TWI813993B - Application of a Ganoderma lucidum liquid fermentation broth for preparing compositions for preventing, improving or treating Alzheimer's disease or related diseases caused by Alzheimer's disease - Google Patents

Abstract

The invention provides a use of Ganoderma lucidum liquid fermentation broth for preparing a composition for preventing, improving or treating Alzheimer's disease or related diseases caused by Alzheimer's disease, wherein the preparation method of the Ganoderma lucidum liquid fermentation broth includes: The Ganoderma lucidum strain is cultured in a liquid medium to obtain a Ganoderma lucidum liquid fermentation product. The Ganoderma lucidum liquid fermentation product is separated from solid and liquid, and the mycelium is removed to obtain the Ganoderma lucidum liquid fermentation liquid. The prevention and improvement of Alzheimer's disease include improving memory impairment, improving anxiety state changes, reducing GFAP (Glial fibrillary acidic protein) activation in the brain, β-amyloid accumulation, or reducing the occurrence of oxidative damage in brain tissue.

Description

Use of Ganoderma lucidum liquid fermentation broth for preparing a composition for preventing, improving or treating Alzheimer's disease or related diseases caused by Alzheimer's disease

The present invention relates to the efficacy of Ganoderma lucidum liquid fermentation products in preventing and treating Alzheimer's disease or related diseases caused by Alzheimer's disease.

According to the 2019 Global Dementia Report of Alzheimer's Disease International (ADI), it is estimated that there are more than 50 million people with dementia worldwide, which is expected to grow to 152 million by 2050, with every three people suffering from dementia. One person every second develops dementia; the current cost of dementia is US$1 trillion per year and is expected to double by 2030. Because the medical and social costs related to Alzheimer's disease (AD) are quite high and will affect the quality of life of patients' families, the prevention of Alzheimer's disease has become an issue that has received considerable attention in recent years.

Alzheimer's disease is an irreversible, progressive neurodegenerative disease that causes the death of brain nerve cells. The symptoms come from the neurons in the areas of the brain that control thinking, memory and language, resulting in degenerative effects, which in turn affects language ability and Loss of spatial memory discrimination ability.

Pathology shows brain tissue atrophy, cortical senile plaques (Senile plaques) and neurofibrillary tangles (Neurofibrillary tangles) and other phenomena. The pathological mechanism of Alzheimer's disease is not clear yet. However, current research indicates that β-amyloid (β The formation of -amyloid peptide; Aβ) is related to the precursor (Amyloid precursor protein; APP), which participates in the proteolysis process of the enzymes β-secretase (BACE1) and γ-secretase (γ-secretase) (see non-patent document 1, 2).

App ^{NL -GF/NL-GF} Mice is a relatively new AD genotype mouse in recent years. Related studies have been published since 2014 (refer to non-patent document 3). App ^{NL -GF/NL-GF} Mice is Swedish, Iberian and Arctic mutant genes, the Swedish mutant gene will increase the accumulation of Aβ, the Iberian mutant gene will increase the ratio of Aβ42/Aβ40, and the Arctic mutant gene will promote oligomerization and reduce proteolysis to promote the accumulation of Aβ.

How to find health foods, medicines or Chinese herbal medicines for treating Alzheimer's disease is the problem to be solved by the present invention.

Ganoderma lucidum is mainly a traditional Chinese medicine used in China. Research on Ganoderma lucidum points out that Ganoderma has multiple effects, including improving immunity, improving cardiovascular disease, reducing toxins in the liver, anti-tumor, and improving cognitive impairment. (Refer to non-patent documents 4, 5, 6, 7, 8, and 9).

Compared with the Ganoderma fruiting bodies of traditional Chinese medicine, the Ganoderma liquid fermentation product obtained by deep liquid fermentation technology is different in location, shape, chemically active ingredients and pharmacological effects. Currently, in the research on using Ganoderma to improve or prevent Alzheimer's disease, there is no use of Ganoderma liquid fermentation products that do not contain Ganoderma mycelium to improve or prevent Alzheimer's disease.

