An essential oil composition for resisting hypoxia, fatigue, and improving cold resistance, and its preparation method and uses.

Abstract

This invention provides an essential oil composition that combats hypoxia, fatigue, and enhances cold resistance. It is prepared from raw materials in the following weight ratios: 4.0–8.0 parts Rhodiola rosea essential oil, 1.5–2.5 parts ginseng essential oil, 0.5–1.5 parts Angelica sinensis essential oil, and 0.5–1.5 parts Ligusticum chuanxiong essential oil. This invention also provides a method for preparing this composition and its uses. The essential oil composition of this invention can combat hypoxia, fatigue, and enhance cold resistance. It has a fragrant aroma and works rapidly and noticeably. The combination of various essential oils can exert a synergistic effect, making it suitable for special environments such as high altitudes, diving, and aviation, as well as for daily health maintenance and disease prevention.

Description

Technical Field

[0001] This invention relates to an essential oil composition that has anti-hypoxia, anti-fatigue, and cold-resistance-enhancing effects, as well as its preparation method and uses, belonging to the field of pharmaceuticals or health products. Background Technology

[0002] Hypoxia refers to a pathological process in which insufficient oxygen supply or impaired oxygen utilization in the body's tissues or cells leads to a series of compensatory changes in their morphology, physiological function, and metabolism. Severe hypoxia can cause dysfunction or even failure of vital organs such as the brain, lungs, and heart. Clinically, diseases such as heart disease and severe asthma can cause hypoxia. Strenuous exercise and excessive labor can also lead to relative oxygen deficiency. Hypoxia is more likely to occur in special environments such as high altitudes, diving, and aviation, causing damage to vital organs and manifesting as high-altitude headaches, acute mountain sickness, and high-altitude pulmonary edema. Acute mountain sickness is the most common hypoxia syndrome, typically presenting as headache, accompanied by loss of appetite, nausea, vomiting, sleep disturbances, fatigue, and dizziness. Low temperatures can worsen the condition, increasing energy consumption and burdening the body; they also affect blood circulation, impairing the transport of oxygen and nutrients, and exacerbating tissue hypoxia. Currently, nimoxicillin, sulfadiazine, edaravone, coenzyme Q10, vitamin C, ligustrazine, and ginkgo biloba extract are commonly used in clinical practice as anti-hypoxia drugs. Although these drugs have a certain relieving effect on hypoxia, they are prone to causing adverse reactions such as gastrointestinal discomfort and allergies.

[0003] Fatigue is a physical or psychological state caused by overwork, physical stress response, negative emotions, malnutrition, chronic diseases, etc., manifested as decreased physical function, muscle soreness, drowsiness, poor concentration, lethargy, and reduced efficiency. Clinically, nutritional supplements, coenzyme Q10, tonifying traditional Chinese medicine, and psychological adjustment can be used to relieve fatigue.

[0004] As early as ancient China, through practical experience and medical theory, people recognized the close connection between hypoxia and fatigue. The ancient medical text *Suwen* states, "Exercise depletes qi…exercise leads to panting and sweating, both internally and externally, thus depleting qi." Excessive exertion easily depletes vital energy (qi), a crucial substance and driving force for maintaining human life. After qi is depleted, the body exhibits a series of signs of weakness, such as fatigue, lethargy, lack of energy, and spontaneous sweating. In the late 19th and early 20th centuries, with the rapid development of physiology and biochemistry, people gained a deeper understanding of cellular respiration, oxygen utilization, and energy metabolism, leading to a more systematic study and understanding of the intrinsic link between fatigue and hypoxia. Oxygen is essential for maintaining human life. Hypoxia can cause abnormal bodily functions, resulting in fatigue and various metabolic disorders, and in severe cases, disease in vital organs such as the heart, brain, and lungs. Hypoxia produces large amounts of harmful substances such as lactic acid and blood ammonia, increasing oxygen free radicals, reducing metabolic capacity, and failing to meet the body's needs for normal activity, thus triggering fatigue. As our understanding of the mechanisms of fatigue deepens, it has been found that fatigue is largely related to insufficient oxygen supply, and prolonged fatigue can indirectly lead to hypoxia. For example, excessive fatigue can affect cardiopulmonary function, reducing the efficiency of blood circulation and oxygen exchange in the heart and lungs, resulting in relative hypoxia in body tissues. Hypoxia caused by respiratory diseases, cardiovascular diseases, and other illnesses can also make patients more prone to fatigue symptoms. Fatigue, in turn, can worsen these conditions, creating a vicious cycle.

[0005] Traditional Chinese medicine essential oils typically possess strong pharmacological activity, are mostly aromatic, pungent, and warming, have fine molecules, high volatility, and strong lipid solubility, easily penetrate biological membranes in the body, and have high bioavailability. They can be rapidly absorbed and take effect through the gastrointestinal tract, nasal and oral mucosa, and skin. Traditional Chinese medicine essential oils have significant advantages in combating hypoxia and fatigue. They can not only quickly relieve symptoms of hypoxia and fatigue but also regulate bodily functions holistically, enhance immunity and blood circulation, and strengthen the body's resistance to hypoxia and fatigue. Summary of the Invention

[0006] The technical solution of the present invention is to provide an essential oil composition that resists hypoxia, fatigue, and improves cold resistance, as well as its preparation method and uses.

[0007] This invention provides an essential oil composition that resists hypoxia, fatigue, and improves cold resistance. It is prepared from raw materials containing the following weight ratios:

[0008] 4.0–8.0 parts of Rhodiola rosea essential oil, 1.5–2.5 parts of ginseng essential oil, 0.5–1.5 parts of Angelica sinensis essential oil, and 0.5–1.5 parts of Ligusticum chuanxiong essential oil.

