CN106692206A - Application of Xylaria nigripes mycelium extract in protecting nerve cells and composition for protecting nerve cells - Google Patents

Abstract

The present invention discloses a use of a mycelium extract of Xylaria nigrostriatus and a composition based on the use. The mycelium extract of Xylaria nigrostriatus has the activity of protecting nerve cells and can be used to prepare a medical composition for protecting nerve cells. The mycelium extract of Xylaria nigrostriatus is prepared by the following steps: providing a substrate; performing a fermentation process, inoculating Xylaria nigrostriatus in the substrate, and culturing for a fermentation time at a fermentation temperature to obtain a fermentation substance; separating the fermentation substance to obtain mycelium of Xylaria nigrostriatus; performing an extraction process, in which a solvent is mixed with mycelium of Xylaria nigrostriatus to form a mixture, and then the mixture is stirred for an extraction time; and removing the solid part of the mixture to obtain a supernatant, wherein the supernatant contains the mycelium extract of Xylaria nigrostriatus, and the mycelium extract of Xylaria nigrostriatus has the activity of protecting nerve cells.

Description

Application of mycelia extract of C. nigricans to protect nerve cells and composition for protecting nerve cells

technical field

The present invention relates to a use of mycelium extract of C. nigriscens and a composition based on the use, in particular to a use of mycelium extract of C. nigriscens to protect nerve cells, and a combination for protecting nerve cells thing.

Background technique

With the advancement of medical treatment, food technology and economic success, the average life expectancy of human beings has been extended, and people have gradually entered into an elderly society. Aging-derived diseases include immune disorders, cancer, cardiovascular diseases, neurodegenerative disorders and metabolic syndromes, etc. Among them, the huge medical and social costs and the damage to the quality of family life brought about by the latter two cannot be ignored.

The human nervous system can be divided into the central nervous system and the peripheral nervous system, in which the central nervous system includes the brain, cerebellum and spinal cord. The basic structural unit of the nervous system is neurons, or nerve cells, and the nerve cells of the adult central nervous system are differentiated nerve cells, which cannot divide and proliferate. Once damaged or dead, they will cause permanent loss of function. Therefore, there is a need for a way to protect nerve cells from injury, degeneration or death.

Xylaria nigripe belongs to the order Xylariales, the family Xylariaceae and the genus Xylaria. It is a traditional Chinese medicinal fungus that usually grows on abandoned termites. The nest, the sclerotium formed by its mycelium is the traditional Chinese medicine material "Wulingshen", also known as Wuling bacteria, Lei Zhenzi and so on. According to Sichuan "Chinese Medicine Chronicles", Wuling ginseng has the description of "invigorating the heart and kidneys and treating insomnia".

Due to the gradual scarcity of wild resources, C. nigritiformis has the problems of difficult collection and unstable quality. At present, it is a method to solve the problems of insufficient yield and unstable quality of natural C. In the process of liquid culture, in addition to synthesizing the organic substances needed for its own growth, C. nigriscens will also secrete metabolites, which may also contain components that can protect nerve cells. Has the potential to develop natural neurocyte-protecting substances. However, different culture methods will lead to different growth patterns of the bacteria, and different gene expressions. The metabolites produced will also be different, and the efficacy will also be different, or new efficacy and use may be produced.

Contents of the invention

One aspect of the present invention is to provide a use of mycelia extract of Xylaria nigripes for preparing a pharmaceutical composition for protecting nerve cells. The mycelium extract of C. nigritiformis is prepared by the following steps: a substrate is provided, and the substance of the substrate consists of 0.5 to 5% by weight of brown rice flour, 0.1 to 1% by weight of malt extract and 99.4 to 94% by weight than water. Then carry out the fermentation process, which inoculates C. nigricans in the substrate, and cultivates it at the fermentation temperature for a fermentation time, so as to obtain the fermented substance. The fermented material is further separated to obtain the mycelia of C. nigritiformis. Then the extraction process is carried out. After the solvent is mixed with the mycelium of C. nigristifolia to form a mixture, the mixture is stirred for an extraction time. Finally, the solid part of the mixture is removed to obtain a supernatant liquid, wherein the supernatant liquid contains the mycelia extract of C. nigeriota, and the mycelia extract of C. nigristipe has the activity of protecting nerve cells.

