JP2009114093A - Carcinogenesis prevention agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a carcinogenesis prevention agent safely administrable over a long time and exhibiting carcinogenesis prevention effect by oral administration and to provide foods and drinks containing the agent and having carcinogenesis suppressing action. <P>SOLUTION: The carcinogenesis prevention agent contains Sparassis crispa, or a substance obtained by the extraction treatment of Sparassis crispa or the treatment of the fungus with an enzyme such as endo-1, 4-glucanase, as an active component. Also disclosed are oral administration agents, foods and drinks containing the carcinogenesis prevention agent. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a carcinogenic preventive agent used for preventing carcinogenesis in humans and animals. The present invention also relates to an orally-administered agent and a food and drink characterized by containing this carcinogenesis-preventing agent.

  In recent years, the incidence of cancer has increased rapidly in Japan due to factors such as the aging of the population, westernization of dietary habits, and smoking. Among them, the incidence of colorectal cancer, lung cancer, liver cancer, pancreatic cancer, etc. And death rates are increasing year by year for both men and women. Currently, cancer accounts for about 30% of all Japanese deaths, and reducing deaths from cancer is an important issue as a national measure.

  In order to reduce cancer deaths, it is essential to take preventive measures not only for early detection and treatment, but also for carcinogenesis itself. The recent westernization of dietary habits is thought to have a significant impact on the increase in cancer patients, and by incorporating foods and food ingredients that are effective in preventing cancer into the diet, cancer risk is reduced. It is considered possible. In particular, regarding colorectal cancer, due to the nature of digestive organ cancer, it is expected that such an improvement in dietary habits is very effective in reducing the risk of developing colorectal cancer.

  In fact, vegetable oils containing conjugated linolenic acid (Non-patent document 1), broccoli sprout (Non-patent document 2), freeze-dried beer powder (Non-patent document 3) and the like have been confirmed to have a colon carcinogenesis-preventing action in animal experiments, Potential for clinical application is expected.

  On the other hand, mushrooms have been used for medicinal / edible use since ancient times, and β-glucan and the like contained in them have been shown to exhibit an antitumor effect since the 1970s. Kaletake-derived krestin and shiitake-derived lentinan In addition, β-glucan-containing preparations such as Schizophyllan derived from Suehirotake (both trade names) have been put to practical use as anti-neoplastic agents. In recent years, β-glucan contained in Agaricus (Kawariharatake), Meshimakobu, Ganoderma, Hanabiratake, etc. is expected to have an antitumor effect, and mushroom fruit bodies and mycelium dried products and extracts have been developed. It has come to be used as a health food material (for example, Patent Documents 1-3).

  Among the above mushrooms and components contained therein, Kaletake-derived Krestin, Bunaharitake, Tombaimaitake, Kawariharatake, Matsutake, etc. (Non-patent Documents 4-6, Patent Documents 4-6) have a carcinogenic inhibitory effect even at the level of animal experiments. Has been confirmed.

  Among the mushrooms mentioned above, Hanabiratake has a very high β-glucan content of 43% by mass or more per fruit body dry powder. It has been clarified that the fruit body of Hanabiratake and its purified β-glucan are administered to cancer-bearing mice, and it has been shown to exhibit very strong antitumor activity, and its use in the pharmaceutical field has also been proposed (Patent Document 7- 8, Non-Patent Documents 7-9).

