KR20150003968A - Composition for preventing or treating obesity, endotoxemia or metabolic disease containing fermented Lonicerae Flos - Google Patents

Abstract

The present invention relates to a composition for preventing or treating obesity, endotoxemia, or metabolic diseases comprising fermented lonicerae flos as an active ingredient. The fermented lonicerae flos of the present invention has excellent treatment effects on endotoxemia including: improvement of intestinal permeability; inhibition of inflammation by inhibiting generation of NO; inhibition of blood serum inflammation biomarkers; and control of intestinal effective microorganisms. Also, the fermented lonicerae flos of the present invention has excellent effects on weight reduction and metabolism improvement including: weight reduction; fat reduction in epididymis, kidney, and liver; and inhibition of blood serum metabolic biomarkers, thereby being useful for treating obesity or metabolic diseases.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or treating obesity, endotoxemia or metabolic diseases,

The present invention relates to a composition for preventing or treating obesity, endotoxemia or metabolic diseases, which comprises fermented gold germ as an active ingredient.

Metabolic disease is a generic term for diseases caused by metabolic disorders in vivo. Metabolic disease refers to diseases caused by imbalance such as carbohydrates, lipids, proteins, vitamins, minerals and water, and is also referred to as metabolic diseases or metabolic disorders. It is difficult to develop a clear treatment method because metabolic diseases act as diverse and complex causes of diseases.

Obesity is a metabolic disorder caused by an imbalance between the consumption of food and energy consumption, which is an excess accumulation of fat in the body. Obesity occurs when the fat contained in the ingested food is decomposed and absorbed by lipase and accumulates in the cells or accumulated fat induces differentiation of lipid precursor cells into adipocytes, thereby increasing the volume or number of adipocytes .

The World Health Organization (WHO) classified obesity as a disease requiring treatment in 1996. Obesity is a cause of various adult diseases and chronic diseases, and is known to be involved in causing diabetes, hypertension, hyperlipidemia, heart disease, arthritis, respiratory disease, sexual dysfunction and cancer. According to the results of the study, worldwide obesity in 2005 was estimated at 400 million people, and by 2015, the global obesity population is expected to increase to 700 million.

The treatment of obesity known to date is drug administration, surgery, diet and exercise therapy, among which the drug administration method is known to have the most excellent effect. However, there is a problem in that administration of a drug having excellent effects, particularly administration of an anti-obesity drug, has side effects such as headache and vomiting, so that it is difficult to administer the drug for a long period of time and the degree of side effects varies depending on the constitution of the individual. In order to solve such a side effect of drug administration, researches on anti-obesity materials based on natural materials or foods with stable stability have been actively conducted.

On the other hand, the bowel is a digestive organs that acts as a barrier to protect the inside of the body while absorbing the nutrients of foods consumed in the body. The chapter is largely known to play a role of a defense wall due to the epidemiological and epidemiological mechanisms. Non-epidermal defense of the intestine is caused by selective intestinal permeation. Excessive intestinal permeability leads to loss of absorption of nutrients needed for life activities and loss of ability to block the entry of harmful substances. Toxic substances introduced into the body interact with the internal immune system, causing a variety of immune responses, and ultimately leading to disease.

On the other hand, when gastrointestinal injury occurs, intestinal bacteria and infectious complications are induced, and the endotoxin is transferred to the organs beyond the intestinal wall. Endotoxemia refers to a phenomenon in which endotoxin, which is a part of intestinal bacteria, is spread. Intestinal bacterial disruption refers to a phenomenon in which intestinal bacteria and various bacterial products are spread beyond the intestinal wall and into the intestines. Endotoxins are known to cause systemic inflammatory response syndrome (SIRS) and sepsis. Intestinal bacterial and intestinal endotoxemia occurs, intestinal bacteria and various products first spread to the mesenteric lymph nodes, travel to the chest tube along the lymphatic system and spread to distant organs such as liver, spleen and lung, Spleen and spread to the systemic bloodstream.

Diseases associated with intestinal bacterial dislocations and endotoxemia include trauma, aortic aneurysm surgery and intestinal transplantation, obstructive jaundice, acute pancreatitis, intestinal obstruction, cirrhosis, heart failure, tumor disease, inflammatory bowel disease, extrapulmonary ARDS ), Systemic inflammatory response syndrome and sepsis, and endotoxemia has been found to correlate with infectious complications of these diseases (Adv.Exp.Med.Biol 1999; 473: 11-30).

On the other hand, Lonicerae Flos or Indong water is a flower of Lonicera japonica Thunberg belonging to Caprifoliaceae, and it is known to be effective in detoxification and detoxification. In addition, gingkohwa has been used for treatment of bruises, wounds, tumors, sore throat, skin diseases, furrows, gait and fever.

Fermented Chinese medicine refers to a medicinal substance which fermented herb medicine as an excellent seed bacterium to increase the medicinal effect of the original herbal medicine or to add a new function. Compared to general Chinese medicine, fermented Chinese medicine has fast action, excellent digestion rate and excellent antioxidant activity. In addition, the fermented Chinese medicine is relatively safe from toxic substances such as pesticides and heavy metals, and has less side effects than general Chinese medicine, but systematic research on it has not been done yet.

The present inventors have conducted studies on natural materials having excellent therapeutic effects of obesity, endotoxemia or metabolic diseases, and found that the fermented product of gold eucharide improves intestinal permeability, inhibits NO generation to suppress inflammation, inhibits serum inflammatory indicator substances And is effective in reducing intestinal microorganisms, reducing body weight, reducing fats in epididymis, kidney and liver, inhibiting serum metabolic indicator substances, and improving weight loss and metabolism, thereby preventing obesity, endometriosis or metabolic diseases And the present invention has been completed.

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating obesity, endotoxemia or metabolic diseases, which comprises a fermented product of gold-silver euroside as an active ingredient.