According to the aforementioned application of the mycelia extract of C. nigritiformis, the aforementioned matrix is composed of 2% by weight of brown rice flour, 0.5% by weight of malt extract and 97.5% by weight of water.

According to the above-mentioned application of the mycelia extract of C. nigritiformis, the fermentation temperature in the fermentation process is 25° C., and the fermentation time is 10 days.

According to the above-mentioned use of the mycelia extract of C. nigritiformis, the solvent in the extraction process may be hot water at 80° C. to 100° C., and the extraction time may be 0.5 hours to 2 hours.

According to the above-mentioned application of the mycelia extract of C. nigritiformis, the solvent in the extraction process may be 70% by weight ethanol, and the extraction time may be 22 hours to 26 hours.

According to the aforementioned use of the mycelia extract of C. nigritiformis, the solid part of the mixture can be removed by centrifugation, filtration or a combination of the above methods.

According to the above-mentioned application of the mycelia extract of C. nigriscens, the protection of nerve cell activity can protect or prevent the nerve cells from being damaged and dying.

According to the above-mentioned use of the mycelia extract of C. nigricans, the protection of nerve cell activity can be achieved by protecting or preventing the DNA of nerve cells from being damaged.

Another aspect of the present invention is to provide a composition for protecting nerve cells, which comprises an effective dose of mycelia extract of C. nigricans.

According to the aforementioned composition for protecting nerve cells, the composition is a form selected from the following groups: pharmaceutical composition, feed composition, beverage composition, nutritional supplement composition, edible composition and health-care edible composition.

Thereby, the mycelia extract of C. nigritiformis of the present invention has the activity of protecting nerve cells, can protect or prevent nerve cells from being damaged and die and can protect or prevent the DNA of nerve cells from being damaged, so the present invention The mycelia extract of C. nigricans of the present invention can be used to prepare a medicinal composition for protecting nerve cells. And the composition comprising the mycelia extract of C. nigriscens can be used as a pharmaceutical composition, a feed composition, a beverage composition, a nutritional supplement composition, an edible composition and a health-care edible composition for the purpose of protecting nerve cells. Use potential in the biomedical healthcare market.

The above summary of the invention aims to provide a simplified abstract of the content of the technical solution, so that readers can have a basic understanding of the content of the technical solution. This summary of the invention is not a complete overview of the content of the technical solutions, and it is not intended to point out the important/key elements of the embodiments of the invention or to define the scope of the invention.

Description of drawings

In order to make the above and other objects, features, advantages and embodiments of the present invention more comprehensible, the accompanying drawings are as follows:

Fig. 1 is the flow chart of the preparation steps of the mycelia extract of C. nigritiformis of the present invention.

detailed description

The disclosure content of this specification proposes a novel use of the mycelium extract of C. nigristiformum, evaluates the protective activity of the mycelium of C. nigriscens mycelium by in vitro test, and further verifies the mycelium of C. nigriscens mycelium The nerve cell protection activity of the extract is to protect or prevent nerve cells from damage and death and to protect or prevent DNA of nerve cells from damage. The mycelia extract of C. nigritiformis proposed in the disclosure content of this specification is a potential supplement for protecting nerve cells, and can be used to prepare a pharmaceutical composition for protecting nerve cells.

The "effective dose" in the present invention refers to the amount of the mycelia extract of C. nigristipe that can effectively protect and/or prevent nerve cells from being damaged. For example, an effective dose of C. nigeriota mycelium extract can prevent nerve cells from damage and/or alleviate to some extent one or more symptoms associated with a disease caused by nerve cell damage.

Now further illustrate the present invention with the following specific test examples, so that those with ordinary knowledge in the technical field of the present invention can fully utilize and practice the present invention without excessive interpretation, and these test examples should not be regarded as To limit the scope of the invention, but to illustrate how to practice the materials and methods of the invention.

<Test example>

1. Preparation of C. nigricans mycelium extract

Please refer to FIG. 1 , which is a flow chart of the preparation steps of the mycelia extract of C. nigritiformis. In FIG. 1 , the method 100 for preparing the mycelia of C. nigritiformis includes step 110 , step 120 , step 130 , step 140 and step 150 .

