JP2014040375A - Nerve regeneration accelerating agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an agent accelerating regeneration of a cut nerve, and a food product containing the agent.SOLUTION: Provided are: (1) a nerve regeneration accelerating agent containing one or more extracts selected from wheat bran, wheat powder and barley rice and one or more substances selected from phosphatidyl serine, theanine, docosahexaenoic acid and ferulic acid; (2) a nerve regeneration accelerating agent according to (1), where the nerve is a peripheral nerve; (3) a nerve regeneration accelerating agent according to (1) or (2), which is an oral agent; (4) a food product having nerve regeneration accelerating function according to any of (1) to (3); and (5) a composition containing one or more extracts selected from wheat bran, wheat powder and barley rice and one or more substances selected from phosphatidyl serine, theanine, docosahexaenoic acid and ferulic acid.

Description

  The present invention relates to an agent that promotes regeneration of severed nerves.

In mature mammals, nerve cells are generally tissues composed of special cells that do not have division ability. Therefore, once a failure occurs, the failure continues for a long time. Especially in the central nervous system such as the brain and spinal cord, there is no regeneration ability.
One of the reasons is that there is no cure for neurodegenerative diseases such as involuntary spinal cord injury due to exogenous injury, Alzheimer's disease, and Parkinson's disease, and the lack of regenerative ability in the central nervous system. On the other hand, the peripheral nerve has a regeneration ability, and even after being cut once, the axon is regenerated and the function is restored. However, even in this case, the period required for the reproduction takes several months to one year or more. For this reason, the pain for the patient is great.
In addition, since nerve regeneration takes a long time, nerve cells die during that time, and function recovery often does not occur. Even in the case of peripheral nerves having regenerative ability in this way, they cannot extend at all in the central nervous system environment such as the brain and spinal cord. For this reason, it has been speculated that there exists a substance in the central nervous system that prevents nerve growth. When a nerve regeneration inhibitor that exists in the central nervous system is suppressed with an antibody or the like, partial nerve regeneration occurs in the center, and functional recovery is also seen. Recently, Nogo was discovered as a central nerve regeneration inhibitor (Non-patent document 1: Nature 403, 434, 2000, Non-patent document 2: Nature 403, 439, 2000). However, by inhibiting Nogo, it is thought that the nerve fibers that regenerate are a part and that there are other regeneration inhibitors, but one of the factors that act to inhibit nerve regeneration in vivo. Cephomarin has also been estimated as one (Non-Patent Document 3: Cell 75, 217, 1993, Non-Patent Document 4: Cell 75, 1389, 1993), but has not been clarified so far.

  By the way, as described above, although the peripheral nerve regenerates, it cannot be easily regenerated due to the influence of an inhibitor in the nerve tissue. Therefore, attempts have been made to regenerate nerves by connecting gaps in peripheral nerves using a junction tube for artificial nerve joining. Patent Document 1 (Japanese Patent Laid-Open No. 5-237139) discloses a nerve regeneration assisting material comprising a bundle of collagen fibers coated with laminin and fibronectin. Patent Document 2 (International Publication No. 98/22155) discloses a tubular body of biodegradable and absorbable material and a collagen having a cavity penetrating the tubular body along the axial line of the tubular body substantially parallel to the axis of the tubular body. An artificial neural tube made of a body and filled with a matrix gel containing collagen, laminin and the like is disclosed. Patent Document 3 (International Publication No. 99/63908) discloses a tubular body of biodegradable absorbent material, and an artificial fiber in which a collagen fiber bundle covered with laminin is inserted in the lumen substantially parallel to the axis of the tubular body. A neural tube is disclosed. Patent Document 4 (Japanese Patent Laid-Open No. 2000-325463) discloses a nerve reconstruction base material having a structure in which fibers made of a bioabsorbable material are bundled. Patent Document 5 (Japanese Patent Laid-Open No. 2001-70436) discloses a support such as a sponge, tube, or coil made of collagen. Patent Document 6 (Japanese Patent Application Laid-Open No. 2002-320630) discloses a support comprising a sponge-like fine matrix made of a biodegradable material or a bioabsorbable material and a linear biological tissue induction path or organ induction path. Is disclosed. Further, Patent Document 7 (Japanese Patent Application Laid-Open No. 2003-19196) includes a sponge made of a biodegradable polymer material and a reinforcing material made of a biodegradable polymer having a longer decomposition and absorption period than the sponge, the inner surface of which is a sponge. A nerve regeneration tube is disclosed.

