Use of ganoderma lucidum spore oil in inhibiting occurrence and development of tumors

Abstract

A use of ganoderma lucidum spore oil in inhibiting occurrence and development of tumors. The ganoderma lucidum spore oil has the effect of improving tumor susceptibility mediated by emotional stress in mice, can also reduce emotional stress in tumor patients, has good safety, and can be used for preparing a drug for inhibiting the occurrence and development of tumors.

Description

An application of Ganoderma lucidum spore oil in inhibiting tumor development and progression Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of Ganoderma lucidum spore oil in inhibiting the occurrence and development of tumors. Background Technology

[0002] Reishi mushroom is a precious treasure in the Chinese medicine treasury, often referred to as the "immortal herb." It can be used to treat restlessness, insomnia, palpitations, cough and asthma due to lung deficiency, shortness of breath due to weakness, and loss of appetite. Reishi spores are extremely small spores ejected from the cap of the mushroom during its mature growth. They are the reproductive cells of Reishi and contain all of its genetically active substances. Modern pharmacological research shows that Reishi spores have effects such as enhancing immune regulation, lowering blood sugar and blood lipids, anti-inflammation, anti-hypoxia ability, and scavenging free radicals. Reishi spore oil is a fat-soluble active substance obtained from Reishi spores through cell wall breaking and supercritical carbon dioxide extraction. Studies have shown that it has effects such as enhancing immunity and protecting the liver. Current research generally considers the main effective components of Reishi to be Reishi polysaccharides and Reishi triterpenoids. Research on the various effects of Reishi has focused more on polysaccharides and triterpenoids. Research and efficacy analysis of other components in spore oil, such as the triglycerides which constitute the largest proportion, are currently less reported.

[0003] In Traditional Chinese Medicine (TCM), psychological activities are referred to as "emotions and mental states." These emotions include joy, anger, worry, pensiveness, grief, fright, and fear, and are an instinctive and comprehensive reflection of human beings when encountering and understanding objective things. Emotions and mental states are important causes of disease. The stress state itself is not a disease, but the resulting decline in bodily physiological functions leads to increased susceptibility to disease. Diseases caused by emotional changes affecting the body's Yin-Yang balance are called emotionally induced diseases. Modern medical research shows that 70% of diseases are related to psychological stress. Due to increased social competition and changes in living environment, emotional stress is increasingly influencing the occurrence and development of diseases. Regarding the relationship between emotional stress and susceptibility to tumors, TCM believes that the occurrence of tumors can be summarized as a deficiency of vital energy, improper diet and environment, external invasion of pathogenic factors, emotional distress leading to internal accumulation of heat toxins, blood stasis, and phlegm accumulation. It particularly emphasizes that emotional factors are an important cause of tumor development and progression. Clinical surveys have revealed that cancer patients commonly experience prolonged emotional distress or have suffered significant psychological trauma before or after the onset of their illness. Animal studies have also shown that chronic restraint stress, simulating psychological stress, can promote tumor growth and metastasis. Therefore, effectively reducing tumor susceptibility caused by emotional stress may be an important strategy for cancer prevention and treatment. Summary of the Invention

[0004] The purpose of this invention is to provide an application of Ganoderma lucidum spore oil in reducing tumor susceptibility. Traditional Chinese medicine theory, clinical surveys, and animal experiments have all found that emotional factors are an important cause of tumor development and progression. Based on the above reasons and needs, it has been discovered that Ganoderma lucidum spore oil possesses a novel property of reducing tumor susceptibility induced by emotional stress, thus providing a new application of Ganoderma lucidum spore oil as a medicine, namely, its application in the preparation of drugs for the prevention or treatment of tumor susceptibility induced by emotional stress.

[0005] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0006] An application of Ganoderma lucidum spore oil in the preparation of a formulation that reduces susceptibility to tumors induced by emotional stress.

[0007] Preferably, it is used to prepare a formulation that inhibits the occurrence and development of tumors under emotional stress.

[0008] This invention, through animal model experiments and patient usage surveys, found that Ganoderma lucidum spore oil has a significant effect on improving tumor susceptibility mediated by emotional stress in mice, and can also reduce emotional stress in cancer patients, with good safety. Therefore, it can be used to prepare drugs that reduce tumor susceptibility induced by emotional stress.

[0009] Preferably, the tumor includes one or more of the following: ovarian cancer, lung cancer, nasopharyngeal carcinoma, gastric cancer, breast cancer, rectal cancer, liver cancer, uterine fibroids, hematologic malignancies, and neuroendocrine tumors.

[0010] Preferably, the Ganoderma lucidum spore oil contains >60% triglycerides, and no Ganoderma lucidum polysaccharides or triterpenoids are detected.

[0011] Preferably, the content of triglyceride components in the Ganoderma lucidum spore oil is >90%.