Rahman et al. tube-fed rats with hot water extract of Ganoderma lucidum fruiting body (200 mg/kg bw) for 8 weeks, which improved the memory and learning ability of AD rats, but it was not confirmed that it could improve Aβ and GFAP (glial) in brain tissue. fibrillary acidic protein) expression amount (see Non-Patent Document 10). Yu et al. tube fed mice Ganoderma lucidum triterpenes (low dose 0.35 g/kg, high dose 1.4 g/kg) for 60 days. In behavioral experiments, it could improve the memory of AD mice and slow down the apoptosis of nerve cells and Oxidative damage, but it has not been proven to improve the expression of Aβ and GFAP in brain tissue (see non-patent document 11). Lai et al. tube-fed mice with petroleum ether and ethanol extracts of Ganoderma lucidum fruiting bodies (100 mg/kg/d) for 6 months, which could reduce the accumulation of Aβ and Tau proteins in the brains of AD mice, but it was not confirmed that it could improve brain function. The amount of GFAP expression in tissues (see Non-Patent Document 12). Huang et al. fed mice Ganoderma lucidum aqueous extract (WGL, 200 mg/kg) and Ganoderma lucidum polysaccharides (GLP, 30 mg/kg) by tube for 90 days, which can reduce the accumulation of Aβ in the brains of AD mice and promote neurological growth. Cell proliferation, but it has not been proven to improve the expression of GFAP in brain tissue (see non-patent document 13).

Although extracts of Ganoderma lucidum fruiting bodies or spores have been used to improve Alzheimer's disease, there has been no research on using Ganoderma lucidum liquid fermentation products to improve or prevent Alzheimer's disease and related diseases caused by Alzheimer's disease. . [Prior Technical Document] [Non-patent literature]

[Non-patent document 1] Spies, P. E., Verbeek, M. M., van Groen, T., & Claassen, J. A. (2012). Reviewing reasons for the decreased CSF Abeta42 concentration in Alzheimer disease. Front Biosci (Landmark Ed), 17, 2024 -2034. [Non-patent document 2] Zhao, H. B., Lin, S. Q., Liu, J. H., & Lin, Z. B. (2004). Polysaccharide extract isolated from ganoderma lucidum protects rat cerebral cortical neurons from hypoxia/reoxygenation injury. J Pharmacol Sci, 95(2 ), 294-298. [Non-patent document 3] Saito, T., Matsuba, Y., Mihira, N., Takano, J., Nilsson, P., Itohara, S., Iwata, N., Saido, T.C., (2014). Single App knock-in mouse models of Alzheimer's disease. Nat. Neurosci. 17(5):661-3. [Non-patent document 4] Gao, Y., Zhou, S., Jiang, W., Huang, M., & Dai, X. (2003). Effects of Ganopoly® (A Ganoderma lucidum Polysaccharide Extract) on the Immune Functions in Advanced‐Stage Cancer Patients. Immunological Investigations, 32(3), 201-215. [Non-patent document 5] Gokce, E. C., Kahveci, R., Atanur, O. M., Gurer, B., Aksoy, N., Gokce, A., Kahveci, O. (2015). Neuroprotective effects of Ganoderma lucidum polysaccharides against traumatic spinal cord injury in rats. Injury, 46(11), 2146-2155. [Non-patent document 6] Jiang, J., Slivova, V., Harvey, K., Valachovicova, T., & Sliva, D. (2004). Ganoderma lucidum suppresses growth of breast cancer cells through the inhibition of Akt/NF -kappaB signaling. Nutr Cancer, 49(2), 209-216. [Non-patent document 7] Kanmatsuse, K., Kajiwara, N., Hayashi, K., Shimogaichi, S., Fukinbara, I., Ishikawa, H., & Tamura, T. (1985). [Studies on Ganoderma lucidum . I. Efficacy against hypertension and side effects]. Yakugaku Zasshi, 105(10), 942-947. [Non-patent document 8] Lin, Z., & Sun, L. (2019). Antitumor Effect of Ganoderma (Lingzhi) Mediated by Immunological Mechanism and Its Clinical Application. Adv Exp Med Biol, 1182, 39-77. [Non-patent document 9] Sun, X. Z., Liao, Y., Li, W., & Guo, L. M. (2017). Neuroprotective effects of ganoderma lucidum polysaccharides against oxidative stress-induced neuronal apoptosis. Neural Regen Res, 12(6) , 953-958. [Non-patent document 10] Rahman, M. A., Hossain, S., Abdullah, N., & Aminudin, N. (2020). Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Ameliorates Nonspatial Learning and Memory Deficits in Rats with Hypercholesterolemia and Alzheimer's Disease. Int J Med Mushrooms, 22(11), 1067-1078. [Non-patent document 11] Yu, N., Huang, Y., Jiang, Y., Zou, L., Liu, X., Liu, S., Zhu, Y. (2020). Ganoderma lucidum Triterpenoids (GLTs) Reduce Neuronal Apoptosis via Inhibition of ROCK Signal Pathway in APP/PS1 Transgenic Alzheimer's Disease Mice. Oxid Med Cell Longev, 2020, 9894037. [Non-patent document 12] Lai, G., Guo, Y., Chen, D., Tang, X., Shuai, O., Yong, T., Wu, Q. (2019). Alcohol Extracts From Ganoderma lucidum Delay the Progress of Alzheimer's Disease by Regulating DNA Methylation in Rodents. Front Pharmacol, 10, 272. [Non-patent document 13] Huang, S., Mao, J., Ding, K., Zhou, Y., Zeng, X., Yang, W., Pei, G. (2017). Polysaccharides from Ganoderma lucidum Promote Cognitive Function and Neural Progenitor Proliferation in Mouse Model of Alzheimer's Disease. Stem Cell Reports, 8(1), 84-94.