It is another object of the present invention to provide a food composition for preventing or ameliorating obesity, endotoxemia or metabolic diseases, which comprises a fermented product of gold-silver europium as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or treating obesity, endotoxemia or metabolic diseases, which comprises a fermented product of giitanium sulphate as an active ingredient.

The present invention provides a food composition for preventing or ameliorating obesity, endotoxemia, or metabolic diseases, which comprises fermented gold germ as an active ingredient.

The gold-silver fermented product of the present invention has an excellent therapeutic effect on endotoxemia such as improving intestinal permeability, inhibiting NO generation to inhibit inflammation, inhibiting serum inflammatory indicator substances, and controlling intestinal useful microorganisms. In addition, the gold-silver fermented product of the present invention is excellent in the effect of reducing body weight, reducing fat in epididymis, kidney and liver, inhibiting serum metabolism indicator substances, and reducing weight loss and metabolism. Can be usefully used.

Figure 1 shows the result of measurement of the TEER change rate of the germinated fermented product according to the present invention immediately after the start of the experiment (1. Control group: no DSS treated group 2. DSS: treated DSS group 3. DSS + LF: And 4. DSS + FLF: DSS and germination fermentation treatment group.
FIG. 2 is a graph showing the change in TEER of the gingival perilla fermented product according to the present invention measured 4 hours after the start of the experiment (1 to 4 are as shown in FIG. 1).
FIG. 3 shows the results of measurement of the TEER change rate of the germanium fermentation product according to the present invention at the elapsed time of 22 hours from the start of the experiment (1 to 4 are as shown in FIG. 1).
FIG. 4 is a graph showing the results of measurement of the rate of change of HRP permeability of the gold-silver fermented product according to the present invention (1 to 4 are as shown in FIG. 1).
5 is a graph showing the results of measurement of the Claudin-1 inhibitory activity of the germinated fermented product according to the present invention (1. normal group; 2. LPS treatment group (control group); 3. LPS + ginger extract extract group: 100 μg / ml treatment group; 4. LPS + gingival extract 400 ㎍ / ㎖ treated group 5. LPS + gold silver fermentation 100 ㎍ / ㎖ treated group and 6. LPS + germanium fermentation 400 ㎍ / ㎖ treated group).
(1) normal group; (2) LPS alone treatment group (control group); (3) 100 ㎍ / ml of LPS / LPS LPS-treated group; Eq. (200 ㎍ / ㎖) + LPS treated group; Epsonol 400 ㎍ / ml + LPS treated group / Ml < / RTI > + LPS treated group; and 8. gingivalized fermented product 400 占 퐂 / ml + LPS treated group).
Figure 7 shows the result of the time-lapse measurement of the weight loss effect of the experimental animals on oral administration of the germinated fermented product according to the present invention (1. Normal diet; 2. High fat diet method; 3. High fat diet + LPS 4. LPS + Ginseng Extract Treated Group, 4. LPS + Ginseng Fermentation Treatment Group, and 6. Highland LPS + Colostrum Treatment Group.
FIG. 8 is a graph showing the results of measuring lipid weight-reducing effects of epididymal tissues of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7).
FIG. 9 is a graph showing the results of measurement of the fat weight-reducing effect of renal tissue of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7).
FIG. 10 is a graph showing the results of measurement of serum total cholesterol content changes in an experimental animal to which the gold-silver fermentation product according to the present invention was orally administered (1 to 6 are as shown in FIG. 7).
FIG. 11 is a graph showing changes in serum glucose content of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7).
FIG. 12 is a graph showing changes in serum triglyceride content of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7).
FIG. 13 is a graph showing changes in serum HDL-cholesterol content of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7).
FIG. 14 is a graph showing the results of measurement of changes in serum GOT content of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7).
FIG. 15 is a graph showing the results of measurement of TNF-.alpha. Concentration in serum of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7).
FIG. 16 is a graph showing the results of measurement of the concentration of CRP in serum of an experimental animal to which the gold-silver fermentation product according to the present invention was orally administered (1 to 6 are as shown in FIG. 7).
FIG. 17 is a graph showing the results of Q-PCR of Bifidobacterium, one of intestinal microorganisms, of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7). ).
18 is a graph showing the results of Q-PCR of Lactobacillus, one of intestinal microorganisms, of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7). .
FIG. 19 is a graph showing the results of Q-PCR of bacteriodet, one of intestinal microorganisms of an experimental animal to which gold-silver fermentation according to the present invention is orally administered (1 to 6 are as shown in FIG. 7). .
FIG. 20 is a graph showing the results of Q-PCR of pirimicut, one of intestinal microorganisms, of an experimental animal in which gold-silver fermentation according to the present invention was orally administered (1 to 6 are as shown in FIG. 7). ).
FIG. 21 is a graph showing the results of an analysis of the ratio of bacteriodet / pyromycetes after performing Q-PCR on intestinal microorganisms of an experimental animal orally administered with the germinated fermented product according to the present invention (1 to 6 7).
FIG. 22 is a graph showing the activity of inhibiting the differentiation of adipocyte differentiation in an experimental animal by oral gavage fermentation according to the present invention through oil red O staining. FIG.
FIG. 23 is a diagram showing the result of group interim analysis on intestinal microorganisms of an experimental animal orally administered with the germinated fermented product according to the present invention. FIG.

The present invention provides a composition for preventing or treating obesity, endotoxemia or metabolic diseases, which comprises a fermented product of gingkoferment as an active ingredient.

The composition comprises a pharmaceutical composition and a food composition.

Hereinafter, the present invention will be described in detail.

In the composition according to the present invention, the gold-silver fermentation product as an active ingredient can be obtained by fermenting Ganoderma lucidum hot-water extract inoculated with a strain of the genus Lactobacillus, preferably Lactobacillus planta, but is not limited thereto.

The above-described gingival sausage fermented product can be obtained by a conventional extraction and fermentation method, and commercially available ones can be purchased and used. Conventional extraction methods include, but are not limited to, ultrasonic extraction, filtration, and reflux extraction.

