KR101031605B1 - Manufacturing method of culture extract of ceriporia lacerata for the therapy of diabetic diseases and culture extract of ceriporia lacerata using the same - Google Patents

Abstract

PURPOSE: A method for preparing Ceriporia lacerate culture extract is provided to enhance the content of protocatechualdehyde and to prevent and treat diabetes. CONSTITUTION: A method for preparing Ceriporia lacerate culture extract comprises: a step of culturing mycelium of Ceriporia lacerate to obtain mycelium culture liquid at 20-30°C for 5-16 days; a step of freeze-drying the culture liquid and pulverizing; and a step of extracting the culture liquid powder of the mycelium with a solvent of water, ethanol, methanol, acetone, butanol, or ethyl acetate. A medium for liquid culture contains 0.2-3 weight% of glucose, 0.2-3 weight% of sugar, 0.2-4 weight% of starch, 0.2-3 weight% of soybean powder, 0.05-0.1 weight% of magnesium sulfate, 0.05-0.1 weight% of monobasic potassium phosphate(KH_2PO_4), and 90-99 weight% of water. A pharmaceutical composition for preventing and treating diabetes contains Ceriporia lacerate culture extract.

Description

Manufacturing Method of Culture Extract of Ceriporia lacerata For The Therapy of Diabetic Diseases And Culture Extract of Ceriporia lacerata Using The Same}

The present invention three other repositories Ria Rock Serra (Ceriporia lacerata) a method for producing a liquid culture, and the culture extracts using, whereby three reports Ria lock Sera other culture extract, the three reports Ria lock Sera other pharmaceutical compositions and functional food for the prevention and treatment of diabetic diseases, including culture liquid extract according It is about.

Three other repositories Ria Rock Serra (ceriporia lacerata ) is a type of white fungus known to perform cometabolism called lignin degradation in order to use carbon sources such as cellulose, hemicellulose, other polysaccharides, and glycerol in the ecosystem. In addition, the rapid lignin part of wood and the organization is efficiently decomposed known, but reports of anti-diabetic effect of the three repositories Ria Rock Sera other is not known.

Diabetes is a representative adult disease, affecting 5% of the world's population, resulting in huge human and economic losses. Diabetes mellitus is classified into two types, of which insulin dependent diabetes (IDDM) is caused by a deficiency of insulin, a glucose-regulating hormone in the blood, mainly in young people in their 10s to 20s. It is also called pediatric diabetes because it develops. In addition, type II diabetes (non-insulin dependent diabetes, NIDDM) occurs mainly after the age of 40, and occupies most of the diabetic patients in our country. Unlike type I diabetes, it is called adult-type diabetes and the cause of the disease is not clear yet, but it is known to be caused by genetic factors and environmental factors.

The etiology of type II diabetes has been observed in both pancreatic beta cells and in insulin secretion and in target insulin cells (insulin resistance).

Diabetic peripheral neuropathy, the most common complication of diabetes, begins to manifest in about 50% of diabetes over 10 years. The initial symptoms begin with paresthesia of the hands, especially the feet, and continue with severe pain, resulting in impaired sensation and impairment of the peripheral autonomic nerves resulting in blood flow disorders. Further progression may result in painless ulcers and infections, and in severe cases, amputation of the damaged limbs may be necessary, and acute complications can lead to sepsis and life-threatening cases.

The mechanism of development of diabetic peripheral neuropathy is not yet clear despite many studies. In terms of treatment, there is ultimately no cure for many diabetic complications, including peripheral neuropathy. Active glycemic control and elimination of risk factors such as hyperlipidemia, high cholesterol, overweight, smoking and drinking have been known to prevent or delay the occurrence of complications to some extent, but there are no drugs to treat complications once they occur. On the other hand, the development of drugs that can delay or prevent the occurrence of complications are in progress at various research institutes at home and abroad, but there is no clinical use.

Therefore, there are no side effects, the three repositories containing safety is ensured with natural materials in three repositories Ria Rock Sera other using, Prototype category weight aldehydes (Protocatechualdehyde), including known that the superior efficacy in the prevention and treatment of diabetes disease in bulk Leah has been developing the requirements for the preparation Rock Serra another culture extracts.

