Application of aceratrol in preparation of medicine for preventing or treating severe pneumonia

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of aceratrol in preparation of a medicine for preventing or treating severe pneumonia. According to the application disclosed by the invention, experimental research is carried out on prevention or treatment of severe pneumonia by constructing a lipopolysaccharide (LPS)-induced mouse severe pneumonia model, and a result shows that the aceratrol has a good protection effect on the LPS-induced mouse severe pneumonia model, and can be used for remarkably relieving pathological morphological injury of LPS-induced mouse lung tissues; moreover, the compound has small side effects and can be used for preparing protective medicines for severe pneumonia. The invention provides an application of aceratrol in preparation of a medicine for preventing or treating severe pneumonia, and the aceratrol has obvious effects in improving acute severe pneumonia and playing an anti-inflammatory effect, small side effects and considerable clinical application prospect and commercial value.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to the use of turmeric alcohol in preparing a medicine for preventing or treating severe pneumonia. Background Art

[0002] Pneumonia is an infectious disease caused by pathogenic bacteria or other microorganisms such as viruses infecting the lower respiratory tract and proliferating, causing lung inflammation. Acute severe pneumonia is a common clinical critical illness caused by a variety of reasons including bacterial, viral infection or trauma, with high morbidity and mortality, seriously threatening the patient's life safety. Severe pneumonia can cause acute lung injury (ALI), which is a clinical syndrome characterized by dyspnea, increased respiratory work, inflammatory cell infiltration in the lungs, increased pulmonary microvascular permeability, severe edema of the alveoli and lung parenchyma, and progressive hypoxemia. In severe cases, it can develop into acute respiratory distress syndrome (ARDS). In addition to severe pneumonia, the causes of ALI also include mechanical stimulation, inhalation of toxic liquids or gases, systemic infection and shock. Although there are many measures and drugs for the treatment of ALI / ARDS, such as antibiotics and glucocorticoids, the mortality rate of ALI / ARDS patients is still as high as 40%, and some surviving patients still have residual lung dysfunction and are accompanied by a variety of other adverse reactions. Currently, there is no clinically available curative drug that can be used for ALI / ARDS patients, which results in a low overall survival rate and quality of life for ALI / ARDS patients. Therefore, there is a great demand for drugs that can effectively prevent and treat severe pneumonia (especially ALI / ARDS).

[0003] Atractylodes is the rhizome of Atractylodes lancea (Thunb.) DC. of the Asteraceae family. Atractylodes is warm in nature, pungent and bitter in taste. It enters the spleen meridian, stomach meridian, and liver meridian. It has the effects of drying dampness and strengthening the spleen, dispelling wind and cold, and improving eyesight, and is a dampness-removing medicine. It is used to treat abdominal distension, diarrhea, edema, beriberi, rheumatic pain, colds, and night blindness. Modern pharmacological studies have shown that Atractylodes has anti-gastritis and anti-gastric ulcer effects; regulates gastrointestinal motility; resists liver damage; resists hypoxia; and has a blocking effect on nicotinic receptors. The decoction can resist experimental gastritis and gastric ulcers, and has a significant inhibitory effect on gastric juice volume, gastric acidity, and pepsin activity. Serum ulcers are ulcers formed by tissue necrosis caused by ischemia and circulatory disorders of the gastric mucosa, and Atractylodes has a significant effect on this. It also has pharmacological effects such as protecting the liver and promoting bile secretion, and resisting hypoxia. Therefore, Atractylodes lancea is often used clinically to treat colds, fever, pneumonia, bronchitis, nephritis, hepatitis, gastroenteritis, cholecystitis, peptic ulcer, cirrhosis, hypertension and other diseases. However, there are few research reports on the active ingredient cypermethrin in Atractylodes lancea, and there are no relevant patents and non-patent literature reporting that cypermethrin has an improving effect on severe pneumonia (especially ALI / ARDS). Summary of the invention

[0004] The purpose of the present invention is to solve the deficiencies of the prior art, to realize the development and utilization of natural products by adopting modern drug research methods, and to provide the use of cypermethrin in the preparation of drugs for preventing or treating severe pneumonia in combination with a large number of pharmacodynamic experimental screenings.