In one embodiment of the present invention, it was obtained in the following manner.

First, the lactic acid bacteria culture solution is added to the germanium powder, and the mixture is subjected to ultrasonic treatment and hot water extraction. After high-pressure sterilization of Ganghwa hot-water extract, cool well at room temperature. The formula is fermented by inoculating Lactobacillus planta with sterilized Ganoderma lucidum extract. The obtained fermentation product is autoclaved at high pressure and centrifuged to obtain a supernatant, thereby obtaining a fermented silver microcrystalline product.

The gold-silver fermented product of the present invention has an excellent therapeutic effect on endotoxemia such as improving intestinal permeability, inhibiting NO generation to inhibit inflammation, inhibiting serum inflammatory indicator substances, and controlling intestinal useful microorganisms. In addition, the gold-silver fermented product of the present invention is excellent in the effect of reducing body weight, reducing fat in epididymis, kidney and liver, inhibiting serum metabolism indicator substances, and reducing weight loss and metabolism. Can be usefully used.

The obesity includes but is not limited to simple obesity, symptomatic obesity, childhood obesity, adult obesity, cell proliferative obesity, cell non-large obesity, upper body obesity, lower body obesity, visceral fat obesity and subcutaneous fat obesity.

The metabolic diseases include, but are not limited to, metabolic obesity, metabolic hyperlipidemia, metabolic diabetes, metabolic dyslipidemia, metabolic arteriosclerosis and metabolic hypertension.

The composition of the present invention may further contain one or more known active ingredients having an effect of preventing or treating obesity, endotoxemia or metabolic diseases together with the fermented gold europae.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral formulations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient 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, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and 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 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. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the individual, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. For the desired effect, the gold-silver fermented product of the present invention may be administered in an amount of 1 mg / kg to 10000 mg / kg per day, or may be administered once a day or divided into several times.

The pharmaceutical composition of the present invention may be administered to a subject in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers for the prevention and treatment of obesity, endotoxemia and metabolic diseases.

In the present invention, the term "health functional food" refers to a food having a biological control function such as prevention and improvement of disease, bio-defense, immunity, recovery after disease and aging inhibition.

The composition of the present invention may be added to a health functional food for the purpose of preventing or improving obesity, endotoxemia or metabolic diseases. When the ferrous sulfide fermentation product of the present invention is used as a food additive, the fermented germanium fermentation product can be used as it is or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the gold or silver fermentation product of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, when the food or beverage is produced. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred examples, experimental examples, and formulation examples are provided to facilitate understanding of the present invention. However, the following examples, experimental examples and preparation examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples, experimental examples and preparation examples.

[Example 1: preparation of fermented gangrenous]

1. Manufacture of Fermented Gold Eunhwa

Lonicerae Flos powder was supplied from Ilsan Hospital of Dongguk University and sealed and stored in a cow. 30 g of the gold germanium powder was placed in a 250 ml beaker and 150 ml of a MRS broth for lactic acid bacteria was added and sonication was performed at room temperature (15 to 20 ° C) for 30 minutes. Hot water extraction was carried out in a shaking water bath at a temperature of 70 ° C and a flow rate of 70 rpm for 3 hours. The obtained gingkochose hot-water extract was autoclaved at 121 ° C for 15 minutes under high pressure, and allowed to stand at room temperature for cooling. Lactobacillus plantarum was inoculated at a concentration of 2 × 10 ⁷ CFU / ml in a sterilized ginger powdery extract and fermented in an incubator at 37 ° C. for 24 hours. The fermented product was autoclaved at 121 ° C for 15 minutes, centrifuged at a flow rate of 3000 rpm for 5 minutes, and the separated supernatant was used in the experiment. The obtained gingylum hot-water extract was referred to as "gingko ginger extract" and was provided as a comparative object of the gingko fermentation product in the following experimental examples.

[Experimental Example 1: Measurement of improving effect on permeability of intestinal membrane of fermented gangrene]

In order to examine the effect of improving the permeability of intestinal germinated fermented product according to the present invention, the permeability test for the germinated fermented product prepared in Example 1 was performed. The measurement of permeability of the intestinal permeability was performed as follows: TEER (Transepithelial Electrical Resistance) measurement test, HRP (Horseradish peroxide) permeability measurement test, and tight junction protein measurement test.

1. Production of HCT-116 cells with enhanced permeability

First, when the differentiation of HCT-116 cells reached a maximum, 24-cell / well culture insert plates (apical) were inoculated with McCoys 5A medium, 10% FBS, 1% penicillin streptococci The cells were cultured at a concentration of 2 x 105 cells / well using ⁵⁰⁰ μl of the culture medium supplemented with the mycin. In a 24-well well receiver plate (basolateral), only 800 μl of culture medium was added and cultured for 24 hours at 37 ° C., 5% CO ₂ , 90% humidity. 10 μl of each strain was added to the above insert plate and added to the culture medium of HCT-116 so that the number of lactic acid bacteria calculated by measuring the absorbance corresponding to the Macfarland scale was 10 ⁹ CFU / 10 μl, Lt; / RTI > After confirming that the cells were well adhered, 3% dextran sodium sulphate (DSS) was added to induce an increase in permeability of the intestinal membrane, followed by culturing for another 24 hours. In the experiment, the Eucalyptus japonica extract and the Ganoderma lucidum fermented product were respectively treated at the time of culture of HCT-116 cells, and the change in the permeability of the intestinal membrane was measured after the addition of DSS.

2. TEER measurement test of fermentation product

In order to carry out the measurement of the permeability measurement of the germanium fermentation product of the present invention, the germanium fermentation product prepared in Example 1 was treated with HCT-116 cells having increased mucosal permeability prepared in the above 1 to determine the TEER (Transepithelial Electrical Resistance) Were measured.