In view of the shortcomings of the prior art, the inventor of this case hopes to provide a health food that can improve and prevent Alzheimer's disease, and provide an important research and development direction for the future development of improving, treating and preventing dementia, which is what the present invention is here. important issues to be resolved.

The object of the present invention is to provide a use of Ganoderma lucidum liquid fermentation broth for preparing a composition for preventing, improving or treating Alzheimer's disease or related diseases caused by Alzheimer's disease, wherein the preparation method of the Ganoderma lucidum liquid fermentation broth, The method includes: cultivating the Ganoderma lucidum strain in a liquid medium to obtain a Ganoderma lucidum liquid fermentation product, separating the solid and liquid of the Ganoderma lucidum liquid fermentation product, and removing the mycelium to obtain the Ganoderma lucidum liquid fermentation liquid.

In order to achieve the aforementioned purpose of the invention, the registration number of the Ganoderma lucidum strain is BCRC930215.

To achieve the purpose of the aforementioned invention, the related disease caused by Alzheimer's disease is memory impairment caused by Alzheimer's disease.

In order to achieve the purpose of the aforementioned invention, the related disease caused by Alzheimer's disease is a change in anxiety state caused by Alzheimer's disease.

In order to achieve the object of the aforementioned invention, the prevention, improvement or treatment of Alzheimer's disease is to reduce the activation of GFAP in the brain.

In order to achieve the object of the aforementioned invention, the prevention, improvement or treatment of Alzheimer's disease is to reduce the accumulation of β-amyloid protein.

In order to achieve the object of the aforementioned invention, the prevention, improvement or treatment of Alzheimer's disease is to reduce the occurrence of oxidative damage in brain tissue.

In order to achieve the aforementioned object of the invention, the Ganoderma lucidum liquid fermentation liquid can be freeze-dried to obtain a Ganoderma lucidum liquid fermentation liquid freeze-dried powder.

In order to achieve the foregoing object of the invention, the effective human dose of the Ganoderma lucidum liquid fermentation broth or the Ganoderma lucidum liquid fermentation broth freeze-dried powder is 8 to 200 mg per kilogram of body weight.

In order to achieve the above-mentioned object of the invention, the optimal dose for human body of the Ganoderma lucidum liquid fermentation broth or the Ganoderma lucidum liquid fermentation broth freeze-dried powder is 15 to 100 mg per kilogram of body weight.

To sum up, Ganoderma lucidum liquid fermentation broth can improve memory impairment and anxiety state changes, reduce GFAP activation and β-amyloid accumulation in the brain, reduce the occurrence of oxidative damage in brain tissue, and can effectively prevent and improve Alzheimer's disease. and related conditions caused by Alzheimer’s disease.

The novel technical features of the present invention, including specific features, are disclosed in the scope of the patent application. With regard to the technical features of the present invention, it is better to understand the preferred embodiments of the present invention in conjunction with the description, the embodiments based on the principles of the present invention, and the drawings. Detailed description.

