JPH07109223A - Improver for activating liver function - Google Patents

Abstract

PURPOSE:To obtain a novel improver for activating liver function, especially an improver for activating liver function useful for fishes. CONSTITUTION:An improver for activating liver function contains yeast belonging to the genus Phaffia rhodozyma, especially an improver for activating liver function for fishes contains yeast belonging to the genus Phaffia rhodozyma.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel liver function activation improving agent.

[0002]

BACKGROUND OF THE INVENTION The liver is one of the most important organs in the body, which plays various important roles such as detoxification, regulation of metabolism of sugars, proteins and lipids, storage, regardless of mammals or fish. is there. For this reason, if the liver is damaged, the result is often fatal to the living body. As the cause of such liver damage, viral infection, ingestion of harmful substances, etc., and particularly in humans, excessive intake of alcohol is considered, and it is known that various liver diseases such as fatty liver, hepatitis, cirrhosis are caused by these. ing.

Such liver diseases have become a serious problem not only in humans but also in livestock and cultured fish in recent years. For example, due to recent advances in aquaculture techniques, many types of fish such as silver salmon, rainbow trout, yellowtail, sea bream, eel, and ayu have been cultivated. In an actual aquaculture site, there is a tendency for a large amount of food to be given to cultured fish in large quantities in a short period of time under overcrowding conditions, which causes stress accumulation in the fish and liver dysfunction due to liver enlargement and the like. It happens easily and is a big problem.

[0004] In humans, the liver damage is often accompanied by no subjective symptom, the discovery is delayed, and as a result, severe liver damage often occurs. Therefore, as part of the health checkup, the liver function is grasped by measuring GOT (glutamate oxaloacetate aminotransferase), (GPT (glutamate pyruvate aminotransferase), etc. in the blood, and the liver disease described above The method of predicting is widely used.

At present, antihistamines, ATP, arginine-hydrochloride, glutathione and the like are used as liver function improving agents. Furthermore, utilization of tea leaf extract (Japanese Patent Laid-Open No. 2-300836) and Streptococcus cells (Japanese Patent Laid-Open No. 61-221124) as liver function improving agents has also been investigated. However, the effect is not sufficient in any of the above.

Therefore, development of a drug that activates liver function and a drug that prevents liver function decline is expected from various fields.

[0007]

In view of the above-mentioned conventional techniques, the problem to be solved by the present invention is to provide a new liver function activation improving agent.

[0008]

Means for Solving the Problems The present inventor has conducted extensive studies in order to solve the above problems. As a result, administration of yeast cells belonging to the genus Phaffia rhodozyma suppresses liver hypertrophy, and at the same time, GOT level in the blood of the recipient is used as a main indicator of the state of liver function. ,
It was found that the GPT value and the TBA value (a numerical value showing the amount of peroxide of lipid, and that a large amount of peroxide is considered to indicate that the liver function is deteriorated) are remarkably improved. Was completed.

That is, the present invention is characterized by containing a yeast belonging to the genus Phaffia rhodozyma, characterized in that it contains a yeast function activation improving agent, in particular a yeast belonging to the genus Phaffia rhodozyma. It is to provide a liver function activation improving agent for fish. The present invention will be described below. Yeasts belonging to the genus Phaffia rhodozyma belong to basidiomycetes, which are known to accumulate large amounts of carotenoid pigments, especially astaxanthin in their cells (Intern.
ational Jornal of Bacteriology Vol.26, p286-291 (197
6)). And, such yeasts are currently used in various microorganism depository institutions such as American Type Culture Collection (ATC).
C), Institute For Fermentation, Osaka (IFO) and the like can be obtained as known yeast strains. In the present invention, the yeast strain belonging to the genus Phaffia rhodozyma obtained from the above depository can be used as it is. Examples of the yeast strain include IFO 10129.
Examples of the strain include ATCC 24202 strain. Furthermore, a yeast strain that has been mutated or transformed by a genetic engineering or protein engineering technique can be used.

As the means for the mutation, generally known means such as irradiation with ultraviolet rays; irradiation with radiation; use of agents such as nitrosoguanidine for inducing gene mutations can be widely used. Also, it is of course possible to use a plurality of these mutation means in combination. Further, as the genetic engineering method,
Segment-directed mutagenesis (Hir
ose, S., Takeuch, K. & Suzuki, Y., Proc, Natl.Acad.Sci.U
SA, 79, 7258-7262 (1982)); Site-directed mutagenesis by synthetic oligonucleotides (Hirose, S., Takeuch, K.,
Hori, H., Hirose, T., Inayama, S. & Suzuki, Y., Proc, Natl.
Acad. Sci. USA, 81, 1394-1397 (1984); Kramer, W. & Fritz,
H.-J., Methods in Enzymol., 154,350-367 (1987); Kunke
l, TA, Roberts, JD & Zakour, RA, Methods in Enzymo
l., 154, 367-382 (1987)).