Specifically, the HCT-116 cells were cultured on an apical plate, treated with DSS, and further treated with Ganoderma lucidum extract or Ganoderma lucidum fermented. After 4 hours and 22 hours, the cells were treated with Millicol-ERS machine (Millipore, USA) was used to measure the TEER value. The TEER measurement electrode was checked for zero before use and the electrode tip was inserted into the hole in the inner side of each transwell to immerse it in the media of the basolateral plate, The short side was inserted only to reach the culture medium of the apical plate, and the damage of the monolayer was noted. As the experimental group, DSS-treated group, DSS-treated group, DSS-treated group and DSS-treated group were set according to the mucosal treatment of HCT-116 cells. As the control group, the DSS non-treated group was set to measure the monolayer TEER value of the wells containing the medium not treated with DSS. The TEER values of the control and experimental groups were measured immediately after the start of the experiment, 4 hours after the start of the experiment, and 22 hours after the start of the experiment. At this time, all the measured values mean the average value calculated from a total of 3 wells treated in the same manner. From the TEER values of the control group and each experimental group, the TEER change ratio was calculated to be 100% for the control group. The percent change in TEER of the control and experimental groups was calculated as a percentage of control (%) using the following equation (1), and the results are shown in FIGS.

As shown in FIGS. 1 to 3, the gold-silver fermentation treatment group of the present invention exhibited a higher TEER change ratio than the DSS-treated group treated with only DSS in HCT-116 cells. Based on the TEER values at four different timepoints except the measured values immediately after the start of the experiment, the percentage of the control group was calculated as 100.96% at 2 hours, 95.58% at 4 hours, 93.87% at 20 hours, And 90.5% after 24 hours, indicating that the permeability of the intestinal mucosa was significantly increased compared with the control.

Therefore, it can be seen that the gold-silver fermentation product of the present invention significantly increases the TEER value, and therefore, the effect of improving the mucosal permeability is excellent.

3. HRP permeability measurement of fermented gold eu

In order to perform the measurement of the permeability measurement of intestinal germinated fermented product of the present invention, the fermented ginseng fermented product prepared in Example 1 was subjected to HRP (Horseradish peroxide) permeability measurement test to the HCT-116 cells prepared in 1 above Respectively.

Specifically, DSS was added to the HCT-116 cells. After 22 hours, the existing medium was removed from the 24-well culture insert plate (top type), and then 60 μl of HRP (peroxide from Horseradish Type VI-A, Sigma) And incubated for 1 hour at 37 ° C, 5% CO ₂ , 90% humidity. Subsequently, 100 μl of the culture medium diluted 10000 times and 100 μl of HRP substrate solution TMB SOL (3,3'5,5'-tetramethylbenzidine) were added to the medium of the receptor plate (outer base type) Lt; / RTI > When the color of the solution on the HRP-treated plate is confirmed to change sharply, the reaction is terminated by treating 100 μl of Stop solution (2M H ₂ SO ₄ ), and a UV ELISA microplate reader Absorbance was measured at a wavelength of 450 nm. At this time, the measurement of HRP means that the mucosal permeability increases as the activity of HRP increases and the concentration of the receptor plate increases. Also, all the measured values mean an average value calculated from a total of three wells treated in the same manner. From the HRP values of the control group and each experimental group, the percent change in HRP permeability was calculated from the control group as 100%. The percent change in HRP permeability of the control and experimental groups was calculated as a percentage of control (%) relative to the control using the following equation (2), and the results are shown in FIG.

As shown in FIG. 4, in the gold-silver fermentation treated group of the present invention, the rate of change of HRP permeability was measured to be 174.01%, which was significantly lower than that of the control group, and it was confirmed that the cell mucosal permeability was remarkably reduced.

Thus, it can be seen that the ferrous sulfide fermented product of the present invention significantly reduces HRP permeability, and thus has an excellent effect of reducing cell mucosal permeability.

4. Measurement of Claudin-1 gene level inhibitory activity of fermented Ganoderma lucidum

In order to perform the measurement of the permeability measurement of the germanium fermentation product of the present invention, HCT-116 cells prepared in the above 1 were treated with the germanium fermentation product and gold silver (nonfermented) prepared in Example 1, The level of the tight gap junction protein, claudin-1, was measured as follows.

Specifically, the HCT-116 cells (1 × 10 ⁷ cells / well), respectively for 2 hours in the honeysuckle and honeysuckle concentration of the fermentation 0, 100, 200, or 400 ㎍ / ml in the following, 0.2 ㎍ / ml < / RTI > LPS for 6 hours. Total RNA of HCT-116 cells treated with LPS was isolated using TRIsure reagent (Bioline, UK) and cDNA was synthesized using AccuPower RT premix kit (Bioneer, Daejeon, Korea). The primer sequences are as follows: glyceraldehydes-3-phosphate dehydrogenase (GAPDH) is a "TGA TGA CAT CAA GAA GGT GGT GAA G and TCC TTG GAG GCC ATG TAG GCC AT (SEQ ID NO: 1) ", and claudin-1 is "CTT CTA CAA ATG GAC CTC T and AGC CTA CAC TAC CTC CTA A (SEQ ID NO: 2). The quantification of the associated gene expression was expressed by the 2-ΔCt equation (ΔCt = Ct-target gene-Ct-GAPDH) and then normalized or normalized by the housekeeping gene GAPDH. The results are shown in Fig.

As shown in FIG. 5, the Claudin-1 gene was significantly increased in all treated groups by LPS treatment as compared with the normal group. The gold-silver fermentation treatment group of the present invention markedly reduced the clodin-1 gene by LPS treatment, especially at a concentration of 400 ug / ml.

Therefore, it can be seen that the gold-eutectic fermented product of the present invention remarkably inhibits the claudin-1 gene, and thus the intestinal permeability is improved.

[Experimental Example 2: Measurement of anti-inflammatory effect of fermented gangrenous]

In order to examine the anti-inflammatory effect of the fermented product of the present invention, the activity of inhibiting nitric oxide (NO) production of the fermented product prepared in Example 1 was measured.