[0009] Preferably, it is prepared from raw materials containing the following weight ratio:

[0010] Rhodiola rosea essential oil 6.0 parts, ginseng essential oil 2.0 parts, angelica essential oil 1.0 part, chuanxiong essential oil 1.0 part.

[0011] The ingredients also include Schisandra chinensis essential oil, with the following weight ratio:

[0012] Rhodiola rosea essential oil 4.0-8.0 parts, ginseng essential oil 1.5-2.5 parts, angelica essential oil 0.5-1.5 parts, chuanxiong essential oil 0.5-1.5 parts, schisandra essential oil 0.25-0.75 parts.

[0013] Preferably, it is prepared from raw materials containing the following weight ratio:

[0014] Rhodiola rosea essential oil 6.0 parts, ginseng essential oil 2.0 parts, angelica essential oil 1.0 part, chuanxiong essential oil 1.0 part, schisandra essential oil 0.5 parts.

[0015] The ingredients also include one or more of the following essential oils: rose essential oil, palmarosa essential oil, bergamot essential oil, jasmine essential oil, and bitter orange essential oil.

[0016] Its weight ratio is:

[0017] 4.0–8.0 parts of Rhodiola rosea essential oil, 1.5–2.5 parts of ginseng essential oil, 0.5–1.5 parts of angelica essential oil, 0.5–1.5 parts of chuanxiong essential oil, 0.25–0.75 parts of schisandra essential oil, and 0.5–2.5 parts of mixed essential oils.

[0018] Preferably, it is prepared from raw materials in the following weight ratio:

[0019] The mixture comprises 6.0 parts of Rhodiola rosea essential oil, 2.0 parts of ginseng essential oil, 1.0 part of Angelica sinensis essential oil, 1.0 part of Ligusticum chuanxiong essential oil, 0.5 parts of Schisandra chinensis essential oil, and 2 parts of mixed essential oil; preferably, the mixed essential oil is composed of 1.0 part of rose essential oil and 1.0 part of palmarosa essential oil.

[0020] The essential oil water composition of the present invention is prepared into a commonly used inhalation formulation by adding the raw materials as active ingredients and excipients or auxiliary ingredients acceptable in the pharmaceutical or health product fields. The inhalation formulation includes essential oil capsules, inhalation aerosols, inhalation powders, inhalation sprays, inhalation liquid formulations, or formulations that can be converted into vapor.

[0021] This invention provides a method for preparing the aforementioned anti-hypoxia and anti-fatigue essential oil composition, which includes the following steps:

[0022] a. Supercritical fluid extraction was used to extract Rhodiola rosea essential oil and ginseng essential oil; steam distillation was used to extract Angelica sinensis and Ligusticum chuanxiong essential oil; other single essential oils were extracted using steam distillation, pressing, supercritical CO2 extraction, or extraction methods.

[0023] b. Mix essential oils and add pharmaceutical or health supplement-acceptable excipients or auxiliary ingredients to prepare commonly used inhaled formulations;

[0024] The preparation method of Rhodiola rosea and ginseng essential oil is as follows:

[0025] Rhodiola rosea and ginseng raw materials are crushed and sieved, and placed in a supercritical fluid extraction vessel. Supercritical fluid extraction is carried out using 70% to 95% ethanol as an entrainer. The amount of entrainer is 25% to 60% of the raw material. The extraction pressure is 20 to 45 MPa, the extraction temperature is 40 to 60℃, and the extraction time is 1 to 3 hours. After extraction, the extracted oil fraction is separated and concentrated under reduced pressure at 40℃ to 50℃ until there is no alcohol taste.

[0026] Preferably, the preparation method of the Rhodiola rosea essential oil is as follows:

[0027] Take Rhodiola rosea raw material, crush it, sieve it, and place it in a supercritical fluid extraction vessel. Use 90% ethanol as an entrainer for supercritical fluid extraction. The amount of entrainer is 30% of the raw material. The extraction pressure is 30 MPa, the extraction temperature is 50℃, and the extraction time is 1.5 h. After the extraction is completed, separate the oil fraction at a separation pressure of 5.2 MPa. Concentrate the oil fraction under reduced pressure at 40℃ until there is no alcohol smell to obtain Rhodiola rosea essential oil.

[0028] The method for preparing the ginseng essential oil is as follows:

[0029] Ginseng raw material is crushed, sieved, and placed in a supercritical fluid extraction vessel. Supercritical fluid extraction is carried out using 85% ethanol as an entrainer, with the amount of entrainer being 50% of the raw material. The extraction pressure is 35 MPa, the extraction temperature is 55℃, and the extraction time is 2.5 h. After extraction, the oil fraction is separated at a separation pressure of 5.2 MPa. The oil fraction is concentrated under reduced pressure at 40℃ until there is no alcohol odor, thus obtaining ginseng essential oil.

[0030] This invention provides the use of the aforementioned essential oil composition in the preparation of health products for improving hypoxia resistance or relieving physical fatigue.

[0031] This invention provides the use of the aforementioned essential oil composition in the preparation of drugs that combat hypoxia, fatigue, and improve cold resistance.

