KR101468698B1 - Development of composition for improving Intestinal Microflora by using three steps fermentation - Google Patents

Abstract

The present invention relates to a composition for improving metamorphosis or intestinal flora prepared by a three-stage fermentation process and a method for producing the same. More specifically, the present invention relates to a food composition and a pharmaceutical composition, which are effective for improving metamorphosis, improving intestinal function and further for treating constipation by improving intestinal beneficial bacteria or intestinal flora and activating intestinal mucus secretion, and a method for producing the same .

Description

[0002] The present invention relates to a method for improving intestinal microflora using a three-stage fermentation process,

The present invention relates to a composition for amelioration or intestinal microflora improvement produced by a three-stage fermentation process and a method for producing the same. More specifically, the present invention relates to a food composition and a pharmaceutical composition which are effective for improving metamorphosis, improving intestinal flora, and further treating constipation by activating intestinal beneficial bacteria or intestinal flora and activating intestinal mucus secretion, and a method for producing the same .

Recent westernization of dietary habits has resulted in increased intake of high calorie foods such as meat and instant foods containing high fat and high protein, and the lack of exercise and stress accumulation are frequent, leading to decreased bowel function and increasing number of constipated patients.

Constipation refers to a condition in which defecation is not normally performed, and refers to a case in which there is a discomfort or a symptom accompanied by a physiological disorder by decreasing the number of stools and the number of stools when the stool is healthy and dry. Generally, if the number of bowel movements is less than twice a week or the daily bowel movement is less than 30 g, a diagnosis of constipation can be made. When constipation is present, not only is the abdomen swollen all the time, but also the toxins of the absent side are absorbed into the intestines and absorbed into the blood, thereby promoting aging of the skin, causing headache, acne and skin rash. It can cause hemorrhoids such as escape of the hemorrhoids. In addition, constipation is a cause of diverse and serious secondary diseases such as bad breath, colorectal diseases such as colorectal cancer, arteriosclerosis, hypertension, stroke, immune deficiency and so, active prevention and treatment are needed.

To date, various medicines and functional foods for improving intestinal microflora and constipation have been marketed, but the effect is temporary and causes various side effects depending on the kind. For example, when a stimulant containing an anthraquinone derivative such as senna or rhubarb is used as a herbal medicine, there are side effects such as abdominal pain and diarrhea, and it has been pointed out that there is a problem in taking or continuously taking during pregnancy. In addition, various functional foods including kelp, yacon, Saururus chinensis, Culchaem tea, Allium tea, aloe, algae, etc. have not been scientifically proved to be effective enough to solve the fundamental problem of constipation improvement. Reliability is inferior.

Accordingly, the present inventors have focused on research for developing a composition capable of fundamentally preventing or treating constipation by improving intestinal flora, and as a result, rice fermented extract by three-stage fermentation has excellent improving effect on intestinal flora improvement and constipation To complete the present invention.

It is an object of the present invention to provide a food composition for the modification or intestinal flora improvement comprising a rice fermentation extract by a three-stage fermentation method and a method for producing the same.

It is another object of the present invention to provide a pharmaceutical composition for preventing or treating constipation comprising a fermented rice extract by a three-stage fermentation method and a method for producing the same.

In order to achieve the above object, the present invention relates to a composition for improving denatured or intestinal flora containing a fermented rice extract by a three-stage fermentation method and a method for producing the same.

In the present invention, the term "rice fermentation extract" means a rice fermentation product obtained by fermenting rice or a product resulting from the fermentation product. Specifically, in the present invention, the term "rice fermentation extract" means rice fermented product itself, or the fermented rice may be obtained by a method such as extraction of a solvent (organic solvent such as water or ethanol), hot water extraction, cold extraction, reflux cooling extraction, It is meant to include extracted concentrates. The rice fermentation extract may be provided in the form of a solution, an extract or a powder by filtration or centrifugation, and there is no limitation on the form of the delivery. Preferably, the rice fermentation extract in the present invention means a rice fermentation extract prepared by a three-stage fermentation process.

In the present invention, the term " three-stage fermentation method "refers to a method of fermenting rice by applying a three-stage fermentation process with different temperature and time conditions to spread the micelle particles between rice, To maximize rice fermentation method.

Preferably, in the present invention, a method for producing a rice fermentation extract through a three-stage fermentation process may include the following steps:

1) mixing the rice with water and heating,

2) adding 0.05 to 10% by weight of enzyme to the above-mentioned grated rice, and saccharifying at 20 to 40 ° C for 10 to 30 hours;

3) a two-stage fermentation step in which the one-stage fermented rice is cultured at 20 to 50 DEG C for 1 to 10 days,

4) a three-stage fermentation step in which the two-stage fermented rice is cultured at 10 to 40 DEG C for 1 to 10 days, and

5) Sterilization and concentration steps.

Step 1) is a step of mixing rice with water and heat-grinding it. At this time, water is preferably added in an amount of 100 to 500% by weight, preferably 200 to 300% by weight, based on 100% by weight of the rice. It is also preferable to be called at 10 to 40 캜, preferably at 15 to 25 캜 for 1 to 24 hours, preferably for 2 to 6 hours. When rice is cooked in water and contains moderate water and then heated to steam rice, the starch particles of rice are gelatinized to form a wide gap between the micelles of the rice. So that fermentation can be promoted in a subsequent culturing step. The heat aging step can be carried out for about 30 to 120 minutes, preferably for 40 to 60 minutes, with heat of 50 to 120 캜, preferably 70 to 90 캜, but the conditions can be suitably selected within a range obvious to a person skilled in the art .

Step 2) is a one-stage fermentation step in which an enzyme is added to the above-mentioned gorgeous rice to be saccharified. This step is to saccharify the starch of rice to produce a monosaccharide, and to induce fermentation in the next step more easily by the monosaccharide.

In addition, the enzyme may be preferably leu. Nuruk can not only sacrifice the starch of rice but also prevent contamination of germs. In the present saccharification process, a composition containing a yeast mold can be used, and yeast can be usually used. Nuruk, white aspergillus ( Aspergillus kawachii , Aspergillus shirousamii, etc.), Aspergillus oryzae , Aspergillus niger , Aspergillus awamori , Aspergillus usamii Etc.) and Rhizopus sp., But the present invention is not limited thereto. In a specific embodiment of the present invention, white yeast was used as koji

The koji is preferably added in an amount of 0.05 to 10% by weight, preferably 0.1 to 1% by weight, based on 100% by weight of the gorgeous rice. The koji is preferably 10 to 45 캜, After 10 to 48 hours, preferably 10 to 30 hours, at 40 ° C, various enzymes are leached to act on rice starch and glycosylation proceeds.

Step 3) is a two-stage fermentation step in which the first-stage fermented rice is cultured at 20 to 50 DEG C for 1 to 10 days.

In this step, the rice subjected to the first stage fermentation can be cultured at 20 to 50 ° C, preferably 30 to 40 ° C, for 1 to 10 days, preferably 2 to 3 days.

Step 4) is a three-stage fermentation step in which the two-stage fermented rice is cultured at 10 to 40 DEG C for 1 to 10 days.

In this step, the rice subjected to the two-stage fermentation can be cultured at 10 to 40 캜, preferably 25 to 40 캜, for 1 to 10 days, preferably 2 to 3 days. By the cultivation step corresponding to the two-stage fermentation and the three-stage fermentation, the micellar particles of rice are widely diffused, and various enzymes can be smoothly performed, so that the fermentation can be more surely proceeded.

Step 5) is a step of sterilizing and concentrating the fermented rice fermentation extract which has been fermented. The sterilization and concentration can be carried out according to a conventional method known in the art, and the sterilized and concentrated rice fermentation extract can be used as a food composition or a pharmaceutical composition by pulverization by spray drying or freeze drying.