Step 110 is to provide a matrix, which essentially consists of 0.5-5% by weight of brown rice flour, 0.1-1% by weight of malt extract and 99.4-94% by weight of water. Preferably, the matrix consists of 2% by weight of brown rice flour, 0.5% by weight of malt extract and 97.5% by weight of water. Furthermore, the matrix used consisted of 20g/L brown rice flour and 5g/L malt extract. The fermentor used was a stirred-tank fermentor of 5 liters with a working volume of 3 liters. To provide carbon source, nitrogen source and water so that the subsequent C. nigritiformis can be cultured in liquid state.

Step 120 is to carry out the fermentation process, inoculate the C. nigritiformis in the substrate, and cultivate it at the fermentation temperature for a fermentation time, so as to obtain the fermented substance. The strain used is Xylaria nigripes BCRC34219, which is preferably purchased from the Center for Biological Resource Conservation and Research of the Food Industry Development Institute (Hsinchu, Taiwan). Xylaria nigripes BCRC34219 is a strain isolated from Taiwan. However, it should be noted that the above-mentioned C. nigricantis strain (BCRC34219) is only one embodiment of the present invention. During the fermentation process, the conditions such as temperature and ventilation are adjusted to make the mycelium of C. nigritiformis grow. Furthermore, the conditions of the fermentation process are as follows: inoculate 300mL of C. nigriscens into the substrate, and then cultivate it for 10 days under the fermentation conditions such as stirring speed of 100rpm, ventilation rate of 1vvm and fermentation temperature of 25°C. Fermented material is available.

Step 130 is to separate the fermented material, separating the fermented material into a solid part and a liquid part, wherein the solid part contains the mycelia of C. nigeriota. After the fermentation process is completed, since the fermented material is filled with mycelium of C. nigriscens, after separating the liquid part and solid part of the fermented material, the mycelium of C. nigriscens can be obtained in the solid part body. The separation method can be a filtration method, a centrifugal method or a combination of the above methods.

Step 140 is to carry out the extraction process. After the solvent is mixed with the mycelium of C. nigritiformis to form a mixture, the mixture is stirred for an extraction time. During the extraction process, the solvent can be hot water at 80° C. to 100° C., and the extraction time can be 0.5 hour to 2 hours. Or the solvent can be 70% ethanol by weight, and the extraction time can be 22 hours to 26 hours. Furthermore, when preparing the mycelia extract of C. nigristiformis, first use water to wash the C. nigriscens mycelium several times, and then use a freeze dryer to dry the C. nigriscens mycelium to prepare into mycelium powder, and the mycelium powder sample is stored in an airtight container. Then the mycelium powder and 90°C hot water are mixed to form a mixture at a weight ratio of 1:10 to 1:30, and the mixture is stirred for 1 hour to extract functional components therein. Alternatively, after mixing the mycelium powder and 70% by weight ethanol at a weight ratio of 1:10 to 1:30 to form a mixture, the mixture is stirred for 24 hours to extract functional components therein.

Step 150 is to remove the solid portion of the mixture to obtain a supernatant, wherein the supernatant comprises the mycelia extract of C. nigritiformis. After the extraction process, the mycelia extract of C. nigritiformis has been dissolved in a solvent, which contains functional components, so that after removal of the solid part of the mixture, the supernatant obtained contains C. nigritiformis Mycelia extract. The solid portion of the mixture can be removed by centrifugation, filtration or a combination of the above. Furthermore, after the extracted sample is filtered and centrifuged, the supernatant is taken for concentration, and then dried with a freeze dryer to obtain the hot water extract of C. Ethanol extract of horn fungus mycelium.

2. The ability of the mycelium extract of C. nigritiformis to protect nerve cells

PC12 neural cell line is isolated from rat renal medullary pheochromocytoma, which belongs to mesoderm tissue, and is generally widely used to study neural differentiation, neurotoxicity, neuropharmacology, and neuroprotective mechanisms. In the experiment, undifferentiated PC12 nerve cells (nPC-12 cells) were used to simulate undifferentiated nerve cells, and the oxidative damage of simulated nerve cells was treated with the inducer H ₂ O ₂ , and the cell survival rate and the release rate of lactate dehydrogenase were analyzed. , DNA damage analysis of cells to explore the protective ability of the mycelia extract of C. The PC12 nerve cells of the following test examples were purchased from the Bioresource Conservation and Research Center of the Food Industry Development Institute (Hsinchu, Taiwan).

