KR101238836B1 - Phamaceutical or food composition for treating or preventing obesity disease comprising a mixture of lactic acid bacteria - Google Patents

Abstract

The present invention relates to a pharmaceutical composition or a food composition for treating or preventing obesity or obesity-related diseases, including mixed lactic acid bacteria.
The composition of the obesity-related disease comprising the mixed lactic acid bacteria culture medium according to the present invention is a mixture of lactic acid bacteria showing different functions and characteristics, respectively, weight loss, lowering of blood lipid components, improving liver function and inhibiting intestinal harmful enzyme activity It has the effect of increasing the total number of intestinal lactic acid bacteria, and not only activates intestinal function and liver function, but also decreases amylase activity while suppressing appetite, increasing lipase activity and promoting fat breakdown It can be used as a pharmaceutical composition and a functional food for the treatment and prevention of diseases.

Description

Pharmaceutical composition or food composition for the treatment or prevention of obesity or obesity-related disease including mixed lactic acid bacterium {PHAMACEUTICAL OR FOOD COMPOSITION FOR TREATING OR PREVENTING OBESITY DISEASE COMPRISING A MIXTURE OF LACTIC ACID BACTERIA}

The present invention relates to a pharmaceutical composition or a food composition for treating or preventing obesity or obesity-related diseases, including mixed lactic acid bacteria.

Recently, the homeostasis of the human body has been disturbed by the change in diet, the lack of exercise, and the increase of stress, which are mainly caused by high-calorie meals, and accordingly, various obesity and related metabolic diseases are rapidly increasing. Related metabolic diseases include not only type 2 diabetes and hypertension, but also serious diseases that threaten human health and life, including cardiovascular diseases such as stroke and atherosclerosis and various cancers. Compared to the latter half of the 20th century, obesity rates increased by 5 to 10 times. In particular, middle-aged and middle-aged people with obesity can be a direct cause of chronic metabolic syndrome, so always be careful. Fats accumulated in the arms and legs are not decomposed and absorbed well, which makes the appearance unpleasant, but there is no big problem in health, but in the case of abdominal obesity, fats are easily dissolved and absorbed into the blood, which can cause metabolic syndrome.

In 1996, the World Health Organization (WHO) classified obesity as a disease and specified it as a target for prevention or treatment.

Known obesity treatments such as Xenical (Roche Pharmaceuticals, Switzerland), Reductil (Eboth, USA), Exolise (Atopama, France), etc. Classified as inhibitors, most anti-obesity agents are appetite suppressants that suppress appetite by regulating neurotransmitters associated with the hypothalamus. However, conventional therapeutics have low side effects such as heart disease, respiratory disease, and neurological disease, and thus have low sustainability. Therefore, the development of more improved obesity drugs is needed, and the currently developed products have satisfactory therapeutic effects without side effects. Since there is no therapeutic agent, there is a need for the development of a new obesity treatment.

However, data suggest that conventional pharmacological treatments for obesity do not appear to be effective in maintaining long-term weight loss, so future research is essential to discover new drug therapies to reduce the prevalence of obesity.

The term probiotic, on the other hand, was first used by Lilly and Stillwell (Lilly DM et al., Science, 'Probiotics: growth promoting factors produced by microorganism', 147; 747, 1996) and refers to living microbial agents. . Parker (Parker R. B., An Nutr & Health, 29; 4-8, 1974) defined probiotics as 'living microorganisms that affect intestinal microbial balance'.

Probiotics allow the intestinal flora to be well developed, leading to the maturation of beneficial microbial communities in the gut with the concept of competitive exclusion, called the 'Nurmi concept', which can subsequently prevent the proximity of pathogenic microorganisms such as Salmonella (Nurmi E. et al. , Nature, 241; 210, 1973), attracting attention because of the potential effects of the treatment and prevention of immune diseases (Borchers, 2009).

In addition, recent experimental studies have shown the protective effect of several bacterial strains on obesity. Among them, Bifidobacteria is one of the most probiotic agents in the mammalian digestive tract among commensal bacteria belonging to a class of lactic acid bacteria. In one study, strains of Bifidobacterium longgum were isolated from Streptococcus. thermophilus and Lactobacillus Relative to total cholesterol in rats and rats were significantly more effective than delbrueckii subspecies bulgaricus (SL).

In another study, Bifidobacterium strains were positively associated with glucose tolerance, insulin-induced glucose and normalized inflammation (reduced endotoxemia, plasma and fat cell proinflammatory cytokines) in probiotic-treated mice. (Cani et al., 2007).

