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

TECHNICAL FIELD

[0001] The present disclosure relates to the field of medical technology, and specifically, to a use of Ganoderma lucidum spore oil in inhibiting occurrence and development of tumors. BACKGROUND

[0002] Ganoderma lucidum, as a treasure in the traditional Chinese medicine of China, has long been known as “magical herb”. It is indicated for uneasiness, insomnia and palpitation, lung deficiency cough and asthma, consumptive disease with shortness of breath, and poor appetite. Ganoderma lucidum spores are ultrafine spores projected from pileus of Ganoderma lucidum during the mature growth stage. They are reproductive cells of Ganoderma lucidum, and possess all genetic active substances of Ganoderma lucidum. Modern pharmacological studies have shown that Ganoderma lucidum spores have effects such as enhancing immune regulation, lowering blood glucose, lowering blood lipids, anti-inflammation, anti-hypoxia, and scavenging free radicals. Ganoderma lucidum spore oil is a lipid-soluble active substance obtained from Ganoderma lucidum spores through wall-breaking and supercritical carbon dioxide extraction. Studies have shown that Ganoderma lucidum spore oil has effects such as enhancing immunity and protecting the liver. Existing studies generally consider that the main active components of Ganoderma lucidum are Ganoderma lucidum polysaccharides and Ganoderma lucidum triterpenoids. Research on various effects of Ganoderma lucidum is also mainly focused on polysaccharides and triterpenoids. However, there are currently few reports on component studies and efficacy analysis of other components in the spore oil, such as triglyceride compounds that account for the largest proportion.

[0003] In traditional Chinese medicine, psychological activities of human beings are referred to as emotions. Emotions refer to feelings such as joy, anger, anxiety, pensiveness, grief, fear, and fright in life, and are instinctive comprehensive responses generated when human beings contact and understand objective things. Emotions are important pathogenic factors. The stress state caused by emotional stress is not a disease itself, but physiological hypofunction of the body caused by stress can lead to disease “susceptibility”. Diseases caused by imbalance of yin and yang in the body due to emotional changes are referred to as emotional pathogenicity. Modern medical studies have shown that 70% of diseases are associated with psychological stress. Due to increased social competition pressure, changes in living environments and the like, emotional stress increasingly affects the onset and progression of diseases. Regarding the relationship between emotional stress and tumor “susceptibility”, traditional Chinese medicine considers that the occurrence of tumors can be attributed to deficiency of vital qi, improper diet and living environment, invasion of six exogenous pathogenic factors, and emotional injuries, which lead to syndromes such as internal accumulation of toxic heat, blood stasis obstruction, and phlegm turbidity coagulation. In particular, emotional factors are emphasized as major pathogenic causes for the occurrence and progression of tumors. Clinical investigations have found that tumor patients generally have experienced long-term emotional abnormalities or have suffered major psychological trauma before and after disease onset. Animal experimental studies have also found that chronic restraint stress, which mimics psychological stress, can promote tumor growth and metastasis. Therefore, effectively reducing tumor susceptibility caused by emotional stress may become one of the important strategies for prevention and treatment of tumors. SUMMARY

[0004] An objective of the present disclosure is to provide a use of Ganoderma lucidum spore oil in reducing tumor susceptibility. Theories of traditional Chinese medicine, clinical investigations, and animal experimental studies have all found that emotional factors are important pathogenic causes of occurrence and progression of tumors. Based on the above reasons and requirements, it has been discovered that Ganoderma lucidum spore oil has a novel property of reducing emotional stress-induced tumor susceptibility, thereby providing a novel use of Ganoderma lucidum spore oil as a medicament, namely use in the preparation of a medicament for preventing or treating emotional stress-induced tumor susceptibility.

[0005] To solve the above technical problems, the present disclosure adopts the following technical solutions:

[0006] Provided is a use of Ganoderma lucidum spore oil in preparing a formulation for reducing emotional stress-induced tumor susceptibility.

[0007] Preferably, it is used for preparing a formulation that inhibits tumor occurrence and progression under emotional stress.

[0008] Through animal model experiments and patient use investigations, the present disclosure has found that Ganoderma lucidum spore oil has a significant effect of improving emotional stress-mediated tumor susceptibility in mice, and can also reduce emotional stress in tumor patients, with good safety. Therefore, it can be used for preparing a medicament that reduces emotional stress-induced tumor susceptibility.

