Application of ganoderma lucidum spore oil to reduction of tumor susceptibility

Abstract

The invention relates to the technical field of medicines, and discloses an application of ganoderma lucidum spore oil to reduction of tumor susceptibility, animal model tests and patient use condition investigation find that the ganoderma lucidum spore oil has an obvious effect of improving mouse emotional stress mediated tumor susceptibility, can also reduce emotional stress of tumor patients, and has a good application prospect. The compound has good safety, so that the compound can be used for preparing medicines for reducing tumor susceptibility.

Description

Technical Field

[0001] The invention relates to the field of medical technology, and in particular to an application of ganoderma lucidum spore oil in reducing tumor susceptibility. 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 immune regulation, lowering blood sugar, lowering blood lipids, anti-inflammatory, 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 extracting them with supercritical carbon dioxide. 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 traditional Chinese medicine, human psychological activities are called emotions. Emotions refer to emotions such as joy, anger, worry, thought, sadness, shock, and fear in life. They are the instinctive comprehensive reflection of people when they contact and understand objective things. Emotions are an important cause of disease. The stress state caused by emotional stress is not a disease in itself, but the low physiological function of the body caused by stress will lead to "susceptibility" to diseases. Diseases caused by emotional changes affecting the imbalance of yin and yang in the body are called emotional diseases. Modern medical research shows that 70% of diseases are related to psychological stress. Due to the increase in social competition pressure and changes in living environment, emotional stress has increasingly affected the occurrence and development of diseases. Regarding the relationship between emotional stress and the "susceptibility" of tumors, traditional Chinese medicine believes that the occurrence of tumors can be summarized as symptoms such as loss of vital energy, improper diet, water and soil, external attack of the six evils, emotional injury leading to internal heat and toxicity, blood stasis, and phlegm condensation, especially emphasizing that emotional factors are important causes of the occurrence and development of tumors. Clinical research has found that cancer patients generally have long-term emotional abnormalities or have suffered major mental trauma before and after the onset of the disease. Animal experimental studies have also found that chronic restraint stress simulating psychological stress can promote tumor growth and metastasis. Therefore, effectively reducing the susceptibility to tumors caused by emotional stress may become one of the important strategies for preventing and treating tumors. Summary of the invention

[0004] The purpose of the present invention is to provide an application of Ganoderma lucidum spore oil to reduce tumor susceptibility. Traditional Chinese medicine theory, clinical research and animal experimental research have all found that emotional factors are important causes of the occurrence and development of tumors. Based on the above reasons and needs, it is found that Ganoderma lucidum spore oil has a new property of reducing the susceptibility to tumors caused by emotional stress, and then a new application of Ganoderma lucidum spore oil as a medicine is provided, that is, the application in the preparation of a medicine for preventing or treating the susceptibility to tumors caused by emotional stress.

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

[0006] An application of ganoderma lucidum spore oil for preparing a preparation for reducing the susceptibility to tumors caused by emotional stress.

[0007] Preferably, it is used to prepare a preparation for inhibiting the occurrence and development of tumors under emotional stress.

[0008] The present invention finds through animal model experiments and patient usage surveys that Ganoderma lucidum spore oil has a significant effect on improving the emotional stress-mediated tumor susceptibility of mice, and can also reduce the emotional stress of tumor patients, and has good safety. Therefore, it can be used to prepare drugs that reduce the susceptibility of tumors caused by emotional stress.

[0009] Preferably, the tumor includes one or more of ovarian cancer, lung cancer, nasopharyngeal cancer, gastric cancer, breast cancer, colorectal cancer, liver cancer, uterine fibroids, blood tumors, and neuroendocrine tumors.

[0010] Preferably, in the Ganoderma lucidum spore oil, the content of triglycerides is greater than 60%, and Ganoderma lucidum polysaccharides and Ganoderma lucidum triterpenoids are not detected.

[0011] Preferably, the triglyceride content in the Ganoderma lucidum spore oil is greater than 90%.