[0012] Preferably, the Ganoderma lucidum spore oil also includes vitamin E; the vitamin E content includes 0.5% to 4% by mass.

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

[0014] A formulation obtained by application as described above.

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

[0016] Preferably, the drug comprises one of the following: tablets, capsules, lozenges, granules, powders, pills, powders, ointments, pills, suspensions, solutions, emulsions, injections, suppositories, creams, sprays, drops, or patches.

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

[0018] An application of Ganoderma lucidum spore oil in reducing tumor susceptibility is disclosed.

[0019] The Ganoderma lucidum spore oil of this invention contains >60% triglycerides, 0%–0.5% ergosterol, and 0.5%–4% vitamin E. No Ganoderma lucidum polysaccharides or triterpenoids were detected. Numerous studies have found that the main components of Ganoderma lucidum that exert its calming effects (such as anti-anxiety, antidepressant, anticonvulsant, sleep improvement, and memory enhancement) are polysaccharides and triterpenoids. Meanwhile, most studies suggest that the immune-enhancing and anti-tumor effects of Ganoderma lucidum spore oil are also due to its rich content of triterpenoids. Therefore, research on the composition of Ganoderma lucidum spore oil mainly focuses on the enrichment of triterpenoids. Based on existing research results, it should be considered that as the content of triterpenoids decreases, the various effects of spore oil will weaken or even disappear.

[0020] However, through experiments and research on Ganoderma lucidum components other than polysaccharides and triterpenoids, this invention unexpectedly discovered that Ganoderma lucidum spore oil with a triglyceride content >60%, ergosterol content of 0%–0.5%, and vitamin E content of 0.5%–4% significantly reduces tumor susceptibility induced by emotional stress. It can significantly inhibit the occurrence and development of tumors in mice under emotional stress, and when used in various cancer patients, it can also significantly reduce the scores of multiple emotional stress-related symptoms. It has no significant effect on the weight and overall health of mice, nor on the health of cancer patients, and no obvious adverse reactions were observed, suggesting good safety. The Ganoderma lucidum spore oil of this invention has the beneficial effect of being used to prepare drugs for the prevention or treatment of tumor susceptibility induced by emotional stress, and the enriched and isolated triterpenoids also hold promise for developing other new products with higher triterpenoid content. Attached Figure Description

[0021] Figure 1 shows the HPLC component detection results of Ganoderma lucidum spore oil in an embodiment of the present invention;

[0022] Figure 2 shows the TLC polysaccharide detection results of Ganoderma lucidum spore oil in an embodiment of the present invention. Detailed Implementation

[0023] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments are not intended to limit the present invention. Modifications or substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and substance of the present invention are all within 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 analysis of Ganoderma lucidum spore oil

[0026] The method for preparing Ganoderma lucidum spore oil used in this invention is as follows:

[0027] Ganoderma lucidum spore powder was subjected to cell wall disruption, followed by supercritical CO2 extraction to obtain crude Ganoderma lucidum spore oil. An appropriate amount of silica gel was dissolved in a petroleum ether-ethyl acetate mixture (v / v) of 8.5–9.5:0.5–1.5, and packed into a column using a wet packing method. The column diameter to height ratio was 1:10–20, and the mass ratio of crude Ganoderma lucidum spore oil to silica gel was 1:10–40. The crude Ganoderma lucidum spore oil was loaded onto the elution column and eluted with the above petroleum ether-ethyl acetate mixture under normal pressure. The eluent was collected, combined, concentrated under reduced pressure to remove the solvent, and vacuum dried. Vitamin E was added to achieve a content of 2% by mass to obtain the Ganoderma lucidum spore oil. Through separation and purification, a large amount of impurities and water were removed, resulting in a clear and transparent Ganoderma lucidum spore oil with stable component content.

[0028] The contents of ergosterol and triglycerides in Ganoderma lucidum spore oil were determined by high performance liquid chromatography (HPLC), as shown in Figure 1. Peak 1 is ergosterol with a content of 0.42%, and peaks 2-7 are different triglyceride components, with a total content of 94.31%. No peaks representing triterpenoid components were found.

[0029] Thin-layer chromatography (TLC) was used to detect whether Ganoderma lucidum spore oil contained polysaccharides, with starch and glucose selected as positive controls.

[0030] 1. Take 10g of Ganoderma lucidum spore oil, add 5ml of water to a separatory funnel, shake thoroughly, and allow to stand for separation. Separate the aqueous layer and discard the oil layer. Concentrate the aqueous layer under reduced pressure to 2ml, add 6ml of strong acid, shake quickly, and place in a water bath at 85℃ for 30min. Remove and let cool to room temperature before use.

[0031] 2. Take 10g of starch and prepare a 10% starch solution. Take 2ml of the solution, add 6ml of strong acid, shake quickly and mix well. Place in a water bath at 85℃ for 30 minutes. Remove and let cool to room temperature before use.

