KR100527103B1 - Composition comprising microorganism having lipid metabolism-improving activity for prevention and treatement of geriatric diseases and the use thereof - Google Patents

Abstract

The present invention relates to a composition containing a microorganism having an effect of improving lipid metabolism, and specifically, a lipid metabolism containing microorganisms obtained by culturing lactic acid bacteria and yeast in a medium in which the soybean, glutinous rice or rice bran of the present invention is mixed. The present invention relates to a composition for preventing and treating geriatric diseases. The composition containing the microorganism of the present invention significantly inhibits the content of neutral lipids and cholesterol in the blood, effectively inhibits free radicals and lipids, and increases the active oxygen species removal enzyme activity to suppress the adult disease and aging process. Effective medicines and dietary supplements can be provided.

Description

Composition comprising microorganism having lipid metabolism-improving activity for prevention and treatement of geriatric diseases and the use according to the present invention.

The present invention relates to a composition for the prevention and treatment of adult diseases related to anti-aging and lipid metabolism containing microorganisms obtained by culturing lactic acid bacteria and yeast in a medium mixed with soy bean curd, glutinous rice or rice bran.

Insulin-independent diabetes mellitus (type II), which is one of the adult diseases, accounts for 84% of Korean diabetic patients. Especially, type II diabetes is obese and less active among adults in their 40s and older. It has the characteristic that many people develop high. One of the main causes of cardiovascular disorders such as atherosclerosis in diabetes mellitus is a study that promotes lipid peroxidation by the reaction of free radicals [Bastar I, Res. Commun. Mol. Pathol. Pharmacol ., 102 (3), pp265-272, 1998] and also reported that lipid peroxidation occurs easily due to its high susceptibility to oxidative stress [Behrens WA, Madere R, Metabolism , 40 (1), pp72-76 , 1991].

Physiological processes radicals generated during ^{_{(O 2 -, H 2 O}} 2, OH and O ₂₎ is a free radical reaction in which a very strong reactivity caused by these represents the destruction effect of major macromolecules, including lipids [Davies KJ, Goldberg AL, J. Biol. Chem ., 262 (17), pp 8220-8226, 1987]. Their production is increased by a number of factors that cause abnormal metabolic processes, photosensitization, drug metabolism, and other cellular metabolism, and therefore, living organisms are always exposed to the harmful effects of free radical reactions caused by them. [Aust SD, Morehouse LA, Thomas CE, J. Free. Radic. Biol. Med ., 1 (1), pp 3-25, 1985. In particular, if cells continue to accumulate harmful effects due to their harmful radicals as they age, they cause aging of the cells as well as diseases related to various adult diseases such as carcinogenesis, arteriosclerosis, heart disease, and skin aging. It is suggested as a cause of human disease occurrence and aging [Harman D. 2001. Aging: overview. Ann. NY Acad. Sci ., 928, pp 1-21, 2001]. Therefore, by inhibiting free radical reactions in vivo by reactive oxygen metabolites or removing harmful free radicals accumulated in the human body, candidates are expected to be useful for the treatment of diseases such as lipid metabolism, immune diseases and cancer. Research has continued.

Lactobacillus has the effect of intestinal action and constipation [Mizutani T, Mitsuoka T, J. Natl. Cancer. Inst ., 63 (6), pp 1365-1370, 1979; Reuter G, Zentralbl. Bakteriol. Mikrobiol. Hyg ., 187 (4-6), pp564-577, 1989], immune enhancing effect [Hosoi T et al., Can. J. Microbiol ., 46 (10), pp892-897, 2000], Industrialization of Bifidus [Roy DG et al., Mol. Gen. Genet ., 239 (1-2), pp33-40, 1993]. In particular, Lactobacillus fermentum (Lactobacillus fermentum) used in this study has a cholesterol lowering effect [Pereira DI, Gibson GR, Appl. Environ. Microbiol ., 68 (9), pp4689-4693, 2002], inhibited lipid peroxidation and increased antioxidant enzymes [Kullisaar T, et al., Int. J. Food Microbilol ., 72 (3), pp 215-224, 2002; Gonzalez SN, et al., Chem. Pharm. Bull ., 37 (11), pp3026-3028, 1989], Isolation of acarbose, a substance that inhibits postprandial blood sugar [Talamond P, et al., Comp. Biochem. Physiol. B. Biochem. Mol. Biol ., 133 (3), pp 351-360, 2002; Calderon Santoyo M, et al., Int. J. Food. Microbiol ., 80 (1), pp77-87, 2003].

