CN115317525A - Application of sabina przewalskii in preparing medicament for treating chronic obstructive pulmonary disease - Google Patents
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- CN115317525A CN115317525A CN202211041729.4A CN202211041729A CN115317525A CN 115317525 A CN115317525 A CN 115317525A CN 202211041729 A CN202211041729 A CN 202211041729A CN 115317525 A CN115317525 A CN 115317525A
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- A—HUMAN NECESSITIES
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/13—Coniferophyta (gymnosperms)
- A61K36/14—Cupressaceae (Cypress family), e.g. juniper or cypress
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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Abstract
The invention discloses an application of sabina przewalskii in preparation of a medicine for treating chronic obstructive pulmonary disease, wherein sabina przewalskii is an alcohol extract of sabina przewalskii. The Sabina przewalskii alcohol extract has the effects of inhibiting inflammatory reaction of chronic obstructive pulmonary diseases, improving the lung functions of the Sabina przewalskii alcohol extract and treating the chronic obstructive pulmonary diseases.
Description
Technical Field
The invention belongs to the technical field of Sabina przewalskii, and particularly relates to application of an alcohol extract of Sabina przewalskii, namely application in preparing a medicament for treating chronic obstructive pulmonary disease.
Background
Chronic Obstructive Pulmonary Disease (COPD) is a progressive disease that can be prevented and treated, covering emphysema and Chronic bronchitis, and is characterized by airflow limitation that cannot be completely reversed. Airflow limitation is usually progressive and is associated with an abnormal inflammatory response of the lungs to harmful particles or gases. COPD is mainly caused by smoking and is also associated with environmental pollutant concentrations. Although COPD mainly affects lung tissues, it also has severe systemic effects on the body. Since 1990 COPD had increased in number by 23%, COPD was expected to become the third leading cause of death in the world in 2030. Meanwhile, as smoking and population aging increase, the number of people dying from the disease per year may exceed 540 million by 2060. Repeated stimulation and damage to lung tissues and tissue repair by external factors cause the lung tissue structure and physiology to change obviously to generate chronic inflammation, which mainly affects central and peripheral airways, lung parenchyma, alveoli and pulmonary vessels. Inflammation and structural changes in the lungs increase with the severity of the disease and persist after a stimulus is removed. Evidence suggests that the host produces an inflammatory response to inhaled stimuli, with activated macrophages and increased leukocytes being the core cells in the process, which is regulated by multiple inflammatory pathways. Meanwhile, the oxidative stress reaction and excessive protease expression generated by the inhalation stimulus-induced tissues can also amplify the influence of chronic inflammation, and further aggravate the tissue damage. At present, no medicine for curing COPD is researched at home and abroad, so that great attention is paid to the search of a new medicine capable of effectively treating COPD.
Sabina przewalskii (Sabina przewalskii) is evergreen conifer of Thuja of Cupressaceae, and is mainly distributed in northeast of Qinghai-Tibet plateau and in western marginal region of loess plateau. The twig with leaves of sabina przewalskii is bitter and astringent in taste and slightly cold, is a common traditional Chinese and Tibetan medicine, has the effects of stopping bleeding, relieving cough and the like, and is mainly used for clinically treating diseases such as hemoptysis, hematemesis, hematuria, hematochezia, epistaxis, metrorrhagia and the like. The Chinese Tibetan medicine also records that the short branches with leaves and fruits can be used for treating nephritis and arthritis. The chemical components of Sabina Przewalskii mainly comprise monoterpenes, sesquiterpenes and their oxygen-containing derivatives, and other components are small amount of esters. The sabina przewalskii leaves contain abundant phenolic substances, and the phenolic compounds have natural antioxidant activity as one of the most important secondary metabolites in plants. Research shows that the Sabina przewalskii extract has pharmacological effects of resisting inflammation, oxidation, tumor and the like, has killing effect on human breast cancer, lung cancer, oral epithelial cancer and the like, and has an unknown anti-tumor mechanism.