[0009] Preferably, the tumor includes one or more selected from the group consisting of ovarian cancer, lung cancer, nasopharyngeal carcinoma, gastric cancer, breast cancer, rectal cancer, liver cancer, uterine fibroids, hematological tumors, and neuroendocrine tumors.

[0010] Preferably, in the Ganoderma lucidum spore oil, a content of triglyceride components is greater than 60%, and no Ganoderma lucidum polysaccharides or Ganoderma lucidum triterpenoids are detected.

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

[0012] Preferably, the Ganoderma lucidum spore oil further includes vitamin E; and a mass content of the vitamin E ranges from 0.5% to 4%.

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

[0014] Provided is a formulation obtained by the use as described above.

[0015] Preferably, the formulation includes a medicament; and the medicament further contains a pharmaceutically acceptable carrier.

[0016] Preferably, the medicament includes one selected from the group consisting of a tablet, a capsule, a buccal tablet, a granule, a soluble granule, a pellet, a powder, an ointment, a Dan, a suspension, a solution, an emulsion, an injection, a suppository, a cream, a spray, a drop, and a patch.

[0017] Compared with the prior art, the implementation of the present disclosure offers the following beneficial effects:

[0018] A use of Ganoderma lucidum spore oil in reducing tumor susceptibility is disclosed.

[0019] In the Ganoderma lucidum spore oil of the present disclosure, the content of triglycerides is greater than 60%, the content of ergosterol ranges from 0% to 0.5%, the content of vitamin E ranges from 0.5% to 4%, and no Ganoderma lucidum polysaccharides or Ganoderma lucidum triterpenoids are detected. A large number of studies have found that the main components in Ganoderma lucidum that exert mind-calming effects (such as anti-anxiety, anti-depression, anti-convulsion, improving sleep, and learning and memory improvement) are polysaccharides and triterpenoids. Meanwhile, most studies consider that the effects of Ganoderma lucidum spore oil in enhancing immunity and resisting tumors are also attributed to its richness in Ganoderma lucidum triterpenoids. Therefore, the research on components of Ganoderma lucidum spore oil mainly focuses on enrichment of Ganoderma lucidum triterpenoids. According to existing research results, it should be considered that, with the decrease in the content of triterpenoids, various effects of the spore oil would be weakened or even disappear accordingly.

[0020] However, through experiments and studies on components of Ganoderma lucidum other than polysaccharides and triterpenoids, the present disclosure unexpectedly found that Ganoderma lucidum spore oil with a triglyceride content greater than 60%, an ergosterol content ranging from 0% to 0.5%, and a vitamin E content ranging from 0.5% to 4% has a significant effect of reducing emotional stress-induced tumor susceptibility, can significantly inhibit the in vivo occurrence and progression of tumors in mice under emotional stress, and can also significantly reduce scores of multiple emotional stress-related symptoms when used in patients with various types of tumors. It has no significant impact on body weight and overall health status of mice, as well as health status of tumor patients, and no obvious adverse reactions are observed, indicating good safety. The Ganoderma lucidum spore oil of the present disclosure has the beneficial effect of being usable for preparing a medicament for preventing or treating emotional stress-induced tumor susceptibility. Meanwhile, the triterpenoids enriched and separated at the same time are also expected to be developed into other new products with higher triterpenoid contents. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] FIG. 1 shows an HPLC component detection result of Ganoderma lucidum spore oil according to an embodiment of the present disclosure; and

[0022] FIG. 2 shows a TLC polysaccharide detection result of Ganoderma lucidum spore oil according to an embodiment of the present disclosure. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0023] To better illustrate the objectives, technical solutions, and advantages of the present disclosure, the present disclosure will be further described below with reference to the drawings and specific embodiments, so that those skilled in the art could better understand and implement the present disclosure. However, the embodiments provided are not intended to limit the present disclosure. Modifications or replacements made to the methods, steps, or conditions of the present disclosure without departing from the spirit and essence of the present disclosure shall fall within the scope of the present disclosure. 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] A method for preparing the Ganoderma lucidum spore oil used in the present disclosure is as follows:

