Application of qi-regulating and blood-activating preparation in preparation of medicine for preventing or treating altitude stress

Abstract

The invention relates to application of a qi-regulating and blood-activating preparation in a medicine for preventing or treating altitude stress, and belongs to the technical field of medicine application. The qi-regulating and blood-activating preparation is prepared by extracting and processing fructus cinnamoii, borneol, ligusticum wallichii and allium macrostemon. Experimental research shows that the qi-regulating and blood-activating preparation can effectively improve plateau hypoxia athletic ability and heart functions, can effectively improve plateau hypoxia lung tissue inflammation infiltration, reduces alveolar wall thickness, reduces lung tissue fibrosis, and reduces myocardial cell LDH release, thereby achieving the effect of effectively preventing and treating acute altitude stress.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical application technology, specifically to the application of qi-regulating and blood-activating preparations in the prevention and treatment of altitude sickness. Background Technology

[0002] High altitude sickness, also known as altitude sickness or mountain sickness, is a common and unique illness in high-altitude regions. It is caused by the rapid ascent to altitudes above 3,000 meters, where the body is exposed to low-pressure, low-oxygen environments. Based on the speed of onset, it is classified into acute and chronic high altitude sickness. Modern medicine believes that high altitude sickness is caused by lower atmospheric pressure and oxygen partial pressure at high altitudes, leading to varying degrees of adaptive changes in the body's respiratory, circulatory, digestive, blood, urinary, endocrine, and central nervous systems. For well-adapted individuals, a series of compensatory mechanisms are quickly established upon entering high altitude, allowing the various systems to reach a new dynamic balance. For those who are poorly adapted, hypoxia can reduce arterial blood oxygen partial pressure and the alveolar-arterial blood oxygen partial pressure difference, resulting in pulmonary hypertension, right ventricular enlargement, myocardial degeneration and necrosis, erythrocyte proliferation, and activation of the sympathetic nervous system, leading to an increased heart rate. Low air pressure can cause cell swelling and increased vascular permeability, leading to cerebral edema, pulmonary edema, gastric mucosal edema, and elevated blood pressure. Acute altitude sickness can present with different symptoms. Mild cases may only experience rapid breathing, rapid heartbeat, and dizziness, without affecting activity and requiring no treatment. Moderate cases may experience headache, nausea, vomiting, palpitations, shortness of breath, chest tightness, blurred vision, insomnia, drowsiness, loss of appetite, abdominal distension, diarrhea, constipation, cyanosis of the lips, and numbness in the hands and feet, making it impossible for them to work and requiring rest and symptomatic treatment. Severe cases may experience difficulty breathing, hemoptysis, rapid heartbeat, elevated blood pressure, and even convulsions, coma, and pulmonary edema, making them unable to care for themselves and requiring oxygen therapy or emergency treatment.

[0003] Currently, treatment for altitude sickness mainly involves oxygen therapy and symptomatic relief. Commonly used medications include aspirin and ibuprofen for headaches, metoclopramide for nausea and vomiting, and propranolol for rapid heartbeat. However, it is well known that Western medicines have significant side effects, highlighting the urgent need to find a drug with fewer side effects that can quickly alleviate altitude sickness.

[0004] Qi-regulating and blood-activating pills are a pure traditional Chinese medicine preparation composed of volatile oil from Litsea cubeba, Allium macrostemon, Ligusticum chuanxiong, and borneol. They possess aromatic and warming properties, regulate qi, and relieve pain. Clinically, they are used to prevent and treat chest pain and angina pectoris (coronary heart disease) caused by insufficient heart yang and blood stasis, with symptoms such as chest tightness, chest pain, palpitations, shortness of breath, and cold limbs. Extensive clinical use has shown that qi-regulating and blood-activating preparations are effective in treating angina pectoris. However, there are no reports on their effects in treating altitude sickness. Therefore, research on the prevention or treatment of altitude sickness and related diseases using qi-regulating and blood-activating preparations is of great significance. Summary of the Invention:

[0005] The purpose of this invention is to provide the application of qi-regulating and blood-activating preparations in the preparation of drugs for the prevention or treatment of altitude sickness.

[0006] The qi-regulating and blood-activating preparation of the present invention is made from the following raw materials in the following weight proportions: 450 parts of Litsea cubeba, 9 parts of borneol, 360 parts of Ligusticum chuanxiong, and 36 parts of Allium macrostemon.