[0032] 3. Take 10 mg of anhydrous glucose and prepare a glucose solution containing 1 mg per ml. Take 2 ml of the solution, add 6 ml of strong acid, shake quickly, and place in a water bath at 85°C for 30 minutes. Remove and let cool to room temperature before use.

[0033] Five μl of each of the solutions obtained after the reactions with the three strong acids was spotted onto the same silica gel thin-layer plate. Methanol was used as the developing solvent. After development, the plates were dried at 105°C and the spots were observed, as shown in Figure 2. In the positive control group, spots representing monosaccharides appeared after hydrolysis of the starch and glucose solutions, while no spots were observed at the corresponding positions in the Ganoderma lucidum spore oil, indicating that no polysaccharides or monosaccharides were detected in the Ganoderma lucidum spore oil.

[0034] Example 2

[0035] Ganoderma lucidum spore oil alleviates tumor development and progression in mice with emotional stress-induced tumor susceptibility.

[0036] 1. Laboratory animals

[0037] SPF-grade C57BL / 6 mice (female, 3-4 weeks old) were housed in an SPF-grade animal room with 12h light / 12h darkness, at a temperature of 22±2℃ and a 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 administration of a mouse model of tumor susceptibility induced by emotional stress

[0043] After two weeks of acclimatization, mice were randomly divided into three groups: a tumor control group, a stress model group, and a stress + Ganoderma lucidum spore oil group. Starting seven days before tumor injection, the tumor control group and the stress model group were given sterile water by gavage daily, while the stress + Ganoderma lucidum spore oil group was given Ganoderma lucidum spore oil by gavage daily at a dose of 5 g / kg for 28 consecutive days.

[0044] Mice in each group were intraperitoneally injected with ID8-Luc tumor cells on day 0, with each mouse receiving 0.5 mL containing 1×10⁻⁶ cells. 7 Each group of mice was tested for tumor cells. Except for the tumor control group, the other groups of mice were subjected to acute restraint stress for 18 hours on days 3 before injection, 3 after injection, 9 after injection, and 15 after injection to simulate emotional stress and establish an emotional stress-mediated tumor-susceptible mouse model.

[0045] The specific procedure for restraint stress in mice is as follows: Use a soldering iron to drill small holes in the tip, body, and cap of a 50mL centrifuge tube to allow the mouse to breathe. Place the mouse head-first into the 50mL centrifuge tube, with its tail protruding through the small hole in the cap. During the restraint period, the mice are deprived of water and food.

[0046] Mice were sacrificed 22 days after tumor injection.

[0047] (2) In vivo imaging monitoring of tumor growth

[0048] Tumor growth in mice was monitored by in vivo imaging on days 7 and 21 post-tumor injection. Each mouse was anesthetized by intraperitoneal injection of 0.2 mL of 4% chloral hydrate solution and then injected with 0.2 mL of D-fluorescein. A 15-minute interval was allowed after injection to allow anesthesia to take effect and fluorescence intensity to reach its peak. The pre-anesthetized mice were then placed in the center of an imaging chamber, the door was gently closed, and fluorescence intensity was measured.

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

[0050] After extracting peritoneal lavage fluid from mice, the cells were centrifuged at 4°C for 5 min, and the supernatant was discarded. Cells were fixed, permeabilized, and blocked. ID8-Luc tumor cells labeled with PE anti-Firefly luciferase antibody were then incubated at room temperature and analyzed by flow cytometry.

[0051] 4. Experimental Results

[0052] During the experiment, the animals in all groups were in good general condition. The results of animal weight measurement are shown in Table 1. Before tumor injection, some animals experienced a decrease in weight due to restraint stress, but there was no significant difference in weight among the groups.

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

[0054] The results of the effects of Ganoderma lucidum spore oil on tumor growth in mice susceptible to tumors due to emotional stress are shown in Table 2. On day 21 after tumor implantation, compared with the tumor control group, stress significantly increased the fluorescence intensity in mice (P < 0.001). The results indicate that emotional stress can promote tumor growth. Compared with the stress model group, Ganoderma lucidum spore oil significantly reduced the fluorescence intensity in mice (P < 0.001). These results suggest that Ganoderma lucidum spore oil can alleviate tumor development and progression in mice susceptible to tumors due to emotional stress.

[0055] Table 2. Effects of Ganoderma lucidum spore oil on tumor growth in mice subjected to emotional stress and tumor bearing. Note: Compared with the tumor control group, *** P < 0.001, compared with the stress model group, ### P < 0.001

[0056] The effects of Ganoderma lucidum spore oil on the number of tumor cells in mice subjected to emotional stress and bearing tumors are shown in Table 3. Compared with the tumor control group, stress significantly increased the number of tumor cells in the model group mice (P < 0.001). Compared with the stress model group, Ganoderma lucidum spore oil significantly reduced the number of tumor cells in stressed mice (P < 0.01).