As a result, the mixture of lyophilized bifidobacteria, lactobacilli and Streptococcus thermophilus improved dietary obesity and increased fatty liver and insulin resistance, which increased liver natural killer T-cells and reduced inflammation signals in mice. Related (Ma et al., 2008).

These findings suggest that Bifidobacterial formulations are safe and can be used in various ways. However, in relation to obesity, Bifidobacterial strains are associated with fat metabolism and body weight and suggested as potential therapeutics for obesity management. There is a lack of specific and various uses.

Accordingly, there is a need for development of an agent for the treatment or prevention of obesity-related diseases through the confirmation of activity as a probiotic using a conventional Bifidobacteria.

In order to solve the above problems, when the mixed lactobacillus culture solution is administered to a high fat diet-induced group, it shows a weight loss effect as well as an excellent inhibitory effect on harmful enzyme activity against intestinal bacteria. It is an object to provide a pharmaceutical composition and functional food for the treatment and prevention of obesity-related diseases comprising a.

The present invention provides a pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising a mixed lactic acid bacteria as an active ingredient for the above purpose.

In the present invention, the mixed lactic acid bacteria is Bifidobacterium pseudocatenuratum SPM 1204 (Accession No. KCTC 10616BP), Bifidobacterium long gum SPM 1205 (Accession No. KCTC 10630BP), and Bifidobacterium long gum SPM 1207 (Accession No. KCTC 11506 BP).

In the present invention, the mixed lactic acid bacteria is Bifidobacterium pseudocatenuratum SPM 1204 (Accession No. KCTC 10616BP), Bifidobacterium long gum SPM 1205 (Accession No. KCTC 10630BP), and Bifidobacterium long gum SPM 1207 (Accession No. KCTC 11506 BP) is characterized in that each culture solution containing 10 ⁸ to 10 ⁹ cfu / ml at the same volume ratio.

In the present invention, the pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising the mixed lactic acid bacteria as an active ingredient is the level of aminotransferase (AST), aspartate aminotransferase (ALT) in the blood It is characterized by reducing.

In the present invention, the pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising the mixed lactic acid bacteria as an active ingredient is characterized by reducing the activity of alpha amylase.

In the present invention, the pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising the mixed lactic acid bacteria as an active ingredient is characterized by increasing the activity of lipase.

The present invention also provides a food composition comprising a pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising the mixed lactic acid bacteria according to the present invention as an active ingredient.

Bifidobacterium sudocatechinulatum SPM 1204, one of the mixed lactic acid bacteria of the present invention, is a novel Bifidobacterium spp. Microorganism isolated from humans, which promotes cross-presentation of dendritic cells to exogenous antigens, It has an immune enhancing activity that induces NO and TNF-α production and activates macrophages, and was deposited by the inventors on March 31, 2004 (Accession No .: KCTC 10616BP).

Bifidobacterium longgum SPM 1205, a mixed lactic acid bacterium of the present invention, is a novel Bifidobacterium strain from human feces, and has a function of activating intestinal function and enhancing immune function. Deposited on the 10th of January (Accession Number: KCTC 10630BP).

Bifidobacterium long gum SPM 1207, a mixed lactic acid bacterium of the present invention, increases high density lipoprotein (HDL) levels, and decreases total cholesterol and low density lipoprotein (LDL) levels. As a Leeum long gum strain, it was deposited by the present inventors with the Korea Microorganism Conservation Center on May 12, 2009 (Accession No .: KCTC 11506BP).

The present invention is to provide a use as a composition for the treatment and prevention of obesity-related diseases by mixing the lactic acid bacteria known to have immune enhancing activity, intestinal function activity and cholesterol level lowering activity individually. It can also be used to reduce intake and reduce body fat.

Mixed lactic acid bacteria contained in the composition of the present invention can be useful in the treatment of obesity-related diseases by reducing amylase activity, inhibiting appetite, causing continuous food intake inhibitory effect, and increasing lipase activity to break down fat to be absorbed. Can be. In addition, the food intake inhibitory effect may be useful for reducing body fat such as cholesterol or fat cells.

