Application of ganoderma lucidum spore oil in prevention and treatment of pulmonary nodules

Abstract

The invention relates to the technical field of medicines, and discloses an application of ganoderma lucidum spore oil in preparation of a medicine for preventing and treating pulmonary nodules, animal model tests find that the ganoderma lucidum spore oil has an obvious effect of inhibiting generation and development of benign granuloma nodules in the lung of a mouse and also has good safety, and the ganoderma lucidum spore oil can be used for preparing medicines for preventing and treating pulmonary nodules. Therefore, the compound can be used for preparing medicines for preventing and treating pulmonary nodules.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to application of ganoderma lucidum spore oil in preventing and treating pulmonary nodules. Background Art

[0002] Ganoderma lucidum is a treasure in the treasure house of traditional Chinese medicine. It is known as the "immortal grass" and can be used for restlessness, insomnia, palpitations, lung deficiency cough, shortness of breath, and loss of appetite. Ganoderma lucidum spores are extremely tiny spores ejected from the cap during the growth and maturity of Ganoderma lucidum. They are the reproductive cells of Ganoderma lucidum and have all the genetic active substances of Ganoderma lucidum. Modern pharmacological studies have shown that Ganoderma lucidum spores have the effects of enhancing immunity, lowering blood sugar, lowering blood lipids, anti-tumor, anti-hypoxia, and scavenging free radicals. Ganoderma lucidum spore oil is a fat-soluble active substance obtained by breaking the wall of Ganoderma lucidum spores and supercritical carbon dioxide extraction. Studies have shown that it has the effects of enhancing immunity and protecting the liver. Existing studies generally believe that the main active ingredients of Ganoderma lucidum are Ganoderma lucidum polysaccharides and Ganoderma lucidum triterpenoids. The various effects of Ganoderma lucidum are also more focused on polysaccharides and triterpenes. For other components in spore oil, such as the composition research and efficacy analysis of triglyceride compounds, which account for the largest proportion, there are currently few reports.

[0003] In recent years, with the widespread application of multi-slice computed tomography and the widespread implementation of lung cancer screening, the detection rate of lung nodules has increased significantly worldwide. An article in the authoritative journal JAMA found that at least 95% of lung nodules are benign and there is no possibility of canceration through observation of patients with lung nodules. Most patients with lung nodules can heal themselves and have a good prognosis, but one-third of patients will still progress to chronic diseases, eventually leading to pulmonary fibrosis, pulmonary hypertension, respiratory failure and even death. Combined with the current environment of frequent respiratory infections caused by viruses and mycoplasmas, the prevention and treatment of lung nodules has become the focus of media and public attention in recent years. Clinically, benign lung nodules that progress slowly or do not change for a long time are mainly followed up and observed, and the guidelines lack effective prevention and treatment drug recommendations. At present, the clinical treatment of pulmonary sarcoidosis is limited, and glucocorticoids are the first choice, but long-term and large-scale use has serious adverse reactions and is prone to relapse after discontinuation of the drug. It is still necessary to continue to find safe and effective drugs for the treatment of lung nodules. Summary of the invention

[0004] The purpose of the present invention is to provide an application of Ganoderma lucidum spore oil in the preparation of a drug for preventing and treating the occurrence and development of pulmonary nodules. The prevention and treatment of pulmonary nodules has become the focus of media and public attention in recent years, but the guidelines still lack effective preventive and therapeutic drug recommendations. Based on the above reasons and needs, it is found that Ganoderma lucidum spore oil has a new property of preventing and treating the occurrence and development of pulmonary nodules, and further provides a new application of Ganoderma lucidum spore oil as a drug, that is, the application in the preparation of a drug for preventing or treating pulmonary nodules.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] An application of ganoderma spore oil is used for preparing a preparation for preventing and treating sarcoidosis.

[0007] Animal model experiments have shown that Ganoderma lucidum spore oil has a significant inhibitory effect on the development of benign granulomatous nodules in the lungs of mice, and has good safety. Therefore, it can be used to prepare drugs to prevent and treat the development of lung nodules.