At present, no literature report on the use of sabina przewalskii or sabina przewalskii extract for treating COPD is found.
Disclosure of Invention
In order to solve the problems, the invention mainly aims to provide the application of sabina przewalskii in preparing the medicine for treating chronic obstructive pulmonary disease, and experiments prove that the sabina przewalskii extract can obviously improve the lung function of a chronic obstructive pulmonary disease model mouse, and inhibit inflammatory cell infiltration of lung tissues and release of inflammatory factors, so that airway inflammatory response and the degree of lung tissue injury are reduced. The Sabina przewalskii ethanol extract has the effects of improving the lung function and lung tissue injury of mice and treating chronic obstructive pulmonary diseases of the mice.
The invention also aims to provide the application of sabina przewalskii in preparing the medicine for treating the chronic obstructive pulmonary disease, which widens the effective method for treating the chronic obstructive pulmonary disease and solves the problem of adverse reaction of patients in the prior art for treating the chronic obstructive pulmonary disease.
In order to achieve the above object, the technical solution of the present invention is as follows.
An application of Sabina przewalskii in preparing medicine for treating chronic obstructive pulmonary disease is disclosed, wherein the Sabina przewalskii is alcohol extract of Sabina przewalskii.
Further, the preparation of the alcohol extract of sabina przewalskii comprises the following steps: crushing Sabina przewalskii leaves, weighing crushed leaf samples, and mixing the crushed leaf samples according to a material-liquid ratio of 1:10 to 1:50, adding 10 to 100 percent ethanol, heating the water bath kettle to 40 to 100 ℃, extracting for 20 to 120min in the water bath kettle, and performing suction filtration after extracting for one to three times to obtain ethanol extract. Concentrating the extractive solution under reduced pressure with rotary evaporator to obtain ethanol extract, drying to constant weight to obtain Sabina przewalskii ethanol extract.
Further, crushing Sabina przewalskii Makino leaves, weighing crushed leaf samples, and mixing the crushed leaf samples according to a material-liquid ratio of 1:20, adding 60% ethanol, heating the water bath to 70 ℃, extracting for 60min in the water bath, performing suction filtration after twice extraction to obtain an ethanol extract, performing reduced pressure concentration on the extract by using a rotary evaporator to obtain an ethanol extract concentrate, and drying to constant weight to obtain the Sabina przewalskii ethanol extract.
The extract can be mixed with pharmaceutically acceptable carrier, and made into various common preparations such as powder, tablet, granule, capsule, syrup, etc. by conventional method.
The alcohol extract of sabina przewalskii kom in pharmaceutically acceptable carriers includes but is not limited to the following:
diluent agent: starch, powdered sugar, lactose, dextrin, microcrystalline cellulose, inorganic salts, sugar alcohols, etc.
Wetting agent and binder: purified water, ethanol, gelatin, polyethylene glycol, cellulose derivatives, and the like.
Disintegrating agent: starch, sodium carboxymethyl starch, cellulose derivatives, crospovidone, and the like.
Lubricant: magnesium stearate, aerosil, talcum powder, polyethylene glycol and the like.
Cosolvent: water, ethanol, glycerol, propylene glycol, liquid paraffin, vegetable oil, etc.
Flavoring agent: sucrose, monosaccharide, aromatic, etc.
Preservative: benzoic acid, sorbic acid, methyl esters, ethyl esters, propyl esters, and the like.