The rice fermented extract of the present invention produced as described above is characterized by being free of fat (crude fat, saturated fat, and trans fat), and little or no cholesterol. Preferably, the rice fermented extract of the present invention may comprise the following nutritional composition:

0 to 10 wt% of water, 0 to 3 wt% of ash, 60 to 95 wt% of carbohydrate, 5 to 30 wt% of saccharides, 0 to 10 wt% of crude protein, 0 to 0.1 wt% of crude fat, 0 to 0.1 wt% 0 to 0.1% by weight of trans fat, 0 to 0.1 mg / 100 g of cholesterol, and 0 to 500 mg / 100 g of sodium.

More preferably, the rice fermented extract of the present invention prepared as described above may include, but is not limited to, the following nutritional compositions:

A trans fat of 0.0% by weight, a cholesterol of 0 mg / 100 g, and a sodium of 47.08% by weight, water content of 7.0% by weight, ash content of 1.1% by weight, carbohydrate of 83.4% by weight, saccharides of 20.7% by weight, crude protein of 8.5% by weight, crude fat of 0.0% mg / 100g.

Preferably, the rice fermented extract of the present invention may contain 2 to 4% by weight of beta-glucan.

The composition containing the fermented rice extract of the present invention can be used as a food composition or a pharmaceutical composition for activating enteric bacteria in the intestines to improve intestinal flora, thereby improving denaturation and improving, preventing or treating constipation.

When used as a food composition, the fermented rice extract of the present invention can be applied to various foods such as beverages, soups, soups, frozen foods, and other processed foods such as breads, cookies, sweets, candies, gums, And can be provided as a health functional food. As used herein, the term " health functional food "means food prepared and processed using raw materials or ingredients having useful functions in accordance with the Act on Health Functional Foods, and" functional " To be used for the purpose of obtaining a beneficial effect for health use such as controlling nutrients or physiological action.

The amount of the composition of the present invention added to each health functional food may be set based on the effective intake amount per day for an adult, but is usually preferably 1 to 50% by weight. The method of addition is not particularly limited, and it may be added from the beginning together with other raw materials used for each food and drink. The amount of the extract in the food or beverage may generally be comprised between 0.01 and 80% by weight of the total weight of the food composition of the present invention.

In addition, the food composition of the present invention may further comprise a food-acceptable food-aid additive. Preferably, natural carbohydrates or various flavors and the like may be contained as additional components. For example, natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the flavor, natural flavoring agents (tau martin, stevia extract, for example, rebaudioside A and glycyrrhizin) and synthetic flavors (saccharin, aspartame, etc.) can be used. In addition, the food composition of the present invention may further contain various nutrients, flavors such as vitamins, electrolytes, dietary components, synthetic flavors and natural flavors, colorants and aging agents, pectic acid and its salts, alginic acid and its salts, Protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the present invention can be used as a lactic acid bacterial drink such as yogurt, or as a mixed agent such as paste. These components may be used independently or in combination.

In another aspect, the present invention relates to a pharmaceutical composition for preventing or treating constipation comprising a fermented rice extract.

The pharmaceutical composition comprising the fermented rice extract according to the present invention can be administered orally or parenterally in the form of powders, granules, solutions, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories, Examples of the carrier, excipient and diluent which can be contained in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, The present invention relates to a process for the preparation of a pharmaceutical composition for the preparation of a medicament for the preparation of a medicament for the preparation of a medicament for the preparation of a medicament for the preparation of a medicament for the preparation of a medicament for the preparation of a medicament for the preparation of a medicament, .

In the case of formulation, it can be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The amount of the pharmaceutical composition of the present invention to be used may vary depending on the age, sex, and body weight of the patient, but it is generally administered in an amount of 0.01 to 3 g / kg, preferably 1 to 3 g / can do. Further, the dose of the extract may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like, and therefore, the dose does not limit the scope of the present invention.

In addition, the composition containing the fermented rice extract of the present invention can be used for the purpose of further enhancing the effect of activating the enteric bacterium in the intestines to improve intestinal flora, thereby improving denaturation and improving, preventing or treating constipation. Yoghurt. ≪ / RTI >

Yogurt refers to lactic acid bacteria or yeast that multiply in the milk of mammals such as milk or soybean oil, and the milk protein formed by solidification of the milk protein by the acid produced at this time may be liquid or solid. Yogurt is fermented to enhance the acidity and flavor, lactic acid, lactic acid bacteria, peptones, peptides and lactic acid bacteria are contained in addition to the nutritional components of milk as the main raw material. Preferably, the liquid yogurt of the present invention preferably contains 10 million or more lactic acid bacteria per ml including Lactobacillus bifidus.

In a specific embodiment of the present invention, it is known to promote the growth and proliferation of bifidobacteria through HPLC analysis [Kor.J.Appl.Microbiol.Biotechnol. Vol.20, no.3, 237-242 (1992)] It has been confirmed that Panose is contained as a major component in rice fermented extract (FIGS. 1A and 1B). In order to confirm that the fermented rice fermented extract of the present invention containing Panus actually enhances the growth and proliferative activity of intestinal benefit bacteria such as bifidus bacteria and has the effect of improving intestinal flora and denaturation, Experiments were performed.

First, the ability of the bifidobacteria to proliferate was examined in vitro for fractions obtained by fractionating the fermented rice extract of the present invention by molecular weight (50 KD or less, 50-150 KD, 150 KD or more) and before the fractionation. As a result, The activity of rice fermented extracts in the presence of complex components such as Panus, yeast, protein, essential amino acids, vitamin B, and minerals, (See Figs. 2A and 2B).

In addition, when the rice fermentation extract was tested for its ability to proliferate against bifidobacteria and lactic acid bacteria, it was confirmed that the medium containing rice fermentation extract had superior ability to proliferate as compared with a medium without rice fermentation extract (see FIGS. 3 and 4). Further, as a result of confirming the ability of rice fermentation extract to multiply bifidobacteria and mixed bacteria of lactic acid bacteria under various medium conditions, excellent growth performance was also confirmed (see FIGS. 5A, 5B and 5C).

As a result of comparison with the medium (distilled water) control group in the normal rat model and the constipation-inducing rat model, in the rice fermented extract administration group according to the present invention, the fecal count, the water content in feces, the number of remaining feces in the colon, Diameter and surface mucosal thickness, small intestinal mobility, colonic mucosal thickness and number of mucus - producing cells were observed. In addition, in the case of the conventional sodium hypochlorite-treated sodium hypochlorite, a significant reduction in the amount of weight gain due to the excessive subcutaneous effect was observed, whereas in the group treated with the fermented rice extract of the present invention, there was little change in body weight and body weight, Respectively.

In addition, the comparison of the effect of lactic acid bacteria proliferation with the medium (distilled water) control group in the constipation inducing rat model showed that the lactic acid bacterial growth effect was excellent in the rice fermented extract administration group according to the present invention. This is a result indicating that the rice fermented extract composition of the present invention has a function of improving beneficial bacteria or intestinal flora in the intestines.

Furthermore, this proliferation effect was more remarkable when liquid yogurt containing rice fermented extract was administered.

Therefore, the composition for preventing intestinal flora improvement, improvement of metamorphosis and prevention and treatment of constipation diseases can be used for patients with intestinal dysfunction and constipation, and it is preferable to provide fundamental treatment.

The three-stage fermentation method of the present invention is effective in improving intestinal microflora and activating intestinal mucus through the activation of beneficial bacteria or intestinal microflora in the fermentation field of rice and thus has an excellent effect in improving constipation. Treatment can be provided.