[0008] Furthermore, the current experimental modeling methods are not completely targeted at the lungs and may have similar effects in preventing and treating sarcoidosis in organs other than the lungs, which can be further developed and utilized.

[0009] Preferably, the sarcoidosis comprises pulmonary nodules.

[0010] Preferably, the sarcoidosis comprises benign granulomatous nodules.

[0011] Preferably, the causes of sarcoidosis include one or more of microorganisms and their related derivatives, and exogenous particles.

[0012] Preferably, the microorganisms include one or more of bacteria, viruses, and mycoplasma; the exogenous particles include one or more of carbon nanoparticles, metal particles, and smoke.

[0013] Preferably, the Ganoderma lucidum spore oil includes one or more components of triglyceride, ergosterol and vitamin E.

[0014] Preferably, in the Ganoderma lucidum spore oil, the content of triglycerides is greater than 60%, the content of ergosterol is 0.1% to 0.5%, the content of vitamin E is 0.5% to 4%, and no Ganoderma lucidum polysaccharide compounds are detected.

[0015] Preferably, the triglyceride content in the Ganoderma lucidum spore oil is greater than 90%.

[0016] Preferably, the mass content of vitamin E in the Ganoderma lucidum spore oil is 2%.

[0017] More preferably, no Ganoderma triterpenoid compounds are detected in the Ganoderma spore oil.

[0018] A preparation obtained by the above application.

[0019] Preferably, the preparation comprises a drug; the drug further comprises a pharmaceutically acceptable carrier.

[0020] Preferably, the medicine comprises one of tablets, capsules, lozenges, granules, granules, pills, powders, ointments, pills, suspensions, solutions, emulsions, injections, suppositories, creams, sprays, drops or patches.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] It is reported that the immune characteristic of sarcoidosis is CD4+T cell-mediated overimmunity, so corticosteroids are still used as first-line treatment drugs in clinical practice. Ganoderma lucidum spore oil is generally believed to have the effect of enhancing immunity. Many studies have reported that Ganoderma lucidum spore oil can increase the ratio of CD4+T cells in peripheral blood lymphocytes, thereby enhancing the body's immune ability. However, the present invention unexpectedly found that Ganoderma lucidum spore oil with a triglyceride content of >60%, an ergosterol content of 0.1% to 0.5%, and a vitamin E content of 0.5% to 4% has a significant effect on preventing and treating pulmonary nodules, and can significantly inhibit the occurrence and development of pulmonary granulomas in mice with benign granulomatous pulmonary nodules, while having no significant effect on the weight and overall health of the mice, and no obvious adverse reactions were observed, indicating good safety. The Ganoderma lucidum spore oil of the present invention has the beneficial effect of being used to prepare drugs for preventing and treating pulmonary nodules. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is the HPLC component detection result of the Ganoderma lucidum spore oil in the embodiment of the present invention;

[0024] Figure 2 This is the TLC polysaccharide detection result of Ganoderma lucidum spore oil in the embodiment of the present invention;

[0025] Figure 3 is the Micro-CT tomography test result of the mouse lung in an embodiment of the present invention;

[0026] Figure 4 4 is a graph showing the results of pathological examination of mouse lung tissue according to an embodiment of the present invention. DETAILED DESCRIPTION

[0027] In order to better illustrate the purpose, technical scheme and advantages of the present invention, the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention. Without departing from the spirit and essence of the present invention, modifications or replacements made to the method, steps or conditions of the present invention all belong to the scope of the present invention. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0028] Example 1

[0029] Preparation and component detection of Ganoderma lucidum spore oil

[0030] The preparation method of the ganoderma spore oil used in the present invention is as follows:

[0031] Take the ganoderma lucidum spore powder, break the wall, and then extract it with supercritical CO2 to obtain the crude ganoderma lucidum spore oil. Take an appropriate amount of silica gel, dissolve it with a petroleum ether-ethyl acetate mixed solution with a volume ratio of 8.5-9.5:0.5-1.5, wet-load it into a column to obtain an elution column, the ratio of the column diameter to the column height of the elution column is 1:10-20, and the mass ratio of the crude ganoderma lucidum spore oil to the silica gel is 1:10-40. Take the crude ganoderma lucidum spore oil and load it onto the elution column, elute it with the above petroleum ether-ethyl acetate mixed solution at normal pressure, collect the eluate, combine it, concentrate it under reduced pressure to remove the solvent, dry it in vacuum, add vitamin E to make its mass content 2%, and obtain the ganoderma lucidum spore oil. Through separation and refining, a large amount of impurities and moisture are removed, and the obtained ganoderma lucidum spore oil is clear and transparent, and the component content is stable.