  When attempting to regenerate peripheral nerves with such a junction tube, the provision of a drug that promotes nerve regeneration should dramatically improve the regeneration efficiency, but no such drug is provided.

JP-A-5-237139 International Publication No. 98/22155 International Publication No. 99/63908 JP 2000-325463 A JP 2001-70436 A JP 2002-320630 A Japanese Patent Laid-Open No. 2003-19196

Nature 403, 434, 2000 Nature 403, 439, 2000 Cell 75, 217, 1993 Cell 75, 1389, 1993

  An object of the present invention is to provide an agent having a peripheral nerve regeneration promoting action. Moreover, let it be a subject to provide the food-drinks containing this agent.

  As a result of testing by focusing on wheat powder, the present inventors have found that an extract of unused resources of multiple wheats containing wheat powder has an effect of promoting neurite generation and nerve regeneration. Based on the above, the present invention is proposed.

That is, the present invention mainly comprises the following solution means.
(1) A nerve regeneration promoter containing one or more extracts selected from wheat bran, wheat powder, and barley bran and one or more substances selected from phosphatidylserine, theanine, docosahexaenoic acid, and ferulic acid.
(2) The nerve regeneration promoting agent according to (1), wherein the nerve is a peripheral nerve.
(3) The nerve regeneration promoter according to (1) or (2), which is an oral preparation.
(4) A food and drink having a nerve regeneration promoting function according to any one of (1) to (3).
(5) A composition comprising one or more extracts selected from wheat bran, wheat powder and barley bran and one or more substances selected from phosphatidylserine, theanine, docosahexaenoic acid and ferulic acid.

  The present invention can provide a new nerve regeneration-promoting agent. Moreover, the food / beverage products which accelerate nerve regeneration can be provided.

The graph which shows the neurite extension effect | action with respect to rat pheochromocytoma-derived PC12 cell by the wheat bran, wheat powder, and the water extract of barley bran. The schematic diagram which shows the location which cut | disconnected the reproduction | regeneration site | part for dyeing | staining sample preparation in order to confirm reproduction | regeneration of a sciatic nerve. The graph which shows the result of having measured the reproduced nerve bundle cross-sectional area by image analysis. The graph which shows the result of having measured the number of reproduced nerve axons by image analysis. The result of having measured the reproduction | regeneration nerve axotomy total area by image analysis is shown. The graph which shows the increase | augmentation of the nerve regeneration promotion effect by using together the water extract of wheat flour, and phosphatidylserine, theanine, docosahexaenoic acid, and ferulic acid.

  One or more extracts selected from wheat bran, wheat powder, barley bran and one or more substances selected from phosphatidylserine, theanine, docosahexaenoic acid, and ferulic acid having a nerve regeneration promoting function are provided. It consists of a composition to contain.