The alcohol extract of Sabina przewalskii adopted by the invention is obtained from Sabina przewalskii of Cupressaceae, is safe and nontoxic, and can be taken for a long time. Experimental results show that the sabina przewalskii alcohol extract can obviously improve lung function indexes such as dynamic lung compliance (Cdyn), airway Resistance (RI) and expiratory peak flow rate (PEF) of a chronic obstructive pulmonary disease model mouse, obviously reduce the proportion of lymphocytes, macrophages and neutrophilic granulocytes (P is less than 0.05 or P is less than 0.01) in alveolar lavage fluid of the mouse, reduce the contents of NF-kB, IL-8 and TNF-alpha (P is less than 0.05 or P is less than 0.01), and relieve lung tissue pathological changes such as alveolar wall collapse, tracheal wall thickening, inflammatory cell aggregation and tissue bleeding, thereby showing that the sabina przewalskii alcohol extract has the effects of inhibiting inflammatory reaction of the chronic obstructive pulmonary disease, improving lung function and treating the chronic obstructive pulmonary disease.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The invention discloses application of sabina przewalskii in preparation of a medicament for treating chronic obstructive pulmonary disease.
The preparation method of the Sabina przewalskii ethanol extract comprises the following steps: crushing Sabina przewalskii leaves, weighing crushed leaf samples, and mixing the crushed leaf samples according to a material-liquid ratio of 1:10 to 1:50, adding 10 to 100 percent ethanol, heating the water bath kettle to 40 to 100 ℃, extracting for 20 to 120min in the water bath kettle, and performing suction filtration after extracting for one to three times to obtain ethanol extract. Concentrating the extractive solution under reduced pressure with rotary evaporator to obtain ethanol extract, drying to constant weight to obtain Sabina przewalskii ethanol extract.
The extract can be mixed with pharmaceutically acceptable carrier, and made into various common preparations such as powder, tablet, granule, capsule, syrup, etc. by conventional method.
Specifically, with reference to the following examples, the specific implementation steps of the present invention are as follows:
example 1 modulation of Lung function in mice by Sabina przewalskii
1. And (4) experimental animals.
ICR mice, SPF grade, male, weight 20g, total 40, purchased from Guangdong province medical laboratory animal center. The samples were randomly divided into four groups of 10. A normal control group, a model group, an example group and a positive control group are set.
2. Sample source and processing
Mice were acclimatized for 1 week before they received a smoking regimen, the smoking method being described in the literature. Smoking for 2 times per day, every 10 cigarettes/1 hr, smoking interval of the same day is more than 4 hr, and smoking is continued for 5 days/week. Each cigarette contains 11mg of tar, 1.0mg of nicotine and 13mg of carbon monoxide. The temperature in the smoking box is maintained at about 26 ℃. Normal groups were left untreated and were raised in a smokeless environment. The administration is performed by intragastric administration (the intragastric volume is 0.2 mL) 1h before each fumigation, the dose of the examples is 100mg/kg (calculated by the prepared dry powder), the dose of the roflumilast is 5mg/kg, the blank control group and the model group are intragastric administered with physiological saline with the same volume, and the animals are respectively taken for index detection after being smoked for 30 days.
3. Pulmonary function detection
The dynamic lung compliance (Cdyn), airway Resistance (RI) and peak expiratory flow rate (PEF) of the mice were determined using a Buxco small animal lung function analysis system.
4. Results of the experiment
Regulation and control of lung function of mice by sabina przewalskii
Compared with a normal control group, the Cdyn and PEF of the model group are obviously reduced, and the RI is obviously increased (P is less than 0.01); the Cdyn, PEF and RI of the roflumilast group are obviously improved, and have obvious difference (P is less than 0.05 or P is less than 0.01) compared with a model group; the example groups Cdyn, PEF and RI were improved compared to the model group and were significantly different (P < 0.05 or P < 0.01) compared to the model group. The results are shown in Table 1.
TABLE 1 changes in pulmonary function in groups of mice: (
n＝10)
Note: compared with the normal group, # # P is less than 0.01; p < 0.05 or P < 0.01 compared to the model group.