All technical and scientific terms described in this specification, unless otherwise defined, have meanings commonly understood by those with ordinary knowledge in the field; the materials used in the present invention, unless otherwise specified, are commercially available. Sell easily available materials.

The terms "effective amount" or "therapeutically effective amount" used in this manual refer to a sufficient amount of a compound or drug that can alleviate one or more disease symptoms or physiological conditions after the user takes the drug; the result To reduce and/or alleviate signs, symptoms, or causes, or to intentionally change other physiological systems. For example, a therapeutic "effective amount" includes a dose of a compound provided by the invention that can clinically significantly reduce disease symptoms. An appropriate effective dose, the effective value of which depends on common pharmaceutical techniques, such as Dose escalation methods.

The terms "increase," "increase," "enhancement," or similar terms used in this package insert refer to increasing or prolonging the intended effectiveness or duration of a drug. Therefore, to enhance the efficacy of therapeutic drugs, "enhancement" refers to the ability to increase or prolong the efficacy or duration of the drug in the rest of the treatment system. The term "increased effective amount" used in this specification refers to an appropriate amount used to increase the effect of another therapeutic drug in the treatment system.

The terms "treatment", "treatment" and "therapy" in this manual include alleviating, alleviating, or improving at least one disease symptom or physiological condition, preventing new symptoms, and inhibiting the disease or physiological condition in a therapeutic or preventive manner. , Prevent or slow down the development of diseases, cause the recovery of diseases or physiological conditions, slow down the physiological conditions caused by diseases, and stop the symptoms of diseases or physiological conditions.

The invention provides a composition containing Ganoderma lucidum liquid fermentation liquid. The Ganoderma lucidum liquid fermentation liquid is obtained by carrying out liquid fermentation culture of Ganoderma lucidum strains, separating the solid and liquid of the fermentation culture product, removing the fruiting bodies, and retaining the fermentation liquid, wherein, the The composition is a pharmaceutical composition, dietary supplement composition, food composition or health food composition.

Example 1

Preparation of Ganoderma Liquid Fermentation Broth

As shown in Figure 1, the preparation process of Ganoderma lucidum liquid fermentation broth is as follows, S01: Cultivate Ganoderma lucidum strains in a liquid medium to obtain Ganoderma lucidum liquid fermentation product; S02: Solid-liquid separation, remove the mycelium, and obtain Ganoderma lucidum liquid fermentation broth; S03 : Lyophilize Ganoderma lucidum liquid fermentation broth to obtain Ganoderma lucidum fermentation broth freeze-dried powder, which is convenient for use and extends the shelf life.

In a preferred embodiment, the preparation process of Ganoderma liquid fermentation broth is shown in Figure 2. A certain amount of Ganoderma lucidum (GANO99) is taken and deposited at the Food Industry Development Research Institute in Taiwan with the registration number BCRC 930215. Biological Resource Preservation and Research Center, and was deposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH under the registration number DSM 33211. It was first inoculated into solid medium for plate culture, and then inoculated into liquid medium. The bacteria are cultured, and then cultured in a fermentation tank for 3 to 15 days under stirring and aeration conditions to obtain a liquid fermentation product of Ganoderma lucidum, in which the liquid culture medium contains 1 to 10% of comprehensive carbon sources, 0.1 to 2% of comprehensive nitrogen sources, and inorganic Salts are 0.01~0.3%, trace minerals are 0.01~0.3%, and after sterilization, the Ganoderma liquid fermentation product includes Ganoderma mycelium and Ganoderma liquid fermentation broth. The liquid fermentation product is subjected to solid-liquid separation. The solid-liquid separation procedure is to add 3-15% diatomaceous earth to the fermentation product, and then press and filter to obtain the liquid Ganoderma fermentation broth; the comprehensive carbon sources include but are not limited to glucose, sucrose, Fructose, maltose, maltodextrin, starch, etc.; comprehensive nitrogen sources include but are not limited to casein, whey protein, milk powder, plant-based protein, yeast and soy protein; solid-liquid separation procedures include but are not limited to adding silicon Algae pressing, filtration, centrifugation, etc.

AD animal mode

The AD experimental animals in this case are C57BL/B6-App ^NL-GF/NL-GF mice. Mice were kept in a room with constant temperature (21°C-22°C), with light and dark cycles adjusted every 12 hours. The experimental period was from 9:00 to 13:00. During the non-experimental period, the mice had free access to food and water.