(Patent application 2002-69983) Lactobacillus fementum (vegetable lactobacillus beverage) developed by Choi et al., That is, unlike conventional lactic acid bacteria, has a completely new property of decomposing ammonia, which is resistant to heat and acid and is impossible for theories. As a bacterium, it is a bacterium cultured by coexisting the advantages of lactic acid bacteria and the benefits of yeast.

Therefore, the present inventors administered the microorganism-containing composition of the present invention to determine the neutral lipid and cholesterol content change in order to scientifically investigate the treatment and prevention of adult diseases related to lipid metabolism according to the ingestion of lactic acid bacteria and yeast, active oxygen and lipid peroxide As a result of measuring free radical scavenging enzyme activity, it was confirmed that the composition of the present invention is effective in treating adult diseases related to lipid metabolism.

An object of the present invention is to provide a microorganism-containing composition made by culturing lactic acid bacteria and yeast effective in preventing and treating geriatric diseases related to aging and lipid metabolism.

It is also an object of the present invention to provide a pharmaceutical composition and a functional vegetable lactic acid bacteria product effective in preventing and treating geriatric diseases related to anti-aging and lipid metabolism.

In order to carry out the above object, the present invention provides a composition for the prevention and treatment of adult diseases related to anti-aging and lipid metabolism containing microorganisms obtained by culturing lactic acid bacteria or yeast in a medium mixed with soy bean curd, glutinous rice or rice bran to provide.

In the present invention, soybean is used to make tofu to remove the soy milk from the soy juice. That is, soybean oil is not used as it is, soybean oil is used so that the microbial-containing composition can be provided in an economical way.

In addition, since glutinous rice usually contains more amino pectin than non-glutinous rice, it is possible to improve the efficiency of lactic acid bacteria and yeast. Therefore, the use of glutinous rice is more effective in preparing microbial-containing compositions.

The blending ratio of bean curd and glutinous rice produced during tofu production is about 70% by weight to about 95% by weight and about 30% by weight to about 5% by weight of glutinous rice, based on the total medium composition, so that the total weight is 100%.

In addition, rice bran may be added to the medium, in which case the mass ratio of soybean, glutinous rice and rice bran relative to the total medium composition is about 80% to about 90%, about 15% to about 7% by weight and about From 5% to about 3% by weight, totaling 100% by weight.

As the lactic acid bacteria used in the present invention, for example, Lactobacillus Fermentum, Lactobacillus Acidophilus, Lactobacillus Casei, Lactobacillus Casebru (Lactobacillus Delbrueckii) , Lactobacillus Arabinosus, Lactobacillus Caucasicus, Lactobacillus Lactis, Lactobacillus Leishmanni, Lactobacillus mucusus Lactobacillus Music Lactobacillus Thermophilus, Lactobacillus Plantarum, Streptococcus Thermophillus, Streptococcus Faecalis, Streptococcus Lactobacillus Streptococcus Bulgaricus (Streptococcus Bulgaricus), and Leukonostock Me One or more of the genus of Leuconostoc Mesenteroides can be selected and used.

As the lactic acid bacteria used in the present invention, those produced by freeze-drying or refrigerated can be used as compared to the usual method. In addition, since general animal lactic acid bacteria are weak to heat or acid, it is preferable to use vegetable lactic acid bacteria, for example, Lactobacillus Fermentum.