The invention three reports Ria lock Sera other extract lyophilized product obtained from a liquid culture of the mycelium by extraction solvent, and was filtered and concentrated to three reports Ria lock Sera other culture extract of the mycelia (hereinafter referred to as "three lipoic Ria lock sera obtained other referred to as the culture extracts') to provide a manufacturing method and thus three repositories Ria Rock Sera other culture extracts.

The invention increases the amount of validity those such as the protocol category weight aldehydes (Protocatechualdehyde) known to be effective in the treatment of diabetes, a method of preventing and three reports Ria therapeutic effects superior lock Sera other culture extract for diabetic disease and accordingly It is intended to provide a extract of Ceriporia laccerata culture.

The present invention is preferred as the first embodiment (S1) comprising: a liquid culture to produce an aged lipoic Ria mycelium culture mycelia of three reports Ria lock Sera other (Ceriporia lacerata); (S2) comprising: a powdered freeze-dried three reports Ria lock Sera other mycelium culture liquid; And (S3) the three reports Ria lock Sera other that after the mycelium culture medium powder, extracted with water, ethanol, methanol, acetone, butanol and ethyl acetate, one solvent or two or mixture of two or more solvents selected from the group consisting of filtered and concentrated under reduced pressure It provides a method for producing a culture extract of the seriporia lac serrata comprising the step.

In the production method of the three reports Ria lock Sera other culture extract according to this embodiment, the liquid culture of the (S1) step can be performed 5-16 days at 20 ℃ ~ 30 ℃.

In the production method of the three reports Ria lock Sera other culture extract according to this embodiment, the liquid culture of the (S1) step can be carried out while injecting the air of 1 ~ 3 vvm (air volume / medium volume / minute) .

At this time, the medium composition for the culture of the liquid is 0.2 to 3% by weight of glucose, 0.2 to 3% by weight of sugar, 0.2 to 4% by weight of starch, 0.2 to 3% by weight of soy flour, magnesium sulfate (MgSO ₄ ) 0.05 to 0.1% by weight , Potassium dihydrogen phosphate (KH ₂ PO ₄ ) It may be one containing 0.05 to 0.1% by weight and water 90 to 99% by weight.

In the production method of the three reports Ria lock Sera other culture extract according to this embodiment, the freeze-dried at (S2) step may be performed for 48 hours ~ 72 hours.

Solvent in the producing method of the three reports Ria lock Sera other culture extract according to this embodiment, (S3) step may be aqueous ethanol solution in water or 50% (w / w) ~ 80% (w / w).

Be those in the production method of the three reports Ria lock Sera other culture extract according to this embodiment, the extraction in the (S3) phase, three reports Ria lock Sera performed by adding a polar solvent of 20 ~ 50mL / g with respect to the other powder have.

In the production method of the three reports Ria lock Sera other culture extract according to this embodiment, the extraction in the (S3) step may be performed for 5 to 7 hours at 20 ℃ ~ 30 ℃.

The invention in a preferred second embodiment provides a three reports Ria lock Sera other culture extract produced by the method for producing the.

The first three reports Ria lock Sera other culture extract according to the second embodiment, may be one which includes a protocol category weight aldehydes (Protocatechualdehyde) 10 ~ 25㎍ / g .

The first three reports Ria lock Sera other culture extract according to the second embodiment, may be one which includes a protocol category weight aldehydes (Protocatechualdehyde) 17 ~ 25㎍ / g .

The invention as a third preferred embodiment provides a pharmaceutical composition for the prevention and treatment of diabetic diseases, including three reports Ria lock Sera other culture extract of said.

The invention as a fourth embodiment provides a health functional food comprising a three reports Ria lock Sera other culture extract of said.

1 is a graph showing the glucose tolerance analysis of the three reports Ria lock Sera other culture extract according to an embodiment of the present invention,
2 is a graph showing blood glucose levels changes over time of the administration of three reports Ria lock Sera other culture extract according to an embodiment of the present invention an animal model.

Hereinafter, the present invention will be described in more detail.

The present invention (S1) three other repositories Ria Rock Serra (Ceriporia liquid culture of the mycelium of lacerata ) to prepare a culture medium of the seriporia laccerata mycelia; (S2) comprising: a powdered freeze-dried three reports Ria lock Sera other mycelium culture liquid; And (S3) the three reports Ria lock Sera other that after the mycelium culture medium powder, extracted with water, ethanol, methanol, acetone, butanol and ethyl acetate, one solvent or two or mixture of two or more solvents selected from the group consisting of filtered and concentrated under reduced pressure It provides a method for producing a culture extract of the seriporia lac serrata comprising the step.