Animal experiment design

Experimental groups: 5-month-old mice were randomly divided into 4 groups after domestication: (1) WT group: Wild type mice (N=9), given normal drinking water, served as a normal control group. (2) AD group: C57BL/B6-App ^NL-GF/NL-GF mice (N=6), given normal drinking water. (3) GL1_L group: C57BL/B6-App ^NL-GF/NL-GF mice (N=8), fed low-dose Ganoderma lucidum (1X; 205 mg/kg Ganoderma lucidum fermentation broth freeze-dried powder). (4) GL1_H group: C57BL/B6-App ^NL-GF/NL-GF mice (N=9), fed high-dose Ganoderma lucidum (5X; 1025 mg/kg Ganoderma lucidum fermentation broth freeze-dried powder).

Mice were tube-fed with Ganoderma lucidum (GANO99) samples when they reached 5 months of age. Mix Ganoderma lucidum freeze-dried powder with normal drinking water and tube-fed them 5 days a week, once a day, and continued until The mice were sacrificed when they reached 9 months of age. Animal behavior experiments were conducted one month before sacrifice, during which the samples were continued to be fed by tube until the mice were sacrificed at 9 months of age. The mice were sacrificed by euthanasia, brain tissue, heart blood and fecal samples were collected, and the samples were analyzed by histopathology.

According to the "Guidance for Industry Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers" proposed by the U.S. Food and Drug Administration in 2005, the conversion factor between humans and mice is 12.3 times, so this experiment was conducted in accordance with Mice were given 205 mg (low dose) and 1025 mg (high dose) of Ganoderma lucidum fermented liquid freeze-dried powder per kilogram of body weight. After coefficient conversion, the human dosage was 16.6 mg and 83.3 mg of Ganoderma lucidum fermented liquid freeze-dried powder per kilogram of body weight. pink.

Experimental indicators

Memory impairment: Animal behavioral experiments, Morris water maze test (MWM) and Y-maze (Y maze) were used to evaluate the memory impairment of AD mice. The MWM experiment is to divide the pool in which mice swim into four quadrants: TQ = target quadrant; OQ = quadrant opposite the target quadrant; LQ = quadrant to the left of the target quadrant; RQ = quadrant to the right of the target quadrant. Observe mice in TQ Quadrant swimming time and distance. The longer the time and distance, the better the memory. The Y maze experiment is to put food in the Novel zone and let the mice explore in the Novel zone. Then remove the food and let the mice explore freely in the Y maze. If the mice stay in the Novel zone for a long time, it means better memory. .

Anxiety state: The anxiety state of AD mice was assessed using the animal behavior test plus maze (Elevated plus maze test; EPMT). The cross maze is composed of two open arms and two closed arms, which are crossed in a cross shape. The intersection is the central area. The entire cross maze is at a certain height from the ground. The mouse faces the open arm and generates People are curious and want to explore, but at the same time they have the nature to seek darkness and avoid light. The conflict between exploration and avoidance occurs between the two, resulting in anxiety. Anxious behavior in mice can be assessed by comparing the time and distance the mice spend in the open and closed arms. It is generally believed that the more time and distance a mouse stays in the open arm, the lower the mouse's anxiety level.

Brain histopathology: Immunohistochemical staining is performed, and the pathological sections of the brain tissue are based on the staining degree, distribution range and percentage of the tissue.

Statistical methods

The results of this experiment are expressed in the form of mean ± standard deviation (Mean ± SD). SPSS software was used for statistical analysis, One-way ANOVA was used for testing, and Duncan’s Multiple Range test was used to test the differences between the samples in each group. When p-value <0.05, it means there is a significant difference, and different English letters (a, b or c) are used to mark the results.

Example 1

Ganoderma lucidum liquid fermentation broth improves memory impairment

Figure 3A and Figure 3B are the Probe test results of the MWM experiment to observe the effect of Ganoderma lucidum liquid fermentation products on memory damage caused by AD. The results show that the time and distance in the target quadrant (TQ) of the AD group are significantly lower than that of the WT group; however, Ganoderma lucidum liquid fermentation products have a significant impact on memory damage caused by AD. The time and distance of TQ in the high- and low-dose groups were significantly higher than those in the AD group, and there was no significant difference from the WT group. It has been shown that supplementing high and low doses of Ganoderma lucidum can significantly improve the memory impairment of AD mice. MWM experimental results confirmed that Ganoderma lucidum liquid fermentation products can restore the memory impairment of AD mice to close to normal.