[0005] To achieve the purpose of the present invention, the present invention adopts the following technical solutions:

[0006] The present invention provides the use of turmeric alcohol in preparing a medicine for preventing or treating severe pneumonia.

[0007] The present invention also provides the use of turmeric alcohol as the only active ingredient in preparing a medicine for preventing or treating severe pneumonia.

[0008] As an optional method, in the above use, the CAS number of the cypermethrin is 186376-50-1, and the chemical structural formula of the cypermethrin is

[0009] As an optional mode, in the above-mentioned use, the severe pneumonia is acute severe pneumonia.

[0010] As an alternative, in the above use, the acute severe pneumonia is acute lung injury (ALI) and / or acute respiratory distress syndrome (ARDS).

[0011] Preferably, the acute pneumonia is ALI / ARDS.

[0012] As an optional mode, in the above-mentioned use, the severe pneumonia includes lung function damage, lung tissue oxidative stress, lung tissue damage, pulmonary edema and lung tissue inflammation.

[0013] As an optional mode, in the above-mentioned use, the medicine further comprises a pharmaceutically acceptable carrier.

[0014] As an optional mode, in the above-mentioned use, the dosage form of the drug is an oral dosage form.

[0015] As an optional mode, in the above-mentioned use, the oral dosage form is a tablet, a capsule, a granule, an oral liquid, a pill, an inhaler or an aerosol.

[0016] The above-mentioned various dosage forms can be prepared according to conventional processes in the field of pharmaceutical preparations.

[0017] Atractylodes macrocephala can be extracted and separated from plants such as Atractylodes lancea and Atractylodes macrocephala containing these active ingredients by conventional extraction and purification methods in the field of natural medicinal chemistry, or can be purchased from commercially available products.

[0018] In the medical uses described above, the administration method, administration time, administration times and administration frequency of the cypermethrin of the present invention need to be determined according to the specific diagnosis results of the disease, which is within the technical scope mastered by those skilled in the art.

[0019] The treatment regimen for mice is applied to humans, and the effective dose of all drugs for humans can be converted by the effective dose of the drug for mice, which is also easy to achieve for those of ordinary skill in the art. It is understood by those skilled in the art that the actual dosage may be lower than the converted dosage range. The effective amount and specific treatment regimen for a subject may be affected by many factors, including the age, weight, sex, diet, administration time, disease susceptibility, disease progression, doctor's judgment, etc. of the subject.

[0020] In order to better understand the essence of the present invention, the pharmacodynamics experiments and their results are used in the following specific implementation section to further illustrate the new use of the tangerine glycol of the present invention in the pharmaceutical field.

[0021] Through the above technical solution, the present invention has at least the following advantages:

[0022] (1) The present invention constructs a lipopolysaccharide (LPS)-induced mouse severe pneumonia model to conduct experimental research on the prevention or treatment of severe pneumonia by fennel. The results show that fennel has a good protective effect on the LPS-induced mouse severe pneumonia model, can significantly reduce the LPS-induced pathological morphological damage of mouse lung tissue, inhibit lung tissue inflammatory response and oxidative stress, and has little side effects. It can be used to prepare protective drugs for severe pneumonia.

[0023] (2) The present invention provides the use of turmeric alcohol in the preparation of a drug for preventing or treating severe pneumonia. The drug has significant effects in improving acute severe pneumonia and exerting anti-inflammatory effects, with few side effects, and has considerable clinical application prospects and commercial value. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 : H&E staining results of lung tissue damage in mice in each group. A: blank control group; B: model group; C: atractylodes alcohol group; D: atractylodes ketone group; D: dexamethasone group, magnification 40x. DETAILED DESCRIPTION

[0025] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0026] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0027] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0028] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0029] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0030] The present invention will be further described with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0031] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0032] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0033] Example:

[0034] The present invention studies the use of cypermethrin in preparing a drug for preventing or treating severe pneumonia through animal experiments. The present invention uses C57BL / 6 mice as experimental subjects and constructs a lipopolysaccharide (LPS)-induced mouse severe pneumonia (acute lung injury) model to conduct experimental research on the use of cypermethrin in preventing or treating severe pneumonia.