The three reports Ria lock Sera other mycelium culture in (S1) phase, three reports Ria lock Sera other (Ceriporia lacerata ) mycelium liquid culture, the medium composition for the culture of the liquid is 0.2 to 3% by weight of glucose, 0.2 to 3% by weight of sugar, 0.2 to 4% by weight of starch, 0.2 to 3% by weight of soy flour, magnesium sulfate ( MgSO ₄ ) 0.05 to 0.1% by weight, potassium dihydrogen phosphate (KH ₂ PO ₄ ) may be to include 0.05 to 0.1% by weight and water 90 to 99% by weight.

At this time, the liquid culture is preferably carried out for 5 to 16 days while injecting 1 to 3 vvm (air volume / medium volume / minute) at 20 ℃ ~ 30 ℃, it is carried out at 25 ℃ 10 days is mycelial growth Most preferred because it is the most active and has a high content of exopolysacchraid.

More specifically, the (S1) phase three reports Ria lock Sera other mycelium broth is then obtained a two or three times subculture after precipitation species three reports Ria lock Sera other parent strain on PDA medium, PDB medium in It is more preferable to maintain the constant temperature of 20 ~ 25 ℃ in the incubator and liquid culture of the mycelium after a small liquid culture process for 7-9 days because the mycelial growth is the most active.

And, (S2) phase, the three reports Ria lock Sera other culture solution prepared in the step (S1) phase and powdered by lyophilization. The freeze-dried and aged lipoic Ria lock Sera other mycelial culture the case of drying at a high temperature effective because material can be destroyed significantly lower temperature of below 30 ℃, preferably at a temperature of 20 ℃ ~ 30 ℃ 48 sigan ~ 72 sigan Is preferably carried out during the process. In the step of freeze-drying in step (S2), it is more preferable to use the vacuum lyophilizer instead of the lyophilizer which is generally used since the change of the indicator material content is minimized.

(S3) and the step (S2) and extracted three reports Ria lock Sera other powdered mycelium culture obtained in step with a solvent, and filtered to prepare a three reports Ria lock Sera other culture extract according to the present invention was concentrated under reduced pressure. The solvent is preferably one solvent or two or more mixed solvents selected from the group consisting of water, ethanol, methanol, acetone, butanol and ethyl acetate. In particular, the extraction solvent is water or 50wt% ~ 80wt% ethanol aqueous solution is more preferable because the antidiabetic effect is the most excellent in vitro experiments.

When extracted with the three reports Ria lock Sera other solvents to the culture mycelium powder, the solvent is preferable since it is easy to extract, which is added in an amount of 20 ~ 50mL / g with respect to the three reports Ria lock Sera other mycelium broth powder. And, after addition of the solvent in the three reports Ria lock Sera other mycelium broth powder it is desirable to extract for for 5 to 7 hours at a temperature of 20 ℃ ~ 30 ℃, at more preferably 25 ℃ 6 hours. The extract is filtered and concentrated under reduced pressure.

Thus, the three repositories Ria Rock Serra another culture extracts prepared in accordance with the present invention, the content validity of those that is effective in the prevention and treatment of diabetes increases significantly, the prevention and treatment of diabetes disease is very excellent. More specifically, there are three reports Ria lock Sera other broth extract is anti-diabetic efficacy protocol category weight aldehydes (Protocatechualdehyde) known to be in accordance with the present invention is contained in a very high content of 10 ~ 25㎍ / g.

The present invention also provides a three repositories Ria Rock Serra another culture extracts pharmaceutical for the prevention and treatment of diabetes and diseases compositions comprising the dietary supplement according to the present invention.

Hereinafter, the configuration of the present invention in more detail through examples, but the scope of the present invention is not limited to the following examples and experimental examples.

Example  One: Serifonia Lakserata  Culture Preparation

After the increase the physiological activity of the three separated from the fruiting bodies collected in Ontario Repo Ria Rock Sera other long-term preservation strains the parent strain 2-3 in the PDA medium to starting a healthy strains times subculture to obtain a superior species. In addition, mycelium cultured in small liquid culture for 7 days was used as a starter while maintaining constant temperature in the incubator using PDB medium.