[0032] The ergosterol and triglyceride contents in Ganoderma lucidum spore oil were determined by high performance liquid chromatography (HPLC). Figure 1 As shown, peak 1 is ergosterol with a content of 0.37%, peaks 2-7 are different triglyceride components with a total content of 93.29%, and no peak representing triterpenoid components was found.

[0033] Thin layer chromatography (TLC) was used to detect whether Ganoderma lucidum spore oil contained polysaccharide components, and starch and glucose were selected as positive controls:

[0034] 1. Take 10g of Ganoderma lucidum spore oil, add 5ml of water, shake thoroughly in a separatory funnel, let stand for stratification, separate the water layer, and discard the oil layer. Concentrate the water layer to 2ml under reduced pressure, add 6ml of strong acid, shake well quickly, and place in a water bath at 85℃ for 30min. Take out and let it cool to room temperature for use.

[0035] 2. Take 10g of starch and prepare a 10% starch solution. Take 2ml of the solution and add 6ml of strong acid. Shake well and place in a water bath at 85℃ for 30min. Take out and let it cool to room temperature for later use.

[0036] 3. Take 10 mg of anhydrous glucose and prepare a glucose solution containing 1 mg per ml. Take 2 ml, add 6 ml of strong acid, shake well quickly, and place in a water bath at 85°C for 30 minutes. Take out and let it cool to room temperature for later use.

[0037] Take 5 μl of the above three strong acid reaction solutions respectively, spot them on the same silica gel thin layer plate, develop them with methanol as the developing agent, dry them at 105℃ and observe the spots. Figure 2 After hydrolysis of starch solution and glucose solution in the positive control group, spots representing monosaccharides appeared, while no spots were shown at the corresponding positions of Ganoderma lucidum spore oil, indicating that no sugar components (including polysaccharides, disaccharides and monosaccharides, etc.) were detected in Ganoderma lucidum spore oil.

[0038] Example 2

[0039] Study on the efficacy of Ganoderma lucidum spore oil on mice with benign granulomatous pulmonary nodules

[0040] 1. Experimental Animals

[0041] SPF C57BL / 6J mice (male, 6-8 weeks old) were housed in an SPF animal room with 12 h light / 12 h dark, a temperature of 22±2°C, and a humidity of 50±5%.

[0042] 2. Experimental Materials

[0043] Freund's incomplete adjuvant, Mycobacterium tuberculosis superoxide dismutase A (SodA), agarose beads, etc. were used to establish the model.

[0044] Ganoderma lucidum spore oil: the Ganoderma lucidum spore oil obtained in Example 1 was used.

[0045] 3. Experimental Methods

[0046] (1) Establishment and drug administration of a mouse model of benign granulomatous pulmonary nodules

[0047] Mice were randomly divided into 4 groups: blank group, model group, low-dose Ganoderma lucidum spore oil group, and high-dose Ganoderma lucidum spore oil group, with 12 mice in each group. Drug administration began 30 days before modeling. The blank group and model group were given solvent soybean oil, and the Ganoderma lucidum spore oil group was given Ganoderma lucidum spore oil by gavage. The low-dose and high-dose groups were given doses of 0.3 g / kg and 1.2 g / kg, respectively. The drug volume of each group was 5 mL / kg, once a day for 51 consecutive days, and the weight changes were recorded.