[0027] Ganoderma lucidum spore powder was taken and subjected to wall-breaking, and then extracted with supercritical CO2 to obtain a crude Ganoderma lucidum spore oil. An appropriate amount of silica gel was taken and dissolved in a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8.5-9.5:0.5-1.5, and the resultant was wet-packed into a column to obtain an elution column. The ratio of column diameter to column height of the elution column was 1:10-20, and the mass ratio of the crude Ganoderma lucidum spore oil to the silica gel was 1:10-40. The crude Ganoderma lucidum spore oil was taken and loaded onto the elution column, eluted with the above mixed solution of petroleum ether and ethyl acetate under atmospheric pressure. Eluates were collected, combined, concentrated under reduced pressure to remove the solvents, and vacuum-dried. Vitamin E was added to make its mass content 2%, thereby obtaining the Ganoderma lucidum spore oil. Through separation and purification, a large amount of impurities and moisture were removed, and the obtained Ganoderma lucidum spore oil was clear and transparent, with stable component contents.

[0028] The contents of ergosterol and triglycerides in the Ganoderma lucidum spore oil were detected by high-performance liquid chromatography (HPLC). As shown in FIG. 1, peak 1 represents ergosterol, with a content of 0.42%; peaks 2-7 respectively represent different triglyceride components, with a total content of 94.31%; and no peak representing triterpenoid components was found.

[0029] Whether the Ganoderma lucidum spore oil contained polysaccharide components was detected by thin-layer chromatography (TLC), with starch and glucose selected as positive controls:

[0030] 1. 10 g of the Ganoderma lucidum spore oil and 5 mL of water were added into a separatory funnel, thoroughly shaken, and allowed to stand for phase separation. An aqueous layer was separated, and an oil layer was discarded. The aqueous layer was concentrated under reduced pressure to 2 mL, and 6 mL of a strong acid was added. The resulting mixture was rapidly shaken, and placed in a water bath at 85 °C for 30 min. The resulting mixture was removed, and cooled to room temperature for subsequent use.

[0031] 2. 10 g of starch was taken to prepare a 10% starch solution. 2 mL of the starch solution was mixed with 6 mL of a strong acid, rapidly shaken, and placed in a water bath at 85 °C for 30 min. The resulting mixture was removed, and cooled to room temperature for subsequent use.

[0032] 3. 10 mg of anhydrous glucose was taken to prepare a glucose solution containing 1 mg of glucose per 1 mL. 2 mL of the glucose solution was mixed with 6 mL of a strong acid, rapidly shaken, and placed in a water bath at 85 °C for 30 min. The resulting mixture was removed, and cooled to room temperature for subsequent use.

[0033] 5 uL of each of the above three solutions after reaction with the strong acid was separately taken and spotted onto the same silica gel thin-layer plate, developed with methanol as a developing solvent, and dried at 105 °C, and the spots thereof were observed, as shown in FIG. 2. After hydrolysis, both the starch solution and the glucose solution in the positive control groups exhibited spots representing monosaccharides, while no spot was showed at the corresponding position of the Ganoderma lucidum spore oil, indicating that no polysaccharide or monosaccharide components were detected in the Ganoderma lucidum spore oil.

[0034] Example 2

[0035] Ganoderma lucidum Spore Oil Alleviates Tumor Occurrence and Progression in Mice with Emotional Stress-Induced Tumor Susceptibility

[0036] 1. Experimental animals

[0037] Specific pathogen-free (SPF)-grade C57BL / 6 mice (female, 3-4 weeks old), housed in an SPF-grade animal facility, with 12 h light / 12 h darkness, at a temperature of 22 ± 2 °C, 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 of mouse models of emotional stress-induced tumor susceptibility and administration

[0043] After 2 weeks of acclimatization, the mice were randomly divided into 3 groups: a tumor control group, a stress model group, and a stress + Ganoderma lucidum spore oil group. Beginning from 7 days before tumor cell injection, animals in the tumor control group and the stress model group were intragastrically administered sterile water daily, and animals in the stress + Ganoderma lucidum spore oil group were intragastrically administered the Ganoderma lucidum spore oil daily, at a dose of 5 g / kg for 28 consecutive days.

[0044] On day 0, mice in each group were intraperitoneally injected with ID8-Luc tumor cells. Each mouse received 0.5 mL containing 1*107 tumor cells. Except for the tumor control group, mice in the remaining groups were subjected to acute restraint stress for 18 h on day 3 before injection and on days 3, 9, and 15 after injection to mimic emotional stress, establishing mouse models of emotional stress-mediated tumor susceptibility.