Barley and wheat are widely used as major food resources. Usually, the endosperm portion of seeds is milled and used for various foods. In the process of milling, the seed coat is screened and used for livestock feed as non-edible.
Wheat is classified into bran and wheat flour in the milling process. Wheat bran is the outer skin part of wheat, mostly composed of insoluble dietary fibers such as cellulose and lignin, and is a coarser material than wheat flour. Also contains endosperm.
The bran used in the present invention is obtained when white wheat is milled.
There are two main methods for classifying wheat flour. One is classified by type, and the other is classified by grade. The classification by grade comes from the form of the raw wheat grain.
Even in the endosperm portion of the same wheat grain, the central part has less ash, white color, and the amount of protein tends to be small. In the milling process, the powder taken mainly from this central part is classified and referred to as advanced powder. The fine powder has a low ash content and has a pale white or pale yellow color. On the contrary, the lower grade powder taken from near the epidermis has more protein, and the color becomes dull and brownish. In general, those with an ash content of 0.3 to 0.35% are special powder, those with 0.35 to 0.45% are 1st powder, those with 0.45 to 0.65% are 2nd powder, those with 0.7 to 1.0% are 3rd powder, and 1.2 ~ 2.0% is classified as powder. Powdered powder is not usually suitable for food, so it is used together with bran as animal feed or as an industrial paste material. The present invention uses this powder as a raw material.
Barley bran is a stage where white barley is refined for food use, and what is generated as a by-product is collectively called “barley bran”. “Barley bran” is classified into whether it is rough, finished or mixed depending on the particle size. “Barley bran” includes a seed coat and a portion derived from a germ, and is also referred to as “Bary Bran”, and is rich in β-glucan, which is a kind of dietary fiber.

In order to prepare the agent of the present invention from barley bran, the bran is extracted with water. The bran can be rough, finished, or mixed. In the present invention, these are collectively called barley bran.
In order to prepare the agent of the present invention from wheat bran, bran is extracted with water. In order to prepare the agent of the present invention from wheat powder, the powder is extracted with water. Hereinafter, when it is described as “powder”, it means wheat powder.
Although the powder may be commercially available for feed, it is preferable because it does not oxidize the lipid contained in the powder immediately after milling.

Add 5 to 10 parts by weight of water to 1 part by weight of wheat bran, wheat flour, or barley bran, and extract at room temperature for 0.5 to 1 hour with stirring. In order to shorten the extraction time, a physical vibration such as an ultrasonic vibration is applied to extract a water-soluble component. For small-scale extraction operation, after adding 5 parts by weight of water to 1 part by weight of wheat bran, powder, barley bran and stirring and dispersing, the active ingredient can be obtained by performing ultrasonic treatment for 3 to 10 minutes. It elutes in water.
Next, solid-liquid separation is performed by centrifugation or filter filtration, and the filtrate or supernatant is collected. The collected supernatant or filtrate is dried and powdered. The drying treatment can be applied by any method such as spray drying, drum dryer or freeze drying as long as it is a method for drying a normal liquid, but it is preferable to dry at a low temperature as much as possible in order not to reduce the activity.
By further adding phosphatidylserine, theanine, docosahexaenoic acid, ferulic acid and the like to the extract from wheat bran, wheat powder and barley bran thus obtained, the nerve growth promoting action is further enhanced. The blending amount of phosphatidylserine, theanine, docosahexaenoic acid and ferulic acid is preferably 10 to 200% based on the extract from wheat bran, wheat powder and barley bran, and particularly preferably wheat bran and wheat powder. In addition, the nerve regeneration effect can be most enhanced by blending the same amount of the nerve regeneration promoter comprising an extract from barley bran.
Phosphatidylserine, theanine, docosahexaenoic acid and ferulic acid are commercially available pharmaceutical raw materials or food raw materials which can be used as long as they contain 10% or more of the above components. Moreover, the safety | security of these components is also well-known, and the safety | security of the nerve regeneration promoter of this invention has no problem at all.

The nerve regeneration-promoting agent of the present invention is obtained by further pulverizing a dried product composed of an extract from wheat bran, wheat powder, and barley bran obtained by the above method to obtain ultrafine particles, phosphatidylserine, theanine, docosahexaene. Mix with acid, ferulic acid.
The composition and nerve regeneration-promoting agent of the present invention can be commercialized as powders, granules, tablets, and capsules by conventional methods. It can also be dissolved in water or fruit juice and commercialized as a liquid or jelly-like beverage. Furthermore, it can also be used by adding to various foods and drinks.
Such a food or drink is not particularly limited.
The effective dose of the composition of the present invention and the nerve regeneration-promoting agent is 1 to 10 g per day for an adult when taken orally.
The following examples further illustrate the present invention.