In addition, the yeast used in the present invention, for example, Pichia Kluyveri, Saccharomyces Cerevisiae, Saccharomyces Pastorianus, Saccharomyces inter Saccharomyces Intermedius, Saccharomyces Validus, Saccharomyces Ellipsoideus, Saccharomyces Malisisler, Saccharomyces Mend Shuriscus Mandshuricus, Saccharomyces Vordermannii, Saccharomyces Peka, Saccharomyces Shasshing, Saccharomyces Piriformis, Saccharomyces Piriformis Saccharomyces Anamensis, Saccharomyces Cartilaginosus, Saccharomyces Anamensis, Saccharo myces Awamori, Saccharomyces Batatae, Saccharomyces Coreanus, Saccharomyces Robustus, Saccharomyces Carlsbergensis, Saccharomyces Carlsbergensis Saccharomyces Monacensis, Saccharomyces Marxianus, Zygo Saccharomyces Major, Saccharomyces Lactis, Saccharomyces One or more may be selected from the group consisting of Saccharomyces Rouxii and Hansenula Anomala, and it is preferable to use Pichia kluyveri.

In addition, the ratio of the number of bacteria between lactic acid bacteria and yeast when inoculating in the medium can be set in the range of about 1:10 to 10: 1.

After incubating the lactic acid bacteria and yeast in the medium, it is incubated at a relatively low temperature, for example, about 10 ° C. to 20 ° C. for several days to tens of days, for example, 10 days to 50 days.

The coexistence of the lactic acid bacteria and yeast cultured in this way is referred to as a mother strain in the present specification. The mother cells (including the medium) contain about 10 ⁸ cells in 1 g. When the mother cells are kept at about 4 ° C, they can be stored for a long time. In this case, it is better to keep it in a vacuum pack (or deoxygenation pack) with air (or oxygen) removed. Also, the parent cells may contain appropriate oxygen.

As described below, since the parental fungus is a very useful composition, it is not always necessary to use the parental fungus as it is.

Herein, the primary bacteria can be prepared by secondary culturing the primary bacteria, and the primary bacteria can be used. The stirring is carried out while adding about 5 to 20 times water, and preferably about 8 to 12 times the weight with respect to 1 weight of the mother cells. The mixed solution is then incubated at about 35 ° C. to 45 ° C. for about 2 to 5 days, preferably about 3 to 4 days.

The bacterium thus obtained is referred to as primary progeny. Since the primary protozoa contains several kinds of bacteria, it is called a primary protococcus without limiting its name. Primary probiotics contain about 2 × 10 ⁷ lactic acid bacteria per ml.

Then, the mixture is stirred by adding about 2 to 8 times by weight of water, preferably about 4 to 6 times by weight, to the weight 1 of the primary progenitor, and then the mixed solution is about 2 days at about 35 ° C to 45 ° C. Incubate for 5 days, preferably about 3 to 4 days. The bacterium thus obtained is called secondary progeny. The secondary progeny contains about 1.5 × 10 ⁷ lactic acid bacteria per ml.

By culturing the seedlings twice, the turbidity of soybeans and rice bran contained in the seedlings becomes thinner, which improves the transparency, thereby extending the use range.

 As a means for evaluating the geriatric efficacy associated with the lipid metabolism of the microorganism-containing composition prepared by the present invention, arteriosclerosis index, total cholesterol, triglyceride, free radicals and antioxidant enzymes confirmed by animal experiments, Adult disease was superior to control and animal lactic acid beverages.

In addition, the present invention provides a pharmaceutical composition for preventing and treating geriatric diseases associated with lipid metabolism containing the microorganisms (parents or probiotics) prepared by the above method as an active ingredient.

The adult disease includes hypertension, coronary artery disease, fatty liver or liver injury, arteriosclerosis, complications caused by arteriosclerosis, various hyperlipidemia, myocardial infarction, obesity, hypertriglyceridemia, osteoporosis, menopausal symptoms, degenerative arthritis, paralytic disease, etc. do.