Then, 1.8% by weight of glucose, 1% by weight of sugar, 1.1% by weight of starch, 1% by weight of soy flour, 0.05% by weight of magnesium sulfate (MgSO ₄ ), 0.05% by weight of potassium dihydrogen phosphate (KH ₂ PO ₄ ) and 95% by weight of water After sterilizing the liquid culture medium containing 20% at 121 ° C. for 20 minutes, the medium was infused with 3 vvm (air volume / medium volume / min) in a 700L fermenter using the medium. by 10 days in liquid culture ℃ was prepared in three reports Ria lock Sera other medium.

In addition, the three sera lipoic Ria lock the other culture medium at a low temperature of 25 ℃ using a vacuum freeze-dryer was powdered by lyophilization for 72 hours.

In addition, the sera were three reports Ria lock other addition of 70wt% aqueous ethanol solution of 100 mL in the culture medium and extracted from the powder 3g 25 ℃ for 6 hours. The extract was filtered and concentrated under reduced pressure to prepare a extract of Ceriporia laccerata culture.

Example  2: Serifonia Lakserata  Preparation of culture extract

Three lipoic Ria lock Sera other by making the distilled water to the culture as a powder 3g in the same manner as in Example 1 except that the addition in an amount of 100 mL to obtain a three reports Ria lock Sera other culture extract.

Experimental Example  One: In in vitro Of antidiabetic activity in

Experimental Example  1-1: α- Glucosidase (α- Glucosidase Inhibitory activity

The α-glucosidase inhibitory activity of the sample indicates that the α-glucosidase recognizes the glucoside portion of the p-nitrophenyl (pNP) glucoside as a substrate, and the pNP and the glucoside are separated by an enzymatic reaction. The amount of pNP broken out here was measured and measured at 405 nm.

That is, the extracts prepared in Examples 1 and 2 were lyophilized to be easily prepared for each concentration, and then prepared into powder, and then distilled water was used for each concentration (5 mg / mL, 10 mg / mL, 50 mg / mL, 100 mg / mL). 100 μl of each dilution diluted with 150 mg / mL) was mixed with 50 μL of 0.75 U / mL of α-glucosidase enzyme solution and 50 mM potassium phosphate buffer (pH 6.5), followed by 2 mM p-nitro. 250 µl of phenyl α-D-glucopyranoside (pNPG) was added and reacted at 37 ° C for 30 minutes. The reaction solution was quenched with 500 μl of 0.1 M sodium carbonate (Na ₂ CO ₃ ), the absorbance was measured at 405 nm, and acarbose was used as a positive control (comparative example).

α-glucosidase is a digestive enzyme present in the brush-border membrane of the small intestine, and hydrolyzes into monosaccharides in which disaccharides and polysaccharide forms of carbohydrate are digested and absorbed. α-glucosidase inhibitors can inhibit blood glucose levels after carbohydrate diets. The results of measuring the α-glucosidase inhibitory activity of the 70 wt% ethanol extract (Example 1) and the water extract (Example 2) of the culture medium of the three S. liporia lacserata mycelium are shown in Table 1. IC ₅₀ of water extract (Example 2) Value is 28.90 mg / mL, IC ₅₀ of 70 wt% ethanol extract (Example 1) The value was 101.00 mg / mL and the activity of the water extract (Example 2) was much better.

sample Concentration (mg / mL) Enzyme inhibitory activity (%) IC ₅₀ (mg / mL) 70% Ethanol Extract Example 1 50 13.06 ± 1.15 101.00 100 47.55 ± 0.94 150 81.44 ± 1.58 Water extract Example 2 5 10.86 ± 0.91 28.90 10 31.51 ± 3.39 50 80.58 ± 5.67 Akabozu Comparative example 0.1 4.55 ± 2.24 4.00 One 47.54 ± 1.42 10 91.34 ± 1.13