[0007] The qi-regulating and blood-activating preparation of this invention is prepared according to existing patent or non-patent literature technology and pharmaceutical standards. Its preparation method is prior art and can be obtained from the literature disclosed in existing patents, or prepared according to the technology and pharmaceutical standards also disclosed therein. For example, it can be prepared according to the methods disclosed in YBZ00782012, CN1515303A, and CN114073740A.

[0008] The qi-regulating and blood-activating preparation is prepared into the required dosage form by adding one or more pharmaceutically acceptable diluents, excipients, solubilizers, sustained-release agents, absorption promoters, disintegrants, wetting agents, and binders as needed, in accordance with conventional methods in the pharmaceutical field.

[0009] The dosage form of the qi-regulating and blood-activating preparation is drop pills, oral liquid, capsules, granules, or tablets; preferably drop pills.

[0010] The dosage and administration of the Qi-regulating and blood-activating preparations described in this invention are determined according to the patient's condition. For example, the commercially available Qi-regulating and blood-activating dripping pills have the following specifications: each pill weighs 25mg, and the dosage and administration are: 10 pills orally three times a day.

[0011] The altitude sickness described in this invention is an acute altitude sickness caused by hypoxia at high altitudes, with specific symptoms including headache, dizziness, nausea, vomiting, palpitations, shortness of breath, chest tightness, cyanosis of the lips, numbness in the hands and feet, etc.

[0012] Studies have shown that the blood-activating preparations described in this invention can improve exercise capacity and cardiac function in high-altitude hypoxia; improve inflammatory infiltration of lung tissue in high-altitude hypoxia, reduce alveolar wall thickness, and reduce pulmonary fibrosis; and improve myocardial cell viability and reduce LDH release from myocardial cells in high-altitude hypoxia, thereby achieving the purpose of effectively preventing and treating acute altitude sickness.

[0013] Acute altitude sickness is a manifestation of hypoxia in the body. When moving from a plain to a high-altitude area in a short period of time, the rapid increase in altitude causes a series of hypoxia symptoms, often resulting in headaches, insomnia, and loss of appetite. Fatigue and difficulty breathing are also prominent symptoms. In severe cases, it can cause hypoxia damage to the body's organ systems or directly endanger life.

[0014] Traditional Chinese medicine believes that altitude sickness is caused by qi deficiency, qi stagnation, or blood deficiency, and the main treatment principle of traditional Chinese medicine is to invigorate qi and promote blood circulation.

[0015] Litsea cubeba is the dried fruit of a plant in the genus Cinnamomum of the Lauraceae family. It is pungent and warm in nature, possessing properties of dispelling cold and dampness, promoting qi circulation, and relieving pain. Modern pharmacological studies have demonstrated its ability to relax smooth muscle, increase coronary blood flow, reduce myocardial oxygen consumption, improve hemodynamics, resist experimental arrhythmias, and exhibit antibacterial, antiviral, anti-inflammatory, anti-swelling, and analgesic effects. It also provides good protection against acute experimental myocardial ischemia and infarction in cats.

[0016] Ligusticum chuanxiong is the rhizome of Ligusticum chuanxiong, a plant belonging to the genus Ligustrum of the family Apiaceae. It is warm in nature, pungent and slightly bitter in taste, and is a commonly used traditional Chinese medicine for promoting blood circulation and removing blood stasis. It contains volatile oils, alkaloids, phenolic substances, organic acids, phthalyl lactones, and other components; it has the effects of promoting blood circulation, regulating qi, dispelling wind, and relieving pain. Clinical pharmacological studies have shown that Ligusticum chuanxiong has the effects of relaxing smooth muscle, increasing coronary blood flow, reducing myocardial oxygen consumption, improving myocardial metabolism, dilating peripheral blood vessels, inhibiting platelet aggregation, preventing thrombosis, increasing blood flow, improving microcirculation, and providing sedation and analgesia.

[0017] Allium macrostemon B. or Allium chinensis G. Don, belonging to the genus Allium of the Liliaceae family, is a dried bulbous plant. It is pungent and bitter in nature, and has the effects of warming the middle jiao and promoting yang, regulating qi and relieving chest tightness, and dispersing stagnation. Clinically, Allium macrostemon is often used for chest pain, heart pain radiating to the back, etc. Current clinical research shows that Allium macrostemon has the effects of inhibiting platelet aggregation, lowering lipids and preventing atherosclerosis, antibacterial and antioxidant properties, anticancer effects, antispasmodic and antiasthmatic effects, analgesic effects, and hypoxia tolerance effects.