As described above, Hanabiratake has strong antitumor activity against tumor-bearing mice, but its carcinogenesis-preventing action has never been studied.
Takuji Tanaka, Rikako Suzuki, “Cancer Prevention by Food Ingredients: Inhibition of Colon Carcinogenesis by Conjugated Linolenic Acid-Containing Plant Seed Oil”, Journal of Complementary and Alternative Medicine, (2005), 2 (2), 91-100 R. Suzuki, H. Kohno, S. Sugie, T. Okada and T. Tanaka, "Azoxymethane-induced rat colonic Aberrant Crypto Forsy Preventive effect of powdered broccoli sprout on azoxymethane-induced rat colonic abberant crypt foci ”, Journal of Toxicologic Pathology, (2004), 17: 119-126 H. Nozawa, A. Yoshida, O. Tajima, M. Katayama, H. Sonobe, K. Wakabayashi and K. Kondo, " Intake of beer inhibits azoxymethane-induced colonic carcinogenesis in male sischer 344 rats ”, International Journal of Cancer (Int. J. Cancer), (2004), 108 : 404-411 JP 2001-10970 A JP 2001-131083 A JP 2003-183176 A H. Kobayashi, K. Matsunaga and M. Fujii, “PSK as a chemopreventive agent”, Cancer Epidemology Biomarkers and Pre Bension (Cancer Epidemiology, Biomarkers & Prevention), (1993), 2: 271-276 Mikio Matayama, Osamu Tajima, Taku Sato, Tomomi Sonobe, "Inhibition of colon carcinogenesis by Beech Haritake", Annual Meeting of the Japan Society of Nutrition and Food Science, (2003), p.208 Fuyuki Hagiwara, Tomio Narusawa, “Inhibition of N-methylnitrosourea-induced colorectal carcinogenesis in rats fed dry powder (Meripilus giganteus)”, Journal of Mushroom Society of Japan, (2004), 12 (3): 125-130 Table WO 2004/004748 Japanese Patent Laid-Open No. 2004-210695 JP 2006-232719 A JP 2000-217543 A Japanese Patent No. 3509736 Akihiko Hasegawa, Munori Yamada, Munehiko Bluho, Ryuichi Fukushima, Naruaki Matsuura, Akio Sugidate, "On the immunomodulating action of Hanabiratake", Cancer and Chemotherapy, (2004), 31 (11): 1761-1763 N. Ohno, NN Miura, M. Nakajima and T. Yadomae, “Anti-tumor 1,3-betaglucan from cultured spalcis crispa fruiting bodies (Antitumor1, 3-beta-glucan from cultured fruit body of Sparassiscrispa). ”, Biological and Pharmaceutical Bulletin (Biol. Pharm. Bull.), (2000), 23 (7): 866-872 N. Ohno, S. Nameda, T. Harada, NN Miura, Y. Adachi, M. Nakajma, K. Yoshida, H. Yoshida and T. Yadomae, “Immunomodulating activity of a β-glucan preparation, SCG, using β-glucan preparation, SCG, extracted from a culinary-medicinal mushroom, Sparassis crispa Wulf.:Fr (Aphyllophoromycetidae), and application to cancer patients). "International Journal of Medicinal Mushroom (Int. J. Med. Mushr.), (2003), 5, 359- 368

  As described above, the carcinogenesis-preventing action of various food-derived ingredients has been clarified so far, and it has been proposed to incorporate them into the diet. Although these materials are considered to be effective in preventing carcinogenesis, the effect is not necessarily remarkable, and there are materials that are not guaranteed for safety when ingested for a long period of time.

  The present invention has been made in view of such circumstances, and is a carcinogenic agent that is safe even if taken for a long period of time and that exhibits a remarkable carcinogenic preventing effect even by oral administration, and a food and drink having a carcinogenic preventing effect. It is intended to provide.

  As a result of diligent research for the development of a safe and novel material in order to solve the above-mentioned problems, the present inventors have found the fact that Hanabiratake has a remarkable carcinogenic prevention effect and have reached the present invention.

  That is, the present invention is summarized as a carcinogenesis-preventing agent characterized by containing a fruit body, mycelium, or a processed product thereof as an active ingredient, and preferably has a carcinogenesis-preventing action as a colorectal carcinogenesis. It is a preventive agent for carcinogenesis.

  The second aspect of the present invention is a gist of an orally-administered agent containing the above-described carcinogenesis-preventing agent of the present invention as an active ingredient.

  The third aspect of the present invention is a food or drink product having a carcinogenesis-preventing effect containing the above-described carcinogenesis-preventing agent of the present invention as an active ingredient.