[0012] Preferably, the Ganoderma lucidum spore oil further comprises vitamin E; the mass content of the vitamin E comprises 0.5% to 4%.

[0013] Preferably, the mass content of vitamin E in the Ganoderma lucidum spore oil is 2%.

[0014] A preparation obtained by use as described above.

[0015] Preferably, the preparation comprises a drug; the drug further comprises a pharmaceutically acceptable carrier.

[0016] 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.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] Disclosed is a use of ganoderma lucidum spore oil for reducing tumor susceptibility,

[0019] The triglyceride content in the ganoderma spore oil of the present invention is greater than 60%, the ergosterol content is 0% to 0.5%, the vitamin E content is 0.5% to 4%, and ganoderma polysaccharides and ganoderma triterpenoids are not detected. A large number of studies have found that the main components of ganoderma that play a calming effect (such as anti-anxiety, anti-depression, anticonvulsant, improving sleep, improving learning and memory, etc.) are polysaccharides and triterpenoids. At the same time, most studies believe that ganoderma spore oil has the effect of enhancing immunity and anti-tumor because it is rich in ganoderma triterpenoids. Therefore, the research on the composition of ganoderma spore oil mainly focuses on the enrichment of ganoderma triterpenoids. According to the existing research results, it should be considered that with the decrease in the content of triterpenoids, the various effects of spore oil will be weakened or even disappear.

[0020] However, the present invention has conducted experiments and research on Ganoderma lucidum components other than polysaccharides and triterpenoid compounds, and unexpectedly found that Ganoderma lucidum spore oil with a triglyceride content of more than 60%, an ergosterol content of 0% to 0.5%, and a vitamin E content of 0.5% to 4% has a significant effect of reducing the susceptibility to tumors caused by emotional stress, and can significantly inhibit the occurrence and development of tumors in mice under emotional stress. When used in a variety of different tumor patients, it can also significantly reduce the scores of multiple emotional stress-related symptoms, while having no significant effect on the weight and overall health of mice, as well as the health of tumor patients, and no obvious adverse reactions are observed, indicating that it has good safety. The Ganoderma lucidum spore oil of the present invention has the beneficial effect of being used to prepare drugs for preventing or treating emotional stress-induced tumor susceptibility, and at the same time, the triterpenoid compounds enriched and separated are also expected to develop other new products with higher triterpenoid content. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is the HPLC component detection result of the Ganoderma lucidum spore oil in the embodiment of the present invention;

[0022] Figure 2 This is the TLC polysaccharide detection result of the Ganoderma lucidum spore oil in the embodiment of the present invention. DETAILED DESCRIPTION

[0023] 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.

[0024] Example 1

[0025] Preparation and component detection of Ganoderma lucidum spore oil

[0026] The preparation method of the ganoderma spore oil used in the present invention is as follows:

[0027] 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.

[0028] 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 0.42%, peaks 2-7 are different triglyceride components with a total content of 94.31%, and no peak representing triterpenoid components was found.

[0029] 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:

[0030] 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.

[0031] 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.

[0032] 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.

[0033] 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 appeared at the corresponding positions of Ganoderma lucidum spore oil, indicating that no polysaccharide or monosaccharide components were detected in Ganoderma lucidum spore oil.

[0034] Example 2

[0035] Ganoderma lucidum spore oil alleviates tumor development in tumor-susceptible mice induced by emotional stress

[0036] 1. Experimental Animals

[0037] SPF C57BL / 6 mice (female, 3-4 weeks old) were housed in an SPF animal room with 12 h light / 12 h dark, temperature of 22±2°C, and humidity of 50±5%.

[0038] 2. Experimental Materials

[0039] Tumor cells: luciferase-labeled mouse ovarian epithelial cancer cells (ID8-Luc cells).

[0040] Ganoderma lucidum spore oil: the Ganoderma lucidum spore oil obtained in Example 1 was used.