[0032] Ancient Chinese and Tibetan medicine texts such as *Shennong Bencao Jing*, *Bencao Gangmu*, *Sibu Yidian*, and *Jingzhu Bencao* all record that Rhodiola rosea has tonic effects. In the formula of this invention, Rhodiola rosea is the principal ingredient, which invigorates qi and blood, unblocks meridians and relieves asthma, benefits intelligence and nourishes the heart, regulates qi and nourishes blood, and promotes longevity. Ginseng is the assistant ingredient, which greatly replenishes vital energy, restores pulse and consolidates the body, tonifies the spleen and lungs, calms the mind and improves intelligence, and nourishes blood and generates fluids. Angelica sinensis is mild and moistening, which not only replenishes blood and nourishes blood, but also invigorates blood and relieves pain. Ligusticum chuanxiong is pungent, warm, and aromatic, and can move without lingering. It can ascend to the top of the head, descend to the blood sea, penetrate the skin and hair, and connect with the limbs. It is a qi-regulating herb in the blood, which can invigorate blood and promote qi circulation, dispel wind and relieve pain. The combination of these two herbs, one moistening and one drying, mutually restrains each other's weaknesses and enhances their strengths, taking into account both qi and blood, nourishing blood and promoting qi circulation. Schisandra chinensis can astringe and consolidate, invigorate qi and generate fluids, tonify the kidneys and calm the heart, and is the guiding ingredient. The fragrant aromas of rose, bergamot, jasmine, and bitter orange blossoms not only harmonize the overall scent but also soothe the liver and regulate qi, serving as guiding herbs. The entire formula works synergistically to replenish qi and blood, nourish blood, strengthen the spleen and lungs, and calm the mind and spirit.

[0033] The essential oil composition of this invention can resist hypoxia, fatigue, and improve cold resistance. It has a fragrant aroma and works quickly and obviously. The combination of various raw material essential oils can play a synergistic role. It is suitable for special environments such as high altitude, diving and aviation, as well as for daily health care and disease prevention and treatment. Detailed Implementation

[0034] Example 1: Preparation of the essential oil composition of the present invention

[0035] 1. Essential oil preparation:

[0036] Single essential oils are extracted using various methods, including supercritical CO2 extraction, steam distillation, molecular distillation, extraction, soaking, and pressing.

[0037] 1.1 Supercritical CO2 extraction method was used to extract essential oils from Rhodiola rosea and ginseng.

[0038] Rhodiola rosea and ginseng raw materials are crushed and sieved, and placed in a supercritical fluid extraction vessel. Supercritical fluid extraction is carried out using 70% to 95% ethanol as an entrainer. The amount of entrainer is 25% to 60% of the raw material. The extraction pressure is 20 to 45 MPa, the extraction temperature is 40 to 60℃, and the extraction time is 1 to 3 hours. After extraction, the extracted oil fraction is separated and concentrated under reduced pressure at 40℃ to 50℃ until there is no alcohol taste.

[0039] 2. Optimal process

[0040] ① Rhodiola Rosea essential oil: Take Rhodiola Rosea raw material, crush it, pass it through an 80-mesh sieve, place it in a supercritical fluid extraction vessel, and use 90% ethanol as an entrainer for supercritical fluid extraction. The amount of entrainer is 30% of the raw material. The extraction pressure is 30 MPa, the extraction temperature is 50℃, and the extraction time is 1.5 h. After extraction, separate the extracted oil fraction at a separation pressure of 5.2 MPa. Concentrate the oil fraction under reduced pressure at 40℃ until there is no alcohol smell.

[0041] ②Ginseng essential oil: Take ginseng raw material, crush it, pass it through an 80-mesh sieve, place it in a supercritical fluid extraction vessel, and carry out supercritical fluid extraction with 85% ethanol as the entrainer. The amount of entrainer is 50% of the amount of raw material. The extraction pressure is 35 MPa, the extraction temperature is 55℃, and the extraction time is 2.5 h. After the extraction is completed, separate the extracted oil fraction at a separation pressure of 5.2 MPa. Concentrate the oil fraction under reduced pressure at 40℃ until there is no alcohol taste.

[0042] 1.2 The essential oils of Angelica sinensis and Ligusticum chuanxiong were extracted by steam distillation.

[0043] Angelica sinensis and Ligusticum chuanxiong raw materials are crushed into the coarsest powder, placed in a volatile oil extraction tank, and steam distilled for 2-4 hours. After standing for a period of time until the oil and water are completely separated in the oil-water separator, the essential oil is separated out.

[0044] Other single essential oils are extracted using methods such as steam distillation, pressing, supercritical CO2 extraction, and extraction, or can be purchased as commercially available products.

[0045] Example 2: Preparation of Aromatherapy Essential Oils

[0046] Take the single essential oils according to the specified proportions, blend them, and mix them evenly. The mixture can be diffused and inhaled using an aromatherapy diffuser, diffuser stone, or diffuser wood.

[0047] Example 3: Preparation of essential oil burst beads

[0048] Compound essential oils are diluted to a concentration of 15% to 50% using medium-chain triglycerides (MCT), grape seed oil, peony seed oil, safflower seed oil, olive oil, or rice bran oil. Appropriate amounts of Tween or glycerol can be added to solubilize them. The mixture is then prepared into popping beads and added to an aromatherapy diffuser for inhalation, or combined with an oxygen cylinder to use oxygen as a fluid for aromatherapy inhalation.

[0049] Example 4: Preparation of essential oil nasal aspirator

[0050] The compound essential oil is added to the adsorption medium of the nasal inhaler to obtain the product.

[0051] Example 5: Preparation of Electronic Atomized Essential Oil Inhaler

[0052] Microcapsule fragrance was prepared using compound essential oils, fat-soluble vitamin E, and soybean phosphatidylcholine as core materials and cyclodextrin as wall material; 50% microcapsule fragrance, 30% glycerol, and 20% propylene glycol were mixed and stirred to obtain the final product.

[0053] The following specific efficacy tests demonstrate the beneficial effects of the present invention.