  ADVANTAGE OF THE INVENTION According to this invention, the carcinogenic preventive agent which shows the outstanding carcinogenic prevention effect can be provided. In addition, since it is made from Hanabiratake, which has been used for medicinal and edible use for a long time, it is extremely safe, so it can be used in orally administered preparations, and can also be used in food and drink. . Therefore, a suitable administration method can be selected for each ingestor.

  Hereinafter, the present invention will be described in detail.

  Hanabiratake is a very rare mushroom that grows on conifers such as larch. An edible mushroom that is crunchy and features a pure white color and a leaf-peony shape. Up until now, it has been said that this bamboo shoot has slow growth and is extremely difficult to artificially cultivate, but recently, a new cultivation method that can be cultivated in a relatively short period of time has been established and can be supplied on a commercial scale. It has become.

  The fruit body used in the present invention may be natural or cultivated. As a method of artificial cultivation, it can be performed by creating a conventionally known fungal bed for artificial cultivation (for example, Japanese Patent Laid-Open No. 11-56098, Japanese Patent No. 3746440, Japanese Patent No. 3509736). Issue).

  In the present invention, a mycelium of agaricus can also be used. The mycelium can be obtained by a liquid culture method. As a carbon source used in the medium, a commonly used carbon source such as dextrin and glycerol can be used in addition to a monosaccharide such as glucose. Moreover, although an inorganic or organic nitrogen source can be used as the nitrogen source, it is preferable to use an organic nitrogen source from the viewpoint of the growth rate. Moreover, adding growth factors such as trace elements and vitamins as needed is no different from normal culture. The culture temperature is 15 ° C to 30 ° C, preferably 18 ° C to 28 ° C, and most preferably 20 ° C to 25 ° C. The pH is 2.5 to 8.0, preferably 3.0 to 7.0, and most preferably 3.5 to 5.0. It is preferable to add an insoluble component to the medium component because it can be grown uniformly. The culture period can be set from several days to several weeks depending on the medium composition and strain. After completion of the culture, the mycelium and the culture filtrate are separated by centrifuging or filtering the culture liquid. Centrifugation can be performed by a centrifugal operation that gives a gravitational acceleration of 100 to 5000 G, preferably 800 to 3000 G. The filtration is performed using a membrane filter of 3.5 to 200 mesh, particularly preferably 4 to 16 mesh.

  The fruit body or mycelium of the bamboo shoot obtained in this way can be used as it is as the carcinogenesis-preventing agent of the present invention. In addition, a solvent extract extracted from a fruit body or mycelium of Hanabira bamboo using a solvent, or an enzyme-treated product obtained by treating a fruit body or mycelium of Hanabira bamboo with an enzyme agent is used as a carcinogenesis-preventing agent of the present invention. In order to obtain such a solvent extract or an enzyme-treated product, the fruit body or mycelium of Hanabiratake may be transferred to the treatment process as it is, or may be dried and processed into powder if necessary. Then, you may move to a processing step.

  Examples of the solvent used for obtaining the solvent extract from the fruit body or mycelium of the bamboo shoot include aqueous solutions and organic solvents. As the aqueous solution, pure water, acid aqueous solution, alkaline aqueous solution, salt solution, etc. can be used, and organic solvents include alcohol, acetonitrile, acetate ester, dimethyl sulfoxide (DMSO), dioxane, ether, glycol, THF, acetone, methylene chloride. , Chloroform, hexane, cyclohexane and the like can be used.

  Although there is no restriction | limiting in particular in the quantity of the solvent used for extraction, It is preferable to use 2-100 times amount with respect to the weight of Hanabira bamboo, and 5-50 times amount is further more preferable. The operability is less than twice the amount, and the work efficiency is poor if the amount is more than 100 times. The extraction can also be performed multiple times using one or more solvents. In the case of performing multiple times, the extraction may be performed from Hanabira bamboo, or the extracted fraction obtained from Hanabira bamboo may be further extracted. Moreover, it can carry out combining them.