1. Cell preparation and culture

RAW 264.7 cells, a murine leukemic monocyte-macrophage cell, were purchased from Korea Cell Line Bank (KCLB, Seoul, Korea) and cultured in 10% fetal bovine serum (FBS) (Dulbecco's modified Eagle's medium, GIBCO, USA). The RAW 264.7 cell line was heat-inactivated by heating to 10%, and DMEM medium supplemented with Fetal bovine serum (FBS), 2 mM L-glutamine, 100 U / ml penicillin, and 100 μg / ml streptomycin And the cells were incubated at 37 ° C. and 5% CO ₂ .

2. Measurement of NO generation inhibitory activity of fermented germanium

The germinated fermented product prepared in Example 1 was treated with the RAW 264.7 cells prepared in Example 1 to measure the inhibitory activity against NO production.

Specifically, the RAW 264.7 cells were cultured in a 24-well plate at a concentration of 2 × 10 ⁵ per well. The unfermented Eucalyptus and the above fermented Gingivalaceae were pretreated for 2 hours at a concentration of 0, 100, 200 or 400 / / ml, respectively, and treated with 0.2 / / ml LPS for 24 hours. The supernatant was separated from the culture of RAW 264.7 cells treated with LPS and NO was measured using a Griess reagent system kit (Promeag, Wis., USA). The results are shown in Fig.

As shown in FIG. 6, the gold-silver fermentation treatment group of the present invention inhibited LPS-induced NO production in a concentration-dependent manner (p < 0.01).

Therefore, it can be seen that the gold-eutectic fermented product of the present invention is excellent in the effect of suppressing inflammation and endotoxin reaction because of its excellent activity of inhibiting NO generation.

[Experimental Example 3: Measurement of weight loss effect of fermented gangrenous]

In order to examine the effect of controlling the weight of the fermented product of the present invention, the fermented product prepared in Example 1 was orally administered to experimental animals, and then the rate of change of body weight with time was measured.

1. Purchase and breeding of experimental animals

Forty-two healthy male rats weighing 200 ± 20 g at 8 weeks of age were purchased from Orient Bio (Seongnam-si, Korea) as experimental animals. The purchased animals were adapted for one week while being freely fed with solid feed (Soyagreentec, Hwaseong-si. Korea) and water in an animal breeding room where temperature and humidity were controlled appropriately. Solid feeds consisted of 20% protein, 4.5% fat, and 63% carbohydrates. The environment of the breeding room was maintained at a temperature of 22 ± 2 ° C and a relative humidity of 40% ~ 60%. Breeding was carried out in the animal breeding room where the intensity was constantly maintained at a constant rate of 12 hours. During the test period, the temperature and humidity of the animal laboratory were constantly maintained by constant temperature and humidity.

Forty-two adult male rats were randomly selected from three to four rats per cage, and were divided into groups according to diet as follows: Normal group (normal group, normal group or N, n = 7); High Fat Diet Group (control or HFD, n = 7); High fat diet + LPS treated group (High Fat Diet + LPS Group or negative control group n = 7); High Fat Diet + LPS + Lonicerae Flos Group, Liliaceae extract treatment group or LF, n = 7); High fat Diet + LPS + fermented Lonicerae Flos, germinated fermented water treatment group or FLF, n = 7); (High fat diet + LPS + colostrum group, colostrum treatment group or Colostrum group, n = 7), and the high fat diet + LPS + colostrum group.

2. Measurement of weight change in experimental animals

The body weight of the experimental animals was measured for 8 weeks while controlling the diet according to the group described in 1 above. The gilt silver obtained in Experimental Example 1 and the gilt-silver fermentation product prepared in Example 1 and the colostrum purchased in the market were placed in a 5 ml syringe (Korea Vaccine, Seoul, Korea) and an oral zonde (Dongseo Scientific, Seongnam-si, Korea) were orally administered to experimental animals at regular intervals on a daily basis. The body weights of the experimental animals were measured once a week for 8 weeks on a certain day and time, and were measured using an electronic balance (Sartorius, Goettingen, Germany). The difference between the weight measured before the start of the experiment and the weight measured after 8 weeks was expressed as an average. The results are shown in Fig.

As shown in Fig. 7, the difference between the normal group and the average was 355 ± 26.74 g. The difference in mean + LPS treated group (negative control group) was 456.71 ± 18.49g, and the body weight was increased for 8 weeks compared to the normal group (p <0.05). In the case of the high fat diet of the present invention, the + LPS + germanium fermented product group had a difference of 359.43 ± 26.11 g from the mean, and it was confirmed that the body weight was reduced compared to the negative control group.

Therefore, it can be seen that the gold-silver fermented product of the present invention is excellent in weight loss effect.

5. Statistical Analysis

All statistical analyzes were performed using the SPSS software package (version 17.0, Statistical package for social science, New York, United states), and the measurements of each treatment group were expressed as mean ± SD . One Way ANOVA. The difference between the groups was postmarked by Tukey HSD and significance was recognized when the significance (p-value) was 0.05 or less.

[Experimental Example 4: Measurement of effect of reducing the weight of epididymal tissue fat of fermented gangrenosum]

To investigate the effect of reducing the epididymal tissue fat weight of the fermented product of the present invention by oral administration to the experimental animals, the change in the fat mass of the epididymal peripheral tissues was measured over time .

1. Sampling of samples and tissues

In Experimental Example 3, samples and tissues were collected from experimental animals whose experimental period of 8 weeks was completed. After the completion of the experiment, the stool was fasted for 6 hours, and a feces and a urine sample were collected. Then, 1 ml of a solution containing 66 mg / ml of lactulose and 50 mg / ml of mannitol was administered. LPS was injected (0.75 mg / kg), and after 18 hours, feces and urine were collected again. Thereafter, anesthesia was performed, followed by laparotomy along the abdominal midline, and blood was drawn from the heart using a syringe. After collecting blood, epididymal fat and kidney fat were extracted. Each organs were washed several times in saline to remove blood, remove water, and stored in formaldehyde (10%).

