Bu Yang Huan Wu decoction in the preparation of treatment of high altitude pulmonary injury of qi deficiency and blood stasis syndrome products in the application

Abstract

The application provides an application of Buyang Huanwu Decoction in the preparation of a product for treating high altitude pulmonary injury of qi deficiency and blood stasis syndrome, and belongs to the technical field of medicines. After the prescription of the Buyang Huanwu Decoction is decocted with water, the decoction is used for treating high altitude pulmonary injury, can reduce pathological injury of lung tissues, reduce thickening of alveolar walls, make lung tissue structure clearer, and close to normal lung tissue morphology; meanwhile, the decoction can enhance exercise capacity, reduce asthenia symptoms of high altitude pulmonary injury of qi deficiency and blood stasis syndrome, and effectively treat high altitude pulmonary injury of qi deficiency and blood stasis syndrome.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to the application of Buyang Huanwu Decoction in the preparation of products for treating high-altitude lung injury with qi deficiency and blood stasis syndrome. Background Technology

[0002] High-altitude lung injury (HAPI) refers to a series of pathological changes in the lungs caused by decreased air pressure and insufficient oxygen content in a hypoxic environment. HAPI is a type of acute high-altitude lung injury, particularly affecting tourists and workers newly arrived at high altitudes. Typical manifestations include high-altitude pulmonary edema (HAPE), a non-cardiac pulmonary edema caused by hypoxia, which may appear within 24 to 96 hours after rapid ascent to altitudes above 2500 meters. Common symptoms include shortness of breath, cough, pink or white frothy sputum, and cyanosis; in severe cases, it can be fatal. In addition, a series of symptoms of qi deficiency and blood stasis may appear, classifying it as qi deficiency and blood stasis type HAPI. Traditional Chinese medicine believes that altitude sickness is mainly caused by a lack of clear qi in the external environment and insufficient ancestral qi in the body, leading to qi deficiency and blood stasis damage in the lungs and other organs. Therefore, altitude sickness is mainly characterized by qi deficiency and blood stasis syndrome, making qi deficiency and blood stasis type HAPI relatively common. Currently, there is insufficient clinical research on drugs for treating qi deficiency and blood stasis type HAPI, making research on effective drugs for the prevention and treatment of altitude sickness essential. Western medicine is effective in preventing and treating high-altitude lung injury caused by qi deficiency and blood stasis, but it has the drawback of relatively large side effects. Traditional Chinese medicine, with its multi-target, lower cost, multiple components, and fewer side effects, holds promise for providing new treatment strategies for high-altitude lung injury with complex pathogenesis.

[0003] Buyang Huanwu Decoction is a traditional Chinese medicine formula composed of Astragalus membranaceus 120g, Angelica sinensis (tail) 6g, Paeonia lactiflora 5g, and Pheretima aspergillum, Ligusticum chuanxiong, Carthamus tinctorius, and Prunus persica 3g each. It is a representative formula for invigorating qi and promoting blood circulation, and a commonly used formula for treating sequelae of stroke. Clinical application focuses on hemiplegia, facial paralysis, pale tongue, white tongue coating, and weak, slow pulse as key diagnostic points. Studies have shown that Buyang Huanwu Decoction has significant neuroprotective effects, but there are currently no reports on its effects on high-altitude lung injury. Summary of the Invention

[0004] In view of this, the purpose of this invention is to provide the application of Buyang Huanwu Decoction in the preparation of products for treating high-altitude lung injury, to reduce pathological damage to lung tissue and bring it closer to the normal state of lung tissue, while relieving the fatigue state of high-altitude lung injury with qi deficiency and blood stasis syndrome.

[0005] This invention provides the application of Buyang Huanwu Decoction in the preparation of products for treating high-altitude lung injury.

[0006] Preferably, the high-altitude lung injury includes high-altitude lung injury due to qi deficiency and blood stasis.

[0007] Preferably, the Buyang Huanwu Decoction can alleviate pathological damage to lung tissue.

[0008] Preferably, the Buyang Huanwu Decoction can alleviate the fatigue symptoms of high-altitude lung injury caused by qi deficiency and blood stasis.