Figure 4 is the Y maze result, observing the effect of Ganoderma liquid fermentation products on memory impairment caused by AD. The results show that the time the AD group stayed in the Novel arm was significantly shorter than that of the WT group, while both the Ganoderma high- and low-dose groups had significantly higher phenomenon in the AD group. It was shown that supplementing high and low doses of Ganoderma lucidum can significantly improve the memory impairment of AD mice. Y maze results also confirmed that Ganoderma lucidum liquid fermentation products can restore the memory impairment of AD mice to normal.

Example 2

Ganoderma fermentation liquid improves anxiety state

Figure 5 is the EPMT result, observing the effect of Ganoderma lucidum liquid fermentation products on the anxiety state caused by AD. The results show that compared with the WT group, the time spent in the close arm of the AD group was significantly shorter; while the time spent in the open arm was significantly shorter. The phenomenon that the high-dose and low-dose groups of Ganoderma lucidum stayed in the close arm was significantly longer than that of the AD group; however, the time that the high-dose and low-dose Ganoderma lucidum groups stayed in the open arm was significantly shorter than that of the AD group, showing that AD is smaller. The anxiety state of the mice was less significant, and the anxiety state of the high- and low-dose groups of Ganoderma lucidum was similar to that of WT mice. The EPMT results showed that the liquid fermentation product of Ganoderma lucidum could restore the anxiety state of AD mice to normal.

Example 3

Ganoderma lucidum liquid fermentation broth reduces GFAP activation and Aβ accumulation in the brain

GFAP is a protein normally found in the brain, and overactivation of GFAP is thought to be associated with Alzheimer's disease. Figure 6 shows the GFAP immunohistochemical staining results of AD mouse brain specimens. The quantitative results are shown in Figure 7A (hippocampus) and Figure 7B (cortex). The results show that the expression amount of GFAP in the hippocampus and cortex of the AD group Significantly higher than that of the WT group; long-term supplementation of high or low doses of Ganoderma lucidum can reduce the activation of GFAP in AD rats, showing that long-term supplementation of Ganoderma lucidum has an inhibitory effect on the activation of GFAP in the brains of AD rats, among which the high-dose group has a greater effect on the cortex. good.

Figure 8 shows the Aβ immunohistochemical staining results of AD mouse brain specimens. The quantitative results are shown in Figure 9A (hippocampus) and Figure 9B (cortex). The results show that the amount of Aβ expression in the hippocampus and cortex of the AD group Significantly higher than that in the WT group, indicating that long-term supplementation of Ganoderma lucidum has an inhibitory effect on Aβ accumulation in AD mice. Long-term supplementation of high or low doses of Ganoderma lucidum can reduce the activation of GFAP in AD mice and cause the accumulation of Aβ. Among them, the high-dose group has an inhibitory effect on the accumulation of Aβ in the cortex. The effect is better.

Example 4

Ganoderma lucidum liquid fermentation broth reduces the occurrence of oxidative damage in brain tissue

Figure 10 shows the SOD immunohistochemical staining results of AD mouse brain specimens. The quantitative results are shown in Figure 11A (hippocampus) and Figure 11B (cortex). The results show that the amount of SOD expression in the hippocampus and cortex is , the SOD expression amount in the AD group was significantly lower than that in the WT group; the SOD expression amount in the hippocampus of the low-dose Ganoderma lucidum group was significantly higher than that of the AD group, while the high-dose Ganoderma lucidum group tended to be higher than the AD group; the SOD expression amount in the cortex , both the high-dose and low-dose groups of Ganoderma lucidum were significantly higher than those of the AD group, indicating that the antioxidant effect of Ganoderma lucidum is more significant in the cortex.

In summary, the present invention is the first to use mycelium-free Ganoderma lucidum liquid fermentation broth for the treatment of Alzheimer's disease. Ganoderma lucidum liquid fermentation broth can improve memory impairment and anxiety state changes, reduce GFAP activation and Aβ accumulation and reducing the occurrence of oxidative damage in brain tissue can effectively prevent and improve Alzheimer's disease.