The present invention also provides a pharmaceutical composition for inhibiting aging containing the microorganism prepared by the above method as an active ingredient.

Compositions containing microorganisms of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Carriers, excipients, and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, trehalose, polydextrose (polydextrin) and mineral oil.

The compositions of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. .

The amount of the composition of the present invention may vary depending on the age, sex, and weight of the patient, but the amount of 10 to 500 ml may be administered once to several times daily. In addition, the dosage of the extract can be increased or decreased depending on the route of administration, the degree of disease, sex, weight, age and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

In addition, the composition of the present invention can be used as a main, secondary ingredient and food additive of other foods.

The present invention provides a food additive containing a microorganism obtained by culturing lactic acid bacteria and yeast in a medium mixed with soy bean curd and glutinous rice, the food additive characterized in that the food additive is used as a food material (additive) Provides a method of use.

In another aspect, the present invention provides a health functional food comprising the microorganism-containing composition and food acceptable food supplement additives for the treatment and prevention of geriatric diseases associated with lipid metabolism.

The composition of the present invention can be used in a variety of drugs, food and beverages for the prevention and treatment of geriatric diseases associated with lipid metabolism. Examples of the food to which the present composition can be added include various foods, beverages, gums, teas, vitamin complexes, and health supplement foods.

Lactobacillus itself of the present invention has little toxicity and side effects, so it is a drug that can be used safely even for long-term administration for the purpose of prevention.

The composition of the present invention may be added to food or beverage for the purpose of preventing adult diseases associated with lipid metabolism. At this time, the amount of the extract in the food or beverage is generally added to the health food composition of the present invention 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 10 g based on 100 ml, preferably Can be added in a ratio of 0.3 to 1 g.

The health beverage composition of the present invention has no particular limitations on the liquid component except for containing the extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides such as conventional sugars such as dextrin, cyclodextrin, trehalose and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention is described in more detail based on the following examples, although the present invention is not limited thereto.

Example 1 Preparation of Mother Cells (1)

Lactobacillus fermentum (0.2 g) and yeast ( pichia kluyveri ) 0.2 g were inoculated into the medium containing 180 kg of bean curd and 20 kg of glutinous rice. The total number of bacteria between lactic acid bacteria and yeast is about 10 ⁵ to The number of bacteria was changed to 10 ⁷ , and the bacteria ratio was appropriately set between about 1:10 and 10: 1.

The medium inoculated with the lactic acid bacteria and yeast was incubated in a cold room set between about 10 ° C. to 20 ° C. for about 30 days (the culture days were appropriately changed between about 10 days and 50 days depending on the state of the medium). 200 kg of the parent strain was prepared. The parent cells can be stored for a long period of time, for example, months to years by sealing in a vacuum pack that blocks the passage of air and then storing at about 4 ° C.

When producing the progeny of Example 3 described below, the mother cells stored at low temperature for a long time were used. This bacterium also contains about 10 ⁸ lactic acid bacteria and yeast per gram.

Example 2 Preparation of Mother Cells (2)

Lactobacillus fermentum and yeast ( Pichia kluyveri ) were inoculated into a medium containing 170 kg of bean curd, 22 kg of glutinous rice, and 8 kg of rice bran. The bacterial cell ratio between lactic acid bacteria and yeast is the same as in the production of the above-mentioned parent bacteria (1).

In this way, the cultured medium was treated as in (1) to prepare the seedlings 200kg. And as described above, about 10 ⁸ lactic acid bacteria and yeast per 1g.

Example 3. Preparation of Primary Probiotics

To 1 kg of the mother cells prepared as described above, about 8 to 12 times the mass of groundwater was added and the mixture was stirred and incubated at about 40 ° C. for 3 to 4 days. About 10 ⁷ lactic acid bacteria in 1 ml of the primary probiotics thus obtained, Microorganisms were obtained in amounts of 10 ⁶ yeasts.