<Test Example 1>
1. Neurite outgrowth promotion test The neurite outgrowth action on neuroblasts is tested.
(1) Cells used and culture method
Rat pheochromocytoma-derived PC12 cells (passage number X + 14) purchased from DS Pharma Biomedical Co., Ltd. were used. The normal culture medium is fetal bovine serum (Life Technologies Japan) 5% inactivated in D-MEM (GIBCO (Life Technology Japan)), equine fetal serum (DS Pharma Biomedical), penicillin streptomycin ( Sigma Aldrich Japan Co., Ltd.) 0.05pot. Cultivation was performed in a collagen type I-coated 10 cm dish (IWAKI (AGC Techno Glass Co., Ltd.). When 80% confluent, cells were detached by pipetting, and a syringe with a 10 mL needle (21G x 1 1/2 "; Terumo The cells were pipetted about 20 times to make jingle cells, and cultured at 37 ° C. in the presence of 5% carbon dioxide and 95% air.

(2) Reagent preparation
Preparation of Nerve growth factor As a neurite growth factor, nerve growth factor (NGF, 2.5S Nerve Growth Factor, Mouse; Wako Pure Chemical Industries, Ltd.) was used. NGF is bovine serum albumin (BSA; Albumin, from Bovine Serum, Fraction V, Fatty acid-free, Nuclease- and Protease-Free; Merck Calbiochem (Merck Co., Ltd.)) 1 mg / mL Saline to 10 μg / mL It was prepared and added to a serum-free medium (D-MEM / 1% penicillin streptomycin / 1% N-2 supplement (GIBCO (Life Technology Japan)) to a final concentration of 10 μg / mL.

(3) Preparation of wheat powder extract 5 ml of water was added to 1 g of wheat powder manufactured by Nippon Flour Milling Co., Ltd., and subjected to ultrasonic treatment for 1 minute 30 seconds. Subsequently, after centrifugation at 1,000 rpm × 10 minutes with a centrifuge, the supernatant was recovered, and water was removed with a freeze dryer to obtain a water extract. This is hereinafter referred to as powdered powder extract.
This powder was stored at −20 ° C. and returned to room temperature when necessary.

(4) Preparation of wheat bran extract 5 mL of water was added to 1 g of wheat bran manufactured by Nippon Flour Milling Co., Ltd., and sonicated for 4 minutes. Thereafter, the mixture was centrifuged at 10,000 rpm for 15 minutes with a centrifuge, the supernatant was collected, and the water extract was made into a water extract by removing water with a freeze dryer. This is hereinafter referred to as a bran extract.
Similarly, it was stored at -20 ° C.

(5) Preparation of barley bran extract 5 mL of water was added to 1 g of barley bran prepared by the National Agricultural Research Organization, National Institute of Agriculture and Food Science, Kinki Shikoku Agricultural Research Center, and sonicated for 4 minutes. After centrifuging at 10,000 rpm × 10 minutes with a centrifuge, the supernatant was recovered and the water extract was obtained by removing water with a freeze dryer.
This is hereinafter referred to as barley bran extract.
Similarly, it was stored and used at -20 ° C.

(6) Preparation of test solution Each extract was prepared with distilled water to be 11.1100 mg / mL, 33.3100 mg / mL and 100 mg / mL, respectively.

(7) Plate seeding of test cells Passaged PC12 cells (passage number X + 17-20) were adjusted to 5.0 × 10 4 cells / mL with serum-free NGF-added medium. To this cell dispersion, add each extract preparation to medium (cell-free serum-free NGF-added medium) to a final concentration of 11.1 μg / mL, 33.3 μg / mL, and 100 μg / mL. Seeding was carried out on a plate (IWAKI (AGC Techno Glass Co., Ltd.)).