[0018] Borneol is a processed product of the resin of the Dipterocarpus borneolaceae plant, or a crystal extracted from the leaves of the Asteraceae plant, or a refined product synthesized chemically using turpentine oil, camphor, and other raw materials. The main component of borneol-based borneol is dextrorotatory borneol, while that of borneol-based borneol is mainly levorotatory borneol. Borneol is pungent, bitter, and slightly cold in nature. It enters the heart, spleen, and lung meridians, and has the functions of opening the orifices, refreshing the mind, clearing heat, and relieving pain. Modern pharmacological studies have proven that borneol has both anti-myocardial ischemia effects and can significantly increase coronary blood flow; in addition, borneol can increase the permeability of the blood-brain barrier and enhance the ability of drugs to cross the barrier.

[0019] The preparation made from Litsea cubeba, Allium macrostemon, Ligusticum chuanxiong, and borneol has the effects of warming the yang, relieving chest congestion, regulating qi and promoting blood circulation. It is used for angina pectoris in coronary heart disease and also conforms to the treatment principles of traditional Chinese medicine for altitude sickness.

[0020] To explore the application of Qi-regulating and blood-activating preparations in the prevention and treatment of altitude sickness, this invention selected mice to establish a mouse model of high-altitude hypoxia by housing them in an animal hypoxic chamber with 7.8% oxygen (simulating an altitude of 7000m). After successful model establishment, mice were treated with Qi-regulating and blood-activating pills at doses of 120mg / kg / d and 240mg / kg / d for 5 days. Tanshinone B (40mg / kg / d) was selected as a positive control. After administration, the mice's exercise capacity, cardiac function, blood pressure, serum CK levels, complete blood count, myocardial hypertrophy indicators, collagen deposition, and lung tissue morphology and collagen deposition were measured. The results showed that compared with the normal control group, the high-altitude hypoxic mice exhibited significantly reduced exercise capacity and cardiac function, accompanied by increased serum CK levels, increased red blood cell, white blood cell, and lymphocyte counts, myocardial hypertrophy, increased inflammatory infiltration in lung tissue, increased alveolar wall thickness, and increased pulmonary fibrosis, while tail vein blood pressure and myocardial fibrosis showed no significant changes. After intervention with Qi-regulating and blood-activating dripping pills, mice showed significantly enhanced motor function and cardiac function, decreased serum CK levels, reduced red blood cell, white blood cell, and lymphocyte counts, reversed cardiomyocyte hypertrophy, reduced pulmonary inflammatory infiltration, decreased alveolar wall thickness, and reduced pulmonary fibrosis, with the effects being dose-dependent. These experimental results indicate that Qi-regulating and blood-activating preparations have significant efficacy in preventing and treating high-altitude hypoxia (i.e., altitude sickness) and can be used for the prevention and treatment of altitude sickness. Attached Figure Description

[0021] Figure 1 Effects of Liqi Huoxue Dripping Pills on Exercise Capacity and Myocardial Hypertrophy in High-Altitude Hypoxic Mice. A. Treadmill Exhaustion Time on Day 5 after Hypoxia. B. Systolic and Diastolic Blood Pressure in the Tail Vein of Mice (Left) and Right. C. Representative Image of Wheat Germ Agglutinin (WBA) Staining in Mouse Heart Tissue, Scale Bar, 50 μm. D. Cross-sectional Area of ​​Mouse Cardiac Cells. Data are expressed as mean ± standard error, n = 6. *P < 0.05, **P < 0.01 vs. Normal Control Group; +P < 0.05, ++P < 0.01 vs. High-Altitude Hypoxia Group.

[0022] Figure 2 Effects of Liqi Huoxue Dripping Pills on Cardiac Function and Serum CK Levels in High-Altitude Hypoxic Mice. A. Representative echocardiographic images of mouse heart tissue. B. Left ventricular ejection fraction (LVEF). C. Shortened left ventricular ejection fraction (LVFS). D. Heart rate. E. Left ventricular systolic volume (LV Vol; s). F. Left ventricular diastolic volume (LV Vol; d). G. Serum creatine kinase (CK) level. Data are expressed as mean ± standard error, n = 6. *P < 0.05, **P < 0.01, ***P < 0.001 vs. normal control group; +P < 0.05, ++P < 0.01, +++P < 0.001 vs. high-altitude hypoxic group.

