raw materials obtained in step S1, distilled water is added for decoction, and the mixture is filtered to obtain the first extract and the residue; wherein the mass ratio of the total mass of the raw materials in step S2 to the mass of the added distilled water is 1:8-16; the decoction time in step S2 is 1-4 hours. Step S3: Add distilled water to the dregs from step S2 and decoct at least once. After each decoction, filter and collect the filtrate and dregs. If multiple decoctions are performed, add distilled water to the dregs collected after the previous decoction and decoct again. Combine all the filtrates to obtain the second extract. The mass ratio of distilled water to dregs added in step S3 is 10-20:

1. The decoction time in step S3 is 1-4 hours. Step S4: Mix the first extract obtained in step S2 and the second extract obtained in step S3 to obtain an aqueous extract of the traditional Chinese medicine composition; Step S5: Filter the aqueous extract of the traditional Chinese medicine composition obtained in step S4 to obtain the target filtrate, and concentrate the target filtrate under vacuum to obtain the traditional Chinese medicine composition.

6. The method according to claim 4, characterized in that, The decoction time in step S2 is 2 hours; the mass ratio of the total mass of the raw materials in step S2 to the mass of the added distilled water is 1:8; the total number of decoctions in step S3 is 1; the time of each decoction in step S3 is 1 hour; the mass ratio of the added distilled water to the dregs in each step S3 is 10:

1.

7. The method according to claim 5, characterized in that, In step S5, the aqueous extract of the traditional Chinese medicine composition obtained in step S4 is filtered to obtain the target filtrate. The target filtrate is then concentrated under vacuum to 0.455 g crude drug / ml - 2.73 g crude drug / ml to obtain the traditional Chinese medicine composition.

8. A pharmaceutical preparation for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance, characterized in that, The pharmaceutical preparation comprises the traditional Chinese medicine composition for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance as described in any one of claims 1-4, as well as pharmaceutically acceptable excipients.

9. The pharmaceutical preparation according to claim 8, characterized in that, The dosage form of the pharmaceutical preparation is tablets, capsules, granules, pills, powders, syrups, mixtures, or oral liquids; the pharmaceutically acceptable excipients are one or more of fillers, disintegrants, binders, lubricants, flavoring agents, preservatives, or suspending agents.

10. The use of a traditional Chinese medicine composition according to any one of claims 1-4 or a pharmaceutical preparation according to claim 8 or 9 in a drug for preventing and treating acute high-altitude pulmonary edema and improving hypoxia tolerance.

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