Experimental Example  1-2: α-amylase (α- Amylase Inhibitory activity

Inhibitory activity of α-amylase derived from Pancreatin from the samples was measured using starch as a substrate. The extracts prepared in Examples 1 and 2 were lyophilized to make them easy to prepare by concentration, and then prepared into powders, and then each extract diluted by concentration (50 mg / mL, 100 mg / mL, 200 mg / mL) with distilled water. 200 μl of 0.7 U / mL α-amylase enzyme solution and 100 μl of 50 mL potassium phosphate buffer (pH 6.5) were mixed at 25 ° C. for 10 minutes, and 200 μl of 1.0% starch was mixed for 5 minutes at 37 ° C. Then, 200 μl of a 48 mM DNS (3,5-dinitrosalicylic acid) solution was added. After heating at 100 ° C. for 15 minutes to develop color and cooling sufficiently, distilled water was added and stirred, and then the absorbance was measured at 540 nm. Acarbose was used as a positive control.

Result the plate three reports Ria lock inhibitory activity of the sera other culture extract for α- amylase derived from a measure of the creatine is shown in Table 2 below. IC ₅₀ of 70 wt% ethanol extract (Example 1) and water extract (Example 2) The values were 102.44 mg / mL and 89.65 mg / mL, respectively, indicating that the inhibitory effect of the water extract was slightly higher. These results are similar to the α-glucosidase inhibitory activity, and when extracted with water, it can be expected that many indicator substances which are effective for diabetes are extracted.

sample Concentration (mg / mL) Enzyme inhibitory activity (%) IC ₅₀ (mg / mL) 70% Ethanol Extract Example 1 50 13.56 ± 2.03 102.44 100 50.23 ± 5.81 200 95.26 ± 1.05 Water extract Example 2 50 21.55 ± 7.23 89.65 100 63.02 ± 7.81 200 95.26 ± 1.05 Akabozu Comparative example 0.01 25.50 ± 3.54 0.28 0.1 47.97 ± 0.45 One 92.90 ± 3.42

Experimental Example  2: oral Glucose tolerance  inspection( OGTT , Oral Glucose Tolerance Test Through) Serifonia Lakserata  Of culture extract Antidiabetic  effect

12 hours or more fasted that normal rats (rat) after (4 weeks old) was measured fasting glucose, and then, the second embodiment to easily lyophilized three reports Ria diluting the lock Sera other culture extract produced in the made into powder 200mg / kg Diluted with distilled water so that the concentration of 5 samples in each group was orally administered first. At this time, the control group was administered the same amount of physiological saline. In addition, except for the control group, 40wt% glucose was orally administered at a concentration of 2g / kg bw, and blood was collected from the vein at 30, 60, 90, and 120 minutes, respectively, and blood glucose change was observed. The blood glucose increase curve was calculated by calculating the increase in blood glucose at each time point, and is shown in FIG. 1.

As shown in FIG. 1, the glucose tolerance test of rats showed the highest blood glucose level in all experimental groups after 30 minutes of glucose administration, and the blood glucose level decreased with time. In addition, compared to the sample treatment group, the control group administered with glucose only showed the highest blood sugar level without treating the sample, and after 60 minutes, the blood glucose reduction was greater in the group administered with the extract of Example 2, but after 120 minutes in all groups Glucose was reduced to almost fasting levels at. Therefore, the extract of Example 2 is considered to have a blood sugar synergistic inhibitory effect.

Experimental Example  3: using animal models Serifonia Lakserata  Of culture extract Antidiabetic  Effect verification

Breeding experimental animals

The experimental animals were Sprague-Dawley male rats with an average body weight of 140 ± 3.2g, and 24 rats of 8 rats were purchased for each experimental group, and the environmental conditions were 23 ± 2 ℃, 60 ± 5% humidity, and 12 hours light cycle. In order to adapt to the preliminary breeding for 1 week with commercial commercial feed (Purina Co., Korea) purchased for 2 weeks experiment was carried out.

Diabetes induction in normal rats

Experimental Type 1 diabetes induction was performed by streptozotocin (STZ, Sigma Chemical Co., St. Louis, MO, USA), which is known to selectively destroy β-cells of the pancreas, with 0.01M citrate buffer (pH). 4.5) was injected intraperitoneally at 60 mg / kg. Diabetes mellitus was collected from the tail vein 24 hours after STZ administration, and animals with fasting blood glucose of 200 mg / dL or more were used in the experiment.