Example 2 Regulation of leukocyte counts in mouse alveolar lavage fluid by Sabina przewalskii
1. Experimental animals and sample source and treatment were the same as in example 1.
2. Mouse alveolar lavage fluid collection and leukocyte differential counting
After completion of the lung function test, the mice were sacrificed by cervical dislocation. The thoracic cavity was opened to separate lung tissue, the left collateral trachea was ligated, 3mL of physiological saline was injected from the main bronchus into the right lung, the lung was gently kneaded with a hand for about 30s, and then lavage fluid was withdrawn and repeated 2 times. The left lung was fixed in paraformaldehyde for pathological examination. Centrifuging alveolar lavage fluid at 4 deg.C and 1500rpm for 15min, freezing supernatant at-80 deg.C for inspection, collecting cell precipitate, performing Swiss-Giemsa staining smear, observing under optical microscope, and classifying and counting leukocytes.
3. Results of the experiment
Regulation of leukocyte amount in mouse alveolar lavage fluid by Sabina przewalskii
Compared with the normal group, the proportion of macrophage and neutrophil in the model group mouse is obviously increased, and the proportion of lymphocyte is obviously reduced (P is less than 0.01). The roflumilast has obvious effect on inflammatory cells in alveolar lavage fluid, and compared with a model group, the roflumilast has the advantages that the proportion of macrophages and neutrophils is obviously reduced, and the proportion of lymphocytes is obviously increased (P is less than 0.05 or P is less than 0.01). Compared with the model group, the proportion of the macrophages and the neutrophils in the example group is obviously reduced, and the proportion of the lymphocytes is obviously increased (P < 0.05 or P < 0.01). The results are shown in Table 2.
TABLE 2 comparison of leukocytes in alveolar lavage fluid of mice in each group: (
n＝10)
Note: compared with the normal group, # # P is less than 0.01; p < 0.05 or P < 0.01 compared to model groups.
Example 3 Effect of Sabina przewalskii on Lung tissue inflammation in mice
1. Experimental animals and sample source and treatment were the same as in example 1.
2. Alveolar lavage fluid supernatant inflammatory factor detection
And (3) detecting the contents of NF-kB, IL-8 and TNF-alpha in the supernatant of the alveolar lavage fluid of each group of mice by an ELISA method, wherein each index is strictly determined according to the operation of a kit instruction.
3. Results of the experiment
Effect of Sabina przewalskii on lung tissue inflammation of mice
Compared with the normal group, the NF-kB, IL-8 and TNF-alpha contents of the model group mice are obviously increased (P is less than 0.01). Compared with the model group, the content of NF-kB, IL-8 and TNF-alpha of the mice in the roflumilast group is obviously reduced (P is less than 0.01). The mice of the example group had significantly reduced levels of NF-. Kappa.B, IL-8 and TNF-. Alpha.compared with the model group (P < 0.05 or P < 0.01). The results are shown in Table 3.
TABLE 3 comparison of inflammatory factor content in alveolar lavage fluid of mice in each group: (
n＝10)
Note: compared with the normal group, # # P is less than 0.01; p < 0.05 or P < 0.01 compared to the model group.
Example 4 Effect of Sabina przewalskii on mouse Lung tissue Structure Change
1. Experimental animals and sample sources and treatments were the same as in example 1.
2. Pathological observation of lung tissue in mice
Fixing lung tissue in 4% paraformaldehyde solution; after being embedded and sliced by paraffin, the tissues are placed at 60 ℃ for baking for 2 hours; storing at 4 deg.C; dewaxing in xylene for 5-10 min; soaking in absolute ethanol for 5min, 95% ethanol for 2 min, and 75% ethanol for 2 min; washing with water for 2 times; dyeing with hematoxylin for 10 minutes, and washing with water; soaking in 1% hydrochloric acid alcohol for 10 s; returning the running water to blue for 10 minutes; washing with water after dyeing for 1 minute; soaking in 95% ethanol for 2 min, and soaking in anhydrous ethanol for 5 min; xylene soak 5 minutes. The hematoxylin staining solution can stain basophilic structures (such as ribosomes, nucleuses, ribonucleic acids in cytoplasm and the like) of tissues into bluish purple; eosin stains the tissue eosinophilic structures (e.g., intracellular and intercellular proteins) pink, making the morphology of the whole cell tissue clearly visible.