[0035] The modeling method refers to CN118593471A.

[0036] 1. Experimental methods:

[0037] 1.1 Animal husbandry:

[0038] Six-week-old SPF male C57BL / 6 mice (purchased from Liaoning Changsheng Biotechnology Co., Ltd.) weighing 20-22 g were selected. The experiment was carried out after a one-week adaptation period. Six mice were placed in a cage, and standard mouse feed and normal temperature drinking water that had been strictly sterilized were regularly replaced.

[0039] The mice were randomly divided into 5 groups: blank control group, model group, atractylodes alcohol group, atractylodes ketone group, and dexamethasone group, with 14 mice in each group.

[0040] 1.2 Animal medication and modeling:

[0041] Atractylodesinol and atractylodesone (purchased from Shenyang Sandu Pharmaceutical Technology Co., Ltd.) were dissolved in a solvent containing 5% ethanol, 40% PEG400 and 55% normal saline.

[0042] The mice in the blank group and the model group were intragastrically administered with a solvent containing 5% ethanol, 40% PEG400 and 55% saline every day for 3 consecutive days.

[0043] For the mice in the turmeric alcohol group and the atractylodes ketone group, 20 mg / kg of turmeric alcohol and atractylodes ketone were given by gavage every day for 3 consecutive days.

[0044] For the mice in the dexamethasone group, 2 mg / kg was intraperitoneally injected daily, diluted with normal saline, for 3 consecutive days.

[0045] One hour after the intragastric administration on the third day, an acute lung injury model was established by tracheal instillation of LPS solution. The specific experimental procedures are as follows:

[0046] (1) Before surgery, mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (30 mg / kg) according to their body weight.

[0047] (2) Fix the limbs of the anesthetized mouse with rubber bands, put a thin wire around its teeth, and disinfect the neck skin with 75% alcohol.

[0048] (3) Make a longitudinal incision along the middle of the mouse’s neck and separate the subcutaneous tissue until the trachea is exposed and the cartilage ring appears.

[0049] (4) The microinjector was inserted into the trachea about 1 cm in parallel with the proximal end. Sterile saline was slowly dripped into the trachea of ​​the control group, and LPS was dripped into the trachea of ​​the other groups.

[0050] (5) After removing the injector, immediately stand the mouse upright, rotate and shake it evenly for 2 minutes, then suture the neck wound. Observe the condition of the mouse and feed it as usual after it wakes up.

[0051] 1.3 Experimental animal treatment and lung tissue specimen collection

[0052] 24 hours after modeling, samples were collected, and 7 mice were killed in each group after weighing, and lung tissues were collected, and the wet weight of the whole lung was weighed. The lung tissues were used for H&E staining and MDA and SOD content determination. The specific operation method is as follows:

[0053] (1) Weigh the 7 mice in each group and kill them by cervical dislocation;

[0054] (2) Fix the mice, wipe the abdomen with 75% alcohol, cut the chest cavity, remove the lung tissue, organize and take pictures, remove the trachea and other excess tissues, weigh the whole lung wet weight, and calculate the lung coefficient;

[0055] (3) The left upper lobe of the lung was fixed in 4% neutral paraformaldehyde for 24 h, routinely paraffin-embedded and sectioned for H&E staining;

[0056] (4) The remaining lung tissues were placed in centrifuge tubes and quickly frozen in liquid nitrogen. After the experiment, they were transferred to -80°C for storage and used for MDA and SOD index detection.

[0057] Lung coefficient calculation formula: Lung coefficient = lung wet weight / body weight × 100%.

[0058] After H&E staining of lung tissue, the pathological morphology of lung tissue was observed.

[0059] The MDA and SOD contents in lung tissues were measured using commercially available kits.