In general, in the treatment and prevention of obesity and its related diseases, a decrease in body fat is required but a decrease in lean fat, ie a decrease in protein, is undesirable. The composition comprising the mixed lactic acid bacteria according to the present invention can act as an excellent component as a therapeutic agent for obesity-related diseases through the reduction of body fat such as fat cells, intestinal harmful enzyme inhibitory activity. In addition, the mixed lactic acid bacteria included in the composition of the present invention reduce the levels of aminotransferase (AST) and aspartate aminotransferase (ALT) in the blood to reduce body fat and activate the liver function while showing the effect of inhibiting obesity As a result, it shows excellent obesity-related treatment or preventive effect without any side effects of body fat reduction.

Obesity-related diseases of the present invention can be selected from the group consisting of obesity, hyperlipidemia, insulin resistance syndrome, obesity-related gastroesophageal reflux, fatty liver, cardiovascular disease and metabolic syndrome.

The composition for treating obesity-related diseases including the mixed lactic acid bacteria culture medium of the present invention may further include appropriate carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The compositions according to the invention can 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. Carriers, excipients and diluents that may be included in the composition comprising the compound or extract 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 and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in the composition. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral use 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 include sterilized 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. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the mixed lactic acid bacteria of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the mixed lactic acid bacteria of the present invention is preferably administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. The administration may be carried out once a day or divided into several doses. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The composition of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

When the mixed lactic acid bacteria of the present invention is used as a food, the mixed lactic acid bacteria can be added as it is or used together with other foods or food ingredients, and can be suitably used according to a conventional method. Functional food for the prevention of obesity-related diseases, including the mixed lactic acid bacteria culture medium of the present invention may be in the form of powder, granules, tablets, capsules or beverages, but is not limited thereto.

There is no particular limitation on the kind of the food. Examples of foods to which the strain can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, dairy products including other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like, and are preferably prepared with fermented milk, yoghurt, cheese, milk drink or milk powder, and more preferably fermented milk, yoghurt, beverage, milk powder, food additives or health supplements.

The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment).

In general, 1 x 10 ⁷ to 10 x 10 ⁸ (bacteria) of the Bifidobacterium long gum strain of the present invention per 1 g (or ml) of the specific food material is added at the time of preparing the food or beverage. And preferably in an amount of 1 × 10 ⁸ to 5 × 10 ⁸ .

The effective dose of the strain of the present invention may be used in accordance with the effective dose of the pharmaceutical composition, but may be less than the above range in the case of long-term intake for the purpose of health and hygiene or health control, Since there is no problem in terms of safety, it is evident that it can be used in an amount above the above range.

There is no restriction | limiting in particular except the said food composition containing the said mixed lactic acid bacteria as an essential component in the ratio which is indicated, and it can contain various flavors etc. as an additional component like a normal food.

The composition of the obesity-related disease comprising the mixed lactic acid bacteria culture medium according to the present invention is a mixture of lactic acid bacteria showing different functions and characteristics, respectively, weight loss, lowering of blood lipid components, improving liver function and inhibiting intestinal harmful enzyme activity It has the effect of increasing the total number of intestinal lactic acid bacteria, and not only activates intestinal function and liver function, but also decreases amylase activity while suppressing appetite, increasing lipase activity and promoting fat breakdown It can be used as a pharmaceutical composition and a functional food for the treatment and prevention of diseases.

1 is a view showing the change in food intake during the experiment period,
2 is a view showing the total number of lactic acid bacteria changes during the experiment period,
Figure 3 is a diagram showing the inhibition of intestinal harmful enzyme activity when ingested mixed lactic acid bacteria of the present invention.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the present invention is not limited to the following Examples and Experimental Examples.

< Example  1. Preparation of Mixed Lactic Acid Bacteria>

Mixed lactic acid bacteria of the present invention is Bifidobacterium sudocatechinulatum SPM 1204, Bifidobacterium longgum SPM 1205 and Bifidobacterium longgum SPM identified from feces of 20 to 30 years old healthy adults present in Korean intestines 1207 and cultured using the medium shown in Table 1 below.

Media Main enumerated microorganisms Incubation time (day) BL agar medium ^b Predominant anaerobes 3 GAM agar medium ^c Bifidobacterium spp. 1-2

^b BL, Blood Liver (Nissui Pharm. Co. Ltd., Japan).

^c GAM, General Anaerobic Medium (Nissui Pharm. Co. Ltd., Japan).