[0057] Table 3. Effects of Ganoderma lucidum spore oil on the number of tumor cells in mice subjected to emotional stress and tumor bearing. Note: Compared with the tumor control group, *** P < 0.001, compared with the stress model group, ## P < 0.01

[0058] The above studies, through the establishment of a mouse model of emotional stress coupled with tumor susceptibility, and with the aid of small animal in vivo imaging and flow cytometry, have shown that Ganoderma lucidum spore oil with triglyceride content >90% and ergosterol content of 0%–0.5% can alleviate tumor progression by reducing the number of tumor cells in emotional stress-bearing tumor mice.

[0059] Example 3

[0060] Observation on the effectiveness of Ganoderma lucidum spore oil in relieving emotional stress in cancer patients

[0061] 1. Research Subjects

[0062] Among the 88 cancer patients who took Ganoderma lucidum spore oil, 17 had symptoms related to emotional stress before taking it. The types of cancer they had mainly included lung cancer, nasopharyngeal carcinoma, gastric cancer, breast cancer, rectal cancer, liver cancer, uterine fibroids, hematologic malignancies, and neuroendocrine tumors.

[0063] 2. Research Materials

[0064] Ganoderma lucidum spore oil: The Ganoderma lucidum spore oil obtained in Example 1 was used.

[0065] 3. Research Methods

[0066] The questionnaire was used to score the emotional stress-related symptoms of cancer patients. The questionnaires of 17 patients who had related symptoms before taking the medication were selected for statistical analysis.

[0067] 4. Research Results

[0068] The symptom scores and statistical analysis results before and after administration are shown in Table 4. No adverse reactions related to this product were observed in any of the cancer patients during the course of treatment, and their general condition was good.

[0069] Table 4 Symptom scores before and after administration (only including cancer patients with symptoms before administration)

[0070] The results showed that after taking Ganoderma lucidum spore oil, symptomatic cancer patients experienced improvement on most emotional stress-related scores, with a significant decrease in symptom scores. Significant differences were observed in the degree of improvement for "depressed mood and decreased interest" and "insomnia or somnolence," while there was also a trend towards improvement in "feeling unrested after sleep" and "feeling fatigue due to emotional reactions." These results indicate that Ganoderma lucidum spore oil has an effect in relieving emotional stress in cancer patients.

[0071] In summary, this invention, through experiments and research on Ganoderma lucidum spore oil with a triglyceride content >90%, ergosterol content 0%–0.5%, and vitamin E content 0.5%–4%, found that Ganoderma lucidum spore oil has the effect of reducing tumor susceptibility induced by emotional stress. It can significantly reduce tumor growth in mice under emotional stress and significantly reduce the scores of multiple emotional stress-related symptoms in various cancer patients. However, it has no significant effect on the weight and overall health of mice, nor on the health of cancer patients, suggesting that Ganoderma lucidum spore oil has good safety. The Ganoderma lucidum spore oil of this invention has the beneficial effect of being used to prepare drugs for the prevention or treatment of tumor susceptibility induced by emotional stress.

[0072] The above-disclosed embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention. Therefore, any equivalent variations made in accordance with 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 agents that reduce susceptibility to tumors caused by emotional stress.

2. The application as described in claim 1, characterized in that, This is used to prepare agents that inhibit tumor development and progression under emotional stress.

3. The application as described in claim 1, characterized in that, The tumors include one or more of the following: ovarian cancer, lung cancer, nasopharyngeal carcinoma, gastric cancer, breast cancer, rectal cancer, liver cancer, uterine fibroids, hematologic malignancies, and neuroendocrine tumors.

4. The application as described in claim 1, characterized in that, The Ganoderma lucidum spore oil contained >60% triglycerides, while Ganoderma lucidum polysaccharides and Ganoderma lucidum triterpenoids were not detected.

5. The application as described in claim 4, characterized in that, The Ganoderma lucidum spore oil contains more than 90% triglycerides.

6. The application as described in claim 1, characterized in that, The Ganoderma lucidum spore oil also includes vitamin E; the vitamin E content is 0.5% to 4% by mass.

7. The application as described in claim 6, characterized in that, The vitamin E content in the Ganoderma lucidum spore oil is 2% by mass.

8. A formulation obtained by the application as described in claim 1.

9. The formulation as described in claim 8, characterized in that, The formulation includes a drug; the drug further contains a pharmaceutically acceptable carrier.

10. The formulation of claim 9, characterized in that, The drug includes one of the following: tablets, capsules, lozenges, granules, powders, pills, powders, ointments, pills, suspensions, solutions, emulsions, injections, suppositories, creams, sprays, drops, or patches.

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