[0009] Preferably, the Buyang Huanwu Decoction treats high-altitude lung injury by reducing the expression of HIF-1α.

[0010] Preferably, the preparation method of Buyang Huanwu Decoction includes the following steps: taking all the medicinal materials of Buyang Huanwu Decoction and mixing them with water, and soaking them; extracting, wherein the extraction is first boiling over high heat for 18-22 minutes, and then simmering over low heat for 0.8-1.2 hours; collecting the extracted medicinal liquid, filtering, and collecting the filtrate to obtain Buyang Huanwu Decoction.

[0011] Preferably, the mass-to-volume ratio of all the medicinal materials in Buyang Huanwu Decoction to water is (1g:8mL) to (1g:12mL).

[0012] Preferably, the soaking time is 10 to 14 hours.

[0013] Preferably, the product includes a drug.

[0014] Preferably, the drug comprises a pharmaceutically acceptable carrier.

[0015] The beneficial effects of this invention are:

[0016] This invention uses the Buyang Huanwu Decoction formula, after being decocted in water, to treat high-altitude lung injury. It can reduce pathological damage to lung tissue, decrease alveolar wall thickening, and make the lung tissue structure clearer, closer to the normal lung tissue morphology. At the same time, it can enhance exercise capacity, alleviate fatigue symptoms of high-altitude lung injury with qi deficiency and blood stasis syndrome, and effectively treat high-altitude lung injury with qi deficiency and blood stasis syndrome. Attached Figure Description

[0017] Figure 1 The distance traveled by mice in each group in the central region (central area) during the OFT experiment; where ** indicates a significant difference compared to the normal group (P < 0.01), # indicates a significant difference compared to the model group (P < 0.05), and ## indicates an extremely significant difference compared to the model group (P < 0.01).

[0018] Figure 2 The number of times each group of mice entered the central region (central area) in the OFT experiment; where ** indicates a significant difference compared with the normal group (P < 0.01), # indicates a significant difference compared with the model group (P < 0.05), and ## indicates a highly significant difference compared with the model group (P < 0.01).

[0019] Figure 3 Images showing HE staining of lung tissue from mice in each group.

[0020] Figure 4 The graph shows the HIF-1α content in the lung tissue of mice in each group. In the graph, ** indicates a significant difference compared with the normal group (P < 0.01), # indicates a significant difference compared with the model group (P < 0.05), and ## indicates a highly significant difference compared with the model group (P < 0.01). Detailed Implementation

[0021] This invention provides the application of Buyang Huanwu Decoction in the preparation of products for treating high-altitude lung injury.

[0022] The Buyang Huanwu Decoction used in this invention is a formula composed of 120 parts Astragalus membranaceus, 6 parts Angelica sinensis (tail), 5 parts Paeonia lactiflora, 3 parts Pheretima aspergillum, 3 parts Ligusticum chuanxiong, 3 parts Carthamus tinctorius, and 3 parts Prunus persica. This invention does not have any special limitations on the sources of Astragalus membranaceus, Angelica sinensis (tail), Paeonia lactiflora, Pheretima aspergillum, Ligusticum chuanxiong, Carthamus tinctorius, and Prunus persica; conventional commercially available products in this field are acceptable.

[0023] The Buyang Huanwu Decoction of the present invention is used to treat high-altitude lung injury. Preferably, it can reduce pathological damage to lung tissue, decrease alveolar wall thickening, and make the lung tissue structure clearer, approaching the morphology of normal lung tissue. Simultaneously, it preferably also enhances exercise capacity, alleviates fatigue symptoms in high-altitude lung injury due to qi deficiency and blood stasis, and effectively treats high-altitude lung injury due to qi deficiency and blood stasis. Preferably, the Buyang Huanwu Decoction treats high-altitude lung injury by reducing HIF-1α expression.