Each of the lactic acid bacteria was inoculated in a GAM agar medium, followed by incubation at 37 ° C., followed by inoculation of 1% of each of these spawn cultures in sterilized GAM (General anaerobic medium) (Nissui Pharm. Co. Ltd., Japan) broth. 18 hours at 37 ° C. under an anaerobic system filled with a mixed gas consisting of N ₂ (90%), H ₂ (5%) and CO ₂ (5%) (Bactron Anaerobic Chamber, Sheldon Manufacturing Inc., USA) Incubated. Thereafter, the culture solution was centrifuged at 4000 rpm for 5 minutes to remove the supernatant and only the cells were recovered.

Each of the cells obtained by culturing the three strains were mixed at a concentration of 1.0 × 10 ⁸ cfu / ml, and then the mixed cultures were orally administered to the rats for the test subjects in the following experimental examples.

< Reference Example  1.Preparation of experimental animals and setting up administration groups>

The experimental animals purchased 36 male SD (Sprague-Dauri) rats of 3 weeks of age from the central experimental animals, and the standard diet (AIN-76A # 100000, Dyets Inc. for 1 week to stabilize metabolic status). , Bethlehem, PA, USA).

Two animals per cage were fed and water and feed were randomly fed. Rats were exposed to light and dark at 12 hour cycles, maintained at 22 ± 2 ° C and humidity at 55 ± 5%, and all animal experiments were conducted according to experimental animal breeding guidelines approved by Sahmyook University.

Rats were randomly selected and set up in 3 groups (Example (HFD-LAB), Comparative Example 1 (SD), and Comparative Example 2 (HFD)) of 12 animals per group.

< Example  2. Measurement of weight change according to diet

As shown in Table 2, Comparative Example 1 (SD) group was fed a standard diet and PBS as a control, Comparative Example 2 (HFD) group was fed a high-fat diet (40% beef tallow modified AIN-76A purified rodent diet # 101556, Dyets Inc., Bethlehem, PA, USA) and PBS, and the Example (HFD-LAB) group fed 0.2 ml of the high fat diet and the mixed lactic acid bacteria prepared in Example 1 of the present invention.

Groups Rats (n) Food Treatment (oral administration for 5 weeks) Comparative Example 1
(SD) group 12 Standard Diet 0.2 ml of sterilized PBS Comparative Example 2
(HFD) group 12 40% High Fat Diet 0.2 ml of sterilized PBS Example
(HFD-LAB) Group 12 40% High Fat Diet 0.2 ml of LAB in sterilized PBS

The compositions of the standard diet and the high fat diet are shown in Table 3 below. All rats were adapted to their diet for a week before the start of the experiment. Rats were then orally administered 0.2 ml of PBS or mixed lactic acid bacteria of Example 1 (1 × 10 ⁸ CFU / ml) once daily for 5 weeks and body weight and dietary intake were measured daily.

Ingredients Standard Diet High fat diet (AIN-76A diet # 100000) (AIN-76A diet # 101556) g kcal g kcal Casein 200 720 200 720 Beef tallow 0 0 400 3600 Methionine 3 12 3 12 Starch 150 540 150 540 Sucrose 500 2000 150 600 Cellulose 50 0 50 0 Corn oil 50 450 0 0 Salt mixture 35 30.8 35 30.8 Vitamine mixture 10 39 10 39 Choline bitartrate 2 0 2 0 Total 1000 3791.8 1000 5541.8

< Reference Example  2. Statistical Analysis

All measurements are expressed as mean ± standard deviation. Tests were performed using one-way or two-way batch analysis, and data analysis was performed using StatView for Macintosh (version 5.0; SAS Institute, Cary, NC, USA).

< Experimental Example  1.weight measurement>

The change in body weight for each group was measured at three-day intervals during the seven week experiment period, and the results are shown in Table 4 below.

In Table 4, the Comparative Example 1 (SD) group is 315.86 ± 12.42 g, the high fat diet only Comparative Example 2 (HFD) group was fed 349.14 ± 29.14 g, the HFD-mixed lactic acid bacteria (HFD-LAB) group was 339.70 The weight gain of the Comparative Example 2 (HFD) group fed only the high fat diet was shown to be ± 26.75 g, whereas the Example (HFD-LAB) group fed the HFD-mixed lactic acid bacteria according to the embodiment of the present invention had a high fat diet. It can be seen that the weight gain is reduced compared to Comparative Example 2 (HFD) fed only.