(8) Activity measurement of neurite outgrowth promoting action 48 hours after plate seeding (test substance exposure), photographs were taken with a stereomicroscope (4 locations / well), and neurites were elongated more than twice as much as the cell body. Number of cells / total number of cells (n = 4).

(10) Measurement results The neurite elongation action of wheat powder extract, bran extract and barley bran extract is shown in FIG.
The neurite outgrowth action of wheat powder extract was stronger than that of wheat bran extract. When the concentration of each extract was 11.1 and 33.3 μg / mL, the percentage of neurite-extended cells was 19.6% and 22.9% in wheat powder, respectively, but 15.9 and 20% until wheat bran . The wheat powder extract also had neurite outgrowth even at low concentrations (11.1 μg / mL). The barley bran extract was found to have almost the same activity as wheat flour.

2. Test to confirm sciatic nerve regeneration promotion effect.
In this test, the above 1. In this test, the nerve regeneration promoting action of the wheat powder extract, which has achieved good results in this test, was tested.
(1) Test sample The wheat flour powder extract prepared in (1) was used.

(2) Preparation of administration sample The test sample was weighed and dispensed in advance for each day, and the administration date and group were described and stored at -20 ° C. The dosing solution was prepared at the time of use, and dissolved by inversion mixing to 3.33 mg / mL and 10 mg / mL.

(3) Model animal rats, Slc: SD, 33 males: 5 weeks old Surgery: 6 weeks old Dissected: 10 weeks old

At the time of arrival, the species, strain, age, number of animals and sex were confirmed, the general state and appearance were observed, and the body weight was measured. The acclimatization period was 7 days.
Individual identification was performed by writing a number on the tail. Based on the weight of the day after surgery, groups were divided so that there was no difference in each group, and two animals after grouping were housed in cages, and a label clearly stating the group, animal number and individual identification number was placed on the front of the cage It was attached.

(4) Environmental conditions of the test system Set temperature: 24 ° C (allowable range 21-27 ° C), set humidity: 55% (allowable range 35-75%), lighting: 12 hours, 7am on, 7pm off Ventilation frequency: Animal breeding room maintained at 10-20 times / hour Animals were raised in a breeding room in a conventional area. The animals were reared in two per cage using plastic Ekon TPX cages (345 × 403 × 177 mm).
The feed was autoclaved (121 ° C., 25 minutes) for a solid feed (CRF-1, Oriental Yeast Co., Ltd.) within 3 months after being obtained, and placed freely in a feeder.
As drinking water, tap water that had been autoclaved (121 ° C., 25 minutes) was freely taken from a water bottle.

(5) Surgical method for creating test model animals Droperidol (Dololeptan Injection; Daiichi Sankyo Co., Ltd.) 0.25mg / kg and tribromoethanol solution * were administered intraperitoneally to 937.5mg / kg, and muscle relaxation and anesthesia were performed. gave. The outside of the left hind leg thigh was shaved, held in a recumbent position and disinfected with 70% ethanol. The upper part of the femur was cut about 1 to 2 cm, the muscle was bluntly detached, the sciatic nerve was exposed, and an 8 mm wide silicon sheet was laid under the sciatic nerve. A silicone tube (Sansho Co., Ltd.) with an inner diameter of 1 mm, an outer diameter of 2 mm, and a length of 6 mm was inserted into the sciatic nerve so that it could be opened and closed, and the silicone tube and sciatic nerve were sewn 1 mm from both ends. (Silk 7-0; Natsume Seisakusho Co., Ltd.). The silicon tube cut was opened and the sciatic nerve was cut to create a gap of about 1 mm. The gap was filled with physiological saline using a syringe, and the cut of the silicon tube was closed. The sciatic nerve was returned to its original position and the skin was sutured.

(6) Test sample administration method Forced oral administration using a gastric tube was used. The number of administrations, administration period, and liquid volume are once a day 28
Administered for 1 day. The dose was calculated at 10 mL / kg from the nearest measured body weight value.