FIG. 1A shows an analysis result of HPLC of panose sold as a standard material.
1B shows the results of analysis of the fermented rice extract using HPLC.
FIG. 2A shows the results of the fermentation of Bifidobacterium lactis The proliferation inducing effect on the absorbance The results are confirmed by (OD _600). Con1 is a control 1, sample 1 is a sample 1, Con2 is a control 2, sample 2 is a sample 2, Con3 is a control 3, sample 1 is a sample 1, whole is a fraction before fractionation, ~ 50K is fraction 50KD or less, 50K ~ Fraction of 150 K or more indicates a fraction of 150 KD or more.
Figure 2b shows the results of the fermentation of Bifidobacterium The results confirm the proliferative effect induced on longum in absorbance (OD _600). Con1 is a control 1, sample 1 is a sample 1, Con2 is a control 2, sample 2 is a sample 2, Con3 is a control 3, sample 1 is a sample 1, whole is a fraction before fractionation, ~ 50K is fraction 50KD or less, 50K ~ Fraction of 150 K or more indicates a fraction of 150 KD or more.
Fig. 3 is a graph showing the results of the fermentation of Bifidobacterium longum and Bifidobacterium lactis Absorbance a proliferative effect on the The results are confirmed by (OD _600). "Control" shows the result in BL medium without rice fermentation extract, and "intestinal microflora improving material" in BL medium supplemented with rice fermentation extract.
4 shows the results of confirming the proliferation effect of fermented rice extracts (FRe) on the lactic acid bacteria. The horizontal axis represents time (h) and the vertical axis represents OD (600 nm). "Control" represents the results of MRS medium without rice fermentation extract and "Fre" with MRS medium supplemented with rice fermentation extract.
FIG. 5A and the results of confirming the effect of the rice fermentation extract on the growth of bifidobacteria and mixed bacteria of lactic acid bacteria. "MRS" is the MRS medium in which the rice fermentation extract is not added, and "MRS + rice" is the MRS medium in the MRS medium supplemented with the rice fermented extract, and the unit of the horizontal axis is time (h) and the unit of the vertical axis is × 10 ⁴ CFU / Results are shown.
FIG. 5B shows the results of confirming the growth effect of rice fermentation extract on bifid bacteria and mixed bacteria of lactic acid bacteria. The horizontal axis in units of hours (h), the unit of the ordinate is a ^{× 10 4 CFU / ml, "} MRS + Maltose" the rice fermentation extract is not added to MRS + Maltose medium, "MRS + Maltose + rice" is a fermented rice extract The results are shown in the added MRS + Maltose medium.
FIG. 5c shows the results of confirming the growth effect of rice fermentation extract on bifidobacteria and mixed bacteria of lactic acid bacteria. The horizontal axis in units of hours (h), the unit of the ordinate is a ^{× 10 4 CFU / ml, "} RCM" rice fermentation extract is not added RCM medium, "RCM + rice" is in the RCM medium with the fermented rice extract added Results are shown.
Figure 6 shows the histological profile of the colon containing the fecal pellet in the normal rat model in alcian blue staining. (A) and (B) are medium control groups, (C) and (D) are drug control groups, (E) ) Represents the FRe300 group. The arrow indicates the surface mucosal thickness, M indicates the colonic mucosa, and F indicates the fecal pellet. The scale bar is 150 탆.
Figure 7 shows the histological profile of colon containing feces in alopecia induced constipation rat model in alcian blue staining. (A) and (B) are medium control groups, (C) and (D) are looperamide control groups, (E) and (F) are drug control groups, (J) represents FRe200 group, (K) and (L) represents FRe300 group. The arrow indicates the surface mucosal thickness, M indicates the colonic mucosa, and F indicates the fecal pellet. The scale bar is 150 탆.
Fig. 8 shows the results of confirming the change in the number of lactic acid bacteria in the contents of the cecum in a model of rofemide-induced constipation rats. (A), (B), (C), (C) and (D) show the control group, the FRE5 group, the FRe10 group, the Yoghurt group, the BFRe0.05 group, BFRe0.1 group and (H) BFRe1 group.
FIG. 9 shows the result of confirming the change in the number of lactic acid bacteria in feces in a model of rosemide-induced constipation rats. (A), (B), (C), (C) and (D) show the control group, the FRE5 group, the FRe10 group, the Yoghurt group, the BFRe0.05 group, BFRe0.1 group and (H) BFRe1 group.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Example  One. Rice fermentation  Preparation of extract

The rice was cooked at 15 ~ 25 ℃ for 2 ~ 6 hours. The rice was cooked at 80 ℃ using 6 times water. The temperature was lowered to 30 ℃ and the rice flour was added to the rice at 10%. Next, the saccharification process (first stage fermentation) was carried out at 30 ° C. for 12 hours, followed by incubation at 30 ° C. for 48 hours for culturing (second fermentation), followed by incubation at 25 ° C. for 72 hours Stage fermentation), followed by sterilization and filtration, followed by pulverization (spray drying or freeze-drying). Beta-glucan was selected as an index material for the rice fermentation extract prepared above, and analyzed according to the "Health Functional Food Slot II.3.4.3 Beta Glucan ". As a result, beta-glucan was found in the fermented rice extract ~ 4%, and exactly 3%.

In addition, it was confirmed that the nutritional components contained in the fermented rice extract were as follows:

A trans fat of 0.0% by weight, a cholesterol of 0 mg / 100 g, and a sodium of 47.08% by weight, water content of 7.0% by weight, ash content of 1.1% by weight, carbohydrate of 83.4% by weight, saccharides of 20.7% by weight, crude protein of 8.5% by weight, crude fat of 0.0% mg / 100g, calories 368kcal.

Example  2. Rice fermentation  The ability of extracts to proliferate lactic acid bacteria Bifidogenetic growth  stimulator, BGS ) exam

2-1. Rice fermentation  In the extract Panos ( Panose ) Component detection test

Panose (1 mg / ml) and rice fermented extract (0.3 mg / ml), which are sold as standard substances, were tested for the presence of Panose, which has a good effect on the growth and proliferation of Bifidus spp. RTI ID = 0.0 > HPLC < / RTI > under conditions:

Column: WATERS Sugar-PakTM1, 6.5 x 300 mm

Column temperature: 70 ° C

Solvent: water for HPLC

Flow: 0.4 ml / min

Injection: 20ul

As a result, peaks were found at the same time in Panose (FIG. 1A) and rice fermentation extract (FIG. This means that Panose is one of the major components of rice fermented extract. Accordingly, the inventors of the present invention conducted further experiments to ascertain whether the rice fermentation extract of the present invention has an effect of enhancing the growth and proliferative activity of enterobacteriaceae such as bifidobacteria.

2-2. Rice fermentation  Extract By fraction  Bifidus Proliferative ability  exam

In order to confirm whether rice fermented extract proliferates bifidobacteria, which are beneficial bacteria in the intestines, rice fermented extracts were fractionated by membrane separation and fractionated by intestinal microflora Bifidobacterium lactis And Bifidobacterium longum .

Specifically, a sample of 50 KD or less, 50-150 KD, 150 KD or more was dissolved in water for HPLC at a concentration of 40 mg / mL, and the filtrate obtained by filtration with a 0.45 μm (PVDF) syringe filter and the unfiltered solution Bifidobacterium was used as a sample for measurement of proliferative activity. Sample (fraction I, more than 150 KD, 150 ~ 50 KD, 50KD or less) is taken for 100 ㎕ dissolved at 10% in deionized water prior to the experiment, was added to a 5mL RCM (Reinforced Clostridial Medium) culture medium, B. lactis and B. longum were incubated at 37 ° C in an anaerobic condition, and then the absorbance (OD ₆₀₀ ) was measured at the time of 16 hours. The experiment was carried out with three iterations, and the mean value and standard deviation were plotted on the graph. Control was done by adding sterile distilled water to RCM broth as the primary medium.