Body weight (g) Comparative Example 1 (SD) Group Comparative Example 2 (HFD) Group Example (HFD-LAB)
group 0 day 60.38 ± 3.07 ^d 60.14 ± 3.72 59.70 ± 3.40 3 day 81.88 ± 3.09 80.64 ± 4.48 81.58 ± 4.56 6 day 106.42 ± 3.40 105.44 ± 5.98 105.83 ± 5.51 9 day 125.86 ± 3.58 123.30 ± 7.89 124.83 ± 6.56 13 day 157.13 ± 4.64 156.36 ± 11.94 158.08 ± 8.97 16 day 178.13 ± 4.58 180.00 ± 14.20 180.75 ± 11.24 20 day 208.00 ± 5.69 215.11 ± 16.56 215.60 ± 13.13 23 day 228.29 ± 4.99 235.20 ± 20.21 236.00 ± 16.92 27 day 259.38 ± 7.19 273.00 ± 21.56 272.36 ± 19.05 30 day 281.86 ± 6.31 296.67 ± 24.02 293.17 ± 24.43 34 day 311.50 ± 8.30 327.88 ± 28.64 323.25 ± 29.19 37 day 325.22 ± 9.67 340.22 ± 29.44 334.91 ± 27.91 41 day 348.38 ± 12.19 364.11 ± 30.64 359.27 ± 29.02 44 day 358.57 ± 11.49 380.00 ± 31.63 371.40 ± 28.91 48 day 376.14 ± 11.20 ^B 409. 29 ± 29.77 ^A 399.40 ± 28.28 ^AB Weight gain 315.86 ± 12.42 ^B 349.14 ± 29.14 ^A 339.70 ± 26.75 ^AB

In addition, the average food intake rate for each group was measured and the results are shown in FIG. 1. As shown in FIG. 1, the average food intake of the Comparative Example 2 (HFD) group was higher than that of the Comparative Example 1 (SD) group, but in the Comparative Example 2 (HFD) group and the Example (HFD-LAB) group No difference was seen.

< Experimental Example  2. Serum analysis and measurement of liver function activation>

At the end of the 7 week experiment period, blood samples from each rat of 3 groups (Example (HFD-LAB), Comparative Example 1 (SD), Comparative Example 2 (HFD)) were collected in tubes and centrifuged. Serum was isolated (3500 rpm / 10 min).

Total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), triglyceride, glucose, leptin, alanine, aminotransferase (AST), aspartate aminotransfer from isolated serum Latase (ALT) levels were analyzed by the Korean Veterinary Association, and the results are shown in Table 5 below.

As shown in Table 5, the HFD-lactic acid bacteria fed Example (HFD-LAB) increased the high-density lipoprotein cholesterol (HDL-C) level compared to the Comparative Example 2 (HFD) group fed the high fat diet, Reduced levels of total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), triglycerides, glucose, leptin, alanine, aminotransferase (AST) and aspartate aminotransferase (ALT) It was found to show an effect of improving liver function.

In particular, it can be seen that the aminotransferase (AST) is reduced by more than 20% compared to Comparative Example 1 and Comparative Example 2.

Comparative Example 1 (SD) Comparative Example 2 (HFD) Example (HFD-LAB) TC (mg / dl) 82.50 ± 13.03 ^{d, B} 99.00 ± 8.51 ^A 85.13 ± 16.33 ^AB HDL-C (mg / dl) 55.36 ± 9.34 ^A 45.33 ± 8.40 ^B 48.64 ± 7.49 ^AB LDL-C (mg / dl) 20.27 ± 2.50 ^A 15.58 ± 4.19 ^B 15.22 ± 4.40 ^B Triglyceride (mg / dl) 146.18 ± 33.76 ^B 278.33 ± 104.61 ^A 246.70 ± 74.18 ^A Glucose (mg / dl) 238.18 ± 46.92 ^A 228.50 ± 37.20 ^A 191.70 ± 24.72 ^B AST (U / L) 113.70 ± 47.11 ^A 107.58 ± 14.51 ^AB 82.50 ± 12.81 ^B ALT (U / L) 38.00 ± 5.25 ^B 63.08 ± 8.64 ^A 62.70 ± 4.35 ^A

< Experimental Example  3. Long-term weight measurement>

Fats (dye testes and posterior peritoneum), liver, spleen after sacrifice of each rat of 3 groups (Example (HFD-LAB), Comparative Example 1 (SD), Comparative Example 2 (HFD)) after the end of the 7 week experiment period. And pancreas were immediately removed and each weighed.