Example 4. Preparation of Secondary Probiotics

With respect to the mass 1 of the primary protozoa prepared as above, ground water was stirred in a state in which about 4 to 6 times the mass was added, and the mixed water was incubated at 40 ° C. for 3 to 4 days.

In 1 mL of the obtained secondary progenitors, about 1.5 × 10 ⁷ lactic acid bacteria were contained.

Experimental Example 1. Experiment of treatment and prevention effect related to lipid metabolism

In order to confirm the efficacy of treating geriatric diseases related to the lipid metabolism of the plant lactic acid bacteria of the present invention, the following animal experiment was performed.

1-1. Preparation of Laboratory Animals

The experimental animal used in this animal experiment was C57BL / 6J-Lep ^ob (ob mouse), which was used for the obesity type diabetes research experiment, and was purchased from The Jackson Laboratory (JAX) in the United States. In the barrier system, the test animals used in this experiment were selected to show obesity at 3-4 weeks of age, and the test animals were 9-year-old males. Body weight and blood glucose (basal blood glucose) were measured immediately before the test, and the average value was similarly adjusted for each group. At this time, the average blood glucose level was about 380 to 430 mg / dl.

1-2. Preparation and Administration of Microbial Containing Compositions

The microorganism-containing composition 1 (Lac.F-100) used in this experiment was 100% of the prokaryote of Example 3, and the number of lactic acid bacteria per 1ml of the microorganisms was about 2 x 10 ⁷ .

Microbial Containing Composition 1, Microbial Containing Composition 2 (Lac.F-50), which is diluted twice with Composition 1, commercially available animal lactobacillus beverage (HYW-100, fermented milk will from Korea Yagurt Co., Ltd.), and A double diluted animal lactic acid bacteria beverage (HYW-50) was placed in a water bottle for free ingestion and administered for 3 weeks. In addition, the control group (Control group) was administered so that only water intake freely.

1-3. Determination of triglyceride, protein and cholesterol content in the blood

The effects of triglyceride (TG), cholesterol and lipoprotein cholesterol, known as the causative agent of adult diseases, on the contents of cholesterol and lipoprotein cholesterol, microorganism-containing compositions 1, 2 (Lac.F-100 and Lac.F-50) and animal Lactobacillus beverages (HYW-100, 50) were each freely consumed, and the effects of administration for about three weeks were compared.

1-3-1. Collection and Separation of Blood

After anesthetizing the experimental animals with ether, blood was collected from the heart, left at low temperature for 2 hours, centrifuged at 700 × g for 10 minutes, and the supernatant (serum) was treated with 0.05 ml of blood per 1.0 ml of anticoagulant. It was placed in a complete CBC (green blood cell count) and used for analysis while cryopreservation at -70 ℃.

1-3-2. Determination of triglyceride, protein and cholesterol content

The content of triglyceride (TG) as a triglyceride in serum obtained above was treated with TG kit reagent (Sigma, USA) for serum triglyceride measurement to quantify the content of triglyceride in serum according to a standard calibration curve.

Protein content in serum [Lowry OH, et al., J. Biol. Chem., 193, pp 265-275, 1951] were measured according to the BSA (standard serum albumin) method.

Content of total cholesterol [Rudel LL and Morris MD, J. Lipid. Res. , 14, pp364-366, 1973] measured the total cholesterol content in the blood by a standard calibration curve by the o -phthalaldhyde method. The contents of low density lipoprotein (LDL) and high density lipoprotein (HDL) -cholesterol in serum were measured according to the method of kit reagent (HDL-C 555, Eiken, Japan), LDL-cholesterol (BLF, Eiken, Japan).

The atherosclerotic index (AI), which is used as an indicator of atherosclerosis, known as an early symptom of adult disease, is described in the method of Haglund O, et al., J. Nutr ., 121, pp165-169, 1991. According to the formula (Total Cholesterol-HDL Cholesterol) / HDL Cholesterol.