[0054] Experimental Example 1: Anti-hypoxia effect of the essential oil composition of the present invention

[0055] 1.1 Effects on a mouse closed hypoxia model

[0056] 1.1.1 Experimental grouping and drug administration methods

[0057] Eighty SPF-grade KM mice, weighing 20±2g, half female and half male, were housed in the laboratory for 5 days and randomly divided into the following groups: blank control group, Rhodiola rosea essential oil group, ginseng essential oil group, Ligusticum chuanxiong-Angelica sinensis essential oil group (Ligusticum chuanxiong-Angelica sinensis 1:1), compound essential oil group 1 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong 6:2:1:1), compound essential oil group 2 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong-Schisandra chinensis 6:2:1:1:0.5), compound essential oil group 3 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong-Schisandra chinensis-Rosa rugosa-Rosa palmatum 6:2:1:1:0.5:1:1) (all raw materials are combinations of essential oils of the raw materials), and positive control Rhodiola rosea capsule group, with 10 mice in each group.

[0058] Mice in the control group and essential oil group underwent 15 minutes of atomized aromatherapy. The control group received 25 ml of water, while the essential oil group received approximately 0.05 g of essential oil in 25 ml of water. The animal aromatherapy chamber had a volume of approximately 50 L (52 cm × 38 cm × 25 cm). The dosage was 0.25 g / kg. The Rhodiola rosea capsule group received 0.5 g / kg. -1 Administer by gavage for 14 consecutive days.

[0059] One hour after the last administration, mice in each group were placed in a 250 mL wide-mouthed bottle containing 10 g of soda lime. The bottle mouth was coated with Vaseline, the stopper was quickly closed, and the bottle was inverted in water to prevent air from entering. Timing was started immediately, and the respiratory survival of the experimental animals was observed until the animals stopped breathing. The recorded time was the mouse's hypoxia tolerance time.

[0060] 1.2 Effects on a mouse model of sodium nitrite poisoning and hypoxia

[0061] 1.2.1 Experimental grouping and drug administration methods

[0062] The experimental grouping and administration methods were the same as in 1.1.1.

[0063] One hour after the last administration, mice in each group were injected intraperitoneally with sodium nitrite (NaNO2) solution at a dose of 230 mg·kg-1. Timing was started immediately after the injection, and the time from injection to cardiac and respiratory arrest was defined as the time of NaNO2 poisoning and hypoxia tolerance in mice.

[0064] 1.3 Effects on a mouse model of simulated high-altitude hypoxia

[0065] 1.3.1 Experimental grouping and drug administration methods

[0066] Ninety SPF-grade KM mice, weighing 20±2g, half male and half female, were housed in the laboratory for 5 days and randomly divided into the following groups: blank group, model group, Rhodiola rosea essential oil group, ginseng essential oil group, Ligusticum chuanxiong-Angelica sinensis essential oil group (Ligusticum chuanxiong-Angelica sinensis 1:1), compound essential oil group 1 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong 6:2:1:1), compound essential oil group 2 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong-Schisandra chinensis 6:2:1:1:0.5), compound essential oil group 3 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong-Schisandra chinensis-Rosa rugosa-Rosa palmatum 6:2:1:1:0.5:1:1), and positive control Rhodiola rosea capsule group, with 10 mice in each group.

[0067] The control group, model group, and essential oil group were subjected to atomized aromatherapy for 15 minutes. The model group and control group were given 25 ml of water, while the essential oil group was given 25 ml of water with approximately 0.05 g of essential oil added. The animal aromatherapy chamber had a volume of approximately 50 L (52 cm × 38 cm × 25 cm). The dosage was 0.25 g / kg. The Rhodiola rosea capsule group was given 0.5 g / kg by gavage. The administration was continued for 14 days.

[0068] Except for the control group, the model group and the essential oil group were placed in a low-pressure, low-oxygen animal experimental chamber simulating a high altitude of 5600m for 7 consecutive days. The chamber temperature was 23±2℃, and the relative humidity was 50-60%. The mice were weighed, given aromatherapy and medication, and given supplemental water and food, and their bedding was changed 1 hour each day. The control group was raised in a plain environment. The mice were removed from the oxygen chamber 7 days after the model was established.

[0069] 1.3.2 Evaluation Indicators

[0070] Mice were euthanized by dislocation, and their heart, liver, spleen, lungs, kidneys, and brain were removed. Fat and connective tissue were removed, and the organs were rinsed three times with physiological saline. After the water was absorbed with filter paper, the organs were weighed and the organ index was calculated. Organ index = organ weight (g) / body weight (g) × 100%.

[0071] 100 mg of lung tissue and myocardial tissue were cut and homogenized. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) in lung tissue were detected by WST-1 method, TBA method and spectrophotometry respectively according to the kit instructions. The expression of HIF-1α, p-AMPK, PGC-1α and Nrf2 in myocardial tissue was measured by Western blotting.

[0072] 2. Test Results

[0073] 2.1 Effects on mouse closed-loop hypoxia model and sodium nitrite poisoning hypoxia model: experimental results

[0074] Compared with the control group, the Rhodiola rosea essential oil group, the positive control group, and the compound essential oil group significantly prolonged the time of mice tolerating closed hypoxia and sodium nitrite poisoning hypoxia. The Angelica sinensis-Ligusticum chuanxiong group significantly prolonged the time of mice tolerating sodium nitrite poisoning hypoxia. Compared with the positive control group, there was no significant difference in the Rhodiola rosea essential oil group, while the compound essential oil group was significantly better than the positive control group. The results are shown in Table 1.