As shown in Table 6, in Comparative Example 2 of the high fat diet, although the weight of the anaerobic and retroperitoneum induced energy accumulation in the form of body fat including the amount of fat, the high fat diet and the practice according to the present invention In the case of the Example group ingested with the lactic acid bacterium composition of the example it can be seen that the weight increase is significantly reduced than in Comparative Example 2.

Comparative Example 1
(SD) Comparative Example 2
(HFD) Example
(HFD-LAB) Body weight (g) 381.00 ± 11.95 ^{d, B} 413.86 ± 30.28 ^A 402.20 ± 29.23 ^AB Fat weight (g)
(epididymal + retroperitoneal) 13.94 ± 2.66 ^B 20.94 ± 4.08 ^A 18.47 ± 2.02 ^A Fat pad weight
(g / 100 g body weight) 3.63 ± 0.65 ^B 5.11 ± 0.60 ^A 4.83 ± 0.40 ^A Spleen (g) 0.66 ± 0.03 0.60 ± 0.07 0.64 ± 0.10 Kidney (g) 2.59 ± 0.15 2.60 ± 0.18 2.61 ± 0.08 Liver (g) 18.47 ± 1.95 ^AB 20.04 ± 2.27 ^A 17.59 ± 2.33 ^B

< Experimental Example  4. Alpha Amylase  And lipase activity measurement>

Alpha-amylase and lipase activity were measured using alpha-amylase assay kit (BioAssay Systems, USA) and lipase assay kit (BioAssay Systems, USA), respectively, and the results are shown in Table 7 below.

As shown in Table 7 below, in the group to which the mixed lactic acid bacteria of the present invention were administered, the activity of amylase was decreased and the activity of lipase was increased as compared to the comparative example 1 group having the standard diet.

Comparative Example 1 (SD) Example (HFD-LAB) Alpha- amylase (U / L) 193.33 ± 9.77 ^d 181.17 ± 11.89 Lipase
(U / L) 121.67 ± 12.44 208.50 ± 15.18

d: Each standard deviation is shown.

Alpha amylase is one of the digestive enzymes that hydrolyze polysaccharides and is contained in human pancreatic (interest) fluid and saliva. Thus, inhibition of α-amylase activity is known to be effective in satisfying appetite to inhibit obesity and also in treating diabetes. In the embodiment of the present invention, in the group administered with the mixed lactic acid bacteria it can be seen that it is effective in inhibiting obesity, diabetes treatment by reducing the activity of α-amylase.

Lipase is one of the pancreatic enzymes that breaks down and absorbs fat from the intestine. It is known to function to induce the utilization of energy in the mitochondria by separating fats in the blood and moving them to the cells and then breaking down fats in the cells. Thus, the mechanism of activating lipases is known as one of the obese drugs. In the case of administering the mixed lactic acid bacteria according to the embodiment of the present invention, as shown in Table 7, it can be seen that the activity of lipase is rapidly increased as compared with the case of Comparative Example 1, which is effective in inhibiting obesity and treating diabetes.

< Experimental Example  5. Measuring whether the chapter function is activated>

5-1. An intestinal gun Lactobacillus  Measure

Fecal samples were collected for determination of total intestinal lactic acid bacteria and harmful enzymes. Fecal samples were taken from the rectum by rectal stimulation and immediately transferred to sterile tubes and stored at 4 ° C. Fecal samples (0.1g) by using a vortex was mixed with 0.1 M phosphate buffer, 0.9ml, 0.1ml is 10 ^-1 to 10 ^- were diluted to a concentration of 10 times from ^7. 1 ml was added to a selective GAM (General Anaerobic Medium) agar medium, and after 48 hours of incubation in anaerobic conditions, the number of Bifidobacterium bacteria was measured (Choi et al., 2005).

The total number of lactic acid bacteria against the number of bacteria is shown in FIG. 2. The mixed lactic acid bacteria count for the total bacterial count was lower in the Example (HFD-LAB) and Comparative Example 2 (HFD) groups than in the Comparative Example 1 (SD) group, but the HFD-LAB group was significantly increased than the HFD group. And it was found.

5-2. β- Glucosidase  Active measurement

1 ml of a reaction solution consisting of 0.8 ml of 2 mM p-nitrophenyl-β-D-glucopyranoside and 0.2 ml of suspended enzyme solution (suspended fecal sample) is reacted at 37 ° C for 30 minutes, and then 1 ml of 0.5N NaOH is added to terminate the reaction. . Thereafter, the reaction solution was centrifuged (3,000 rpm / 10 min) and the supernatant was measured for absorbance at 405 nm. The results are shown in FIG. 3.