[0041] 3. Experimental Methods

[0042] (1) Establishment and drug administration of the emotional stress-induced tumor susceptibility mouse model

[0043] After 2 weeks of adaptive feeding, the mice were randomly divided into 3 groups: tumor control group, stress model group, and stress + Ganoderma lucidum spore oil group. Starting from 7 days before tumor injection, the animals in the tumor control group and stress model group were given sterile water by gavage every day, and the animals in the stress + Ganoderma lucidum spore oil group were given Ganoderma lucidum spore oil by gavage every day, with a dose of 5g / kg, for 28 consecutive days.

[0044] On day 0, mice in each group were intraperitoneally injected with ID8-Luc tumor cells. Each mouse was injected with 0.5 mL of 1×10 7 Except for the tumor control group, mice in other groups were subjected to acute restraint stress for 18 h on the 3rd day before injection, the 3rd day after injection, the 9th day after injection, and the 15th day after injection to simulate emotional stress and establish an emotional stress-mediated tumor-susceptible mouse model.

[0045] The specific operation of restraint stress on mice is as follows: use an electric soldering iron to drill small holes on the tip, body and lid of a 50 mL centrifuge tube for the mouse to breathe, and place the mouse head toward the tip in the 50 mL centrifuge tube, with the tail passing through the small hole in the lid. During the restraint period, the mice were deprived of water and food.

[0046] Mice were sacrificed on day 22 after tumor injection.

[0047] (2) In vivo imaging to monitor tumor growth

[0048] Tumor growth in mice was monitored by in vivo imaging on days 7 and 21 after tumor injection. Each mouse was anesthetized by intraperitoneal injection of 0.2 mL of 4% chloral hydrate solution and injected with 0.2 mL of D-fluorescein. After the injection, wait 15 minutes for the anesthesia to take effect and the fluorescence intensity to reach the highest. The pre-anesthetized mouse was placed in the center of the imaging box, the door was gently closed, and the fluorescence intensity of the mouse was detected.

[0049] (3) Flow cytometry to detect the number of residual tumor cells in vivo

[0050] After extracting the mouse peritoneal lavage fluid, centrifuge at 4°C for 5 minutes and discard the supernatant. After fixing, permeabilizing and blocking the cells, PE anti-Firefly luciferase antibody was used to label ID8-Luc tumor cells at room temperature and detected by flow cytometry.

[0051] 4. Experimental Results

[0052] During the experiment, the animals in each group were generally in good condition. The results of animal weight test are shown in Table 1. Before tumor injection, some animals lost weight due to restraint stress, but there was no significant difference in animal weight among the groups.

[0053] Table 1 Animal weight test results (g)

[0054]

[0055] The results of the effect of Ganoderma lucidum spore oil on tumor growth in mice susceptible to tumors induced by emotional stress are shown in Table 2. On the 21st day after tumor loading, stress significantly increased the fluorescence intensity in mice compared with the tumor control group (P < 0.001). The results showed that emotional stress can promote tumor growth. Compared with the stress model group, Ganoderma lucidum spore oil can significantly reduce the fluorescence intensity in mice (P < 0.001). The results showed that Ganoderma lucidum spore oil can alleviate the occurrence and development of tumors in mice susceptible to tumors induced by emotional stress.

[0056] Table 2 Effect of Ganoderma lucidum spore oil on tumor growth in tumor-bearing mice with emotional stress

[0057]

[0058] Note: Compared with the tumor control group, *** P<0.001, compared with the stress model group, ### P<0.001

[0059] The results of the effect of Ganoderma lucidum spore oil on the number of tumor cells in tumor-bearing mice with emotional stress are shown in Table 3. Compared with the tumor control group, stress can significantly increase the number of tumor cells in the model group mice (P < 0.001). Compared with the stress model group, Ganoderma lucidum spore oil can significantly reduce the number of tumor cells in stressed mice (P < 0.01).