  The temperature during the extraction operation is not particularly limited, but when an aqueous solution is used, it is preferably 4 to 200 ° C, more preferably 20 to 150 ° C, and most preferably 50 to 121 ° C. When the temperature is 4 ° C. or lower, the extraction efficiency is poor. When using an organic solvent, 0-50 degreeC is preferable, 4-40 degreeC is further more preferable, and 10-30 degreeC is the most preferable. If the temperature is 0 ° C. or lower, the extraction efficiency is poor. When using either an aqueous solution or an organic solvent, the extraction time is not particularly limited, but is preferably about 10 minutes to 3 days, more preferably 30 minutes to 2 days, and most preferably 2 hours to 1 day. The amount extracted is less than 10 minutes, and the work efficiency is low after 3 days. Extraction can be performed while standing, but the extraction efficiency can be increased by stirring or shaking.

  In addition, after further diluting or concentrating the extract to further increase the effectiveness and safety, ultrafiltration or reverse osmosis membrane treatment or those treated with activated carbon or various resins, or those treatment solutions are diluted or A concentrated product can also be used.

A processed product can be obtained by treating the fruit body and mycelium of the bamboo shoot with an enzyme agent containing endo-1,4-glucanase, xylanase and endo-1,3-glucanase. For example, preparations containing enzymes derived from Trichoderma viride (trade names “Fancerase”, “Cellulase Onoka” manufactured by Yakult Honsha, etc.) and commercially available enzyme agents such as hemicellulase, cellulase, mannase, arabanase Enzymes such as xylanase and pectinase can also be used.

  Enzymatic treatment conditions are preferably those conventionally known. In other words, the ratio of addition to Hanabira bamboo is 0.01 to 5%, preferably 0.05 to 1%, and most preferably 0.1 to 0.5% when "Cellulase Onoka" is taken as an example. The pH of the treatment liquid is 2.0 to 9.0, preferably 3.0 to 8.0, and most preferably 4.0 to 7.0. The enzyme action temperature is 20 ° C to 70 ° C, preferably 30 ° C to 65 ° C, and most preferably 40 ° C to 60 ° C. The enzyme treatment time may be set to several minutes to several hours depending on the process.

  After the enzyme treatment, the treatment liquid is heated to stop the enzyme reaction. Usually, the enzyme is inactivated by heating at 80 ° C. to 100 ° C. for about 10 minutes. Then, the treatment liquid may be dried by freeze drying or spray drying without filtering the residue to give a dry product such as powder, or the filtrate from which the residue has been filtered is concentrated to dry the extract or powder. It is good also as a thing. The enzyme-treated product of Hanabiratake of the present invention includes these dried products and concentrated extracts.

  The fruit bodies or mycelium of the bamboo shoots obtained as described above, or the processed product obtained by extracting and enzymatically treating them have an anti-carcinogenic effect and also have excellent characteristics such as no side effects. is doing.

  In general, the carcinogenic preventive agent of the present invention preferably contains 0.01 to 100% (w / w) of fruit bodies or mycelia of Hanabiratake or processed products thereof. More preferably, 0.1 to 80% (w / w) is added. If it is this range, formulation will be easy and sufficient effect can be expected.

  The form of the carcinogenic preventive agent of the present invention can be made into various forms depending on the way of application. In the case of oral administration, tablets, capsules, powders, granules, pills, solutions, emulsions, suspensions, solutions, spirits, syrups, extracts, and elixirs can be used.

  Various pharmaceutically acceptable carriers can be added to the formulation. For example, excipients, binders, disintegrants, lubricants, flavoring agents, coloring agents, sweeteners, corrigents, solubilizers, suspending agents, emulsifiers, coating agents can be included. It is not limited to. The carcinogenic preventive agent of the present invention may be sustained or sustained release.

  Cancer prevention in the present invention means to prevent the occurrence of cancer in humans and animals, but the symptoms that are so-called precancerous symptoms progress and the precancerous state becomes cancerous. It also includes the effect of delaying / suppressing. From such a viewpoint, it is desirable to continuously ingest the cancer preventive agent of the present invention on a daily basis.

  The cancer preventive effect in the present invention is not limited to the type of cancer, and is various cancers such as colorectal cancer, lung cancer, stomach cancer, breast cancer, prostate cancer, liver cancer, esophageal cancer, pancreatic cancer and the like. Although effective, it is extremely effective especially for the prevention of colorectal cancer.