5-3. β- Glucuronidase  Active measurement

1 ml of a reaction solution consisting of 0.8 ml of 2 mM p-nitrophenyl-β-D-glucuronide and 0.2 ml of suspended enzyme solution was reacted at 37 ° C. for 30 minutes, and 1 ml of 0.5N NaOH was added to terminate the reaction. Thereafter, the reaction solution was centrifuged (3,000 rpm / 10 min), and the supernatant was measured for absorbance at 405 nm. The results are shown in FIG. 3.

5-4. Tryptopanase  Active measurement

0.2 ml of complete reagent solution (0.25 ml of 20 mM tryptophan) and enzyme solution suspended in 100 ml of 0.05 M potassium phosphate buffer containing 2.75 mg of pyridoxal phosphate, 19.6 mg of disodium EDTA dihydrate, 10 mg of bovine serum albumin and pH 7.5 The reaction solution consisting of ㎖ was reacted at 37 ° C. for one hour, and 2 ml of color reagent (ρ-dimethylaminobezaldehyde 14.7 g, 95% ethanol 948 ml, H ₂ SO ₄ 52 ml) was added to terminate the reaction, followed by centrifugation. The absorbance was measured at 550 nm (3,000 rpm / 10 min) and the results are shown in FIG. 3.

5-5. Urease  Active measurement

A reaction solution consisting of 0.3 ml of urea substrate solution (solution dissolved in 4 mM urea in 20 mM sodium phosphate buffer) and 0.5 ml of suspended enzyme solution was reacted at 37 ° C. for 30 minutes, and 0.1 ml of 1 N (NH ₄ ) ₂ SO _{4 was} added. The reaction was terminated by addition.

Thereafter, 1 ml of a phenolnitroprusside reagent and 1 ml of an alkaline hypochlorite reagent (NaClO) were added to the reaction solution for 20 minutes at 65 ° C. The reaction solution was centrifuged at 3,000 rpm for 10 minutes, and the supernatant was measured for absorbance at 603 nm. The results are shown in FIG. 3.

As shown in Figure 3 intestinal detrimental activity of β-glucosidase, β-glucuronidase, tryptopanase except urease in the group (HFD-LAB) ingested mixed lactic acid bacteria of the present invention is It was confirmed that the reduction was 36%, 30% and 13%, respectively.

Claims (7)

Bifidobacterium sudocatechinulatum SPM 1204 (Accession No. KCTC 10616BP), Bifidobacterium long gum SPM 1205 (Accession No. KCTC 10630BP), and Bifidobacterium long gum SPM 1207 (Accession No. KCTC 11506 BP) A pharmaceutical composition for treating or preventing obesity or obesity-related diseases comprising phosphorus mixed lactic acid bacteria.
delete The method of claim 1,
The mixed lactic acid bacteria are Bifidobacterium pseudocatenuratum SPM 1204 (Accession No. KCTC 10616BP), Bifidobacterium long gum SPM 1205 (Accession No. KCTC 10630BP), and Bifidobacterium long gum SPM 1207 (Accession No. KCTC 11506 BP) A pharmaceutical composition for treating or preventing obesity or obesity-related diseases, including mixed lactic acid bacteria, characterized in that each culture solution containing 10 ⁸ to 10 ⁹ cfu / ml in the same volume ratio.
The method of claim 1,
A pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising the mixed lactic acid bacteria as an active ingredient is characterized by reducing the levels of aminotransferase (AST) and aspartate aminotransferase (ALT) in the blood. A pharmaceutical composition for treating or preventing obesity or obesity-related diseases comprising mixed lactic acid bacteria.
The method of claim 1,
A pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising the mixed lactic acid bacteria as an active ingredient is a pharmaceutical for the treatment or prevention of obesity or obesity-related diseases comprising a mixed lactic acid bacteria, characterized in that to reduce the activity of alpha amylase Composition.
The method of claim 1,
Pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising the mixed lactic acid bacteria as an active ingredient Pharmaceutical composition for the treatment or prevention of obesity or obesity-related diseases comprising a mixed lactic acid bacteria, characterized in that to increase the activity of lipase .
A food composition for the treatment or prevention of obesity or obesity-related diseases comprising the pharmaceutical composition of any one of claims 1 or 3.

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