[0023] Figure 3 Effects of Liqi Huoxue Dripping Pills on Blood Routine of High-Altitude Hypoxic Mice. A. White blood cells (WBC) in anticoagulated whole blood of mice. B. Lymphocytes (Lymph) in anticoagulated whole blood of mice. C. Monocytes (Mon) in anticoagulated whole blood of mice. D. Granulocytes (Gran) in anticoagulated whole blood of mice. E. Red blood cells (RBC) in anticoagulated whole blood of mice. F. Hematocrit (HCT) in anticoagulated whole blood of mice. G. Hemoglobin (HGB) in anticoagulated whole blood of mice. Data are expressed as mean ± standard error, n = 6. ** P < 0.01 vs. normal control group; + P < 0.05 ++ P < 0.01 vs. high-altitude hypoxia group.

[0024] Figure 4 Effects of Liqi Huoxue Dripping Pills on Inflammatory Infiltration and Collagen Deposition in Lung Tissue of High-Altitude Hypoxic Mice. A. Representative image of HE staining of mouse lung tissue, scale bar, 50 μm. B. Representative images of Sirius red staining of mouse heart tissue (top) and lung tissue (bottom), scale bar, 50 μm. C. Sirius red positive area in mouse heart tissue. D. Sirius red positive area in mouse lung tissue. Data are expressed as mean ± standard error, n = 3. ** P < 0.01 vs. normal control group; + P < 0.05 ++ P < 0.01 vs. high-altitude hypoxia group.

[0025] Figure 5 Effects of Qi-regulating and Blood-activating Dripping Pills on the Viability and LDH Release of H9c2 Cells under Hypoxia. A. H9c2 Cell Viability under Normoxic Conditions. B. H9c2 Cell Viability under Hypoxia Conditions. C. LDH Release from H9c2 Cells under Hypoxia Conditions. Data are expressed as mean ± standard error, n = 6. *** P < 0.001 vs. normal control group; + P < 0.05 ++ P < 0.01, +++ P < 0.001 vs. hypoxia group.

[0026] Figure 6 Effects of Qi-regulating and Blood-activating Dripping Pills on the positive rates of Annexin V and PI in hypoxic H9c2 cells. A. Representative flow cytometry graph. B. Annexin V positive rate in H9c2 cells. C. Double positive rate of Annexin V and PI in H9c2 cells. Data are expressed as mean ± standard error, n = 3. ** P < 0.01 vs. normal control group; + P < 0.05 ++ P < 0.01, +++ P < 0.001 vs. hypoxia group.

[0027] Figure 7 Effects of Liqi Huoxue Dripping Pills on necrosis-like apoptosis-related proteins in hypoxic H9c2 cells. A. Representative Western blot bands. BG. Western blot gray values ​​of RIPK1, RIPK3, and MLKL compared to β-actin, and the Western blot gray values ​​of p-RIPK1 and p-RIPK3 compared to p-MLKL and RIPK1, RIPK3, and MLKL. Data are expressed as mean ± standard error, n = 6. * P < 0.05 ** P < 0.01 vs. normal control group; + P < 0.05 ++ P < 0.01 vs. hypoxia group.

[0028] Figure 8 Effects of Liqi Huoxue Dripping Pills on the Viability and LDH Release of Hypoxic HCMEC Cells. A. HCMEC cell viability under normoxic conditions. B. HCMEC cell viability under hypoxic conditions. C. LDH release from HCMEC cells under hypoxic conditions. Data are expressed as mean ± standard error, n = 6. *** P < 0.001 vs. normal control group; + P < 0.05 ++ P < 0.01 vs. hypoxia group.

[0029] Figure 9 Effects of Qi-regulating and Blood-activating Dripping Pills on the positivity rates of Annexin V and PI in hypoxic HCMEC cells. A. Representative flow cytometry image. B. Annexin V positivity rate in HCMEC cells. C. Double positivity rate of Annexin V and PI in HCMEC cells. Data are expressed as mean ± standard error, n = 3. ** P < 0.01 vs. normal control group; + P < 0.05 ++ P < 0.01 vs. hypoxia group.