[0024] The preferred method for preparing the Buyang Huanwu Decoction includes the following steps: mixing all the medicinal materials of Buyang Huanwu Decoction with water, wherein the preferred mass-to-volume ratio of the mixed medicinal materials to water is (1g:8mL) to (1g:12mL), more preferably (1g:9mL) to (1g:11mL), and further preferably 1g:10mL; soaking the mixture, wherein the soaking time is preferably 10-14 hours, more preferably 11-13 hours, and further preferably 12 hours; and extraction after soaking, wherein the extraction is preferably performed by first boiling over high heat for 18-20 minutes. The decoction is prepared by boiling for 2 minutes, followed by simmering over low heat for 0.8–1.2 hours. More preferably, it is boiled over high heat for 19–21 minutes, followed by simmering over low heat for 0.9–1.1 hours. Even more preferably, it is boiled over high heat for 20 minutes, followed by simmering over low heat for 1 hour. The extraction is preferably performed 2–3 times; in one embodiment, it can be performed 2 or 3 times. After extraction, the decoctions obtained from each extraction are collected and combined, filtered, and the filtrate is collected to obtain Buyang Huanwu Decoction. This invention does not specifically limit the filtration method; conventional filtration methods in the art are acceptable. In one embodiment, gauze filtration can be used. In one embodiment, to facilitate preservation, after obtaining Buyang Huanwu Decoction, the solvent can be removed and the powder can be dried. This invention does not specifically limit the solvent removal and drying methods. In one embodiment, rotary evaporation can be used for solvent removal, and freeze drying can be used for drying.

[0025] The products described in this invention preferably include pharmaceuticals. The pharmaceuticals preferably include pharmaceutically acceptable carriers, including but not limited to diluents, binders, lubricants, disintegrants, and preservatives.

[0026] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0027] Unless otherwise specified, the following embodiments are all conventional methods.

[0028] Unless otherwise specified, all materials and reagents used in the following examples are commercially available. The Buyang Huanwu Decoction in the following examples consists entirely of Astragalus membranaceus 120g, Angelica sinensis (tail) 6g, Paeonia lactiflora 5g, Pheretima aspergillum 3g, Ligusticum chuanxiong 3g, Carthamus tinctorius 3g, and Prunus persica 3g.

[0029] Example 1

[0030] All the medicinal materials of Buyang Huanwu Decoction were soaked in 10 times (1g:10mL) of water for 12 hours, boiled over high heat for 20 minutes, and simmered over low heat for 1 hour. The decoction was collected. Eight times (1g:8mL) of water was added to the dregs, boiled over high heat for 20 minutes, and simmered over low heat for 1 hour. The decoction was collected again. Eight times (1g:8mL) of water was added to the dregs again, boiled over high heat for 20 minutes, and simmered over low heat for 1 hour. The decoctions from the three decoctions were combined, filtered through three layers of gauze, and the filtrate was collected and rotary evaporated. The resulting freeze-dried powder was stored at -20℃ for later use. The final yield of the freeze-dried powder was 42%.

[0031] Example 2

[0032] All the medicinal materials of Buyang Huanwu Decoction were soaked in 8 times the amount of water (1g:8mL) for 10 hours, boiled over high heat for 18 minutes, and simmered over low heat for 1.2 hours. The decoction was collected. The dregs were then soaked in 12 times the amount of water (1g:12mL) of the total amount of Buyang Huanwu Decoction, boiled over high heat for 18 minutes, and simmered over low heat for 1.2 hours. The decoctions from both decoctions were combined, filtered through three layers of gauze, and the filtrate was collected and rotary evaporated. The resulting freeze-dried powder was stored at -20℃ for later use. The yield was 39%.

[0033] Example 3

[0034] Add all the medicinal materials of Buyang Huanwu Decoction to 10 times (1 g∶10 mL) the amount of water and soak for 14 h. Boil it vigorously for 22 min, then simmer for 0.8 h, and collect the medicinal liquid; add 10 times (1 g∶10 mL) the amount of water of all the medicinal materials of Buyang Huanwu Decoction to the medicinal residues, boil it vigorously for 22 min, then simmer for 0.8 h, and collect the medicinal liquid; continue to add 10 times (1 g∶10 mL) the amount of water of all the medicinal materials of Buyang Huanwu Decoction to the medicinal residues, boil it vigorously for 22 min, then simmer for 0.8 h, and collect the medicinal liquid; combine the medicinal liquids obtained from the three decoctions, filter with three layers of gauze, collect the filtrate for rotary evaporation, and obtain the freeze-dried powder after freeze-drying, and store it at -20 °C for later use. The yield is 45%.