[0045] The specific operation of mouse restraint stress was as follows: Small holes were drilled in the tip, body, and cap of a 50 mL centrifuge tube with an electric soldering iron to allow the mice to breathe. Each mouse was placed into the 50 mL centrifuge tube, with its head facing the tip and its tail passed through the small hole in the cap. The mice were deprived of food and water during the restraint period.

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

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

[0048] Tumor growth in the 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 then injected with 0.2 mL of D-luciferin. After the injections, wait for 15 min for anesthesia to take effect and fluorescence intensity to reach the maximum. The pre-anesthetized mice were placed in the center of an imaging chamber. The door was gently closed, and fluorescence intensity of the mice was detected.

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

[0050] After extracting the peritoneal lavage fluid from the mice, the fluid was centrifuged at 4 °C for 5 min, and the supernatant was discarded. After fixation, permeabilization, and blocking of the cells, the ID8-Luc tumor cells were labeled by incubation with a PE anti-Firefly luciferase antibody at room temperature, and detected using a flow cytometer.

[0051] 4. Experimental results

[0052] During the experimental period, animals in each group exhibited good general conditions. The detection results of animal body weight are shown in Table 1. Before tumor injection, the body weight of some animals decreased due to restraint stress, while no significant difference in body weight was observed among the groups.

[0053] Table 1 Animal Body Weight Detection Results (g) Before Group                                  Before injection      Before sampling administration Tumor control group        13.82 ± 0.41         13.64 ± 0.43         17.45 ± 0.39 14.23 ± 0.33 12.52 ± 0.30 17.40 ± 0.21 Stress model group Stress + Ganoderma 13.94 ± 0.36          12.78 ± 0.27          17.27 ± 0.23 lucidum spore oil group

[0054] The results of the effect of Ganoderma lucidum spore oil on tumor growth in mice with emotional stress-induced tumor susceptibility are shown in Table 2. On day 21 after tumor bearing, compared with the tumor control group, stress can significantly enhance fluorescence intensity in the mice (P < 0.001). The results show that emotional stress can promote tumor growth. Compared with the stress model group, Ganoderma lucidum spore oil can significantly reduce the fluorescence intensity in the mice (P < 0.001). The results demonstrate that Ganoderma lucidum spore oil can alleviate tumor occurrence and progression in mice with emotional stress-induced tumor susceptibility.

[0055] Table 2 Effect of Ganoderma lucidum Spore Oil on Tumor Growth in Emotionally Stressed Tumor-Bearing Mice Group         _ Fluorescence intensity (x104) (p / s / cm2 / sr) Day 7 after injection Day 21 after injection Tumor control group 0.80 ± 0.18 2.00 ± 0.33 Stress model group 1.18 ± 0.13 4.44 ± 1.05*** Stress + Ganoderma lucidum spore oil group 0.66 ± 0.15 1.25 ± 0.17### Note: ***P < 0.001 compared with the tumor control group; ###P < 0.001 compared with the stress model group.

[0056] The results of the effect of Ganoderma lucidum spore oil on the number of tumor cells in emotionally stressed tumor-bearing mice are shown in Table 3. Compared with the tumor control group, stress can significantly increase the number of tumor cells in mice of the model group (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).

[0057] Table 3 Effect of Ganoderma lucidum Spore Oil on the Number of Tumor Cells in Emotionally Stressed Tumor-Bearing Mice Group                          Cell number (x106) Tumor control group                        1.20 ± 0.18 Stress model group 2.71 ± 0.18*** Stress + Ganoderma lucidum spore oil group 1.69 ± 0.14## Note: ***P < 0.001 compared with the tumor control group; ##P < 0.01 compared with the stress model group.

[0058] The above study, by establishing the mouse models of emotional stress plus tumor susceptibility and using techniques such as small-animal in vivo imaging and flow cytometry, demonstrates that Ganoderma lucidum spore oil with a triglyceride content greater than 90% and an ergosterol content ranging from 0% to 0.5% can alleviate tumor progression by reducing the number of tumor cells in emotionally stressed tumor-bearing mice.