Cultivation of the above yeast strain belonging to the genus Phaffia rhodozyma can be carried out using a medium in which a saccharide used in the culture of ordinary yeast is used as a carbon source and an organic nitrogen source is further added. As the carbon source, glucose, maltose, sucrose, invert sugar, molasses, etc. can be used. As the organic nitrogen source, for example, peptone, yeast extract, dry yeast, soybean flour, cottonseed flour, CSL (corn steep liquor), etc. can be used.

The culturing temperature can be in the range of 20 to 30 ° C., preferably around 24 ° C. The culturing time varies depending on the culturing temperature and other culturing conditions, but is usually preferably about 1 to 8 days. After the culturing, the bacterial cells of Phaffia rhodozyma yeast can be collected by using a generally known collecting method such as centrifugation.

In the present invention, "containing yeast belonging to the genus Phaffia rhodozyma" means not only containing the yeast cells obtained as described above as they are, but also by biological, chemical or physical means. A crushed yeast cell, an extract of the yeast cell, or the like can be used. If yeast cells are used as they are as an active ingredient of the liver function activation-improving agent of the present invention, there is an advantage that the production process of the liver function activation-improving agent can be simplified. Considering the increase in liver function activation improving effect of,
It is preferable to use yeast cells crushed by biological, chemical or physical means or the yeast cell extract as an active ingredient of the liver function activation improving agent of the present invention.

As the means for crushing the above-mentioned microbial cells, generally known yeast cells are treated with an enzyme agent mainly containing β1,3-glucanase; biological means such as about 0.2N sodium hydroxide and the like are treated with an alkaline solution. Chemical means of can be used. Among these treatment means, the alkali treatment is preferable in terms of treatment cost and treatment reliability. Further, as a means for physically crushing yeast cells, a generally known means for crushing the cells, for example, mechanical crushing with glass beads crusher such as wet glass bead crusher or high-pressure crusher such as Mantongarin. Processing means or the like can be adopted. Among these, it is preferable to employ the crushing process using a wet glass bead crusher in that the crushing process is reliable.

As a means for extracting the yeast cell extract, a commonly known extraction means such as solvent extraction can be used. For such solvent extraction, for example, a commonly known solvent extraction means using water, oils and fats, alcohols, acetone, or a mixture thereof can be adopted. When such an extract is used as an active ingredient of the liver function activation-improving agent of the present invention, it is preferable to use a concentrate obtained by concentrating the extract by a generally known method.

The above-mentioned bacterial cells may be wet bacterial cells immediately after culturing, or may be dry bacterial cells. Subjects to be administered with the liver function activation-improving agent of the present invention may be mammals or fish. Further, there is no limitation on the species of animals belonging to mammals or the specific types of fish belonging to fish, but when administered to fish, it is applied to cultured fish such as silver salmon, rainbow trout, hamachi, bream, eel, and ayu. It is realistic to administer.

The dosage form and dosage form can be selected according to the subject to be administered. That is, when the yeast cells are administered to fish, especially cultured fish, the yeast cells belonging to the genus Phaffia rhodozyma obtained as described above, the pulverized product and / or the extract are used in the present invention. Although it may be given as it is as a liver function activation-improving agent, it can be given by adding it to a normal diet. The amount added is 0.01% by weight or more, preferably 0.1% by weight or more based on the diet.

By administering such a fish liver function activation-improving agent to fish, the liver enlarged due to satiation is restored to a normal size, and parameters such as an elevated GOT value indicating liver function are improved to normal values. It is possible to Also,
When yeast belonging to the genus Phaffia rhodozyma is administered to mammals, particularly humans, the yeast can be used as a liver function activation-improving agent with the active ingredient. The liver function activation-improving agent can be used as a liver function activation-improving agent as it is by formulating yeast cells or crushed cells. In addition, a suitable pharmaceutical preparation carrier may be blended with the yeast cell to prepare a pharmaceutical composition for use. As the drug carrier, a filler, a filler, a binder, a moisturizer, which is usually used when a drug is prepared according to the use form,
Any excipient or diluent such as a disintegrant or a surface active agent can be used. The form of the pharmaceutical composition is not particularly limited as long as it contains the above yeast cells and the like in a form that effectively exerts a liver function activation improving action.
For example, it may be a solid preparation such as tablets, powders, granules and pills, or may be in the form of liquid preparations, suspension preparations, emulsions and the like. It is also possible to prepare a dry product which can be made into a liquid by adding a suitable carrier to the liquid agent or the like before use.

Each of the above-mentioned preparations can be prepared by a conventional method.

[0020]

EXAMPLES The present invention will be specifically described below with reference to examples. [Example 1] Yeast culture Phaffia rhodozyma IFO 10129 strain was preliminarily incubated at 121 ° C for 2
Glucose sterilized for 0 minutes 4%, peptone 0.3%, CSL
1%, ammonium sulfate 0.1%, monopotassium phosphate 0.1
One platinum loop was planted in 100 ml of a medium consisting of 10% and cultured at 23 ° C for 5 days. The cultured bacterial cells were collected by centrifugation and then freeze-dried.