[0030] Figure 10 Effects of Liqi Huoxue Dripping Pills on necrosis-like apoptosis-related proteins in hypoxic HCMEC cells. A. Representative Western blot bands. B. Western blot gray values ​​of RIPK1, RIPK3, and MLKL compared to β-actin, and the Western blot gray values ​​of p-RIPK1 and p-RIPK3 compared to p-MLKL and RIPK1, RIPK3, and MLKL. Data are expressed as mean ± standard error, n = 6. * P < 0.05 ** P < 0.01 vs. normal control group; + P < 0.05 ++ P < 0.01 vs. hypoxia group. Detailed implementation method:

[0031] To better understand this invention, the following experimental examples illustrate the application of qi-regulating and blood-activating preparations in the preparation of drugs for preventing and treating altitude sickness. The following experiments are intended to illustrate the invention and not to limit it.

[0032] Experiment Example 1: Evaluation of the protective effect of Liqi Huoxue Dripping Pills against high-altitude hypoxia injury in mice

[0033] 1. Experimental Design

[0034] Mice in the normal control group were raised under normoxic conditions, while mice in the hypoxic group were raised in a small animal hypoxic chamber with 7.8% oxygen (simulating an altitude of 7000m). After 5 days of hypoxic feeding, the ejection fraction (EF) and ejection fraction shortening rate (FS) of the mice were measured using a small animal ultrasound instrument to confirm the successful establishment of the high-altitude hypoxia model.

[0035] Subsequently, appropriate drug treatments were administered. The low-dose group of Liqi Huoxue Diwan (120 mg / kg / d) was administered via gavage, and the high-dose group was administered via gavage (240 mg / kg / d). The positive control group received intraperitoneal injection of Tanshinone B (40 mg / kg / d). After 5 consecutive days of administration, the mice's exercise capacity was assessed using a treadmill. Small animal ultrasound was used to measure ejection fraction (EF), ejection fraction shortening rate (FS), systolic left ventricular volume (LV Vol;s), diastolic left ventricular volume (LV Vol;d), and heart rate. Blood was collected from the orbital fossa to detect complete blood count and serum CK levels. Heart tissue was stained with wheat germ agglutinin (WGA) to detect cardiac cell hypertrophy. Lung tissue was stained with hematoxylin and eosin (HE) to observe morphological changes. Heart and lung tissue were stained with Sirius red to observe fibrosis.

[0036] 2. Experimental Results

[0037] 2.1 Qi-regulating and blood-activating dripping pills improve motor function and myocardial hypertrophy in high-altitude hypoxic mice.

[0038] After 5 days of hypoxic feeding, the mice's locomotor ability ( Figure 1 A) significantly decreased, left ventricular wall myocardial cell area ( Figure 1 C and D) increased significantly, while the systolic and diastolic blood pressure of the tail vein did not change significantly. Figure 1 B). The Qi-regulating and Blood-activating Droplets improved the motor function and cardiomyocyte area of ​​mice in a dose-dependent manner, but had no significant effect on their tail vein systolic and diastolic blood pressure.

[0039] 2.2 Qi-regulating and blood-activating dripping pills improve cardiac function in high-altitude hypoxic mice

[0040] Compared with the normal group, after 5 days of hypoxia, the left ventricular ejection fraction (LVEF) of mice was significantly reduced. Figure 2 B) and left ventricular fractional shortening (LVFS, Figure 2 C) were significantly decreased, and heart rate ( Figure 2 D), left ventricular systolic volume ( Figure 2 E), and diastolic volume ( Figure 2 F) were increased, and serum CK level ( Figure 2 G) was elevated. The Liqi Huoxue Dropping Pills improved the cardiac function of mice and reduced the serum CK level in a dose-dependent manner.

[0041] 2.3 The Liqi Huoxue Dropping Pills decreased the red blood cell and white blood cell counts in mice with plateau hypoxia

[0042] The results of blood routine examination showed that the white blood cell count ( Figure 3 A), lymphocyte count ( Figure 3 B), monocyte count ( Figure 3 C), neutrophil count ( Figure 3 D), red blood cell count ( Figure 3 E), and hematocrit ( Figure 3 F) in mice with plateau hypoxia were significantly increased, while the hemoglobin content ( Figure 3 G) showed no obvious change. The Liqi Huoxue Dropping Pills decreased the white blood cell count, lymphocyte count, monocyte count, neutrophil count, red blood cell count, and hematocrit in mice in a dose-dependent manner, while having no obvious effect on the hemoglobin content.