[0060] 1.4 Bronchoalveolar lavage fluid (BALF) collection and processing

[0061] 24 hours after modeling, bronchoalveolar lavage fluid was collected. First, the remaining 7 mice in each group were weighed, and then BALF was extracted for the determination of TNF-α and IL-6. The specific operation method is as follows:

[0062] (1) After weighing, mice were moderately anesthetized by intraperitoneal injection of sodium pentobarbital according to their body weight;

[0063] (2) When the tail root, hind limbs, eyelash reflexes and other activities of the mouse disappear and the mouse breathes slowly and evenly, fix the mouse's limbs and teeth, disinfect the mouse's neck skin with 75% alcohol, make a longitudinal opening, separate the neck tissue, expose the tracheal cartilage ring, and use a 19G cannula to puncture the trachea. Insert the needle about 1 cm, pull out the needle core, connect a 1mL syringe, and slowly inject pre-cooled saline (0.8mL / time, repeated 2 times). Collect the lavage fluid in a 1.5mL centrifuge tube and store it on ice (BALF recovery rate 85%-95%).

[0064] (3) The BALF lavage fluid was centrifuged at 3000 g for 10 min at 4°C, the supernatant was aliquoted and stored at -80°C, and the levels of TNF-α and IL-6 were detected using ELISA kits.

[0065] 1.5 Data representation and statistical analysis methods:

[0066] The results were analyzed using SPSS 27.0 statistical software, and the data were expressed as mean ± standard deviation. One-way analysis of variance was used for comparison among multiple groups, and t-test was used for comparison between two groups. P < 0.05 indicated statistically significant differences.

[0067] 2. Experimental results:

[0068] 2.1 Gross examination results of mouse lung tissue and lung coefficient

[0069] After dissecting each group of mice, it was found that the lung tissue of the mice in the blank control group was pink and uniform in color, without edema. Compared with the lung tissue of the mice in the blank control group, the lung tissue of the mice in the model group was obviously edematous, the surface color darkened and spots appeared. The pulmonary edema of the mice in the atractylodes alcohol group was significantly improved, the color of the lung tissue became lighter, the spots became fewer, and the glossiness of the lung tissue surface was improved. The degree of improvement of the gross examination results of the lung tissue by atractylodes alcohol was close to that of the positive control drug dexamethasone. However, the improvement of the pulmonary edema and lung tissue damage in the mice in the atractylodes ketone group was not obvious, and its gross examination results were only slightly better than those of the model group.

[0070] The lung coefficient results of each group of mice are shown in Table 1 below. As shown in Table 1, compared with the blank control group, the lung coefficient of the model group mice increased significantly, indicating that the pulmonary edema of the model group mice was more serious. And cypermethrin can significantly improve the pulmonary edema of model mice, and its effect is close to that of the positive control drug dexamethasone group. However, atractylodesone, which is also the main active ingredient in Atractylodes, did not improve the pulmonary edema of mice significantly.

[0071] Table 1: Lung coefficient of mice in each group (%)

[0072] Group Lung coefficient (%) Blank control group 42.11 Model Group 78.34 Atractylodes group 55.48 Atractylodes ketone group 74.92 Dexamethasone group 47.39

[0073] 2.2 Results of pathological morphological examination of mouse lung tissue

[0074] The H&E staining results of lung tissue damage in each group of mice are shown in Figure 1 Compared with the blank control group (see Figure 1 Compared with the model group, the mice in the model group showed pulmonary edema, loss of alveolar structure, obvious inflammatory cell infiltration in the tissues, and thickening of the lung interstitium (see Figure 1 B). Atractylodes alcohol group (see Figure 1 C) and dexamethasone group (see Figure 1 E) It has a significant improvement effect on the lung tissue pathological morphology of model mice.

[0075] However, the atractylodesone group (see Figure 1 D) The mouse lung tissue still had obvious edema, loss of alveolar structure, inflammatory cell infiltration in the tissue and thickening of the pulmonary interstitium, which indicated that atractylodes kenyl ketone had no obvious effect on improving pulmonary edema and lung tissue damage.