[0075] Table 1 Effects on hypoxia time in mice

[0076]

[0077]

[0078] Compared with the blank group, * P < 0.05 ** P < 0.01; compared with the positive group, # P < 0.05 ## P < 0.01;

[0079] 2.2 Effects on the mouse model of simulated high-altitude hypoxia: experimental results

[0080] Compared with the control group, there were no significant differences in the liver and kidney indices of mice in the model group, while the spleen index showed a decreasing trend, and the heart, lung, and brain indices increased. Compared with the model group, the positive control group, Rhodiola rosea essential oil group, and compound essential oil group significantly reduced the heart and brain indices of mice (P < 0.05), the drug administration group significantly reduced the lung index of mice (P < 0.05), and the compound essential oil group significantly increased the spleen index of mice (P < 0.05). Compared with the positive control group, the compound essential oil group significantly reduced the heart index of mice (P < 0.05), with no significant difference compared with the control group; compound essential oil 2 and compound essential oil 3 significantly reduced the lung and brain indices of mice (P < 0.05), with no significant difference compared with the control group. The results are shown in Table 2.

[0081] Table 2. Effects on organ indices in a mouse model of simulated high-altitude hypoxia

[0082]

[0083] Compared with the model group, * P < 0.05 ** P < 0.01; compared with the control group, # P < 0.05 ## P < 0.01;

[0084] Compared with the positive group, & P < 0.05 && P < 0.01.

[0085] Compared with the control group, the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the lung tissue of mice were decreased, while the level of malondialdehyde (MDA) was increased (P < 0.05). Compared with the model group, the drug-treated groups all showed increased SOD and GSH-Px levels and decreased MDA levels (P < 0.05). Compared with the positive control group, the compound essential oil group showed significantly increased SOD and GSH-Px levels and decreased MDA levels (P < 0.05), while there was no significant difference compared with the control group (P > 0.05). Compared with Rhodiola rosea, ginseng, and Angelica sinensis-Ligusticum striatum essential oils, the compound essential oil showed better effects. The results are shown in Table 3.

[0086] Table 3. Effects of SOD, MDA, and GSH-Px content in lung tissue of a mouse model of simulated high-altitude hypoxia.

[0087]

[0088] Compared with the model group, * P < 0.05 ** P < 0.01; compared with the control group, # P < 0.05 ## P < 0.01;

[0089] Compared with the positive group, & P < 0.05 && P < 0.01.

[0090] Compared with the control group, the expression of HIF-1α in the heart tissue of mice was increased, while the expression of p-AMPK, PGC-1α, and Nrf2 was decreased (P < 0.05). Compared with the model group, all treatment groups showed increased expression of HIF-1α, p-AMPK, PGC-1α, and Nrf2 (P < 0.05). Compared with the positive control group, the compound essential oil group showed significantly increased expression of HIF-1α, p-AMPK, PGC-1α, and Nrf2 (P < 0.05). Compared with Rhodiola rosea, ginseng, and Angelica sinensis-Ligusticum striatum essential oils, the compound essential oil showed better effects. The results are shown in Table 4.

[0091] Table 4. Effects of HIF-1α, p-AMPK, PGC-1α, and Nrf2 expression in cardiac tissue of a mouse model of simulated high-altitude hypoxia.

[0092]

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

[0094] Compared with the positive group, & P < 0.05 && P < 0.01.

[0095] Superoxide dismutase (SOD) and glutathione (GSH-Px) can scavenge peroxides in the body and reflect the body's antioxidant capacity. Multivitamin A (MDA) is the end product of lipid free radical metabolism and reflects the body's oxidative stress level. Therefore, SOD, GSH-Px, and MDA can serve as indicators of oxidative stress, directly reflecting the body's antioxidant capacity. Experiments showed that under high-altitude hypoxia exposure, the activities of SOD and GSH-Px in mouse lung tissue were significantly reduced, while MDA was significantly increased. This indicates that hypoxia caused the body to produce a large number of oxygen free radicals, consuming antioxidants such as SOD and inhibiting their activity, leading to lipid peroxidation and the production of large amounts of MDA, inducing oxidative stress damage. After administration, the activities of SOD and GSH-Px were significantly increased, while MDA production was reduced. This indicates that the compound essential oil has a good anti-hypoxia effect at high altitudes, can improve lung tissue damage, improve the body's free radical scavenging ability, reduce lipid peroxidation, improve oxidative stress damage, and thus enhance the body's resistance to hypoxia in high-altitude environments.

[0096] HIF-1α participates in the hypoxia response, mediating extracellular signal transduction, stress response, and energy metabolism in cardiomyocytes. It is an initiating factor for reconstructing cardiomyocyte metabolic pathways and endogenous protective mechanisms. AMPK is a key factor in the regulation of energy metabolism in the body. Increased expression of HIF-1α nucleoprotein can induce activation of the AMPK pathway, reduce cellular ATP consumption, and decrease oxygen demand, helping the body adapt to the hypoxic microenvironment. AMPK phosphorylation can induce PGC-1α transcription and regulate Nrf2 expression. PGC-1α is a major regulator of mitochondrial function, and Nrf2 is a major regulator of oxidative stress. Under high-altitude hypoxic conditions, AMPK phosphorylation levels in the heart, brain, and skeletal muscle are significantly reduced, and the expression of PGC-1α and Nrf2 proteins is significantly decreased. Experiments have shown that compound essential oils can significantly increase the expression of HIF-1α, p-AMPK, PGC-1α, and Nrf2 in mouse myocardial tissue. The above results indicate that compound essential oils may exert their anti-altitude hypoxia effect by activating the HIF-1α / AMPK signaling pathway, further promoting the transcription of PGC-1α and Nrf2 genes, improving myocardial energy metabolism, and reducing oxidative stress damage.