[0043] 2.4 The Liqi Huoxue Dropping Pills improved the inflammatory infiltration and collagen deposition in the lung tissue of mice with plateau hypoxia

[0044] The results of HE staining showed that compared with the normal control group, the inflammatory infiltration in the lung tissue of mice in the plateau hypoxia group was significantly increased, and the alveolar wall thickness was significantly thickened ( Figure 4 A). The results of Sirius red staining showed that compared with the normal control group, there was no obvious change in the cardiac collagen deposition in mice in the plateau hypoxia group ( Figure 4 B and C), while the collagen deposition in the lung tissue was significantly increased ( Figure 4 B and D). The Liqi Huoxue Dropping Pills improved the inflammatory infiltration and alveolar wall thickness in the lung tissue of mice and reduced the pulmonary fibrosis in a dose-dependent manner.

[0045] 3. Conclusion

[0046] 1) The Liqi Huoxue Dropping Pills have the effects of improving the exercise and cardiac function of mice with plateau hypoxia, reversing myocardial hypertrophy, and reducing their serum CK level, red blood cell count, and white blood cell count;

[0047] 2) The Liqi Huoxue Dropping Pills inhibit the inflammatory infiltration and collagen deposition in the lung tissue of mice with plateau hypoxia.

[0048] Experimental Example 2: Evaluation of the protective effect of the Liqi Huoxue Dropping Pills on hypoxic cardiomyocytes

[0049] 1. Experimental protocol

[0050] H9c2 cells in the normal control group were cultured in complete medium under normoxic conditions (95% air, 5% CO2) for 24 h, and H9c2 cells in the hypoxia group were cultured in sugar-free medium under hypoxic conditions (95% N2, 5% CO2) for 24 h. Then, corresponding drug treatments were carried out. Different doses of Rationalizing Qi and Activating Blood Circulation Dropping Pills (10 μg / mL, 20 μg / mL, and 50 μg / mL) and salvianolic acid B (50 μM) were given simultaneously during hypoxia. The drug administration time was 24 h. After drug treatment, the viability of H9c2 cells was detected using the MTS kit; the LDH release of H9c2 cells was detected using the LDH kit; the positive rates of Annexin V and PI in H9c2 cells were detected using flow cytometry; and the expression of proteins related to necroptosis in H9c2 cells was detected using Western blot.

[0051] 2. Experimental results

[0052] 2.1 Rationalizing Qi and Activating Blood Circulation Dropping Pills improve the viability of hypoxic H9c2 cells and reduce the LDH release of hypoxic H9c2 cells

[0053] Under normoxic conditions, Rationalizing Qi and Activating Blood Circulation Dropping Pills at a concentration of 1000 μg / mL and salvianolic acid B at a concentration of 1000 μM had no obvious effect on the viability of H9c2 cells ( Figure 5 A). Under hypoxic conditions, the viability of H9c2 cells was significantly decreased ( Figure 5 B), and the LDH release increased ( Figure 5 C). Rationalizing Qi and Activating Blood Circulation Dropping Pills improved the viability of H9c2 cells in a dose-dependent manner and reduced its LDH release, with the best effect at 50 μg / mL.

[0054] 2.2 Rationalizing Qi and Activating Blood Circulation Dropping Pills reduce the positive rates of Annexin V and PI in hypoxic H9c2 cells

[0055] Flow cytometry results showed that compared with the normal control group, the positive rate of Annexin V in H9c2 cells in the hypoxia group ( Figure 6 A and B) and the double positive rate of Annexin V and PI ( Figure 6 A and C) were significantly increased. Rationalizing Qi and Activating Blood Circulation Dropping Pills reduced the positive rate of Annexin V and the double positive rate of Annexin V and PI in H9c2 cells in a dose-dependent manner.

[0056] 2.3 Rationalizing Qi and Activating Blood Circulation Dropping Pills inhibit the levels of proteins related to necroptosis in hypoxic H9c2 cells

[0057] Western blot results showed that, compared with the normal control group, the levels of necroptosis-related proteins (RIPK1 / p-RIPK1, p-RIPK3, MLKL / p-MLKL) in H9c2 cells of the hypoxia group were significantly increased ( Figure 7 ). Qili Huoxue Dropping Pills decreased the expression of necroptosis-related proteins (p-RIPK1, p-RIPK3, and p-MLKL) in H9c2 cells in a dose-dependent manner.