As a result, as shown in Table 1, the proliferative effect did not differ greatly depending on the molecular weight of the rice fermentation extract fraction. On the other hand, Bipidogenic growth stimulant (BGS) activity was higher in the mixtures than the fractions in samples 1, 2 and 3, which were not separated by molecular weight. Among them, BGS activity of sample 3 was the highest. This means that when the effective factors such as yeast, protein, essential amino acid, vitamin B, and minerals contained in the rice fermentation extract are present in a complex form rather than a partial form, the proliferative activity of the intestinal beneficial bacteria is relatively higher (FIGS. 2A and 2B) 2b).

sample B. lactis absorbance B. longum absorbance Control 1 0.172 ± 0.009 0.153 + 0.028 Sample 1 (whole) 0.618 + 0.013 0.688 ± 0.001 Sample 1 (~ 50KD) 0.210 + 0.022 0.230 0.010 Sample 1 (50K to 150KD) 0.315 + 0.036 0.431 + 0.031 Sample 1 (150 KD) 0.249 + 0.029 0.183 + 0.011 Control group 2 0.172 + 0.010 0.153 + 0.011 Sample 2 (whole) 0.351 + 0.030 0.443 + 0.012 Sample 2 (~ 50KD) 0.327 + 0.034 0.409 + 0.018 Sample 2 (50KD to 150KD) 0.264 + 0.032 0.404 ± 0.001 Sample 2 (150 KD) 0.280 + 0.026 0.338 + 0.014 Control group 3 0.172 + 0.010 0.153 + 0.011 Sample 3 (before fractionation) 0.841 + 0.032 0.796 + 0.024 Sample 3 (~ 50KD) 0.421 + 0.098 0.540 + 0.015 Sample 3 (50KD to 150KD) 0.355 + 0.027 0.488 + 0.029 Sample 3 (150 KD) 0.196 + 0.004 0.235 + 0.011

2-3. Rice fermentation  Bifidus of extract Proliferative ability  exam

After dissolving the fermented rice extract, with 10% in the distilled water prior to the experiment by taking the 100 ㎕, it added to 5mL BL medium ^{(Difco TM BL Broth, Becton,} Dickinson and Company), B. lactis And B. then inoculated by 1%, respectively for 12 hours at 37 ℃ longum culture in anaerobic conditions by measuring the absorbance (OD ₆₀₀₎ was confirmed that proliferation. As a control, 100 쨉 l of sterilized distilled water was added to a BL medium to which no rice fermentation extract as a basic medium was added. As a result, the rice fermentation extract showed a significant proliferative effect on bifidobacteria (FIG. 3).

2-4. Rice fermentation  Lactic acid bacteria of extract Proliferative ability  exam

The rice fermented extract was dissolved in 10% of the third distilled water before the experiment. 100 μl of the extract was added to 5 ml MRS broth (Difco ^™ MRS broth, Becton, Dickinson and Company), and B. lactis was inoculated 1% For 72 hours at 37 ° C., and the absorbance (OD ₆₀₀ ) thereof was measured to confirm its proliferative activity. As a control, 100 μl of sterilized distilled water was added to the MRS medium to which the rice fermentation extract as the basic medium had not been added. As a result, the rice fermented extract showed a significant proliferative effect on the lactic acid bacteria (FIG. 4).

2-5. Rice fermentation  Bifidobacteria and lactic acid bacteria Mixed fungus Proliferative ability  exam

To confirm the effect of rice fermentation extract on the growth of bifidobacteria and lactic acid bacteria in various lactic acid bacteria / bifidus culture media, Streptococcus thermophiles , Bifidobacterium lactis , Lactobacillus acidophilus was confirmed.

The rice fermented extract was dissolved in 10% of the third distilled water before the experiment, and 100 μl of the rice fermentation extract was added to the culture medium of MRS (Difco ^™ MRS broth, Becton, Dickinson and Company) and MRS + maltose medium, (Difco ^™ RCM Broth, Becton, Dickinson and Company) medium. The cells were incubated at 37 ° C. for 16 hours under anaerobic conditions. The control (control) was carried out by adding 100 of sterilized distilled water to three media without rice fermentation extract. As a result, the fermented rice extract showed significant proliferative effect on the mixed bacteria (Table 2, Figs. 5A, 5B and 5C).

time MRS
(CFU / ml) MRS + rice
(CFU / ml) MRS + Maltose
(CFU / ml) MRS + Maltose + rice
(CFU / ml) RCM
(CFU / ml) RCM + rice
(CFU / ml) 0H 0 0 0 0 0 0 6H 0 0 0 0 3.6 × 10 ⁴ 8.3 × 10 ⁴ 12H 5.2 × 10 ⁴ 4.6 × 10 ⁵ 1.0 × 10 ⁶ 1.8 × 10 ⁶ 1.3 × 10 ⁶ 2.5 × 10 ⁶ 16H 2.6 x 10 ⁶ 3.8 × 10 ⁶ 7.0 × 10 ⁶ 1.4 × 10 ⁷ 2.9 × 10 ⁶ 2.2 x 10 ⁷

Rice represents rice fermented extract.

2-6. Rice fermentation  Experimental result of extract powder

In order to confirm the effect of bifidobacteria propagation, the ability of the rice fermentation extract powder to bifidus bacterium BB-12 was confirmed.

The control (control) was a Bifidobacterium enhancer (a kind of sediment derived from Japanese sake, referred to as sakekazu or obaku)) was used as a comparative experimental group, and whey extract powder (Whey extract powder) was used as a comparative test group. Each of the rice fermented extract powder, control, liquid whey and whey extract powder was dissolved in 10% of the third distilled water before the experiment, and 100 μl of the powder was added to 5 ml of the BL medium. Each of the bifidobacteria BB-12 was inoculated at 1% After incubation at 40 ℃ for 12 hours under anaerobic conditions, the growth ability was confirmed. In the no-added test group, 100 μl of sterilized distilled water was added. As a result, the rice fermented extract powder showed a higher proliferative activity than the liquid whey and whey extract powder, and exhibited a similar proliferative activity to the BL medium supplemented with the Bifidobacterium enhancer (Table 3).

Item Number of colonies of BB-12 Control group Bifidobacterium enhancer > 6.9 x 10 < ⁷ > cfu / mL Experimental group No additives ≫ 1.3 x 10 < ⁶ > cfu / mL Liquid whey ≫ 3.4 x 10 < ⁷ > cfu / mL Whey extract powder > 4.7 × 10 ⁷ cfu / mL Rice fermented extract powder > 6.8 × 10 ⁷ cfu / mL

Example  3. Normal In rats Rice fermentation  Subcutaneous extraction ( 泻 下 ) effect

The subchronic effects of rice fermented extracts were evaluated using normal rats. (Fecal count, wet weight, dry weight, and moisture content) for 24 hours, fecal (fecal), and fecal (fecal) fecal extracts were administered orally once a day for 15 days, pellet) and the mean diameter, the migration rate of the intestinal epithelium, and the histopathologic changes of the colonic feces and mucosa were observed. The results were compared with those of 5 mg / kg sodium picosulfate, which is currently used as a subcutaneous agent.

Six 7-week-old normal SD rats (6-week-old male, SLC, Japan) were prepared with 7 groups per group as follows (FRe represents fermented rice extracts)

Medium control: sterilized distilled water administered control

Drug Control: 5 mg / kg pico sodium sulfate administered group

FRe 100 group: Rice fermented extract 100 mg / kg group

FRe 200 group: rice fermented extract 200 mg / kg group

FRe 300 group: rice fermented extract 300 mg / kg group

Sterile distilled water was used as a vehicle for rice fermentation extract and was orally administered once a day for 15 days. The body weight was recorded 1 day before administration, on the day of administration, on days 1, 7, 14 and 15 after administration, and feces index (24 hours of feces, Wet weight, dry weight and moisture content) were respectively measured. In addition, 15 days after the start of the administration, 18 hours of fasting, the carotid transfer rate in the small intestine was measured, and histopathological changes of the colonic feces and mucosa were observed.