As shown in Table 1, the content of the neutral lipid and total cholesterol of the group to which the microorganism-containing compositions 1 and 2 were administered was 8.6 to 12.8% and 6.6 to 15.8% of the control group, respectively. Comparing the inhibitory effect of LDL-cholesterol, which is directly related to the onset of the adult disease, the inhibitory effect was about 18.8 to 25.3% or more compared to the control group. In addition, it can be seen that the atherosclerotic index (AI), which is known as an early onset of adult disease, is significantly suppressed by almost 37.2 to 56.2% in the microorganism-containing composition-administered group of the present invention. It is very interesting that this decrease in arteriosclerosis index is almost consistent with the decrease in LDL-cholesterol content. This is because the increase in LDL-cholesterol content and arteriosclerosis index is closely related to the onset of adult disease. The level of HDL-cholesterol, which is widely known as an anti-cholesterol factor, and particularly associated with exercise and longevity, was also increased by 16.9 to 21.5% in the group containing the microorganism-containing composition of the present invention.

Animal lactobacillus beverages also showed inhibition of triglycerides and LDL-cholesterol levels compared to controls. However, it shows that the degree of inhibition is weak compared to the microorganism-containing composition of the present invention.

In conclusion, the microorganism-containing composition of the present invention is expected to suppress the neutral lipid content, and significantly inhibits the content of blood cholesterol, which plays an important role in lipid metabolism, and is expected to be of great help in the prevention of adult diseases.

Effect of Microorganism-Containing Compositions of the Present Invention on Lipid Metabolism Item Control Microbial Containing Composition 1Lac.F-100 Microbial Containing Composition 2Lac.F-50 Animal Lactic Acid Bacteria Drink-100 Animal Lactic Acid Bacteria Drinks-50 Protein (mg / ml serum) 25.52 ± 0.59 ^* (100.0%) 25.92 ± 0.66 (101.6%) 26.96 ± 0.33 ^b (105.6%) 27.17 ± 0.41 ^c (106.5%) 27.23 ± 0.34 ^c (106.7%) Neutral Lipid (mg / ㎗ Serum) 137.53 ± 1.38 (100.0%) 119.90 ± 0.41 ^c (87.2%) 125.65 ± 7.16 ^b (91.4%) 124.59 ± 4.82 ^c (90.6%) 129.71 ± 2.34 ^c (94.3%) Total Cholesterol (mg / dl Serum) 123.96 ± 6.93 (100.0%) 104.34 ± 2.24 ^c (84.2%) 115.73 ± 3.24 ^a (93.4%) 112.61 ± 5.46 ^a (90.8%) 121.82 ± 4.56 (98.3%) LDL-cholesterol (mg / dl serum) 112.04 ± 5.79 (100%) 83.64 ± 0.48 ^c (74.7%) 90.94 ± 1.79 ^c (81.2%) 91.88 ± 2.28 ^c (82.0%) 95.47 ± 3.45 ^c (85.2%) HDL-cholesterol (mg / dl serum) 56.03 ± 2.91 (100.0%) 68.08 ± 0.97 ^c (121.5%) 65.48 ± 0.39 ^c (116.9%) 59.18 ± 2.13 (105.6%) 58.22 ± 1.16 (103.9%) Arteriosclerosis Index (AI) 1.21 ± 0.13 (100.0%) 0.53 ± 0.10 ^c (43.8%) 0.76 ± 0.08 ^c (62.8%) 0.90 ± 0.16 ^a (74.4%) 1.09 ± 0.17 (90.1%) ^* Mean value ± SE (n = 5). ^{** a} p <0.05 compared to control; ^b p <0.01; ^c p <0.001

Example 2. Determination of free radicals and lipid peroxide (LPO), free radical scavenging enzyme

In order to examine the effects on the lipid peroxidation and the antioxidant effect of the composition containing the microorganism of the present invention, lipid peroxidation, hydroxy radicals, superoxide radicals, superoxide dismutase, glutathione fur in the blood after administration of the composition The content of oxidase was measured.