[0058] 3. Conclusion

[0059] 1) Qili Huoxue Dropping Pills have the effect of improving hypoxia-induced injury in H9c2 cells;

[0060] 2) Qili Huoxue Dropping Pills inhibit the expression of necroptosis-related proteins in hypoxic H9c2 cells.

[0061] Experimental Example 3: Evaluation of the protective effect of Qili Huoxue Dropping Pills on hypoxic vascular endothelial cells

[0062] 1. Experimental protocol <​​​​​​​​​​​​​​​​​

[0067] 2.2 Qi-regulating and blood-activating dripping pills reduce the positive rates of Annexin V and PI in hypoxic HCMEC cells.

[0068] Flow cytometry results showed that, compared with the normal control group, the Annexin V positivity rate of HCMEC cells in the hypoxia group was significantly higher. Figure 9 A and B) and the double positivity rate of Annexin V and PI ( Figure 9 A and C) significantly increased. The Qi-regulating and Blood-activating Droplets reduced the Annexin V positivity rate and the double positivity rate of Annexin V and PI in HCMEC cells in a dose-dependent manner.

[0069] 2.3 Qi-regulating and blood-activating dripping pills inhibit the levels of necrosis-like apoptosis-related proteins in hypoxic HCMEC cells.

[0070] Western blot results showed that, compared with the normal control group, the levels of necrosis-like apoptosis-related proteins (RIPK1 / p-RIPK1, p-RIPK3, p-MLKL) in HCMEC cells were significantly increased in the hypoxia group. Figure 10 The Qi-regulating and Blood-activating Droplets reduce the expression of necrosis-like apoptosis-related proteins (p-RIPK1, p-RIPK3, and p-MLKL) in HCMEC cells in a dose-dependent manner.

[0071] 3. Conclusion

[0072] 1) Qi-regulating and blood-activating dripping pills have the effect of improving hypoxia-induced damage in HCMEC cells;

[0073] 2) Liqi Huoxue Diwan inhibits the expression of necrosis-like apoptosis-related proteins in hypoxic HCMEC cells.

[0074] It should be noted that the above embodiments and test examples are only for further elaboration and understanding of the technical solutions of the present invention, and should not be construed as further limiting the technical solutions of the present invention. Inventions that do not highlight substantive features or make significant progress by those skilled in the art still fall within the protection scope of the present invention.

Claims

1. The use of a Liqi Huoxue preparation in the preparation of a medicine for preventing or treating high altitude reaction, characterized in that, The qi-regulating and blood-activating preparation is made from the following raw materials in the following weight proportions: 450 parts of Litsea cubeba, 9 parts of borneol, 360 parts of Ligusticum chuanxiong, and 36 parts of Allium macrostemon.

2. Use according to claim 1, characterized in that, The qi-regulating and blood-activating preparation is prepared in accordance with existing patent or non-patent literature technology and pharmaceutical standards.

3. The application according to claim 1, characterized in that, The qi-regulating and blood-activating preparation is prepared into the required dosage form by adding one or more pharmaceutically acceptable diluents, excipients, solubilizers, sustained-release agents, absorption promoters, disintegrants, wetting agents, and binders as needed, in accordance with conventional methods in the pharmaceutical field.

4. The application according to claim 3, characterized in that, The dosage forms of the qi-regulating and blood-activating preparations are pills, oral liquids, capsules, granules, and tablets.

5. The application according to claim 4, characterized in that, The dosage form of the qi-regulating and blood-activating preparation is drop pills.

6. The application according to any one of claims 1-5, characterized in that, The altitude sickness mentioned refers to acute altitude sickness caused by hypoxia at high altitudes.

7. The application according to any one of claims 1-5, characterized in that, The symptoms of altitude sickness are: headache, dizziness, nausea, vomiting, palpitations, shortness of breath, chest tightness, cyanosis of the lips, numbness in the hands and feet, or a combination thereof.

8. The application according to any one of claims 1-5, characterized in that, The aforementioned blood-activating preparation is used to improve the ability to exercise under hypoxia at high altitudes and cardiac function.

9. The application according to any one of claims 1-5, characterized in that, The qi-regulating and blood-activating preparation is used to improve inflammatory infiltration of lung tissue in high-altitude hypoxia, reduce alveolar wall thickness, and reduce pulmonary fibrosis.

10. The application according to any one of claims 1-5, characterized in that, The aforementioned blood-activating preparation is used to improve the vitality of myocardial cells in high-altitude hypoxia and reduce the release of LDH from myocardial cells.

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