[0097] In summary, the essential oil composition of this invention can significantly prolong the duration of closed-loop hypoxia and NaNO2 poisoning hypoxia in mice. It reduces the lung, brain, and heart organ indices in a mouse model of simulated high-altitude hypoxia, increases the spleen index, raises SOD and GSH-Px levels in lung tissue, and lowers MDA levels; it also increases the expression of HIF-1α, p-AMPK, PGC-1α, and Nrf2 in heart tissue. The essential oil can improve lung tissue damage and enhance the body's ability to scavenge free radicals, thereby increasing hypoxia resistance. It can further promote PGC-1α and Nrf2 gene transcription by activating the HIF-1α / AMPK signaling pathway, improving myocardial energy metabolism, and reducing oxidative stress damage, thus exerting an anti-high-altitude hypoxia effect. It can also enhance immunity and improve hypoxia tolerance to a certain extent.

[0098] Test Example 2: Anti-fatigue effect test of the essential oil composition of the present invention

[0099] 1. Experimental grouping and administration methods

[0100] Eighty SPF-grade KM mice, weighing 20±2g, half male and half female, were housed in the laboratory for 5 days and randomly divided into the following groups: blank control group, Rhodiola rosea essential oil group, ginseng essential oil group, Ligusticum chuanxiong-Angelica sinensis essential oil group (Ligusticum chuanxiong-Angelica sinensis 1:1), compound essential oil group 1 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong 6:2:1:1), compound essential oil group 2 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong-Schisandra chinensis 6:2:1:1:0.5), compound essential oil group 3 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong-Schisandra chinensis-Rosa rugosa-Rosa palmatum 6:2:1:1:0.5:1:1), and positive control Rhodiola rosea capsule group, with 10 mice in each group.

[0101] Mice in the blank control group and essential oil group were subjected to aromatherapy by atomization for 15 minutes. The blank control group was 25 ml of water, and the essential oil group was 25 ml of water with about 0.05 g of essential oil added. The volume of the animal aromatherapy chamber was about 50 L (52 cm × 38 cm × 25 cm). The dosage was 0.25 g / kg. The Rhodiola rosea capsule group was administered 0.5 g / kg by gavage for 14 consecutive days.

[0102] 2. Mouse pole climbing experiment

[0103] Thirty minutes after the last administration, the pole-climbing test was conducted. The mice were placed on top of a glass rod to induce static muscle tension, and the time it took for the mice to fall off the glass rod due to muscle fatigue was recorded. The average of the three measurements was taken as the pole-climbing time.

[0104] 3. Mouse weighted swimming experiment

[0105] After the pole-climbing experiment, the mice were allowed to recover for 3 days, during which time each group continued to receive nebulized aromatherapy medication. The swimming experiment was conducted 30 minutes after the last administration. A lead wire weighing 6% of the mouse's body weight was placed on the mouse's tail in a bucket of water 40 cm deep at a temperature of 25 ± 2℃. The mice were propelled to swim continuously by stirring the water. The time it took for the mouse to be unable to resurface for 6 seconds after entering the water, from when its head was completely submerged, was recorded as the mouse's exhaustion swimming time. After recording the time, the mice were removed from the water.

[0106] Blood was collected from the eyeballs of mice, and the mice were euthanized by dislocation. The livers were removed, and liver tissue homogenates were prepared. Whole blood lactate (LD) and liver glycogen were determined by colorimetric method.

[0107] 4. Test Results:

[0108] 4.1 Results of the mouse pole climbing experiment and weighted swimming experiment

[0109] Compared with the blank group, the drug treatment group significantly prolonged the time for mice to climb the pole. The Rhodiola rosea essential oil group, the positive control group, and the compound essential oil group significantly prolonged the time for mice to reach exhaustion while swimming under load (P<0.05). Compared with the positive control group, there was no significant difference between the Rhodiola rosea essential oil group and the ginseng essential oil group, while the compound essential oil group was significantly better than the positive control group (P<0.05). The results are shown in Table 5.

[0110] Table 5. Effects on pole climbing time and exhaustion during weighted swimming in mice.

[0111]

[0112] Compared with the blank group, * P < 0.05 ** P < 0.01; compared with the positive group, # P < 0.05 ## P < 0.01;

[0113] 4.2 Results of experiments on the effects of exercise on liver glycogen and lactate in mice

[0114] Compared with the blank group, the drug treatment group significantly reduced the lactate (LD) content in the blood of mice and increased the liver glycogen content (P<0.01). Compared with the positive drug group, there was no significant difference in the Rhodiola rosea essential oil group and the ginseng essential oil group. The Angelica sinensis-Ligusticum chuanxiong essential oil group was weaker than the positive drug group. The compound essential oil group was significantly better than the positive group (P<0.05). The results are shown in Table 6.

[0115] Table 6 Effects on liver glycogen and lactate levels in mice after exercise

[0116]

[0117] Compared with the blank group, * P < 0.05 ** P < 0.01; compared with the positive group, # P < 0.05 ## P < 0.01;

[0118] The essential oil composition significantly prolonged the pole-climbing time and exhaustion time of mice during weight-bearing swimming. It also reduced blood lactose levels and increased liver glycogen levels in mice after exercise, thus exerting an anti-fatigue effect.

[0119] Experimental Example 3: Cold Resistance Test of the Essential Oil Composition of the Present Invention

[0120] 1. Experimental grouping and administration methods

[0121] 160 SPF-grade KM mice, weighing 20±2g, half male and half female, were housed in the laboratory for 5 days and randomly divided into the following groups: blank control group, Rhodiola rosea essential oil group, ginseng essential oil group, Ligusticum chuanxiong-Angelica sinensis essential oil group (Ligusticum chuanxiong-Angelica sinensis 1:1), compound essential oil group 1 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong 6:2:1:1), compound essential oil group 2 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong-Schisandra chinensis 6:2:1:1:0.5), compound essential oil group 3 (Rhodiola rosea-ginseng-Angelica sinensis-Ligusticum chuanxiong-Schisandra chinensis-Rosa rugosa-Rosa palmatum 6:2:1:1:0.5:1:1), and positive control Rhodiola rosea capsule group, with 20 mice in each group.