2. Measurement of tissue fat around the epididymis

The weight of epididymal fat obtained in 1 above was measured. The weight of the measured epididymal fat was statistically analyzed by the same method as described in Experimental Example 1 above. The results are shown in Fig.

As shown in FIG. 8, the weight change ratio of the epididymal surrounding tissue fat was 5.38 ± 0.87 g in the normal group. The lipid weight of the epididymal tissues was higher than that of the normal group (p <0.05), indicating that the high fat diet + LPS treatment group (negative control group) was 15.36 ± 2.91g. It was confirmed that the high fat diet of the present invention was 8.96 ± 1.37 g in the + LPS + fermented germinated fermented food group, and the fat weight of the epididymal tissues was reduced compared to the negative control group.

Therefore, it can be seen that the gold-silver fermented product of the present invention is excellent in reducing fat of epididymal tissues.

[Experimental Example 5: Measurement of weight reduction effect of kidney tissue fat of fermented gangrenous]

In order to examine the effect of reducing the weight of kidney tissue fat in the fermented product of the present invention, the fermented product prepared in Example 1 was orally administered to an experimental animal, and the change in the weight of the fat around the kidney was measured .

After extracting the kidney tissue in the same manner as described in 1 of Experimental Example 4, the fat weight of the tissue around the kidney was measured. The weight of the measured tissue fat around the kidney was analyzed by the same method as described in Experimental Example 1 above. The results are shown in Fig.

As shown in Fig. 9, the weight change ratio of the tissue fat around the kidney was 6.62 ± 1.09 g in the normal group. The high fat diet + LPS treatment group was 25.99 ± 2.61 g, indicating that the fat weight of kidney tissue was increased compared to the normal group (p <0.05). The high fat diet of the present invention was confirmed to be 12.72 ± 3.74 g in the + LPS + germanium fermented water treatment group, indicating that the fat weight of kidney tissues was decreased compared to the negative control group (p <0.001). In addition, it was confirmed that the fat weight of the kidney tissue was decreased in the high fat diet + LPS + fermented group (p <0.05) compared to the high fat diet group.

Therefore, it can be seen that the gold-silver fermented product of the present invention is excellent in reducing the fat of the kidney tissue.

[Experimental Example 6 Control of Serum Metabolic Indicator Activity of Fermented Gold Ears]

In order to examine the effect of controlling the activity of serum metabolic markers of the gingkoferment fermented product according to the present invention, the fermented gingivalae prepared in Example 1 was orally administered to experimental animals, The metabolic indicator activity was measured. The metabolic indicator may be a total cholesterol, glucose, triglyceride, HDL-cholesterol and GOT, as well as assay kits (AM202-k , AM-2011C, AM-101, etc.). All the measured values were statistically analyzed in the same manner as described in Experimental Example 1 above.

1. Obtaining of serum

Blood was collected from the heart of anesthetized rats using a syringe in the same manner as described in 1 of Experimental Example 4. The collected blood was centrifuged at 1000 g for 15 minutes at a temperature of 4 째 C to obtain serum, and then put in a refrigerator maintained at a temperature of -70 째 C and stored until analysis.

2. Measurement of serum metabolic indicator

end. Total cholesterol measurement

As described in the above 1, the change in the total cholesterol content of the stored serum was measured using the assay kit, and the results are shown in FIG.

As shown in FIG. 10, the change rate of total cholesterol in the serum was 48.42 ± 7.93 mg / dl in the normal group. The high - fat diet + LPS - treated group (negative control) was 90.49 ± 16.81 ㎎ / ㎗ and the total cholesterol level was increased compared to the normal group (p <0.05). The high fat diet of the present invention was found to be 56.37 ± 8.45 mg / dL in the + LPS + fermented germinated fermented milk group, indicating that the serum total cholesterol content was decreased compared to the negative control group (p <0.001).

I. Measurement of glucose content

As described in the above 1, the change in glucose content of the stored serum was measured using the assay kit, and the results are shown in FIG.

As shown in Fig. 11, the change rate of glucose content in the serum was found to be 244.22 ± 22.13 mg / dl in the normal group. The high - fat diet + LPS treatment group (negative control group) was 214.05 ± 33.30 ㎎ / ㎗. The high - fat diet of the present invention was confirmed to be 176.20 ± 51.68 mg / dL in the + LPS + fermented ginseng fermented food group, and it was confirmed that the glucose content was reduced compared to the normal or negative control group.

All. Measurement of Triglyceride Content

As described in the above 1, the change in the triglyceride content of the stored serum was measured using the above assay kit, and the results are shown in FIG.

As shown in Fig. 12, the change rate of the content of triglyceride in the serum was found to be 26.53 ± 6.86 mg / dl in the normal group. The high - fat diet + LPS treatment group (negative control) was 59.21 ± 12.20 ㎎ / ㎗. The high fat diet of the present invention was confirmed to be 43.24 ± 7.67 mg / dL in the + LPS + fermented germinated fermented food group, and it was confirmed that the neutral fat content was reduced compared to the negative control group.

la. Determination of HDL-cholesterol content

As described in the above 1, the change in the HDL-cholesterol content of the stored serum was measured using the above assay kit, and the results are shown in FIG.

As shown in Fig. 13, the change rate of the HDL-cholesterol content in the serum was 34.80 ± 7.10 mg / dl in the normal group. The HDL-cholesterol content of the control group was 21.55 ± 4.44 ㎎ / ㎗ compared to the control group (p <0.05). The high fat diet of the present invention was confirmed to be 26.36 ± 6.05 mg / dL in the + LPS + fermented ginseng fermented food group, which was higher than that of the negative control group.

hemp. GOT content measurement

As described in the above 1, the change in the GOT content of the stored serum was measured using the above assay kit, and the results are shown in FIG.