1. Cell Viability Analysis

The viability analysis of PC12 nerve cells was calculated by the cell viability analysis method (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide; MTT assay) to calculate the hyphae of C. Protective rate of body extracts on PC12 nerve cells. MTT is a water-soluble yellow dye. Succinate dehydrogenase (SDH) in the mitochondria of living cells can metabolize and reduce MTT. At the same time, under the action of cytochrome C, it produces blue-purple water-insoluble MTT-formazan. , has an absorbance value at a wavelength of 595nm. In addition to monitoring the reducing ability of mitochondria, this method can also represent the number of viable cells. The higher the absorbance value, the higher the survival rate of cells.

In the experiment, the culture medium of PC12 nerve cells was DMEM (Dulbecco's Modified Eagle Medium) medium containing 10% fetal bovine serum (FBS). When performing cell viability analysis, PC12 nerve cells were first seeded in 96-well microplates at a concentration of 3×10 ⁵ cells/well, and cultured in a 37°C, 5% CO ₂ incubator for 24 hours, then administered to samples or The inducer H ₂ O ₂ , in which only the inducer H ₂ O ₂ was added as the control group, while the inducer H ₂ O ₂ and the samples were added as the experimental group, and the samples were hot water extraction of C. and the ethanol extract of C. nigricans mycelium, and then cultured for 24 hours in a 37°C, 5% CO ₂ incubator. After culturing, suck out the culture solution, wash twice with phosphate buffered solution (PBS), add 150 μl of MTT with a concentration of 0.5 mg/ml, place it in an incubator at 37°C and 5% CO for ₄ hours, suck out the MTT, and then Add 100 μl dimethyl sulfoxide (DMSO) to dissolve MTT-formazan and make the solution color. After standing overnight, measure the absorbance value at a wavelength of 595nm with an enzyme immunoassay reader, which can represent the number of viable cells. The PC12 nerve cell protection rate is calculated as follows:

in is the survival rate of PC12 nerve cells when the inducer H ₂ O ₂ and the sample are added simultaneously; is the survival rate of PC12 nerve cells when only the inducer H ₂ O ₂ was added.

Please refer to Table 1 below for the analysis of the protection rate of C. nigricans mycelium hot water extract and C. nigricans mycelium ethanol extract on PC12 nerve cells by cell viability analysis method.

Table I

The experimental results show that when the hot water extract of C. nigerium mycelium is at a concentration of 10, 50, and 100 μg/mL, the protection rate of the hot water extract of C. nigerium mycelium on PC12 nerve cells is 9.4%. , 24.1% and 29.6%. When the concentration of ethanol extract of C. nigriscens mycelium was 10, 50, and 100 μg/mL, the protection rates of ethanol extract of C. nigriscens mycelium to PC12 nerve cells were 14.3%, 29.2% and 33.2% . It shows that whether it is the ethanol extract of the mycelium of C. nigriscens or the hot water extract of the mycelium of C. nigriscens, it can protect the PC12 nerve cells from being oxidized by H ₂ O ₂ , and protect or prevent the PC12 nerve cells. Cells die from injury in a dose-dependent manner. However, under different sample concentrations, the protection rate of ethanol extract of C. nigriscens mycelium to PC12 nerve cells was higher than that of hot water extract of C. nigriscens mycelium.

2. Analysis of the release rate of lactate dehydrogenase

H ₂ O ₂ causes oxidative damage to the cells, and also leads to the destruction of the cell membrane, thus releasing lactate dehydrogenase (lactate dehydrogenase, LDH) into the medium, so the lower the release rate of LDH in the medium, the more cells are affected. The lower the ratio of damage, the more effective the sample is in protecting cells.

Experimentally, PC12 neurons were first seeded in 96-well microplates at a concentration of 3×10 ⁵ cells/well, and cultured in a 37°C, 5% CO ₂ incubator for 24 hours before giving the samples or the inducer H ₂ O ₂ , in which only adding inducer H ₂ O ₂ was taken as the control group, while adding inducer H ₂ O ₂ and samples was taken as the experimental group, and the samples were the hot water extract of C. The ethanol extract of the horn fungus mycelia was then cultured in a 37°C, 5% CO ₂ incubator for 24 hours. After culturing, the culture solution was aspirated, washed twice with phosphate buffered saline (PBS), and the LDH activity in the culture solution was analyzed using the LDH Cytotoxicity Detection Kit (Cytotoxicity Detection Kit, TaKaRa Bio Laboratories, Japan) of Nippon Treasure Biolabs. , the percentage of toxicity to PC12 nerve cells can be calculated (percentagecytotoxicity), which means that the cells are damaged. When the analysis result shows that the activity of LDH is low, the percentage of toxicity is small, and the damage to the cells is less serious, which means that the sample has a better effect of protecting cells. In this assay kit, lactate dehydrogenase activity is determined by a colorimetric method. After a specific substrate reacts with the enzyme, the absorbance of the reaction solution is linearly positively correlated with lactate dehydrogenase activity. The formula for calculating the toxicity percentage is as follows:

Wherein OD _sample is the amount of LDH released from cells after adding sample treatment. OD _{Low control} is the control group without adding samples, that is, the amount of LDH released spontaneously by cells. OD _{High control} is the amount of LDH released when Triton X-100 is added to the control group without adding samples to make the cells 100% dead.

Please refer to the following table ₂ , in order to analyze the release rate of LDH, when adding the hot water extract of the mycelia of _C. Toxicity percentage of oxidative damage.

Table II

The experimental results showed that when the hot water extract of the mycelia of C. nigeriota was at a concentration of 10, 50, and 100 μg/mL, the hot water extract of the mycelia of C. nigristifolia had a significant effect on H ₂ O ₂ -induced oxidation of PC12 nerve cells. The toxicity percentages of the injuries were 40.5%, 35.7% and 32.9%, respectively. When the concentration of ethanol extract of C. nigriscens mycelia was 10, 50, and 100 μg/mL, the toxicity percentages of ethanol extracts of C. nigriscens mycelium to H ₂ O ₂ induced oxidative injury of PC12 nerve cells were respectively 44.4%, 36.3% and 33.1%. It shows that whether it is the ethanol extract of the mycelia of C. nigritiformis or the hot water extract of the mycelium of C. nigriscens, it has the effect of protecting PC12 nerve cells from the oxidation damage of H ₂ O ₂ , and it has a dose-dependent effect. sex. However, at different sample concentrations, the ethanol extracts of C. nigriscens mycelium protected PC12 nerve cells more than the hot water extracts of C. nigristiform mycelia.

3. DNA damage analysis of cells

Single-cell gel electrophoresis is a method that can detect DNA damage in a single cell. Its principle is to use low-melting point agar gel electrophoresis to separate DNA fragments of different sizes and monitor whether the cell DNA is broken. If the cellular DNA is not damaged, it will move slowly, and the damaged and broken DNA fragments will move outside the nucleus after electrophoresis, and after nucleic acid staining, it will form a phenomenon like a comet tail, also known as Comet assay. Therefore, single-cell gel electrophoresis can be used to observe the DNA damage of PC12 nerve cells after they are oxidatively injured by the inducer H ₂ O ₂ and protected by the mycelium extract of C. nigricans.

Experimentally, the PC12 nerve cells without any treatment were used as the control group, the induction agent H ₂ O ₂ was added only as the control group, and the induction agent H ₂ O ₂ and samples were added simultaneously as the experimental group. The control group was treated with 300 μg/mL H ₂ O ₂ for 2 hours, and then subjected to single-cell gel electrophoresis. The experimental group was first treated with hot water extract of C. nigriscens mycelium or ethanol extract of C. nigriscens mycelium at a concentration of 10, 50, and 100 μg/mL for 24 hours, and then treated with 300 μg/mL H ₂ O ₂ treatment for 2 hr, followed by single-cell gel electrophoresis. In terms of quantitative analysis, image processing software can be used to count the DNA tailing ratio and tailing length.

Please refer to Table 3 below, the DNA damage analysis results of PC12 nerve cells in different experimental groups.

Table three

The experimental results show that the PC12 nerve cells in the control group will not have tailing when performing single-cell gel electrophoresis. When PC12 nerve cells in the control group were subjected to single-cell gel electrophoresis, the ratio of tailed DNA reached 68.3%, and the average tail length was 80. However, the P12 nerve cells in the experimental group were first treated with ethanol extract of C. _nigriscens mycelium or hot water extract of _C. After 2 hours of treatment, it can be found that the ratio of tailed DNA and the tail length are reduced when performing single-cell gel electrophoresis. Especially after treatment with 100 μg/mL hot water extract of C. nigeriota mycelium, the ratio of tailing DNA was reduced to 7.6% and the average length of tailing was reduced to 11.6. It shows that whether it is the ethanol extract of the mycelium of C. nigriscens or the hot water extract of the mycelium of C. nigriscens, it has the effect of protecting PC12 nerve cells from being oxidized by H ₂ O ₂ , and can protect or Prevent the DNA of PC12 nerve cells from damage, and regardless of the sample concentration, the ability of the ethanol extract of C. nigricans mycelium to protect PC12 nerve cells is better than that of the hot water extract of C. nigricans mycelium .