From 5 l of the culture broth, 50 g of dried cells were obtained. [Example 2] Alkali treatment of yeast Phaffia rhodozyma IFO 10129 strain was pre-treated at 121 ° C for 2
Glucose sterilized for 0 minutes 4%, peptone 0.3%, CSL
1%, ammonium sulfate 0.1%, monopotassium phosphate 0.1
One platinum loop was planted in 100 ml of a medium consisting of 10% and cultured at 23 ° C. for 5 days.

The cultured cells were collected by centrifugation and then
Treat with 0.1N sodium hydroxide at room temperature for 1 hour and with sulfuric acid.
After adjusting the pH to 5, it was washed with water and dried with a spray dryer. About 40 g of dried cells was obtained from 5 l of the culture solution. The compositions of the yeasts obtained in Example 1 and this Example 2 were as shown in Table 1 below.

[0023]

[Table 1]

[Example 3] Mechanical treatment of yeast 50 g of the dry yeast cells obtained in Example 1 were crushed with a wet glass bead crusher (manufactured by Dynomill Shinmaru Enterprise Co., Ltd.), washed with water, and then dried. The dried product was dried to obtain 38 g of crushed cells. [Test Example 1] The experimental animals were divided into 4 groups, each containing 15 rainbow trout having a body weight of about 100 g that had been preliminarily bred for 2 weeks.

The diets for breeding used the diets shown in Table 2 as a basic diet, and instead of cellulose, 20 g of the yeast cells prepared in Example 2 was prepared in the third group, and in Example 4 in the fourth group. 20 g of yeast cells were added.

[0026]

[Table 2]

The rearing of the rainbow trout was carried out in a 30 cm × 40 cm × 60 cm aquarium with an average water temperature of 16 ° C. during the rearing period, and the first group was fed with a restricted feed of 1.4% of the fish weight per day. The second, third, and fourth groups have a fish weight of 2 per day.
About 15% of the satiety was fed. 61 days after breeding, fish weight, liver weight, serum GOT, GPT, T for 10 fish in each group
The BA value was measured, and the results are shown in Table 3.

[0028]

[Table 3]

In the table, * indicates a significant difference from the second group (P <
0.05). Compared with the first group that was subjected to the restricted feeding, the second group that was fed with the satiety food was clearly observed to have liver hypertrophy and a lowered liver function value. However, a clear improvement in the liver function value was observed in the third and fourth groups fed the diet supplemented with Phaffia rhodozyma yeast. [Example 4] Yeast culture Phaffia rhodozyma IFO 10129 strain was preliminarily incubated at 121 ° C for 2
Glucose sterilized for 0 minutes 4%, peptone 0.3%, CSL
1%, ammonium sulfate 0.1%, monopotassium phosphate 0.1
One platinum loop was planted in 100 ml of a medium consisting of 10% and cultured at 23 ° C for 5 days.

The cultured cells were collected by centrifugation and then
Treat with 0.1N sodium hydroxide at room temperature for 1 hour and with sulfuric acid.
After adjusting to pH 5, it was washed with water and dried with a spray dryer. About 80 g of dried cells was obtained from 10 l of the culture solution. [Example 5] Preparation of yeast extract To 40 g of the yeast cells obtained in Example 4, 200 ml of a water-methanol mixture (1: 9) was added, and the mixture was pulverized with a homogenizer and further shaken occasionally for 3 hours. I left it. After removing the yeast cells by centrifugation, the supernatant was concentrated under reduced pressure to obtain about 10 g of an oily substance. [Reference Example 1] Preparation of baker's yeast A commercially available moist product of baker's yeast (Saccharomyces cereviciae) was suspended in distilled water, crushed with a wet glass bead crusher (Dynomill), washed with water and dried. .

About 500 g of dry yeast was obtained from 500 g of the wet yeast. [Test Example 2] The experimental animals were divided into 4 groups, each containing 15 rainbow trout, which had been preliminarily bred for 2 weeks and had a body weight of about 100 g. Regarding the diet for breeding, the diet shown in Table 2 above is used as the basic diet, and
20 g of the yeast cells prepared in the above Example 4, 20 g of the yeast extract prepared in the above Example 5 in the third group, and 20 g of the baker's yeast prepared in the above Reference Example 1 in the fourth group. added.

The breeding was carried out in a 30 cm × 45 cm × 60 cm aquarium, and the average water temperature during the breeding period was 14 ° C.
About% of the feed was divided into two times a day, and fed by satiety. After 65 days of rearing, fish weight, liver weight, serum GOT, GPT, and TBA values were measured for 10 fish in each group,
The results are shown in Table 4.

[0033]

[Table 4]

In the table, * indicates a significant difference from the first group (P <
0.05). Compared to the first group to which yeast was not added, and the fourth group to which baker's yeast dry matter was added, in the second and third groups to which the Phaffia rhodozyma yeast cells or their extracts were added,
A clear improvement in the numerical values of liver function was observed.

[0035]

INDUSTRIAL APPLICABILITY The present invention provides a novel liver function activation improving agent, particularly a liver function activation improving agent used for fish.

Claims (2)

[Claims] 1. A liver function activation-improving agent comprising yeast belonging to the genus Phaffia rhodozyma. 2. A liver function activation-improving agent for fish, comprising yeast belonging to the genus Phaffia rhodozyma.