[0048] On the 31st day, the model group and the Ganoderma lucidum spore oil group were subcutaneously injected with an emulsion of SodA and Freund's incomplete adjuvant, and the blank group was subcutaneously injected with Freund's incomplete adjuvant emulsion, and the drug was continued. On the 44th day, the model group and the Ganoderma lucidum spore oil group were injected with agarose beads coupled with SodA through the tail vein, and the blank group was injected with PBS, and the drug was continued until the 51st day.

[0049] (2) Micro-CT imaging examination of mouse lungs

[0050] Before sampling, each group of mice was anesthetized with isoflurane gas, and the mice's breathing was observed. After several minutes, when the mice's breathing became stable, they were placed in the small animal Micro-CT instrument, put on breathing masks, adjusted instrument parameters, set positions, adjusted cameras, and observed the mice's breathing status and body position. After ensuring the mice's chest position and breathing status, a tomography scan was performed to detect the imaging changes in the mice's lungs.

[0051] Micro-CT statistics: The lungs of each mouse were kept at the same window level, and the same-sized area was selected at each window level for each mouse to calculate the CT value for analysis.

[0052] (3) Pathological examination of mouse lung tissue

[0053] Fixation and embedding: Fix the lung tissue with 4% paraformaldehyde, wash and trim the block, cut the tissue into a smooth surface, dehydrate with 50%, 70%, 80%, 95%, and 100% ethanol solutions, immerse in xylene I and II respectively for transparency, then immerse in 60℃ wax I and II respectively for permeation, and finally embed in paraffin.

[0054] Section staining: The embedded tissue was cut into paraffin sections with a thickness of about 4 μm, and immersed in transparent agent I, transparent agent II, and transparent agent III for dewaxing, immersed in 100%, 90%, 80%, 70% and other alcohol solutions for 5 minutes each, washed with water and stained with hematoxylin for 3 minutes, differentiated with 0.5% hydrochloric acid ethanol for a few seconds, washed with tap water for 15 minutes, anti-blued and eosin stained for 2 minutes, immersed in 95% ethanol I, 95% ethanol II, 100% ethanol I, 100% ethanol II, transparent agent I, transparent agent II, and transparent agent III for dehydration and transparency, and finally sealed with neutral gum.

[0055] Granuloma area statistics: Microscope observation and photography were performed, and Image J software was used to perform statistical analysis of the granuloma area in lung pathological images.

[0056] 4. Experimental Results

[0057] During the experiment, the animals in each group were in good condition. The results of animal weight test are shown in Table 1. After SodA injection into mice to establish the model, the weight of the mice decreased to a certain extent one day after the model was established and then gradually recovered. Compared with the blank group, the weight of the model group decreased, but there was no significant difference. Ganoderma lucidum spore oil had no significant improvement on the weight loss of mice with benign granulomatous pulmonary nodules model. There was no significant difference in the weight of the animals in each group when the samples were collected.

[0058] Table 1 Animal weight test results (g)

[0059]

[0060] The results of the effects of Ganoderma lucidum spore oil on lung imaging in mice with granulomatous pulmonary nodules are as follows Figure 3As shown. The lung imaging results showed that compared with the blank group, the model group had gray ground-glass shadows in different positions of the mouse lungs; the gray ground-glass shadows in the low- and high-dose groups of Ganoderma lucidum spore oil were reduced. The CT value is the corresponding value of the attenuation coefficient of each tissue in the CT image and the X-ray. The mouse lungs were scanned by X-rays. Due to the different absorption capacities of different tissue components in the lungs for X-rays, the CT values ​​were different. The CT values ​​of mice with benign granulomatous pulmonary nodules were higher than those of normal mice. The CT value results of each group after statistics are shown in Table 2. Compared with the blank group, the CT value of the model group was significantly increased (P<0.01); compared with the model group, the CT values ​​of the low- and high-dose groups of Ganoderma lucidum spore oil were significantly reduced (P<0.05 or P<0.01), and the difference was statistically significant, indicating that low- and high-dose Ganoderma lucidum spore oil has an improvement effect on granulomatous pulmonary nodules.