As shown in FIG. 14, the change rate of the GOT content in the serum was 23.59 ± 1.34 mg / dl in the normal group. The high - fat diet + LPS treatment group (negative control group) was 124.35 ± 63.38 mg / dl, and the GOT content was decreased compared to the normal group (p <0.001). It was confirmed that the high fat diet of the present invention was confirmed to be 10.55 ± 1.93 ㎎ / ㎗ (p <0.001) in the + LPS + fermented germinated fermented milk group, compared with the normal group and the negative control group.

3. Conclusion

It can be seen that the gold-silver fermented product of the present invention is excellent in the effect of controlling the activity of the serum metabolic indicator.

[Experimental Example 7: Regulation of serum inflammation index substance activity of fermented gold europaea]

In order to examine the effect of controlling the activity of the serum inflammation index substance of the gingkoferment fermented product according to the present invention, the fermented gingivalae prepared in Example 1 was orally administered to experimental animals, and then the activity of the serum inflammation indicator substance Were measured. The inflammatory index substance was measured using a kit (BD ELISA Rat TNF-α & CRP kit) of BD Biosciences (San Diego, CA, USA) as TNF-α and CRP. Serum was obtained and used in the experiment in the same manner as described in Experimental Example 6-1. All the measured values were statistically analyzed in the same manner as described in Experimental Example 1 above.

1. Measurement of TNF-α concentration

The concentration of TNF-α in the serum was measured using the above assay kit, and the results are shown in FIG.

As shown in FIG. 15, the high-fat diet of the present invention showed a significant decrease in TNF-α compared with the + LPS-treated diet (p <0.05).

2. Measurement of CRP concentration

The concentration of CRP in the serum was measured using the above assay kit, and the results are shown in FIG.

As shown in FIG. 16, it was confirmed that the high-fat diet system of the present invention had a significantly decreased CRP concentration compared to the + LPS-treated diet (p <0.05) in the + LPS + germanium fermentation treatment group.

3. Conclusion

From the above results, it can be seen that the gold-silver fermented product of the present invention is excellent in controlling the activity of the inflammatory indicator substance.

[Experimental example 8] Measurement of useful microorganism distribution in the intestine through Q-PCR [

In order to investigate the changes in the distribution of useful microorganisms in the intestinal fermentation product according to the present invention, the intestinal microflora of the intestine-related intestinal flora, namely, Bifidobacteria spp., Lactobacillus spp., Bacteriodetes, The relative DNA levels of Firmicutes were quantified by real-time PCR.

1. Obtaining samples

In order to obtain an intestinal microflora sample of the obesity-related intestine, fecal samples at 6 hours and 18 hours after the end of the experiment, which had been collected and stored in Experimental Example 1, were used, and an AccuPrep stool DNA extraction kit (Bioneer, Daejeon , Korea) were used to extract genomic DNA from rat feces.

2. Q-PCR measurement of internal microflora in obesity-related intestine

end. Bifidobacterium

The relative DNA levels of the Bifidobacterium bacteria obtained in the above 1 were quantified by real-time PCR, and the results are shown in FIG.

As shown in FIG. 17, it was confirmed that the high-fat diet system of the present invention had an effect of changing the relative DNA level of Bifidobacterium bacteria in the + LPS + germanium fermentation treatment group compared to the high fat diet system and high fat diet system + LPS treatment group .

I. Lactobacillus

The relative DNA level of the lactobacillus obtained in 1 above was quantified by real-time PCR, and the results are shown in FIG.

As shown in FIG. 18, it was confirmed that the high-fat diet system of the present invention had an effect of changing the relative DNA level of Lactobacillus in the high fat diet system and the high fat diet system compared to the + LPS-treated group.

All. Bacteriodet

The relative DNA level of the bacteriodetes obtained from 1 above was quantified by real-time PCR, and the results are shown in FIG.

As shown in FIG. 19, the high-fat diet system of the present invention, + LPS + germanium fermentation treatment group, showed a higher relative level of Lactobacillus than the + LPS treatment group (P <0.05), respectively.

la. Pirimikutes

The relative DNA levels of the pyrimucestes obtained in 1 above were quantified by real-time PCR, and the results are shown in FIG.

As shown in FIG. 20, it was confirmed that the high-fat diet system of the present invention increased the relative DNA level of pirimicutes in the + LPS + germanium fermentation treatment group compared to the + LPS treatment group (p <0.05 ).

hemp. Percentage of bacteriodet / pyromycetes

The relative DNA level relative to the proportion of bacteriodetin / pyrimictes obtained in 1 above was quantified by real-time PCR, and the results are shown in FIG.

As shown in FIG. 21, it was confirmed that the high-fat diet method of the present invention increased the ratio of bacteriodet / pyromycetes compared to the + LPS-treated diet (p < 0.05).

3. Conclusion

From the above results, it can be seen that the gold-silver fermented product of the present invention is excellent in the effect of controlling the level of useful microorganisms in intestines.

[Experimental Example 9: Measurement of liver fat weight reduction effect of fermented gangrenosum]

In order to examine the effect of the fermentation product of the present invention on the weight loss of liver tissue fat, the fermented product prepared in Example 1 was orally administered to an experimental animal, and the change of liver fat weight was measured with time. For measurement of liver tissue fat, Oil Red 0 staining was performed, and all values measured were statistically analyzed in the same manner as described in Experimental Example 1 above.

The frozen liver tissues, which had been extracted and stored in Experimental Example 1, were mounted on a silicon-coated slide at a temperature of 60 ° C for 10 minutes with an oil red O solution (Cayman chemical, USA) Mayer ' s hamatoxylin solution for 1 minute. All slides were observed with an Olympus BX61 microscope (Tokyo, Japan) and each slide was photographed using an Olympus DP70 digital camera (Olympus DP70 digital camera, Tokyo, Japan). The area of reddish fat tissue was calculated using Image J software (Image J software, Bethesda, Md., USA) and the results are shown in FIG.

As shown in FIG. 22, the gold-silver fermentation treatment group of the present invention was confirmed to inhibit the differentiation of adipocytes.