The measurement of hydroxy radicals, known as the most powerful free radicals among free radicals, is a measure of the degree of hydroxyl radical generation to the extent of deoxyribose destruction. Ribose was destroyed to form aldehyde, which was measured according to the method of [Halliwell B and Gutteridge JMC, FEBS Lett., 128, pp347-350, 1981], which was developed by reacting with thiobarbituric acid in an acid solution.

Lipid peroxide (LPO) content in serum known to be produced by these oxygen radicals was measured as described [Choi JH and Yu BP, AGE, 17, pp93-97, 1994].

Determination of superoxide dismutase (SOD) activity, which is important as an enzyme for eliminating reactive oxygen species, is described by Oyanagui Y, Anal. Biochem ., 42, pp 290-296, 1984]. In addition, the activity of glutathione peroxidase (GSHPx) in serum was measured as a method for measuring the reduction of NADPH at 340 nm as an important scavenger in vivo [Lawrence RA and Burk RF, Lipid, 19, pp444-452, 1978].

Effect of Administration of Compositions 1 and 2 (Lac.F-100, 50) Containing Microorganisms of the Present Invention on Lipid Peroxidation in Blood of Obese Mice Table 2 compares the production of reported superoxide radicals and hydroxyl radicals, known as the most powerful free radicals, and the lipid peroxide (LPO) content. . In addition, superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) as biodefense enzymes are widely known as powerful antioxidant enzymes present in the body, and their amounts are shown in Table 2. .

As shown in Table 2, the microbial-containing composition significantly inhibited the generation of almost 8.4 to 13.8% of hydroxy radicals, and also effectively inhibited the activities of 10.1 to 17.1% or superoxide radicals. By suppressing the activity of the free radicals due to the administration of the microorganism-containing composition of the present invention, it was found that the content of lipid peroxide produced at this time was also suppressed by 10.6 to 14.3%.

As a result of the above, the microorganism-containing composition of the present invention not only can inhibit the generation of free radicals such as hydroxy radicals and superoxide radicals as strong active oxygen, but also has a close relationship with the onset or aging of adult diseases. It was found that the formation of lipids was also very effective.

In addition, the antioxidant enzyme that deactivates the active oxygen by the administration of the composition containing the microorganism of the present invention is the elimination enzyme, which is 15.2 to 21.7% compared with the control group when comparing the effect on the activity of the most powerful eliminating enzyme SOD As can be seen that the activity is effectively increased. In addition, the activity of GSHPx was about 8.7 to 18.0% compared to the control group, which proved to produce antioxidant enzymes in vivo.

The microorganism-containing composition used in this study not only inhibits the production of reactive oxygen species, but also significantly increases the antioxidant enzymes, which may be involved in the inhibition of the aging process as well as in adult diseases.

Effect of Microorganism-Containing Compositions of the Present Invention on Anti-Aging Effects Item Control Microbial Containing Composition 1Lac.F-100 Microbial Containing Composition 2Lac.F-50 Animal Lactic Acid Bacteria Drink-100 Animal Lactic Acid Bacteria Drinks-50 Lipid Peroxidation (LPO) (nmol / ml Serum) 5.10 ± 0.23 ^* (100.0%) 4.37 ± 0.15 ^c (85.7%) 4.56 ± 0.28 ^b (89.4%) 4.49 ± 0.39 ^a (88.0%) 4.52 ± 0.33 ^a (88.6%) Hydroxy radicals (nmol / mg protein) 4.27 ± 0.42 (100.0%) 3.68 ± 0.27 ^a (86.2%) 3.91 ± 0.13 (91.6%) 3.80 ± 0.13 (89.0%) 3.92 ± 0.10 (91.8%) Superoxide radical (nmol / mg protein) 86.60 ± 3.01 (100.0%) 71.75 ± 3.11 ^c (82.9%) 77.84 ± 4.86 ^b (89.9%) 78.30 ± 4.11 ^b (90.4%) 82.24 ± 0.90 ^a (95.0%) Superoxide Dismutase (SOD) (unit / mg protein) 212.83 ± 7.97 (100.0%) 258.98 ± 5.51 ^c (121.7%) 245.28 ± 3.06 ^c (115.2%) 257.30 ± 8.62 ^c (120.9%) 242.44 ± 5.33 ^c (113.9%) Glutathione Peroxidase (GSHPx) (IU / g Protein) 7.32 ± 0.24 (100.0%) 8.64 ± 0.57 ^b (118.0%) 7.96 ± 0.11 ^b (108.7%) 7.91 ± 0.52 (108.1%) 7.54 ± 0.55 (103.0%) ^* Mean ± SE (n = 5). ^{** a} p <0.05 compared to control; ^b p <0.01; ^c p <0.001