  The 2nd oral administration agent of this invention contains the carcinogenesis preventive agent of above-described this invention. Hanabiratake is an extremely safe material because it has been considered edible and medicinal for a long time. From this point of view, it is considered that the amount of carcinogenic preventive used is not strictly limited, but generally the lower limit is an amount that can exert an effect according to the purpose of prevention, and the upper limit is ease of intake, economics From the viewpoint of sex and the like, a practical amount is used as a standard, and usually about 0.01 g to about 100 g, preferably about 0.1 g to about 10 g, per day for an adult in terms of dried bamboo shoots. Of course, it can be changed according to the age, weight, symptom, administration period, course of treatment, etc. of the ingested person. The daily dose can be taken in several divided doses.

  The oral administration agent of the present invention includes processed foods and drinks, quasi drugs, aqueous components used in pharmaceuticals, oily components, plant extracts, animal extracts, powders, surfactants, oils, alcohol, pH adjusters, antiseptics It is prepared by blending agents, antioxidants, thickeners, pigments, fragrances and the like together with the carcinogenic preventive agent of the present invention into raw materials. Forms can be tablets, capsules, powders, granules, pills, solutions, emulsions, suspensions, solutions, spirits, syrups, extracts, elixirs, but are not limited to these. . In addition, various pharmaceutically acceptable carriers can be added to the preparation. For example, excipients, binders, disintegrants, lubricants, flavoring agents, coloring agents, sweeteners, corrigents, solubilizers, suspending agents, emulsifiers, coating agents can be included. It is not limited to. The carcinogenic preventive agent of the present invention may be sustained or sustained release.

  The oral administration agent of the present invention includes, in addition to the carcinogenic preventive agent of the present invention and the above-described components, further inorganic components such as iron and calcium, various vitamins, oligosaccharides, dietary fibers such as chitosan, soybean extract And the like, lipids such as lecithin, and saccharides such as sucrose and lactose.

  Furthermore, the orally administered agent of the present invention includes, in addition to the above-described carcinogenic preventive agent of the present invention, other carcinogenic components, antioxidants, anti-inflammatory agents, orally administered anti-malignant tumor agents, animal-derived and / or Or it may contain plant-derived lactic acid bacteria preparations and other active ingredients. In such a case, stress related to carcinogenesis such as oxidative stress can be reduced, and the growth of cancerous cells can be suppressed. Therefore, it can be set as an oral administration agent having a higher effect.

  The 3rd food / beverage products of this invention contain the above-mentioned carcinogenesis preventive agent of this invention. Hanabiratake is an extremely safe material because it has been considered edible and medicinal for a long time. From this point of view, it is considered that the amount of carcinogenic preventive used is not strictly limited, but generally the lower limit is an amount that can exert an effect according to the purpose of prevention, and the upper limit is ease of intake, economics From the viewpoint of sex and the like, a practical amount is used as a standard, and usually about 0.01 g to about 100 g, preferably about 0.1 g to about 10 g, per day for an adult in terms of dried bamboo shoots. Of course, it can be changed according to the age, weight, symptom, administration period, course of treatment, etc. of the ingested person. The daily dose can be taken in several divided doses.

  The food and drink of the present invention are processed foods and drinks, quasi drugs, aqueous components used in pharmaceuticals, oily components, plant extracts, animal extracts, powders, surfactants, oils, alcohols, pH adjusters, preservatives. It is prepared by blending an antioxidant, a thickener, a pigment, a fragrance and the like together with the carcinogenesis-preventing agent of the present invention into the raw material. As forms, healthy foods and beverages such as tablets, capsules, powders, granules, pills, liquids, emulsions, suspensions, solutions, spirits, syrups, extracts and elixirs; noodles; breads; Beverages such as fruitless beverages, beverages containing fruit juice, lactic acid bacteria beverages, tea beverages, coffee beverages, soy milk beverages, soups; confectionery such as snacks, cookies, gums and candies; It can be dessert foods such as pudding, bavaria, jelly, yogurt, cake and other instant foods.