[0122] Mice in the blank control group and essential oil group were subjected to aromatherapy by atomization for 15 minutes. The blank control group was 25 ml of water, and the essential oil group was 25 ml of water with about 0.05 g of essential oil added. The volume of the animal aromatherapy chamber was about 50 L (52 cm × 38 cm × 25 cm). The dosage was 0.25 g / kg. The Rhodiola rosea capsule group was administered 0.5 g / kg by gavage for 14 consecutive days.

[0123] 2. Evaluation Indicators

[0124] Thirty minutes after the last administration, the mice were placed in a refrigerator at (-6±2)℃. The refrigerator was opened and checked every 0.5 hours. Dead animals were picked up and the number of animals that died within 5.0 hours was recorded. The mortality rate of each group of animals was used as a comparison index to compare the cold resistance of each group of mice.

[0125] 3. Experimental Results on Improving Cold Resistance

[0126] Compared with the blank group, the drug-treated group reduced the mortality rate of mice. Compared with the positive drug group, the compound essential oil group was significantly better than the positive drug group. The results are shown in Table 7.

[0127] Table 7 Effects on cold resistance in mice

[0128]

[0129] The essential oil composition can significantly reduce the mortality rate of mice under low temperature conditions and enhance the body's ability to resist cold.

[0130] Test Example 4: Clinical Trial of the Essential Oil Composition of the Present Invention

[0131] Compound essential oil 3 was diluted with medium-chain triglycerides (MCT) to a concentration of 25% to prepare essential oil capsules. These capsules were placed in the oxygen generator valve cavity of an oxygen cylinder. When in use, the capsules were popped to release oxygen as a fluid for aromatherapy inhalation, and the anti-hypoxia effect was evaluated.

[0132] 1. Forty-five healthy adult volunteers were selected, including 30 males and 15 females, with an age of (28.75±7.49) years, a height of (165.77±8.32) cm, and a weight of (62.97±10.29) kg. After clinical medical examination, they were found to have no acute or chronic diseases of the circulatory, respiratory, or gastrointestinal systems, and met the requirements for human experiments in a low-pressure chamber.

[0133] 2. Before entering the chamber, the volunteers' heart rate and blood oxygen saturation were measured. The volunteers sat quietly in the low-pressure chamber and breathed the air inside the chamber. The low-pressure chamber simulated an exposure to an altitude of 5km for 45 minutes.

[0134] Thirty-nine volunteers experienced symptoms of hypoxia, including drowsiness, headache, dizziness, chest tightness, fever, blurred vision, nausea, and vomiting. Among them, 27 were male and 12 were female. Heart rate and blood oxygen saturation were immediately measured after exiting the isolation ward, and the volunteers' complaints were also taken into account.

[0135] 3. A randomized, single-blind experiment was conducted, in which volunteers experiencing hypoxia symptoms were divided into two groups: a control group that inhaled only oxygen and an experimental group that inhaled essential oil oxygen. The control group consisted of 19 people (13 males and 6 females), and the experimental group consisted of 20 people (14 males and 6 females). There were no statistically significant differences in general characteristics such as gender, age, weight, heart rate, and blood oxygen saturation between the two groups (P > 0.05), indicating that they were comparable.

[0136] 4. After exiting the isolation chamber, both groups of volunteers immediately received oxygen. Their complaints were inquired about, and the time when they felt their hypoxia symptoms began to lessen or disappear was recorded. Heart rate and blood oxygen saturation were measured at 5, 15, and 30 minutes after oxygen inhalation.

[0137] 5. Clinical efficacy analysis

[0138] Compared with the control group, the experimental group volunteers experienced a significantly shorter time for their hypoxia symptoms to begin to lessen or disappear, and the experimental group was able to improve their heart rate and blood oxygen saturation in a short period of time. The experimental results are shown in Tables 8 and 9.

[0139] Table 8 shows the effects on the time of relief or disappearance of hypoxia symptoms and heart rate.

[0140]

[0141] Compared to before entering the cabin, * P < 0.05 ** P < 0.01; compared with before oxygen inhalation, ▲ P < 0.05 ▲▲ P < 0.01; compared with the control, # P < 0.05 ## P < 0.01.

[0142] Table 9. Effects on blood oxygen saturation

[0143]

[0144] Compared to before entering the cabin, * P < 0.05 ** P < 0.01; compared with before oxygen inhalation, ▲ P < 0.05 ▲▲ P < 0.01; compared with the control, # P < 0.05 ## P < 0.01.

[0145] The essential oil combination can rapidly increase blood oxygen saturation, reduce heart rate, and improve hypoxia symptoms in patients with hypoxia.

Claims

1. An essential oil composition that combats hypoxia, fatigue, and enhances cold resistance, characterized in that: It is an inhaled formulation prepared from the following raw materials in the following weight ratio: The ingredients are: 4.0-8.0 parts of Rhodiola rosea essential oil, 1.5-2.5 parts of ginseng essential oil, 0.5-1.5 parts of angelica essential oil, and 0.5-1.5 parts of chuanxiong essential oil; wherein the weight ratio of Rhodiola rosea essential oil, ginseng essential oil, angelica essential oil, and chuanxiong essential oil is 6:2:1:

1.

2. The essential oil composition for resisting hypoxia, fatigue, and improving cold resistance according to claim 1, characterized in that: It is an inhaled formulation prepared from the following raw materials in the following weight ratio: Rhodiola rosea essential oil 6.0 parts, ginseng essential oil 2.0 parts, angelica essential oil 1.0 part, chuanxiong essential oil 1.0 part.