(7) Group composition and dosage of test group The following dosages were used.
Dose (mg / kg) Number of animals used Control group 0 11
Wheat powder extract 33.3mg / kg group 33.3 11
Wheat powder extract 100 mg / kg group 100 11

(8) Anatomy, preparation of tissue specimen
After oral administration for 4 weeks, it was anesthetized with a tribromoethanol solution, exsanguinated from the abdominal aorta and sacrificed. The sciatic nerve as the surgical site was removed and immersed in a 10% neutral buffered formalin solution.
For paraffin embedding and block preparation, after being immersed in a 10% neutral buffered formalin solution overnight, as shown in Fig. 2, the upper part of the sciatic nerve regeneration site is colored with vermilion and the lower part with black ink, and sealed automatic fixation Degreasing, dehydration, and paraffin infiltration were performed using an embedding device (Tissue Tech VIP5 Junior; Sakura Finetech Japan Co., Ltd.). Blocks were prepared with a paraffin-embedded block preparation device (Tissue Tech TEC5 Embedding Console System; Sakura Finetech Japan Co., Ltd.).

(9) Slicing and immunostaining After excision of the upper part of the sciatic nerve (Vermilion), slice several μm from the upper part of the regeneration part for immunostaining so that there is no difference in each individual sliced part, Slides were made. For immunostaining, the primary antibody is neurofilament protein (Anti-Neurofilament M (145 kDa), C-terminus (NFP); Nihon Millipore Corporation), and the secondary antibody is Alexa F488: goat-anti rabbit F488 (fluorescent FITC-labeled 488; Life Technologies Japan Ltd.) was used.

(10) Image analysis a. Image analysis of nerve bundle cross-sectional area The image taken with increasing exposure time was binarized using photoshop, and the area of the black part was obtained using Image J (NIH Image 1.63).
A. Image analysis of the number of axons and the total area of axotomy
Using slides that were fluorescently stained with NFP (minus), an exposure time that did not emit fluorescence was set and photographed.
As with the nerve bundle cross section, binarization was performed using photoshop, and the number and total area of axons were determined using Image J (NIH Image 1.63).

(11) Results The image analysis results are shown in FIG. 3, FIG. 4, and FIG. As a result of the nerve bundle cross-sectional area (FIG. 3), it was confirmed that the nerve bundle cross-sectional area was significantly increased in the 100 mg / Kg administration group compared to the non-administration group.
Similarly, the number of axons (FIG. 4) was confirmed to be increased in the 100 mg / Kg-administered group compared to the non-administered group.
In addition, it was confirmed that the total area of axotomy (FIG. 5) was significantly increased in the wheat powder extract 100 mg / Kg administration group as compared to the non-administration group.
From the above results, it was clarified that the wheat powder extract promoted the regeneration of the cut nerve by oral administration.
<Test Example 2>
1. Confirmation of nerve regeneration enhancement effect of phosphatidylserine, theanine, docosahexaenoic acid and ferulic acid Neurite outgrowth promotion test The neurite outgrowth action on neuroblasts is tested.
(1) Cells used and culture method
Rat pheochromocytoma-derived PC12 cells (passage number X + 14) purchased from DS Pharma Biomedical Co., Ltd. were used. The normal culture medium is fetal bovine serum (Life Technologies Japan) 5% inactivated in D-MEM (GIBCO (Life Technology Japan)), equine fetal serum (DS Pharma Biomedical), penicillin streptomycin ( Sigma Aldrich Japan Co., Ltd.) 0.05pot. Cultivation was performed in a collagen type I-coated 10 cm dish (IWAKI (AGC Techno Glass Co., Ltd.). When 80% confluent, cells were detached by pipetting, and a syringe with a 10 mL needle (21G x 1 1/2 "; Terumo The cells were pipetted about 20 times to make jingle cells, and cultured at 37 ° C. in the presence of 5% carbon dioxide and 95% air.