[0060] Table 3 Effect of Ganoderma lucidum spore oil on the number of tumor cells in tumor-bearing mice with emotional stress

[0061]

[0062] Note: Compared with the tumor control group, *** P<0.001, compared with the stress model group, ## P<0.01

[0063] The above studies established an emotional stress-induced tumor-susceptible mouse model and, with the help of small animal in vivo imaging and flow cytometry, showed that Ganoderma lucidum spore oil with a triglyceride content of >90% and an ergosterol content of 0% to 0.5% can alleviate tumor progression by reducing the number of tumor cells in emotional stress-induced tumor-bearing mice.

[0064] Example 3

[0065] Observation on the effectiveness of Ganoderma lucidum spore oil in relieving emotional stress in patients with tumors

[0066] 1. Research subjects

[0067] Among the 88 cancer patients who took Ganoderma lucidum spore oil, 17 had emotional stress-related symptoms before taking it. The types of tumors they suffered mainly included lung cancer, nasopharyngeal cancer, gastric cancer, breast cancer, rectal cancer, liver cancer, uterine fibroids, blood tumors, neuroendocrine tumors, etc.

[0068] 2. Research Materials

[0069] Ganoderma lucidum spore oil: the Ganoderma lucidum spore oil obtained in Example 1 was used.

[0070] 3. Research Methods

[0071] The scale in the questionnaire was used to score the emotional stress-related symptoms of tumor patients, and the questionnaires of 17 patients who had relevant symptoms before taking the medicine were selected for statistical analysis.

[0072] 4. Research Results

[0073] Symptom scores before and after administration and statistical analysis results are shown in Table 4. No adverse reactions related to this product were observed in all tumor patients during the period of administration, and they were generally in good condition.

[0074] Table 4 Symptom scores before and after taking the medicine (only tumor patients with symptoms before taking the medicine were included)

[0075]

[0076] The results showed that after taking Ganoderma lucidum spore oil, symptomatic cancer patients showed improvement in most emotional stress-related scores, and the symptom scores were significantly reduced. Among them, there was a significant difference in the degree of improvement in "depression, loss of interest" and "insomnia or drowsiness", and there was also a certain improvement trend in "still feeling energetic after sleep" and "feeling tired due to emotional reactions". The above results show that Ganoderma lucidum spore oil has the effect of relieving emotional stress in cancer patients.

[0077] In summary, the present invention has conducted experiments and research on Ganoderma lucidum spore oil with a triglyceride content of more than 90%, an ergosterol content of 0% to 0.5%, and a vitamin E content of 0.5% to 4%, and found that Ganoderma lucidum spore oil has the effect of reducing the susceptibility to tumors caused by emotional stress, can significantly reduce the growth of tumors in mice under emotional stress, and can also significantly reduce the scores of multiple emotional stress-related symptoms in a variety of different tumor patients, while having no significant effect on the weight and overall health of mice, and the health of tumor patients, 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 drugs for preventing or treating susceptibility to tumors caused by emotional stress.

[0078] 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 to prepare preparations that reduce susceptibility to tumors caused by emotional stress.

2. The use according to claim 1, characterized in that: Used to prepare preparations that inhibit the occurrence and development of tumors under emotional stress conditions.

3. The use according to claim 1, characterized in that: The tumor includes one or more of ovarian cancer, lung cancer, nasopharyngeal cancer, gastric cancer, breast cancer, colorectal cancer, liver cancer, uterine fibroids, blood tumors, and neuroendocrine tumors.

4. The use according to claim 1, characterized in that: In the ganoderma spore oil, the content of triglyceride components is greater than 60%, and ganoderma polysaccharides and ganoderma triterpenoid compounds are not detected.

5. The use according to claim 4, characterized in that: The content of triglyceride components in the Ganoderma lucidum spore oil is greater than 90%.

6. The use according to claim 1, characterized in that: The ganoderma lucidum spore oil also includes vitamin E; the mass content of the vitamin E is 0.5% to 4%.

7. The use according to claim 6, characterized in that The mass content of vitamin E in the ganoderma lucidum spore oil is 2%.

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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