Fecal Moisture Content = [(Wet Wet Weight - Fecal Dry Weight) / Wet Wet Weight] x 100

Carbon shift rate in small intestine = [(total length of the small intestine - moving distance of the intestine) / total length of the small intestine] x 100

Histomorphometry: Fecal surface mucosal thickness and mucin production per mm ² (alcian blue positive cells), mean mucosal thickness

3-1. Weight and Weight gain  change

A significant decrease in body weight (p <0.05) was observed in the sodium picosasulfate group compared to the vehicle control group, but no significant changes in body weight and body weight were observed in the rice-fermented extract-treated group compared to the vehicle control group (Table 4). In addition, the amount of body weight gain during the administration period was -33.60, 1.33, 8.53, and 4.27% in the group administered with 100, 200 and 300 mg / kg of rice picosaproate, rice fermented extract, respectively. That is, a significant reduction in weight gain was observed in the pico sodium sulfate-treated group, but not in the case of the rice fermented extract-treated group, and the adverse effects due to excessive subcutaneous effects were not observed .

group weight Weight gain
[BA]
Starting date of administration [A] After 14 days of administration [B] Last (15th) Administration * Medium control group 323.57 ± 12.96 377.14 ± 17.19 347.57 ± 14.23 53.57 7.68 Drug control 325.00 + - 9.78 360.57 ± 17.38 329.00 ± 15.02 35.57 ± 9.80 ^a FRe 100 group 325.14 + - 9.14 379.43 + - 13.45 345.14 + - 12.81 54.29 + - 7.30 FRe 200 group 322.86 + - 14.11 381.00 ± 29.48 345.86 ± 26.68 58.14 ± 18.93 FRe 300 group 322.71 + - 10.98 378.57 ± 18.91 344.86 ± 18.31 55.86 + - 10.02

Each value represents the mean ± standard deviation of 7 rats (unit: g)

* Overnight fasted

^a p < 0.01, compared to vehicle control by MW test.

3-2. Feces  Change in the indicator

Fecal counts, wet weights, dry weights, and moisture contents similar to those of the vehicle control group were recognized before the start of administration but were significantly (p <0.01 or p < 0.05) An increase in the content was recognized in sodium picosulfate and in all three rice fermented extract-treated groups (Tables 5 to 8).

18, 18.36 and 18.36% in 24, 24, and 24 hours, respectively, compared with the control group, and the fecal numbers in the 100, 200 and 300 mg / kg of rice fermented extracts showed a change of 58.31, 18.11, 18.36 and 18.36% , 16.88, 17.66 and 20.26%, respectively. In addition, the fecal moisture content was 93.59, 17.27, 24.97 and 23.90% in the 100, 200 and 300 mg / kg of rice fermented extract, respectively, after 7 days of administration compared with the vehicle control group. , 18.05, 31.72 and 32.90%, respectively.

Fecal indicators such as the number of feces and the water content in the feces have been used to assess the sub-efficacy of various drugs, as the fecal excretion of constipation is significantly reduced and excessive water absorption has been known to significantly reduce fecal water content (Wintola OA, et al., BMC Gastroenterol. 10:95, 2010; Wu D. et al., J Ethnopharmacol. 134: 406-13, 2011). Therefore, the increase in the fecal water and the increase in the water content in the feces by administration of the fermented rice extract can be regarded as a direct proof of the subcutaneous effect of the fermented rice extract.

group The number of feces after the administration day (24 hours) One day before administration ~ Start date of administration After 6 to 7 days of administration 13-14 days after administration Medium control group 58.43 ± 6.19 57.57 + - 6.95 55.00 ± 5.80 Drug control 58.86 + - 6.44 91.14 ± 11.23 ^a 79.43 ± 12.22 ^a FRe 100 group 59.71 + - 7.87 68.00 ± 4.93 ^b 64.29 + - 7.43 ^b FRe 200 group 60.71 + - 9.05 68.14 + - 6.47 ^b 64.71 + - 7.13 ^b FRe 300 group 59.14 ± 9.89 68.14 + - 4.45 ^b 66.14 + - 4.38 ^b

Each value represents the mean ± standard deviation of 7 rats (unit: dogs)

^a p < 0.01 and ^b p < 0.05, as compared to vehicle control by LSD test.

group After 24 hours of administration, One day before administration ~ Start date of administration After 6 to 7 days of administration 13-14 days after administration Medium control group 10.144 ± 1.128 10.061 + 1.744 10.249 0.736 Drug control 10.273 0.979 28.673 ± 3.226 ^a 27.973 + - 2.850 ^c FRe 100 group 10.247 ± 1.139 13.217 + 1.303 12.594 + - 1.802 ^c FRe 200 group 10.366 + - 0.733 13.362 ± 1.437 13.768 ± 2.526 ^c FRe 300 group 10.465 ± 1.626 13.462 ± 2.261 ^b 13.423 ± 2.693 ^c

Each value represents the mean ± standard deviation of 7 rats (unit: g)

^a p < 0.01 and ^b p < 0.05, as compared to vehicle control by LSD test.

^c p < 0.01, as compared to drug control by MW test.

group After 24 hours of administration, the fecal dry weight One day before administration ~ Start date of administration After 6 to 7 days of administration 13-14 days after administration Medium control group 6.094 + - 0.337 6.934 + 0.653 6.599 + 0.362 Drug control 6.182 + 0.440 8.288 + 0.793 ^a 7.614 ± 0.736 ^a FRe 100 group 6.230 0.609 7.510 +/- 0.656 7.307 + - 0.836 FRe 200 group 6.291 + - 0.350 7.193 + 0.709 7.201 + - 0.812 FRe 300 group 6.182 ± 0.793 7.263 + 1.039 6.889 ± 1.202

Each value represents the mean ± standard deviation of 7 rats (unit: g)

^a p < 0.01, compared to vehicle control by LSD test.

group Fecal moisture content after administration (24 hours) One day before administration ~ Start date of administration After 6 to 7 days of administration 13-14 days after administration Medium control group 39.581 ± 3.801 36.671 ± 5.276 35.372 ± 5.073 Drug control 39.592 + - 4.275 70.992 + 1.967 ^a 72.685 + - 2.442 ^c FRe 100 group 38.981 + - 4.310 43.003 + 4.092 41.758 ± 2.678 ^d FRe 200 group 39.101 + - 4.716 45.828 ± 5.728 ^a 46.592 ± 8.476 ^c FRe 300 group 40.550 + - 4.868 45.433 + - 6.911 ^a 47.009 ± 11.797 ^d

Each value represents the mean ± standard deviation of 7 rats (unit:%)

^a p < 0.01 and ^b p < 0.05, as compared to vehicle control by LSD test.

^c p < 0.01, ^d p < 0.05 MW when compared to drug control by test.

3-3. Colon Fecal  change

The decrease in the number of fecal remnants in the colon compared to the medium control group was observed in the pseudomonasulfate and three doses of all rice fermented extract administration groups and the average diameter of the remaining feces in the colon was also similar to that of the medium control (P <0.01 or p <0.05), respectively, except for the 100 mg / kg rice fermented extract group (Table 9).

The residual fecal numbers in the colon compared to the medium control showed -68.75, -40.63, -46.88 and -43.75% in the picosanoic acid sodium and the rice fermented extract 100, 200 and 300 mg / kg, respectively. The average diameter of the remaining feces in the colon Were -34.44, -2.29, -9.96 and -11.96%, respectively.

group Remaining feces in the colon Count Average Diameter (um) Medium control group 4.57 ± 0.98 4.93 ± 0.31 Drug control 1.43 + 0.98 ^a 3.24 ± 0.48 ^a FRe 100 group 2.71 ± 1.11 ^a 4.82 ± 0.40 FRe 200 group 2.43 ± 1.27 ^a 4.44 ± 0.41 ^b FRe 300 group 2.57 + 0.79 ^a 4.34 ± 0.41 ^b

Each value represents the mean ± standard deviation of 7 rats

^a p < 0.01 and ^b p < 0.05, as compared to vehicle control by LSD test.

3-4. In the collection Tombstone  Change in the moving rate

In the pico sodium sulfate administration group, similar to the media control group, there was a significant increase in the intestinal transport rate (p <0.01 or p <0.05) compared to the medium control group in all three rice fermented extract administration groups 10).

10.00, 10.99, 16.10 and 14.13% in the small intestine, respectively, compared to the control group, in the 100, 200 and 300 mg / kg of rice fermented extract, respectively.