[0076] 2.3 Levels of MDA and SOD in lung tissues of mice in each group

[0077] The results of MDA and SOD levels in the lung tissues of mice in each group are shown in Table 2. Compared with the blank control group, the MDA level in the lung tissues of mice in the model group was significantly increased (p<0.001) and the SOD level was significantly decreased (p<0.001), indicating that the model was successfully established and the lung tissues of mice in the model group showed severe oxidative damage.

[0078] Compared with the model group, the MDA level in the lung group of mice treated with cypermethrin or dexamethasone was significantly reduced ( # p<0.05, ## p<0.01) and SOD levels were significantly increased ( ## p<0.01, ### p<0.001), indicating that cypermethrin can significantly reduce the degree of oxidative damage in the lung tissue of model mice, and its effect is close to that of dexamethasone.

[0079] However, atractylodesone, which is also the main active ingredient in Atractylodes lancea, did not significantly improve oxidative damage in mouse lung tissue.

[0080] Table 2: MDA and SOD levels in lung tissues of mice in each group

[0081]

[0082]

[0083] Note: Compared with the blank control group, *** p<0.001; compared with the model group, # p<0.05, ## p<0.01, ### p<0.001.

[0084] 2.4 Levels of TNF-α and IL-6 in BALF lavage fluid of mice in each group

[0085] The results of TNF-α and IL-6 levels in BALF lavage fluid of mice in each group are shown in Table 3. Compared with the blank control group, the levels of TNF-α and IL-6 in BALF lavage fluid of mice in the model group were significantly increased (p<0.001), indicating that the model was successfully established and severe inflammatory response occurred in the lung tissue of mice in the model group.

[0086] Compared with the model group, after treatment with cypermethrin or dexamethasone, the levels of TNF-α and IL-6 in the BALF lavage fluid of mice were significantly reduced (p<0.001), indicating that cypermethrin can significantly reduce the degree of inflammatory damage in the lung tissue of model mice, and its effect is close to that of dexamethasone.

[0087] However, atractylodesone, which is also the main active ingredient in Atractylodes lancea, did not significantly improve inflammatory damage to the lung tissue of mice.

[0088] Table 3: Levels of TNF-α and IL-6 in bronchoalveolar lavage fluid of mice in each group

[0089] Group TNF-α(pg / mL) IL-6 (pg / mL) Blank control group 48.16±9.27 187.14±35.46 Model Group <![CDATA[133.38±27.51 *** ]]> <![CDATA[357.29±54.18 *** ]]> Atractylodes group <![CDATA[59.45±12.33 ### ]]> <![CDATA[204.73±32.94 ### ]]> Atractylodes ketone group <![CDATA[121.69±30.56 *** ]]> <![CDATA[326.38±62.55 *** ]]> Dexamethasone group <![CDATA[51.13±10.78 ### ]]> <![CDATA[195.52±27.31 ### ]]>

[0090] Note: Compared with the sham operation group, *** p<0.001; compared with the model group, ### p<0.001.

[0091] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. Use of turmeric alcohol in preparing a medicine for preventing or treating severe pneumonia.

2. Use of turmeric alcohol as the sole active ingredient in the preparation of a drug for preventing or treating severe pneumonia.

3. The use according to claim 1 or claim 2, characterized in that: The severe pneumonia is acute severe pneumonia.

4. The use according to claim 3, characterized in that: The acute severe pneumonia is acute lung injury and / or acute respiratory distress syndrome.

5. The use according to claim 1 or claim 2, characterized in that: The severe pneumonia includes lung function impairment, lung tissue oxidative stress, lung tissue damage, pulmonary edema and lung tissue inflammation.

6. The use according to claim 1 or claim 2, characterized in that: The medicament further comprises a pharmaceutically acceptable carrier.

7. The use according to claim 6, characterized in that: The dosage form of the drug is an oral dosage form.

8. The use according to claim 7, characterized in that: The oral dosage form is tablet, capsule, granule, oral liquid, dropping pill, inhalant or aerosol.

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