(2) Reagent preparation
Preparation of Nerve growth factor As a neurite growth factor, nerve growth factor (NGF, 2.5S Nerve Growth Factor, Mouse; Wako Pure Chemical Industries, Ltd.) was used. NGF is bovine serum albumin (BSA; Albumin, from Bovine Serum, Fraction V, Fatty acid-free, Nuclease- and Protease-Free; Merck Calbiochem (Merck Co., Ltd.)) 1 mg / mL Saline to 10 μg / mL It was prepared and added to a serum-free medium (D-MEM / 1% penicillin streptomycin / 1% N-2 supplement (GIBCO (Life Technology Japan)) to a final concentration of 10 μg / mL.

(3) Preparation of wheat powder extract As in Test Example 1, 5 mL of water was added to 1 g of wheat powder manufactured by Nippon Flour Milling Co., Ltd., and subjected to ultrasonic treatment for 1 minute 30 seconds. Subsequently, after centrifugation at 1,000 rpm × 10 minutes with a centrifuge, the supernatant was recovered, and the water from which the water was removed with a freeze dryer was used as the powder extract.
This powder was stored at −20 ° C. and returned to room temperature when necessary.

(4) Phosphatidylserine, theanine, docosahexaenoic acid, ferulic acid Commercially available health food ingredients were used.
Phosphatidylserine (PS)
Product Name: Lipamine PS 90PN
Manufacturer: Cargill
Preparation solvent: Distilled water

L-Theanine Product Name: Suntheanine Manufacturer: Taiyo Chemical Co., Ltd. Solvent: Distilled water

Docosahexaenoic acid (DHA)
Product Name: DD Oil DHA-46
Manufacturer:
Preparation solvent: Dimethyl sulfoxide (DMSO)

Ferulic acid manufacturer: Tsukino Rice Fine Chemicals Co., Ltd. Preparation solvent: 80% ethanol

(6) Preparation of test solution Wheat flour extract, phosphatidylserine, L-theanine is distilled water, DHA is DMSO, and ferulic acid is 80% ethanol, dissolved or 25 mg / ml and 50 mg / ml, respectively. Suspended.

(7) Plate seeding of test cells Passaged PC12 cells (passage number X + 17-20) are 5.0 × 10 ⁴ cells / mL in serum-free NGF supplemented medium
It was. Each test solution is added to this cell dispersion to a medium (cell-free serum-free NGF-added medium) so that the final concentration is as follows, and then on a collagen type I-coated 24-well plate (IWAKI (AGC Techno Glass Co., Ltd.)) Sowing.
Control
Wheat powder 50μg / mL
PS 50μg / mL
Theanine 50μg / mL
DHA 50μg / mL
Ferulic acid 50μg / mL
Wheat powder 25μg / mL + PS 25μg / mL
Wheat powder 25μg / mL + Theanine 25μg / mL
Wheat powder 25μg / mL + DHA 25μg / mL
Wheat powder 25μg / mL + ferulic acid 25μg / mL

(8) Activity measurement of neurite outgrowth promoting action 48 hours after plate seeding (test substance exposure), photographs were taken with a stereomicroscope (4 locations / well), and neurites were elongated more than twice as much as the cell body. Number of cells / total number of cells (n = 4).

(9) Measurement results The measurement results are shown in FIG. It was found that the addition of phosphatidylserine, theanine, docosahexaenoic acid and ferulic acid to the wheat powder extract synergistically showed neurite outgrowth action.

Claims (5)

  A nerve regeneration promoter comprising one or more extracts selected from wheat bran, wheat powder, barley bran and one or more substances selected from phosphatidylserine, theanine, docosahexaenoic acid, and ferulic acid.   The nerve regeneration-promoting agent according to claim 1, wherein the nerve is a peripheral nerve.   The nerve regeneration-promoting agent according to claim 1 or 2, which is an oral agent.   The food-drinks which have the nerve regeneration promotion function in any one of Claims 1-4. A composition comprising one or more extracts selected from wheat bran, wheat flour and barley bran and one or more substances selected from phosphatidylserine, theanine, docosahexaenoic acid and ferulic acid.