Therefore, it can be seen that the gold-silver fermented product of the present invention is excellent in the effect of inhibiting the differentiation of adipocytes in liver tissue and reducing the fat of liver tissue.

[Experimental example 10: Effect of fermentation of gingkoferment on intestinal microorganism]

In order to examine the intestinal microbial control effect of the gold-silver fermentation product of the present invention, intramuscular qualitative analysis of intestinal microorganisms was performed using PCR-DGGE (Denaturing Gradient Gel Electrophoresis). All the measured values were statistically analyzed in the same manner as described in Experimental Example 1 above.

Specifically, faecal and urine samples were collected 6 hours and 18 hours after the end of the experiment, which was being collected and stored in Experimental Example 1, and an AccuPrep stool DNA extraction kit (Bioneer, Daejeon, Korea) was used To extract genomic DNA from rat feces. Thereafter, PCR was performed using a universal bacterial primer (5 '→ 3': AGA GTT TGA TCC TGG CTC AG and AAG GAG GTG ATC CAG CC, SEQ ID NO: 3) PCR) was performed. This PCR amplicon was used as a template to perform nested PCR (nest PCR). The primers used for the double polymerase chain reaction were 341 fGC (5'-CGC CCG CCG CGC GCG GCG GGC GGG GCG GGG GCA CGG GGG GCC TAC GGG AGG CAG CAG-3 ', SEQ ID NO: 4) and 518r GCT GG-3 ', SEQ ID NO: 5). The Nest PCR amplicon samples were electrophoresed at 40 ° C for 15 hours at a temperature of 60 ° C and a voltage of 80V using a Dcode ™ System (BIO-RAD, USA) (40% acrylamide / / bis) (37.5: 1)). After staining with Ethidium Bromide (EtBr) for 15 minutes, the gel was photographed under UV illumination (LAS-3000; Fuji photo film, Tokyo, Japan). The photographed DGGE patterns were analyzed using Bionumerics 3.0 software (Applied Maths, Sint-Martens-Latem, Belgium). The results are shown in Fig.

As shown in FIG. 23, a pattern comparison of intestinal microorganisms using PCR-DGGE revealed that intestinal microorganisms were clustered in the feces of animals in each group.

Therefore, it can be seen that the gold-silver fermented product of the present invention is excellent in the effect of regulating intestinal microflora.

Hereinafter, a pharmaceutical composition for prevention or treatment of obesity, endotoxemia or metabolic diseases containing the gold-silver fermentation product of the present invention, and a food composition for preventing or ameliorating obesity, endotoxemia or metabolic diseases are described, Rather than to limit the invention.

[Preparation Example 1: Preparation of pharmaceutical preparation]

1. Manufacturing of powder

Fermented gangrenous 20 ml

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

2. Preparation of tablets

10 ml of fermented silver

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

3. Preparation of capsules

10 ml of fermented silver

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

4. Preparation of injections

10 ml of fermented silver

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

5. Manufacture of liquids

Fermented gangrenous 20 ml

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

[Formulation example 2. Preparation of food preparation]

1. Manufacture of health food

100 ml fermented gold silver

Vitamin mixture quantity

70 g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 g of vitamin B12

Vitamin C 10 mg

Biotin 10 g

Nicotinic acid amide 1.7 mg

Folate 50 g

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2. Manufacture of health drinks

100 ml fermented gold silver

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (10)

A pharmaceutical composition for the prevention or treatment of obesity, endotoxemia or metabolic diseases, which comprises fermented ginseng (Lonicerae Flos fermentation) as an active ingredient.
The pharmaceutical composition for preventing or treating obesity, endotoxemia or metabolic diseases according to claim 1, wherein the germinated fermented product is obtained by fermenting Ganoderma lucidum hydrothermal extract with a strain of Lactobacillus sp.
The pharmaceutical composition for preventing or treating obesity, endotoxemia or metabolic diseases according to claim 2, wherein the Lactobacillus sp. Strain is Lactobacillus plantarum.
The method of claim 1, wherein the obesity is selected from the group consisting of simple obesity, symptomatic obesity, childhood obesity, adult obesity, cell proliferative obesity, cell non-large obesity, upper body obesity, lower body obesity, visceral fat obesity and subcutaneous fat obesity A pharmaceutical composition for preventing or treating obesity, endotoxemia or metabolic diseases, characterized in that it is at least one kind of obesity.
The method according to claim 1, wherein the metabolic disease is one or more metabolic diseases selected from the group consisting of metabolic obesity, metabolic hyperlipidemia, metabolic diabetes, metabolic dyslipidemia, metabolic arteriosclerosis and metabolic hypertension. A pharmaceutical composition for the prevention or treatment of a disease or metabolic disease.
A food composition for preventing or ameliorating obesity, endotoxinemia or metabolic diseases, which comprises fermented gingkoferment as an active ingredient.
The food composition for preventing or improving obesity, endotoxemia or metabolic diseases according to claim 6, wherein the germinated fermented product is obtained by fermenting Ganoderma lucidum hot-water extract with a strain of Lactobacillus.
8. The food composition according to claim 7, wherein the Lactobacillus sp. Strain is Lactobacillus plantarum. 7. A food composition for preventing or ameliorating obesity, endotoxemia or metabolic diseases.
7. The method of claim 6, wherein the obesity is selected from the group consisting of simple obesity, symptomatic obesity, childhood obesity, adult obesity, cell proliferative obesity, cell non-large obesity, upper body obesity, lower body obesity, visceral fat obesity and subcutaneous fat obesity A food composition for preventing or ameliorating obesity, endotoxemia or metabolic diseases, characterized in that it is at least one kind of obesity.
The method of claim 6, wherein the metabolic disease is one or more metabolic diseases selected from the group consisting of metabolic obesity, metabolic hyperlipidemia, metabolic diabetes mellitus, metabolic dyslipidemia, metabolic arteriosclerosis and metabolic hypertension. A food composition for preventing or ameliorating a disease or metabolic disease.

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