[0035] Experimental Example 1

[0036] Use a low-pressure oxygen chamber to simulate the environment at an altitude of 6000 m on the plateau and conduct low-pressure and hypoxic treatment on mice for 72 h to construct a mouse model of plateau lung injury with qi deficiency and blood stasis syndrome, and intervene with different doses of Buyang Huanwu Decoction (freeze-dried powder) prepared in Example 1. Evaluate the effect of Buyang Huanwu Decoction on the behavior of mice with plateau lung injury with qi deficiency and blood stasis syndrome through the open field test; detect the pathological changes of the lung tissue of mice with plateau lung injury with qi deficiency and blood stasis syndrome by hematoxylin-eosin staining (HE) to clarify the effect of Buyang Huanwu Decoction in resisting plateau lung injury with qi deficiency and blood stasis syndrome. Further clarify the mechanism of action of Buyang Huanwu Decoction in resisting plateau lung injury with qi deficiency and blood stasis syndrome by ELISA detection of lung tissue.

[0037] 1. Experimental animals

[0038] A total of 48 SPF-grade C57BL / 6J male mice, with a body weight of 20 ± 2 g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the license number was: SCXK (Beijing) 2021-0006. The mice were raised in a standard environment, with 4 mice in each cage, and were illuminated with a 12 h / 12 h light-dark cycle (8:00 a.m. - 20:00 p.m.), the feeding temperature was controlled at 22 ± 2 °C, the relative humidity was 50 ± 10%, and the mice were given free diet.

[0039] 1.1 Establishment of a mouse model of plateau lung injury with qi deficiency and blood stasis syndrome

[0040] Randomly divide the mice into 6 groups, with 8 mice in each group. They are the normal group, the model group (low-pressure and hypoxic group), the acetazolamide group, the low-dose group of Buyang Huanwu Decoction (low-dose group), the medium-dose group of Buyang Huanwu Decoction (medium-dose group), and the high-dose group of Buyang Huanwu Decoction (high-dose group). Except for the normal group, the mice in the other groups were placed in a low-pressure oxygen chamber to simulate an altitude of 6000 m and continuously decompressed for 72 h to establish the model.

[0041] 1.2 Administration

[0042] Four days before mouse modeling, except for the normal group and the model group, all other mice were given drug treatment. The acetazolamide group was given acetazolamide, while the low-dose, medium-dose, and high-dose Buyang Huanwu Decoction groups were given different doses of Buyang Huanwu Decoction prepared in Example 1. The drugs in each group were prepared with purified water to the corresponding concentration and administered by gavage at a dose of 0.2 mL / mouse, once daily. Prophylactic administration began four days before placement in the oxygen chamber and continued for 7 days. The dosages for each group are shown in Table 1.

[0043] Table 1. Drug administration details for each group of mice.

[0044] Group Dosage (g / kg / d) normal group 0 Model group 0 Acetazolamide group 0.1 High-dose group of Buyang Huanwu Decoction 8 Medium dose group of Buyang Huanwu Decoction 4 Low-dose group of Buyang Huanwu Decoction 2

[0045] 1.3 Experiment

[0046] After the drug administration was completed, the mice in each group underwent the following experiments:

[0047] 1.3.1 Open Field Analysis Experiment (OFT)

[0048] OFT is primarily used to study the motor abilities of mice. This experiment used an open field test chamber, which was divided into four equal-sized chambers (50cm × 50cm × 40cm). The outer walls of each chamber were black. The area near the bottom of the chamber formed the peripheral activity zone, while the remaining area formed the central activity zone. Before testing, mice were placed in the open field test chamber for acclimatization for 15 seconds. At the start of the experiment, the mice were placed in the center of the open field. Simultaneously with the release of the mice, a camera above the test chamber began synchronously tracking and recording for 5 minutes. The evaluation metrics for this open field test were the distance traveled in the central area and the number of times the mice entered the central area.

[0049] Experimental results are as follows Figure 1 and Figure 2 As shown, compared with the normal group, the model group showed a decreasing trend in central zone distance and number of entries (P < 0.01). Compared with the model group, all dosage groups of Buyang Huanwu Decoction showed a significant increase in central zone distance and number of entries, with the medium dosage group showing the most significant increase and a statistically significant difference.