Experimental Example 3. Acute Toxicity Test

 The ICR-based mouse and Sprague Dawley (cod hyochang science) were divided into four groups of 10 animals each to orally administer the prokaryotic composition of Example 3 of the present invention at doses of 500, 725, 1000 and 5000 mg / kg, respectively. Two weeks after the administration, no toxicities were observed in all four groups, and no symptoms were apparent in the control group.

From the above results, it was confirmed that the composition of the present invention has little acute toxicity.

Hereinafter, an example of the preparation of the pharmaceutical composition will be described, but it is not intended to limit the present invention but merely to explain in detail.

Formulation Example 1 Preparation of Tablet

200 mg of the probiotic dry matter of Example 3

Lactose 100mg

Starch 100mg

Magnesium Stearate

The tablets were prepared by mixing the above components and tableting according to a conventional method for producing tablets.

Formulation Example 2 Preparation of Capsule

100 mg of the probiotic dry matter of Example 3

Lactose 50mg

Starch 50mg

Talc 2mg

Magnesium Stearate

The capsules were prepared by mixing the above components and filling the gelatin capsules according to a conventional method for preparing capsules.

Formulation Example 3 Preparation of Liquid

1000 mg of the probiotic dry matter of Example 3

20 g of sugar

20 g of isomerized sugar

Lemon flavor

Purified water was added to adjust the total volume to 100 ml.

The above-mentioned components were mixed according to a conventional method for preparing a liquid solution, filled into a 100 ml brown bottle, and sterilized to prepare a liquid solution.

Formulation Example 4 Preparation of Health Beverage

Example 3 0.1-80%

5-10% sugar

Citric acid 0.05 ~ 0.3%

Caramel 0.005 ~ 0.02%

Vitamin C 0.1 ~ 1%

79 to 94% of purified water is mixed here to make a syrup. The syrup is sterilized at 85 to 98 ° C. for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and then prepared by injecting carbon dioxide to 0.5 to 0.82%. A carbonated beverage containing microorganisms was prepared.

Example 3 Homogeneous blending of subsidiary bacteria such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%), followed by instant sterilization Healthy drinks were prepared by packing them in small packaging containers such as bottles and plastic bottles.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.

Thus, the composition containing the microorganism obtained by culturing the lactic acid bacteria or yeast in the medium of the soy bean curd, glutinous rice and rice bran of the present invention is a pure vegetable lactic acid bacteria is more effective in improving lipid metabolism than the commercially available animal lactobacillus beverages. Confirmed. Since the microorganism-containing composition of the present invention has an excellent effect on preventing and treating geriatric diseases related to lipid metabolism, it is a very useful invention as a biopharmaceutical industry and a dietary supplement.

Claims (9)

An anti-aging pharmaceutical composition containing a medium containing microorganisms obtained by culturing Lactobacillus fmentum and Peach croissant berry in a medium containing soy bean curd, glutinous rice and rice bran. delete delete delete delete delete delete delete A pharmaceutical composition for the prevention and treatment of hypertension, coronary artery disease, and atherosclerosis comprising a medium containing microorganisms obtained by culturing Lactobacillus fmentum and Peach croissant berry in a medium containing soy bean curd, glutinous rice and rice bran.