The above detailed description is a specific description of possible embodiments of the present invention. However, the embodiments are not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not depart from the technical spirit of the present invention shall be included in this case. within the scope of the patent.

Figure 1 is the preparation process of Ganoderma lucidum liquid fermentation broth.

Figure 2 is a schematic diagram of the Ganoderma liquid fermentation process and test sample preparation.

Figure 3A is the distance result in the Probe test of MWM, observing the effect of Ganoderma lucidum on memory impairment in AD mice.

Figure 3B is the time result in the Probe test of MWM to observe the effect of Ganoderma lucidum on memory impairment in AD mice.

Figure 4 is the Y maze result, observing the effect of Ganoderma lucidum on memory impairment in AD mice.

Figure 5A shows the time of close arm in EPMT to observe the effect of Ganoderma lucidum on the anxiety state of AD mice.

Figure 5B shows the open arm time during EPMT to observe the effect of Ganoderma lucidum on the anxiety state of AD mice.

Figure 6 shows the results of GFAP immunohistological staining of AD mouse brain tissue.

Figure 7A is the quantitative results of GFAP immunohistological staining in the brain tissue (hippocampus) of AD mice.

Figure 7B is the quantitative results of GFAP immunohistological staining of brain tissue (cortical layer) of AD mice.

Figure 8 shows the results of Aβ immunohistological staining of AD mouse brain tissue.

Figure 9A is the quantitative results of Aβ immunohistological staining in the brain tissue (hippocampus) of AD mice.

Figure 9B is the quantitative result of Aβ immunohistological staining in AD mouse brain tissue (cortical layer).

Figure 10 is the SOD immunohistological staining results of AD mouse brain tissue.

Figure 11A is the quantitative results of SOD immunohistological staining of AD mouse brain tissue (hippocampal gyrus).

Figure 11B is the quantitative result of SOD immunohistological staining of AD mouse brain tissue (cortical layer).

Ganoderma lucidum strain GANO99 was deposited at the Biological Resources Preservation and Research Center of the Food Industry Development Institute in Taiwan on November 22, 2019, with the deposit number BCRC 930215.

Ganoderma lucidum strain GANO99 was deposited with the German Microbiology and Cell Culture Collection Center Co., Ltd. (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH) on July 10, 2019, with the deposit number DSM 33211.

Claims (7)

A use of Ganoderma lucidum liquid fermentation broth for preparing a composition for preventing, improving or treating Alzheimer's disease or related diseases caused by Alzheimer's disease, wherein the preparation method of the Ganoderma lucidum liquid fermentation broth includes: adding the Ganoderma lucidum bacteria The seeds are cultured in a liquid culture medium under stirring and aeration conditions to obtain a Ganoderma lucidum liquid fermentation product. The Ganoderma lucidum liquid fermentation product is separated from solid and liquid, and the mycelium is removed to obtain the Ganoderma lucidum liquid fermentation liquid; wherein the Alzheimer's The disease is caused by the accumulation of β-amyloid protein due to the promotion of oligomerization and reduction of protein hydrolysis; the registration number of the Ganoderma lucidum strain is BCRC 930215, and the Ganoderma lucidum liquid fermentation broth or the Ganoderma lucidum liquid fermentation broth freeze-dried powder in humans Effective doses range from 8 to 200 mg per kilogram of body weight. The use described in item 1 of the patent application, wherein the related disease caused by Alzheimer's disease is memory impairment caused by Alzheimer's disease. The use described in item 1 of the patent application, wherein the related disease caused by Alzheimer's disease is a change in anxiety state caused by Alzheimer's disease. The use described in item 1 of the patent application, wherein the prevention, improvement or treatment of Alzheimer's disease is to reduce the activation of GFAP in the brain. The use described in item 1 of the patent application, wherein the prevention, improvement or treatment of Alzheimer's disease is to reduce the occurrence of oxidative damage in brain tissue. For the use described in item 1 of the patent application, the Ganoderma lucidum liquid fermentation liquid can be freeze-dried to obtain a Ganoderma lucidum liquid fermentation liquid freeze-dried powder. For the uses described in Item 1 or 6 of the patent application, the optimal dose for the human body of the Ganoderma lucidum liquid fermentation broth or Ganoderma lucidum liquid fermentation broth freeze-dried powder is 15 to 100 mg per kilogram of body weight.