[0059] Example 3

[0060] Observation of Effectiveness of Ganoderma lucidum Spore Oil in Alleviating Emotional Stress in Patients with Tumors

[0061] 1. Research subjects

[0062] Of 88 patients with tumors who took Ganoderma lucidum spore oil, 17 had emotional stress-related symptoms before administration. The tumor types mainly included lung cancer, nasopharyngeal carcinoma, gastric cancer, breast cancer, rectal cancer, liver cancer, uterine fibroids, hematological tumors, neuroendocrine tumors, and the like.

[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] A questionnaire scale was used to score the emotional stress-related symptoms of the patients with tumors. Questionnaires of 17 patients having related symptoms before administration were selected for data statistical analysis.

[0067] 4. Research results

[0068] Symptom scores before and after administration and statistical analysis results are shown in Table 4. No adverse reactions related to the product were observed in all tumor patients during administration, and they exhibited good general conditions.

[0069] Table 4 Symptom Scores Before and After Administration (Only Tumor Patients Having Symptoms Before Administration Included) Analysis variable Before administration After administration P value Inability to concentrate 0.118 ± 0.485 0.118 ± 0.485 - Low mood and reduced interest 2.412 ± 3.318 1.412 ± 1.906 0.039 Insomnia or hypersomnia 3.412 ± 3.874 1.765 ± 2.658 0.031 Still feeling unrefreshed after sleep 2.059 ± 3.325 1.235 ± 1.985 0.11 Feeling fatigued due to emotional 1.353 ± 2.668 0.647 ± 1.367 0.083 responses

[0070] The results indicate that, after administration of Ganoderma lucidum spore oil, tumor patients with symptoms exhibited improvement in most emotional stress-related scores, with significantly reduced symptom scores. Among them, the improvements in “low mood and reduced interest” and “insomnia or hypersomnia” showed significant differences, while there were also certain trends toward improvement in the items of “still feeling unrefreshed after sleep” and “feeling fatigued due to emotional responses”. The above results indicate that Ganoderma lucidum spore oil has an effect of alleviating emotional stress in tumor patients.

[0071] In summary, through the experiments and studies on Ganoderma lucidum spore oil with a triglyceride content greater than 90%, an ergosterol content ranging from 0% to 0.5%, and a vitamin E content ranging from 0.5% to 4%, the present disclosure has found that Ganoderma lucidum spore oil has the effect of reducing emotional stress-induced tumor susceptibility, can significantly reduce tumor growth in mice under emotional stress, and can also significantly reduce scores of multiple emotional stress-related symptoms in patients with various different tumors. It has no significant impact on body weight and overall health status of the mice, as well as health status of the tumor patients, indicating that Ganoderma lucidum spore oil has good safety. The Ganoderma lucidum spore oil of the present disclosure has the beneficial effect of being usable for preparing a medicament for preventing or treating emotional stress-induced tumor susceptibility.

[0072] The above merely discloses preferred embodiments of the present disclosure and should not be construed as limiting the scope of the present disclosure. Therefore, equivalent changes made according to the claims of the present disclosure shall still fall within the scope of the present disclosure.

Claims

1. A use of Ganoderma lucidum spore oil in preparing a formulation for reducing emotional stress-induced tumor susceptibility.

2. The use according to claim 1, for preparing a formulation for inhibiting occurrence and progression of tumor under emotional stress.

3. The use according to claim 1, wherein the tumor comprises one or more selected from the group consisting of ovarian cancer, lung cancer, nasopharyngeal carcinoma, gastric cancer, breast cancer, rectal cancer, liver cancer, uterine fibroids, hematological tumors, and neuroendocrine tumors.

4. The use according to claim 1, wherein in the Ganoderma lucidum spore oil, a content of triglyceride components is greater than 60%, and no Ganoderma lucidum polysaccharides or Ganoderma lucidum triterpenoids are detected.

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

6. The use according to claim 1, wherein the Ganoderma lucidum spore oil further comprises vitamin E; and a mass content of the vitamin E ranges from 0.5% to 4%.

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

8. A formulation obtained by the use according to claim 1.

9. The formulation according to claim 8, wherein the formulation comprises a medicament; and the medicament further comprises a pharmaceutically acceptable carrier.

10. The formulation according to claim 9, wherein the medicament comprises one selected from the group consisting of a tablet, a capsule, a buccal tablet, a granule, a soluble granule, a pellet, a powder, an ointment, a Dan, a suspension, a solution, an emulsion, an injection, a suppository, a cream, a spray, a drop, and a patch.

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