  In addition to the carcinogenic preventive agent of the present invention and the above-described components, the food and drink of the present invention are further inorganic components such as iron and calcium, various vitamins, dietary fibers such as oligosaccharides and chitosan, soy extract, etc. Proteins, lipids such as lecithin, and sugars such as sucrose and lactose may be included.

  In addition, the food and drink of the present invention contains an anti-carcinogenic agent characterized in that the existing health foods, beverages, confectionery, frozen desserts, desserts and other instant foods contain the above-mentioned Hanabiratake of the present invention. Can also be obtained.

  Furthermore, in addition to the above-described carcinogenic preventive agent of the present invention, the food / beverage product of the present invention includes other carcinogenic preventive components, antioxidant components, anti-inflammatory components, orally administered anti-tumor components, animal-derived and / or plants. Lactic acid bacteria derived from lactic acid bacteria and other active ingredients may be included. In such cases, stress related to carcinogenesis such as oxidative stress can be reduced, and the growth of cancerous cells can be suppressed. It can be set as the food / beverage products with a higher effect.

  Examples of the present invention will be given below, but these do not limit the present invention.

Production Example 1 [Manufacture of Hanabiratake fruit body]
Hanabiratake fruiting bodies were produced as follows. A fungus substrate containing larch sawdust, flour, nutrients (banana, honey, prawns, peptone, calcium chloride, hyponex) and water in a weight ratio of sawdust: flour: nutrient: water = 100: 11.5: 1.9: 51 Got ready. This microbial bed base material (520 g) was put into a 850 ml polypropylene culture bottle, and the culture bottle was sterilized according to a conventional method, followed by inoculation with inoculum of Hanabiratake (16 g). Then, the fruit body of Hanabiratake was harvested by leaving this culture bottle at a temperature of 23 ° C. for 56 days. The fruit body weight was 140 g per culture bottle.

Production Example 2 [Manufacture of agaric mycelium]
Hanabiratake mycelium was produced as follows. Yeast extract 0.4% by mass, glucose 2% by mass, potassium dihydrogen phosphate 0.1% by mass, disodium hydrogen phosphate 0.1% by mass dissolved in water, adjusted to pH 5.0 with 1N hydrogen chloride, 500ml 200 ml each was placed in an Erlenmeyer flask and sterilized according to a conventional method. An agar piece with a diameter of 6 mm was punched out from a flat plate medium on which the seeds of agaric mushrooms were grown in this liquid medium, inoculated with one piece, and cultured under shaking at 100 ° C. for 21 days in the dark at 24 ° C. Harvested. The dry weight of the mycelium was 2 g per Erlenmeyer flask.

Test Example 1 [Inhibition of growth on cancer cells]
The growth inhibitory action against cancer cells was evaluated for the extract of the fruit body of Hanabiratake obtained in Production Example 1 and the extract of the mycelium of Hanabiratake obtained in Production Example 2. Moreover, the growth inhibitory activity was compared with Agaricus. Agaricus purchased commercially available fruit body powder and used it for the test.

  The fruit bodies and mycelium lyophilized powder and Agaricus powder were suspended in RPMI 1640 medium containing 10% FBS at a concentration of 25 mg / ml and extracted at 50 ° C. for 1 hour. The obtained supernatant was sterilized by filtration through a 0.20 μm filter to obtain a test substance.

Human leukemia cell K562, human colon cancer cell Caco-2, human colon cancer cell HCT-116 (distributed from Osaka University School of Health Sciences) in 10% FBS-containing RPMI1640 medium and suspended in a 96-well plate at 6 × 10 ³ / well It dispensed so that it might become. The above test substance was added so that the final concentration was 2 mg / ml in terms of each mushroom powder, and cultured in an incubator under conditions of 37 ° C. and 5% CO ₂ . Cell counting kit-8 (manufactured by Doujin Chemical) was added at 10 μl / well at the start of culture (after 0 hours) and 48 hours after the start of culture, and left at 37 ° C. and 5% CO ₂ for 2 hours. Thereafter, the absorbance (450 nm-620 nm) was measured, and the inhibition rate of cell proliferation by each mushroom extract was calculated according to the following formula 1.