3. An essential oil composition that combats hypoxia, fatigue, and enhances cold resistance, characterized in that: It is an inhaled formulation prepared from the following raw materials in the following weight ratio: The ingredients are: 4.0-8.0 parts of Rhodiola rosea essential oil, 1.5-2.5 parts of ginseng essential oil, 0.5-1.5 parts of angelica sinensis essential oil, 0.5-1.5 parts of chuanxiong essential oil, and 0.25-0.75 parts of schisandra chinensis essential oil; wherein the weight ratio of Rhodiola rosea essential oil, ginseng essential oil, angelica sinensis essential oil, chuanxiong essential oil, and schisandra chinensis essential oil is 6:2:1:1:0.

5.

4. The essential oil composition for resisting hypoxia, fatigue, and improving cold resistance according to claim 3, characterized in that: It is an inhaled formulation prepared from the following raw materials in the following weight ratio: Rhodiola rosea essential oil 6.0 parts, ginseng essential oil 2.0 parts, angelica essential oil 1.0 part, chuanxiong essential oil 1.0 part, schisandra essential oil 0.5 parts.

5. An essential oil composition that resists hypoxia, fatigue, and improves cold resistance, characterized in that: It is an inhaled formulation prepared from the following raw materials in the following weight ratio: The ingredients are: 4.0-8.0 parts Rhodiola rosea essential oil, 1.5-2.5 parts ginseng essential oil, 0.5-1.5 parts Angelica sinensis essential oil, 0.5-1.5 parts Ligusticum chuanxiong essential oil, 0.25-0.75 parts Schisandra chinensis essential oil, and 0.5-2.5 parts mixed essential oil; the mixed essential oil is composed of rose essential oil and palmarosa essential oil; wherein the weight ratio of Rhodiola rosea essential oil, ginseng essential oil, Angelica sinensis essential oil, Ligusticum chuanxiong essential oil, Schisandra chinensis essential oil, rose essential oil, and palmarosa essential oil is 6:2:1:1:0.5:1:

1.

6. The essential oil composition for resisting hypoxia, fatigue, and improving cold resistance according to claim 5, characterized in that: It is an inhaled formulation prepared from the following raw materials in the following weight ratio: The oil consists of 6.0 parts of Rhodiola rosea essential oil, 2.0 parts of ginseng essential oil, 1.0 part of angelica essential oil, 1.0 part of chuanxiong essential oil, 0.5 parts of schisandra essential oil, and 2.0 parts of mixed essential oil; the mixed essential oil is composed of 1.0 part of rose essential oil and 1.0 part of palmarosa essential oil.

7. The essential oil composition for resisting hypoxia, fatigue, and improving cold resistance according to any one of claims 1-6, characterized in that: It is prepared from the raw materials as active ingredients and with the addition of excipients acceptable in the pharmaceutical or health product fields to form commonly used inhaled preparations, wherein the inhaled preparations include inhaled aerosols, inhaled powders, and inhaled sprays.

8. A method for preparing an essential oil composition for anti-hypoxia and anti-fatigue as described in any one of claims 5-7, characterized in that: It includes the following steps: a. Supercritical fluid extraction was used to extract Rhodiola rosea essential oil and ginseng essential oil; steam distillation was used to extract Angelica sinensis and Ligusticum chuanxiong essential oil; other single essential oils were extracted using steam distillation, pressing, supercritical CO2 extraction, or extraction methods. b. Mix essential oils and add pharmaceutically or health supplement-acceptable excipients to prepare commonly used inhaled formulations; The preparation method of Rhodiola rosea and ginseng essential oil is as follows: Rhodiola rosea and ginseng raw materials are crushed and sieved, and placed in a supercritical fluid extraction vessel. Supercritical fluid extraction is carried out using 70% to 95% ethanol as an entrainer. The amount of entrainer is 25% to 60% of the raw material. The extraction pressure is 20 to 45 MPa, the extraction temperature is 40 to 60℃, and the extraction time is 1 to 3 hours. After extraction, the extracted oil fraction is separated and concentrated under reduced pressure at 40℃ to 50℃ until there is no alcohol taste.

9. The method for preparing the anti-hypoxia and anti-fatigue essential oil composition according to claim 8, characterized in that: The preparation method of the Rhodiola rosea essential oil is as follows: Take Rhodiola rosea raw material, crush it, sieve it, and place it in a supercritical fluid extraction vessel. Use 90% ethanol as an entrainer for supercritical fluid extraction. The amount of entrainer is 30% of the raw material. The extraction pressure is 30 MPa, the extraction temperature is 50℃, and the extraction time is 1.5 h. After the extraction is completed, separate the oil fraction at a separation pressure of 5.2 MPa. Concentrate the oil fraction under reduced pressure at 40℃ until there is no alcohol smell to obtain Rhodiola rosea essential oil. The method for preparing the ginseng essential oil is as follows: Ginseng raw material is crushed, sieved, and placed in a supercritical fluid extraction vessel. Supercritical fluid extraction is carried out using 85% ethanol as an entrainer, with the amount of entrainer being 50% of the raw material. The extraction pressure is 35 MPa, the extraction temperature is 55℃, and the extraction time is 2.5 h. After extraction, the oil fraction is separated at a separation pressure of 5.2 MPa. The oil fraction is concentrated under reduced pressure at 40℃ until there is no alcohol odor, thus obtaining ginseng essential oil.

10. Use of the essential oil composition according to any one of claims 1-7 in the preparation of health products for improving resistance to hypoxia or relieving physical fatigue.

11. Use of the essential oil composition according to any one of claims 1-7 in the preparation of a medicament for resisting hypoxia, fatigue, and improving cold resistance.

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