From the above, the analysis results of the mycelium extract of C. nigriscens disclosed in the present invention in protecting nerve cells show that the mycelia extract of C. nigriscens can protect or prevent nerve cells from being damaged and dying. And can protect or prevent the DNA of nerve cells from damage. Therefore, the mycelia extract of C. nigritiformis of the present invention can be used to prepare a pharmaceutical composition for protecting nerve cells. The medicinal composition for protecting nerve cells prepared by the mycelia extract of C. nigristifolia of the present invention can be administered via topical administration, orally (such as oral powder, capsule, suspension or tablet) , or parenterally (eg, injection) and other appropriate methods. Therefore, the composition containing an effective dose of the mycelia extract of C. nigritiformis can be used as a pharmaceutical composition, a feed composition, a beverage composition, a nutritional supplement composition, an edible composition, and a health-care edible composition for the purpose of protecting nerve cells. In terms of materials, it has the potential to be used in the biomedical health care market.

Although the present invention has been disclosed above in terms of implementation, it is not intended to limit the present invention. Any person skilled in the art may make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection of the present invention The scope is to be determined as defined by the claims.

Claims (11)

1. a kind of purposes of Xylaria nigripes mycelium extract, it is characterised in that it is used to prepare protection The medical composition of nerve cell, wherein the Xylaria nigripes mycelium extract is made by following step ：

There is provided matrix, wherein the matrix essence by 0.5~5% weight than coarse rice powder, 0.1~1% weight ratio Malt extract and 99.4~94% weight than water constituted；

Fermentation technology process is carried out, Xylaria nigripes is inoculated with the matrix, and trained under fermentation temperature A fermentation time is supported, to obtain fermented material；

The fermented material is separated, to obtain the Xylaria nigripes mycelium；

Extraction process process is carried out, the extraction process process utilizes solvent and the Xylaria nigripes mycelia After body mixing resulting mixture, the mixture is stirred into an extraction time；And

The solid of the mixture is removed partly, to obtain supernatant, wherein the supernatant is comprising described Xylaria nigripes mycelium extract, and the Xylaria nigripes mycelium extract has protection nerve thin Cytoactive.

2. the purposes of Xylaria nigripes mycelium extract as claimed in claim 1, it is characterised in that The matrix by 2% weight than coarse rice powder, 0.5% weight than malt extract and 97.5% weight The water of ratio is constituted.

3. the purposes of Xylaria nigripes mycelium extract as claimed in claim 1, it is characterised in that The fermentation temperature is 25 DEG C.

4. the purposes of Xylaria nigripes mycelium extract as claimed in claim 1, it is characterised in that The fermentation time is 10 days.

5. the purposes of Xylaria nigripes mycelium extract as claimed in claim 1, it is characterised in that The solvent is 80 DEG C to 100 DEG C of hot water, and the extraction time is 0.5 hour to 2 hours.

6. the purposes of Xylaria nigripes mycelium extract as claimed in claim 1, it is characterised in that The solvent is the ethanol of 70% percentage by weight, and the extraction time is 22 hours to 26 hours.

7. the purposes of Xylaria nigripes mycelium extract as claimed in claim 1, it is characterised in that The solid portion is removed using the combination of centrifugation, filter type or aforesaid way.

8. the purposes of Xylaria nigripes mycelium extract as claimed in claim 1, it is characterised in that It is described to protect neural cell activity to protect or preventing nerve cell dead from damage.

9. the purposes of Xylaria nigripes mycelium extract as claimed in claim 1, it is characterised in that The protection neural cell activity is by protecting or preventing the DNA of nerve cell from damaging.

10. it is a kind of protect nerve cell composition, it is characterised in that the composition include effective dose Xylaria nigripes mycelium extract as claimed in claim 1.

The composition of 11. protection nerve cells as claimed in claim 10, it is characterised in that it is selected from A kind of form in following group：Medical composition, fodder compound, beverage composition for treating dental erosion, nutritional supplementation Composition, edible composition and edible health composition.