It is known that the intestinal permeability of the substance represents the motility of the entire gastrointestinal tract, and that the measurement of the intestinal balloon transport rate is very useful for diagnosing constipation (Wintola et al., BMC Gastroenterol. 10:95, 2010). Thus, a reduction in balloon mobility in the small intestine indicates constipation (Sagar et al., BMC Complement Altern Med. 5:18, 2005; Meite et al., BMC Complement Altern. Med. 10: 7, 2010).

Therefore, the increase of the carbon transfer rate in the small intestine of the rice fermented extract as a result of this experiment can be regarded as indirect evidence of the subterranean effect of the fermented rice extract.

group Camouflage moiety (for 30 minutes) Total length (cm) Carbon shift length (cm) Carbon shift in small intestine (%) Medium control group 105.19 ± 4.99 73.57 ± 7.20 69.88 + - 5.31 Drug control 104.57 + - 4.73 75.14 + - 7.55 71.85 + - 6.55 FRe 100 group 105.57 ± 5.63 81.64 ± 4.86 ^b 77.56 ± 6.82 ^b FRe 200 group 105.07 ± 4.77 85.21 + - 8.14 ^a 81.14 ± 7.29 ^a FRe 300 group 105.93 ± 2.73 84.43 +/- 7.55 ^a 79.76 +/- 7.45 ^b

Each value represents the mean ± standard deviation of 7 rats

^a p < 0.01 and ^b p < 0.05, as compared to vehicle control by LSD test.

3-5. Histopathological change

The number of mucin-producing cells per unit area of the intestinal mucosa within the colon and the number of mucous-producing cells per unit area of the colon were significant (p <0.01) as compared with the medium control group, (P < 0.01 or p < 0.05) (Table 11 and Fig. 6), respectively, in all the treatment groups except for the 100 mg / kg administration group of rice fermented extracts showing similar mucosal thickness to the vehicle control group.

The thickness of the surface mucosa in the colon was 238.85, 47.69, 106.86 and 105.41%, respectively, in the 100, 200 and 300 mg / kg groups of rice picosapesia and rice fermented extracts, respectively. 3], [3], [4], [6], [7], [7], [7], [7] and [ 406-13, 2011), and the increase in the thickness of surface mucus in the colon by administration of fermented rice extract is also a direct evidence of the sub-efficacy of the fermented rice extract.

In addition, the number of cells producing mucus per unit area in the mucosa of the colon showed a change of 128.45, 50.29, 103.43, and 99.58% in the 100, 200 and 300 mg / kg of the rice picosapesia extract and the rice fermented extract, respectively. The thickness of the colon mucosa was 56.65, 5.88, 32.49, and 20.03% in the 100, 200 and 300 mg / kg groups of rice picosapesulfate and rice fermented extract, respectively, as compared with the medium control group. Histopathologically, a decrease in mucus secretion in the colon is directly related to constipation (Yang ZH et al., PLoS One. 3: e3348, 2008) (McCullogh et al., Gut. 42: 799-806, 1998).

Therefore, the increase in the thickness of the mucosa of the colon and the increase in the number of mucus - producing cells caused by the fermentation of the rice fermentation can be regarded as a direct indicator of the subterritorial effect of the fermented rice extract.

group Measurement of tissue morphology (at sacrifice) Fecal surface mucosal thickness (um) Number of mucus-producing cells (cells / mm ² ) Colonic mucosal thickness (um) Medium control group 74.36 ± 7.83 593.50 ± 105.70 473.52 + - 54.78 Drug control 251.95 ± 60.20 ^a 1355.83 ± 247.79 ^a 741.78 ± 119.48 ^a FRe 100 group 109.81 ± 24.96 ^a 892.00 ± 214.31 ^b 501.35 ± 36.65 FRe 200 group 153.81 ± 40.18 ^a 1207.33 + - 340.83 ^a 627.36 ± 74.65 ^a FRe 300 group 152.73 + - 33.26 ^a 1184.50 ± 226.62 ^a 568.35 ± 60.15 ^b

Each value represents the mean ± standard deviation of six samples.

^a p < 0.01 and ^b p < 0.05, as compared to vehicle control by MW test.

3-6. conclusion

In the above, the fermented rice extract showed a significant increase in the number of feces, the water content in the feces, the number of the remaining feces in the colon, the average diameter and surface mucosal thickness, the small intestinal mobility, the thickness of the colon mucosa, Increase, and it was confirmed that it exhibits excellent shaking effect. In the 100 and 200 mg / kg administration groups, a definite dose-dependency was observed, and the optimum effective dose of rice fermented extract was estimated to be around 200 mg / kg. In addition, in the group treated with sodium pico sodium sulfate, a significant decrease in the amount of weight gain due to the excessive subcutaneous effect was observed. However, in the group treated with rice fermented extract, the change in body weight and body weight were similar to those in the medium control group, and no excessive subcutaneous effect was observed. The possibility of side effects due to the effects is low.

Example  4. Rofemide ( Loperamide ) Induced constipation Rat  In the model Rice fermentation  Subtle effect of extract

The sublingual effects of rice fermented extracts were evaluated using a loperamide induced constipation rat model. 100, 200 and 300 mg / kg of fermented extract composition was orally administered for 1 day each time after 3 hours of administration of 3 mg / kg of ropelamide, and the body weight and the fecal indicator (24 hours of feces, wet weight, Weight and water content), number of feces in the colon and average diameter, migration rate of the small intestine, and histopathological changes of the fecal colonic and mucosa were observed. The results were compared with the 5 mg / kg group of sodium picosulfate used currently.

6-week-old SD rats (6-week-old male, SLC, Japan) were prepared with 6 groups per group as follows (FRe represents fermented rice extracts)

Medium control: physiological saline and sterile distilled water administered control

Loperamide Control group: Loperamide 3 mg / kg + sterile distilled water group

Drug Control: Loperamide 3 mg / kg + pico sodium sulfate 5 mg / kg group

FRe100 group: rofemide 3 mg / kg + rice fermented extract 100 mg / kg group

FRe200 group: rofemide 3 mg / kg + rice fermented extract 200 mg / kg group

FRe300 group: Loperamide 3mg / kg + rice fermented extract 300mg / kg group

According to the method of Bustos et al. (Bustos D, et al., Acta Gastroenterol Latinoam. 21: 3-9, 1991) and Wintola et al. (Wintola OA, et al., BMC Gastroenterol. / kg was dissolved in physiological saline and orally administered once a day for 6 days to induce constipation. Rice fermented extracts were either physiological saline or sterile distilled water as a vehicle and were orally administered once a day for 6 days. Body weight was recorded 1 day before administration, on the day of administration, on days 1, 2, 3, 4, and 5 after administration. Four days after administration, fecal indicator (24 hours fecal pellet count, wet weight, Dry weight and water content) were measured, and 5 days after the start of administration, 18 hours of fasting were measured, and the carotid transfer rate in the small intestine was measured and histopathological changes of the colonic feces and mucosa were observed.

Fecal Moisture Content = [(Wet Wet Weight - Fecal Dry Weight) / Wet Wet Weight] x 100

Carbon shift rate in small intestine = [(total length of the small intestine - moving distance of the intestine) / total length of the small intestine] x 100

Histomorphometry: Fecal surface mucosal thickness and mucin production per mm ² (alcian blue positive cells), mean mucosal thickness

4-1. Weight and Weight gain  change

(P < 0.01 or p < 0.05) of body weight and body weight gain was observed in sodium picosulfate administration group, but no change in body weight and weight gain associated with rice fermented extract administration was observed ).