[0061] Table 2 Statistical results of Micro-CT tomography of mouse lungs

[0062]

[0063]

[0064] Note: Compared with the blank group, **P<0.01; Compared with the model group, # P<0.05, ## P<0.01

[0065] The results of the effects of Ganoderma lucidum spore oil on lung pathology in mice with granulomatous pulmonary nodules are as follows Figure 4 As shown. The alveolar tissue of the blank group was intact and normal; multiple granulomas were seen around the agarose beads in the lung tissue of the model group. The center of the granuloma was composed of epithelial cells, and a large number of lymphocytes surrounded the granuloma. Compared with the model group, the area of ​​granulomas in the lungs of the mice in the Ganoderma lucidum spore oil group was improved to a certain extent, and only a thin layer of epithelial cells surrounded the bare beads, and there was less lymphocyte infiltration around them. The results of the statistics of the granuloma area are shown in Table 3. Compared with the model group, the area of ​​granulomas in the lungs of the low-dose Ganoderma lucidum spore oil group was significantly reduced (P<0.05), and the difference was statistically significant. The area of ​​granulomas in the high-dose Ganoderma lucidum spore oil group also showed a trend of reduction to a certain extent, indicating that low and high doses of Ganoderma lucidum spore oil have an improvement effect on granulomatous pulmonary nodules.

[0066] Table 3 Statistical results of granuloma area in mouse lung tissue

[0067]

[0068] Note: Compared with the model group, *P<0.05

[0069] The above study established a mouse model of granulomatous pulmonary nodules and used Micro-CT imaging and pathological analysis techniques to show that Ganoderma lucidum spore oil with a triglyceride content of >60%, an ergosterol content of 0.1% to 0.5%, and a vitamin E content of 0.5% to 4% can alleviate the progression of pulmonary nodules by reducing the formation and growth of granulomas in mice with granulomatous pulmonary nodules.

[0070] In summary, the present invention has conducted experiments and research on Ganoderma lucidum spore oil with a triglyceride content of more than 60%, an ergosterol content of 0.1% to 0.5%, and a vitamin E content of 0.5% to 4%, and found that Ganoderma lucidum spore oil has the effect of preventing and treating the occurrence and development of pulmonary nodules, and can significantly reduce the growth of granulomas in mice with granulomatous pulmonary nodules, while having no significant effect on the weight and overall health of the mice, indicating that Ganoderma lucidum spore oil has good safety. The Ganoderma lucidum spore oil of the present invention has the beneficial effect of being used to prepare a drug for preventing or treating pulmonary nodules.

[0071] The above disclosure is only the preferred embodiment of the present invention, which certainly cannot be used to limit the scope of the present invention. Therefore, equivalent changes made according to the claims of the present invention are still within the scope of the present invention.

Claims

1. An application of Ganoderma lucidum spore oil, characterized in that: Used for preparing preparations for preventing and treating sarcoidosis.

2. The use according to claim 1, characterized in that: The sarcoidosis includes pulmonary nodules.

3. The use according to claim 1, characterized in that: The sarcoidosis includes benign granulomatous nodules.

4. The use according to claim 1, characterized in that: The causes of sarcoidosis include one or more of microorganisms and their related derivatives, and exogenous particles.

5. The use according to claim 4, characterized in that: The microorganisms include one or more of bacteria, viruses, and mycoplasma; the exogenous particles include one or more of carbon nanoparticles, metal particles, and smoke.

6. The use according to claim 1, characterized in that: The ganoderma spore oil comprises one or more components of triglyceride, ergosterol and vitamin E.

7. The use according to claim 1, characterized in that: In the ganoderma spore oil, the content of triglycerides is greater than 60%, the content of ergosterol is 0.1% to 0.5%, the content of vitamin E is 0.5% to 4%, and no ganoderma polysaccharide compounds are detected.

8. A preparation obtained by the use as claimed in claim 1.

9. The preparation according to claim 8, characterized in that The preparation comprises a drug; the drug further comprises a pharmaceutically acceptable carrier.

10. The preparation according to claim 9, characterized in that The medicine includes one of tablets, capsules, lozenges, granules, granules, pills, powders, ointments, pills, suspensions, solutions, emulsions, injections, suppositories, creams, sprays, drops or patches.

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