3. Results of the experiment
Effect of Sabina przewalskii on mouse lung tissue structure change
The normal group of mice has complete lung tissue structure without obvious abnormality. Compared with a normal group of mice, the lung structure of the model group of mice is obviously changed, and the model group of mice has the obvious phenomena of alveolar wall collapse, trachea wall thickening, inflammatory cell aggregation, tissue bleeding and the like. Sabina przewalskii can weaken structural change of lung tissues to a certain extent. The positive control drug roflumilast can also inhibit lung tissue damage to a certain extent.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. An application of Sabina przewalskii in preparing medicament for treating chronic obstructive pulmonary disease is characterized in that the Sabina przewalskii is Sabina przewalskii alcohol extract.
2. The use of sabina przewalskii kom for preparing a medicament for treating chronic obstructive pulmonary disease according to claim 1, wherein the sabina przewalskii kom alcohol extract is prepared by the following steps: crushing Sabina przewalskii leaves, weighing crushed leaf samples, and mixing the materials according to a material-liquid ratio of 1:50, adding 10 to 100 percent ethanol, heating the water bath kettle to 40 to 100 ℃, extracting for 20 to 120min in the water bath kettle, and performing suction filtration after extracting for one to three times to obtain ethanol extract. Concentrating the extractive solution under reduced pressure with rotary evaporator to obtain ethanol extract, drying to constant weight, and making into QILIANYUANBAI ethanol extract.
3. The use of sabina przewalskii as claimed in claim 2, wherein the sabina przewalskii is prepared by pulverizing sabina przewalskii leaves, weighing the pulverized leaves, adding 60% ethanol at a ratio of 1.
4. The use of sabina przewalskii as claimed in claim 1, wherein the alcoholic extract of sabina przewalskii is administered in a pharmaceutically acceptable carrier including but not limited to the following:
diluent agent: starch, powdered sugar, lactose, dextrin, microcrystalline cellulose, inorganic salts, sugar alcohols;
wetting agent and binder: purified water, ethanol, gelatin, polyethylene glycol, cellulose derivatives;
disintegrating agent: starch, sodium carboxymethyl starch, cellulose derivatives, crospovidone;
lubricant: magnesium stearate, superfine silica gel powder, talcum powder and polyethylene glycol;
cosolvent: water, ethanol, glycerol, propylene glycol, liquid paraffin, and vegetable oil;
flavoring agent: sucrose, monosaccharide, flavoring agent;
preservative: benzoic acid, sorbic acid, methyl ester, ethyl ester and propyl ester.
5. The use of sabina przewalskii kom as claimed in claim 1 in the preparation of a medicament for the treatment of chronic obstructive pulmonary disease, wherein the alcoholic extract of sabina przewalskii kom can be combined with a pharmaceutically acceptable carrier and prepared into powders, tablets, granules, capsules, syrups by conventional methods.
6. The use of sabina przewalskii as claimed in claim 5, for the manufacture of a medicament for the treatment of chronic obstructive pulmonary disease, wherein the pharmaceutical formulation for oral or parenteral administration for the treatment of chronic obstructive pulmonary disease can be prepared.
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Non-Patent Citations (3)
| Title |
|---|
| 刘林芳: "不同海拔祁连圆柏的化学成分及生物活性研究", 中国硕士学位论文数据库，硕士电子期刊, no. 08, pages 26 - 27 * |
| 师瑞;李金潇: "基于网络药理学探究祁连圆柏治疗慢性阻塞性肺疾病的潜在作用机制", 青海师范大学学报(自然科学版), vol. 38, no. 02, pages 45 - 46 * |
| 王奇;方罗;姜建伟;: "祁连圆柏叶乙醇提取物挥发性成分GC-MS分析", 浙江中医杂志, no. 01, pages 60 - 61 * |
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