[0050] 1.3.2 HE staining of lung tissue

[0051] After the open field analysis experiment, mice were euthanized, and lung tissue was fixed in 4% paraformaldehyde for at least 24 hours. Subsequently, the tissue was dehydrated and paraffin-embedded in alcohols of different concentrations, then sectioned. The mouse lung tissue sections were dewaxed, cell nuclei were stained with hematoxylin, and cytoplasm was stained with eosin. The sections were then dehydrated again and sealed. Images of these sections were acquired and analyzed using an upright optical microscope (Nikon Eclipseci, Japan) to assess the morphological changes in the mouse lung tissue.

[0052] The results are as follows Figure 3 As shown, HE staining results revealed that the alveolar walls of mice in the model group were significantly thickened, and the alveolar cavities were almost unformed and varied in size. Compared with the model group, the pathological damage to the lung tissue of mice treated with acetazolamide and Buyang Huanwu Decoction was significantly reduced, the lung tissue structure was clearer, the alveolar wall thickening was improved, and the morphology was close to that of normal lung tissue.

[0053] 1.3.3 Lung tissue ELISA detection

[0054] Mouse lung tissue was collected, the weight of the tissue to be tested was accurately weighed, physiological saline was added according to the weight-to-volume ratio, and the tissue was ground on ice to prepare a tissue homogenate. The homogenate was centrifuged at 3000 rpm for 10 min, and then the supernatant of the tissue homogenate was collected. The HIF-1α level of the lung tissue was detected using an ELISA kit (Jiangsu Enzyme Immunosorbent Assay, catalog number: MM-0393M1).

[0055] The results are as follows Figure 4 As shown, the results revealed that compared with the normal group, the expression level of HIF-1α in the lung tissue of the model group mice was significantly increased, and the intervention of Buyang Huanwu Decoction significantly reduced the protein expression of HIF-1α in the lung tissue of stressed mice. Therefore, the above results indicate that Buyang Huanwu Decoction can exert its anti-lung injury effect in high-altitude patients with qi deficiency and blood stasis syndrome by reducing HIF-1α expression and regulating the HIF-1 signaling pathway.

[0056] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of Buyang Huanwu Decoction in the preparation of drugs for treating high-altitude lung injury, wherein Buyang Huanwu Decoction is made from 120 parts of Astragalus membranaceus, 6 parts of Angelica sinensis tail, 5 parts of Paeonia lactiflora, 3 parts of Pheretima aspergillum, 3 parts of Ligusticum chuanxiong, 3 parts of Carthamus tinctorius and 3 parts of Prunus persica.

2. The application according to claim 1, characterized in that, The high-altitude lung injury mentioned includes high-altitude lung injury due to qi deficiency and blood stasis.

3. The application according to claim 1 or 2, characterized in that, The Buyang Huanwu Decoction can reduce pathological damage to lung tissue.

4. The application according to claim 2, characterized in that, The Buyang Huanwu Decoction can alleviate the fatigue symptoms of high-altitude lung injury caused by qi deficiency and blood stasis.

5. The application according to claim 1 or 2, characterized in that, The Buyang Huanwu Decoction treats high-altitude lung injury by reducing the expression of HIF-1α.

6. The application according to claim 1, characterized in that, The preparation method of the Buyang Huanwu Decoction The process includes the following steps: taking all the medicinal materials of Buyang Huanwu Decoction and mixing them with water, then soaking them; extracting the decoction by first boiling it over high heat for 18-22 minutes, then simmering it over low heat for 0.8-1.2 hours; collecting the extracted liquid, filtering it, and collecting the filtrate to obtain Buyang Huanwu Decoction.

7. The application according to claim 6, characterized in that, The mass-to-volume ratio of all the medicinal materials in Buyang Huanwu Decoction to water is (1g:8mL) to (1g:12mL).

8. The application according to claim 6, characterized in that, The soaking time is 10 to 14 hours.

9. The application according to claim 1, characterized in that, The drug includes a pharmaceutically acceptable carrier.

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