Compared with the normal medium control group, the weight gain during the administration period was 7.77% in the rofaperide control group. In the 100, 200 and 300 mg / kg rice fermentation extract group, 0.90, -1.80 and -6.31%, respectively. That is, significant decrease in weight gain due to excessive subcutaneous effect was observed in the sodium picosapthate treated group, but it was observed that in the rice fermented extract administered group, the body weight and the body weight change were similar to those of the medium control group and did not induce excessive subcutaneous effects.

group weight Weight gain
[BA] Starting date of administration [A] Four days after administration Last (6th) Administration [B] Medium control group 250.50 ± 11.67 294.33 ± 14.38 267.67 ± 12.44 17.17 + 1.94 Rofemide control 248.33 8.41 291.17 ± 9.02 266.83 + - 8.52 18.50 ± 2.43 Drug control 250.33 + - 9.35 277.50 ± 12.36 ^ab 253.83 + - 12.27 ^a 3.50 ± 3.56 ^cd FRe 100 group 252.17 + - 5.42 292.00 ± 5.97 270.50 + - 7.06 18.33 + - 6.74 FRe 200 group 250.17 + 9.06 290.83 + - 13.48 268.33 + - 13.02 18.17 + - 6.88 FRe 300 group 247.33 + - 10.27 286.00 ± 10.64 264.67 ± 12.16 17.33 + - 6.65

Each value represents the mean ± standard deviation of 6 rats (unit: g)

* Overnight fasted

^a p < 0.05, compared to vehicle control by LSD test.

^b p < 0.05, compared to rofemide control by LSD test.

^c p < 0.01, as compared to vehicle control by MW test.

^d p < 0.01, compared to rofemide control by MW test.

4-2. Feces  Change in the indicator

The increase in fecal matter and water content was significantly (p <0.01) higher in the rofaperide control group than in the normal medium control group, but the increase in the fecundity and moisture content was significantly (p <0.01) Sodium sulphate, rice fermented extract 200 and 300 mg / kg, respectively.

On the other hand, significant increases in fecal and water contents were observed in the 100 mg / kg rice fermented extract group compared with the rofeferamide control group, but the significant (p <0.01) change was limited to the water content (Table 13).

In comparison with the normal medium control group, the fecal counts at the 4th day after administration and the fermentation at 24 hours showed a change of -21.97% in the fermented control group. In the picofuroxan sodium and 100, 200 and 300mg / kg rice fermented extract groups, , Respectively, which were 67.87, 24.91, 34.30 and 58.12%, respectively.

The water content of the feces for 24 hours after the 4th day of administration compared to the normal medium control was -32.29% in the rofemide control group, and in the group of 100, 200 and 300mg / kg rice fermented extract of rice p 72.24, 97.73 and 91.23%, respectively, compared to the ferramide control group.

group Fecal parameters of the fourth administration day (24 hours) Number of feces Waste weight
(g / 24hr / rat) Fecal dry weight
(g / 24hr / rat) Water content (%) Medium control group 59.17 ± 3.19 10.006 ± 0.954 5.855 + 0.496 41.311 + - 4.106 Rofemide control 46.17 + - 6.40 ^a 7.673 ± 1.110 ^b 5.560 + 0.270 26.320 +/- 10.426 ^a Water control 77.50 ± 23.45 ^c 25.991 + 5.432 ^ac 6.575 ± 1.072 74.394 + 2.769 ^ac FRe 100 group 57.67 ± 9.18 9.964 ± 1.976 5.392 + 0.748 45.307 + - 4.113 ^c FRe 200 group 62.00 7.90 ^c 13.134 ± 2.721 ^bc 6.265 + 1.153 52.042 ± 1.885 ^ac FRe 300 group 73.00 + 5.87 ^ac 13.651 ± 1.791 ^ac 6.725 ± 0.597 ^bc 50.332 + 4.768 ^ac

Each value represents the mean ± standard deviation of 6 rats

^a p < 0.01 and ^b p < 0.01, as compared to vehicle control by MW test.

^c p < 0.01, as compared to rofemide control by MW test.

4-3. Colon Fecal  change

The number of fecal remnants and mean diameter increased in the rofemide control group compared to the normal medium control group (p <0.01), but the number of fecal remnants in the colon was significantly higher than that of the ropelamide control group (p <0.01) Decreases in mean diameter were recognized in sodium picosulfate and in all three rice fermented extract-treated groups (Table 14).

In comparison with the normal medium control group, the residual fecal numbers in the colonic fermented control group were 115.79%. In the 100, 200 and 300 mg / kg rice fermented extract group, the values of -80.49 and -73.17 , -87.80 and -78.05%, respectively.

The mean diameter of the remaining feces in the colon was 30.70% in the rofemide control group compared to the normal medium control group. In the group treated with 100, 200 and 300 mg / kg fermented rice extract of pico sulphate, -40.24 , -28.40, -36.17 and -31.57%, respectively.

group Remaining feces in the colon Count Average Diameter (um) Medium control group 3.17 ± 1.47 4.86 ± 0.59 Rofemide control 6.83 ± 0.98 ^a 6.35 ± 0.50 ^d Drug control 1.33 ± 1.21 ^ac 3.80 ± 0.47 ^ef FRe 100 group 1.83 ± 1.17 ^bc 4.55 ± 0.75 ^f FRe 200 group 0.83 + 0.75 ^ac 4.05 + - 0.35 ^bf FRe 300 group 1.50 ± 0.84 ^bc 4.35 ± 0.25 ^f

Each value represents the mean ± standard deviation of 6 rats

^a p < 0.01 and ^b p < 0.05, as compared to vehicle control by LSD test.

^c p < 0.01, compared to rofemide control by LSD test.

^d p < 0.01 and ^e p < 0.05, as compared to vehicle control by MW test.

^f p < 0.01, compared to rofemide control by MW test.

4-4. In the collection Tombstone  Change in the moving rate

(P <0.01) or less than that of the normal control group (p <0.01). However, in the case of sodium picosapenta sulfate and all three rice fermented extract extracts, <0.05), respectively (Table 15).

Compared with the normal medium control group, the intestinal flotation rate was -20.87% in the rofaperide control group, and 15.10, 16.40, and 10.8% in the 100, 200 and 300 mg / kg rice fermentation extract groups, respectively, 22.65 and 18.74%, respectively.

group Camouflage moiety (for 30 minutes) Total length (cm) Carbon shift length (cm) Carbon shift in small intestine (%) Medium control group 113.17 ± 11.00 78.40 +/- 10.66 69.25 + - 6.42 Rofemide control 112.07 + - 4.67 61.15 + - 5.61 ^a 54.80 + - 7.01 ^a Drug control 111.83 + - 5.45 70.58 + - 9.41 ^c 63.07 ± 7.27 ^c FRe 100 group 110.58 +/- 7.51 70.42 + - 4.75 ^c 63.79 + 4.07 ^c FRe 200 group 111.75 ± 5.18 74.75 ± 6.12 ^a 67.21 + - 8.81 ^b FRe 300 group 112.08 ± 5.15 72.83 + - 6.73 ^c 65.07 + - 6.44 ^c

Each value represents the mean ± standard deviation of 6 rats

^a p < 0.01, compared to vehicle control by LSD test.

^b p < 0.01 and ^c p < 0.05, as compared to rofemide control by LSD test.

4-5. Histopathological change

Significant (p <0.01) reduction of mucus-producing cells and mucosal thickness per unit area of the intestinal mucosa in the colon compared to the normal medium control group was observed in the rofeeramide control group, (p < 0.01 or p < 0.05). Numerical increase in mucus production cells per follicle surface mucosal thickness, colonic mucosa thickness and per unit area was recognized in sodium picosulfate and all rice fermented extract administration groups (Table 16 and Fig. 7).

In comparison with the normal medium control group, the surface mucosal thickness of the fecal colonic feces was -73.71% in the loperamide control group. In the 100, 200 and 300 mg / kg rice fermentation extracts of pico sulphate, 478.17, 677.89, and 506.38%, respectively.

The number of mucin-producing cells per unit area of the colon mucosa was -69.84% in the rofemide control group compared with the normal medium control group, and that of the sodium picosapenta and rice fermented extract 100, 200 and 300 mg / Respectively, and showed a change of 100.10, 42.27, 92.06 and 74.12%, respectively.