  Table 1 shows the growth inhibition rate for the growth of various cancer cells for the extract of Hanabiratake and Agaricus.

  From Table 1, it became clear that the extract of Hanabiratake has a remarkable growth inhibitory effect on various cancer cells. Moreover, the action was stronger than Agaricus.

Test Example 2 [AOM-induced rat precancerous lesion development prevention effect]
Azoxymethane (AOM: manufactured by SIGMA) is a chemical carcinogen that induces colorectal cancer when administered subcutaneously to animals and forms abnormal crypt foci (ACF) that are precancerous lesions. Using AOM-induced ACF formation as a test system, we evaluated the anti-carcinogenic effect of Hanabiratake.

  The fruit body of Hanabiratake obtained in Production Example 1 was dried with hot air and then crushed with a mill to obtain a dry powder. This was mixed with feed AIN-76 for experimental animals so as to be 0.3% (w / w), 1.0% (w / w), and 3.0% (w / w) to prepare a test feed. The control feed was AIN-76 without added Hanabira bamboo.

  The test groups consisted of 4 groups: a control group, an agaric bamboo powder 0.3% (w / w) dietary administration group, an agaric bamboo powder 1.0% (w / w) dietary administration group, and an agaric bamboo powder 3.0% (w / w) dietary administration group. The target animal was a male F344 rat. F344 rats (5 weeks old) were purchased from Japan SLC, and were reared in a light cycle of 7:00 to 19:00 in an environment of temperature 23 ± 2 ° C. and humidity 50 ± 10%. After one week of acclimatization, administration of the test feed described above was started. One week and two weeks after the start of administration, 15 mg / kg AOM was subcutaneously administered to induce colon carcinogenesis.

  Five weeks after the start of administration of the test feed, the rats were sacrificed and the large intestine was removed. This was thoroughly washed with physiological saline, then cut vertically from the anus to the cecum using dissecting scissors, formed into a plate, sandwiched between filter papers, and immersed in a 10% neutral formalin solution for 24 hours or more. Subsequently, formalin was removed by leaving it in running water for 30 minutes or more, and then immersed in a 0.2% (w / v) methylene blue solution for 10 minutes for staining. After staining, the attached pigment was removed in running water, then the number of ACF and abnormal crypt cells (Abberant crypt: AC) was counted using a stereomicroscope, and the average number of AC per ACF was also calculated. These values were tabulated for each group, and the average value and standard deviation of each were obtained.

  On the other hand, the colon tissue after ACF count was sliced into 3 μm thickness after embedding in paraffin, and after deparaffinization, the expression of apoptotic cells and PCNA was detected by immunohistochemical staining. Apoptotic cells were detected by TUNEL method using Apoptosis in situ Detection kit (Wako), and PCNA expression was detected by avidin-biotin complex method using anti-PCNA antibody (Dako). Staining was performed according to a conventional method, and the stained image was observed under a microscope to determine the ratio of positive cells.

  The count results of the ACF number are shown in Table 2 below. Statistical analysis was performed by Dunnett's test. Groups showing statistical significance are indicated by asterisks.

  The results of immunohistochemical staining are shown in Table 3 below. Statistical analysis was performed by Dunnett's test. Groups showing statistical significance are indicated by asterisks.

From Tables 2 and 3, it was clarified that oral intake of Hanabiratake has an inhibitory effect on the occurrence of AOM-induced colorectal precancerous lesions.

Claims (4)

An agent for preventing carcinogenesis, comprising Hanabiratake and / or a processed product thereof. The carcinogenic preventive agent according to claim 1, wherein the carcinogenic preventing action is a colon carcinogenic preventing action. An orally administered agent comprising the carcinogenic preventive agent according to claim 1 or 2. A food or drink product having a carcinogenesis-preventing effect, comprising the carcinogenesis-preventing agent according to claim 1 or 2.