The thickness of the mucosa of the colon was -53.68% in the roperaemide control group compared with the normal medium control group, and 110.91, 50.39, and 50.39%, respectively, in the picosapenta sulfate, rice fermented extract 100, 200 and 300 mg / 86.91 and 85.38%, respectively.

group Measurement of tissue morphology (at sacrifice) Fecal surface mucosal thickness (um) Number of mucus-producing cells
(cells / mm ² ) Colonic mucosal thickness (um) Medium control group 64.71 ± 14.09 643.20 ± 117.17 499.08 ± 102.84 Rofemide control 17.27 ± 2.97 ^d 194.00 ± 14.73 ^d 231.16 + - 42.32 ^a Drug control 157.77 ± 48.75 ^df 388.20 ± 55.56 ^df 487.54 ± 95.24 ^b FRe 100 group 99.85 + - 12.08 ^df 276.00 ± 51.35 ^df 347.64 ± 54.21 ^ac FRe 200 group 134.34 ± 44.04 ^ef 372.60 ± 89.34 ^df 432.08 + - 74.44 ^b FRe 300 group 104.72 + - 14.41 ^df 337.80 ± 60.41 ^df 428.53 + - 102.52 ^b

Each value represents the mean ± standard deviation of 6 rats

^a p < 0.01, compared to vehicle control by LSD test.

^b p < 0.01 and ^c p < 0.05, as compared to rofemide control by LSD test.

^d p < 0.01 and ^e p < 0.05, as compared to vehicle control by MW test.

^f p < 0.01, compared to rofemide control by MW test.

4-6. conclusion

In the above, the fermented rice extract showed significant fecal count, fecal water content, number of fecal pellets remaining in the colon, average diameter and surface mucosal thickness, intestinal mobility, colonic mucosal thickness and mucus production cell , And it was shown that the roperaemide - induced constipation rats had a very good suppression effect. On the other hand, the dependence of the administration dose was confirmed in the administration group of 100 and 200 mg / kg, and the optimum effective dose of the fermented rice extract was determined to be about 200 mg / kg. In addition, in the group treated with sodium pico sodium sulfate, significant decrease in weight gain due to excessive subcutaneous effect was recognized, but in the group treated with rice fermented extract, changes in body weight and body weight were similar to those in the medium control group. Thus, excessive subcutaneous effects were not observed. The possibility of side effects due to the effects is low.

Example  5. Rofemide ( Loperamide ) Induced constipation Rat  In the model Rice fermentation  Intestinal tract of extract Beneficial bacteria  (Lactobacillus; Lactobacillus ) Propagation effect

Lactobacillus proliferation effect of rice fermented extract was evaluated by Loperamide - induced constipation rat model. Liquid yogurt containing liquid yogurt (Bulgarian (apple flavor, Namyang dairy)), compound rice fermented extract composition, or composite rice fermented extract composition is administered orally 1 hour after fermentation 3 mg / kg per day, once daily for 6 days And the changes in the contents of cecum and the number of lactic acid bacteria in the feces were measured.

6-week-old SD rats (6-week-old male, SLC, Japan) were prepared with 6 groups per group as follows (FRe represents fermented rice extracts)

Medium control: physiological saline and sterile distilled water administered control

Loperamide Control group: Loperamide 3 mg / kg + sterile distilled water group

FRe5 group: Loperamide 3 mg / kg + rice fermented extract 5 mg / kg group

FRe10 group: rofemide 3 mg / kg + rice fermented extract 10 mg / kg group

Yoghurt group: rofemide 3 mg / kg + yogurt group

BFRe0.05 group: 10 mg / kg of liquid yogurt containing fermented rice fermented extract of 3 mg / kg + 0.05%

BFRe0.1 group: 10 mg / kg of liquid yogurt containing fermented rice fermented extract of 3 mg / kg + 0.1%

BFRe1 group: 10 mg / kg of liquid yogurt containing 3 mg / kg + 1% rice fermented extract of rofemide

According to the method of Bustos et al. (Bustos D, et al., Acta Gastroenterol Latinoam. 21: 3-9, 1991) and Wintola et al. (Wintola OA, et al., BMC Gastroenterol. / kg was dissolved in physiological saline and orally administered once a day for 6 days to induce constipation. Rice fermented extracts were either physiological saline or sterile distilled water as a vehicle and were orally administered once a day for 6 days.

5-1. Changes in the Number of Lactic Acid Bacteria in the Cecal Contents

Especially, in case of liquid yogurt containing 0.05, 0.1 and 1% rice fermented extract, the same amount of rice fermented extract and liquid phase of rice fermented extract were observed in all cows except for 5 mg / kg of rice fermented extract. The increase in the number of lactic acid bacteria was found to be superior to that of the yogurt single composition administration group.

The number of lactic acid bacteria in the cecum contents was -8.98% in the cecum contents compared with the medium control, and the liquid fermented extracts 5 and 10 mg / kg, liquid yoghurt, 0.05, 0.1 and 1% In the 10 mg / kg administration group, the changes were 19.70, 51.52, 120.45, 193.18, 237.12, and 754.55%, respectively, compared with the rofeelamide control group (FIG.

5-2. Feces  Changes in the number of lactic acid bacteria

In the case of liquid yogurt containing 0.05, 0.1, and 1% rice fermented extract, the same amount of rice fermented extract and liquid yogurt extract of the same dose group were observed in all the groups except the 5 mg / kg rice fermented extract group. An increase in the number of lactic acid bacteria was found to be superior to that of the single composition administration group.

Compared with the medium control, the number of lactic acid bacteria in the feces was -3.40% in the rofemide control, and 10 mg / kg of rice yogurt containing liquid fermentation extract, liquid yogurt, 0.05, 0.1 and 1% rice fermented extract / kg, respectively, compared to the rofemide control group (Fig. 9).

5-3. conclusion

These results suggest that fermented rice extracts showed an increase in intestinal content and fecal lactobacillus compared to rofemide control, and showed excellent lactic acid bacterial growth effect in fermented - induced constipation rats. This is a result indicating that the rice fermented extract composition of the present invention has a function of improving beneficial bacteria or intestinal flora in the intestines. In addition, this proliferation effect was remarkable when liquid yogurt containing rice fermented extract was administered. In particular, the dependence of the dose on the dose was observed in the dose of 10 mg / kg, and the optimum effective concentration of the liquid yogurt containing the fermented rice extract was judged to be around 1%.

Claims (10)

1) mixing the rice with water and heating,
2) a step 1 fermentation step of adding 0.05 to 10% by weight of koji powder to the above-mentioned aged rice and saccharifying at 20 to 40 ° C for 10 to 30 hours,
3) a two-stage fermentation step in which the one-stage fermented rice is cultured at 20 to 50 DEG C for 1 to 10 days,
4) a three-stage fermentation step in which the two-stage fermented rice is cultured at 10 to 40 DEG C for 1 to 10 days, and
5) sterilization and concentration of the obtained three-stage fermented rice,
A method for producing rice fermented product having a function of improving denaturation or improving intestinal flora. The method of claim 1, wherein step (5) further comprises solvent extraction, hot water extraction, cold extraction, reflux cooling extraction or ultrasonic extraction. The method according to claim 1, wherein said step (3) is carried out at 30 to 40 DEG C for 2 to 3 days. The method according to claim 1, wherein said step (4) is carried out at 25 to 40 DEG C for 2 to 3 days. A fermented rice fermented by the method of any one of claims 1 to 4,
A food composition for improving metamorphosis or improving intestinal flora. 6. The food composition of claim 5, further comprising at least one selected from the group consisting of liquid yoghurt and a food acceptable food additive. 6. The food composition according to claim 5, which has intestinal lactic acid bacteria or bifidobacteria. A fermented rice fermented by the method of any one of claims 1 to 4,
A pharmaceutical composition for preventing or treating constipation. The pharmaceutical composition according to claim 8, further comprising liquid yoghurt. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition has intestinal lactic acid bacteria or bifidobacteria.

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