Experimental  Classification

The test group three manufactured after diabetic to view after the normal control (NC) and diabetic supplies the normal diet without diabetic normal diet a diabetic control group (DM), diabetic treatment effect supplying the embodiment 2 weeks Example 2 reports Ria The lacserata culture extract was freeze-dried to prepare a powder, and then divided into three groups of DM-water, orally administered with dilution at a concentration of 200 mg / kg.

Body weight, dietary measure

A one-time, after the administration of STZ as in Example 2. After three reports Ria lock Sera other culture extract produced in a week administered once, and the day of autopsy on body weight immediately before the start of STZ administration were determined for all animals. On the other hand, by feeding a certain amount of diet once a week for each breeding cage and measuring the remaining amount of feed at the same time the next day, the average daily dietary intake (g / day) was calculated.

While oral administration to a three reports Ria lock Sera other culture extract prepared in Example 2 on the diabetic mice induced by STZ to the weight change of the animal breeding two weeks are shown in Table 3 below. In the normal group (NC), the weight was continuously increased over time, while in the diabetic group (DM), the weight was decreased due to the effect of STZ. Meanwhile, in the diabetic treatment group (DM-Water), the weight was reduced than the normal group, but showed a higher weight than the DM group.

STZ administration causes type 1 diabetes due to the destruction of β-cells in the pancreas, which leads to a decrease in insulin production and its effects. Insufficient energy production by glucose metabolism affects growth and development. In the result of this experiment, if the diabetes group (DM-water) will show the weight loss compared to the control group (NC) after a diabetic is contemporary saga somewhat improved three-lipoic Ria Rock Serra another addition of the culture water extracts to increase weight It is estimated.

In addition, the average daily food intake was increased in the diabetic experimental group (DM, DM-water) compared to the control group, which appears to be a result of the symptoms of diabetes, multi-diet, multidose, multidose. Despite the higher dietary intake of the diabetic experimental group (DM, DM-water) compared to the normal group (NC), weight loss continued due to degenerative changes in body metabolism caused by diabetes.

sample Body weight (g / rat) Weight gain (g / day) Feed Intake (g / day) Feed Intake Efficiency NC 340.08 ± 5.42 6.62 ± 0.76 28.54 ± 0.10 0.20 DM 248.21 ± 11.28 0.80 ± 0.10 27.77 ± 0.12 0.02 DM-water 299.04 ± 13.65 2.88 ± 0.43 30.08 ± 0.09 0.08

Blood Sugar and Biochemical Analysis

Blood glucose changes during the breeding period were fasted for 12 hours or more once a week, and blood was collected from the tail vein and measured with a blood glucose meter (ACCU-CHEK Sensor, Germany). Then, rats fasted for 12 hours after 2 weeks of breeding were anaesthetized with ether. Blood was collected with a heparinized syringe from the abdominal vein, and left for 30 minutes after blood collection, and centrifuged at 3,000 rpm for 15 minutes to separate plasma. The separated plasma was quenched with liquid nitrogen and stored at -80 ° C until analysis. The separated plasma was analyzed for blood glucose, insulin, ALT, AST, total cholesterol (TC) content using a blood analyzer.

The results of analyzing blood glucose level changes are shown in FIG. 2. The blood glucose level was significantly increased in the DM group compared to the NC group, and there was no difference between the DM group and the DM-Water group before the administration of the sample, but the blood glucose level was higher than the DM group one week after the addition of the extract of Example 2 Significantly decreased. At 2 weeks after the administration of the sample, the blood glucose of the DM-water group was significantly decreased compared to the DM group. DM group was intraperitoneally injected with STZ to inhibit insulin secretion due to pancreatic destruction, resulting in significantly higher blood glucose than the control group, whereas DM-water group appeared to have reduced blood glucose due to the administration of the extract of Example 2.

As a result of observing changes in the activity of ALT and AST in the blood, the enzyme activity was increased in the DM group compared to the NC group, but the activity of ALT and AST was decreased in the group to which the extract of Example 2 was added. In other words, ALT and AST are known to be enzymes that increase their activity when hepatic damage caused by drugs, the activity of these enzymes increased by the addition of the extract of Example 2 is reduced by the addition of the extract of Example 2 The extract can be seen to somewhat improve the liver damage caused by STZ.

Plasma total cholesterol concentration was increased in DM group compared to NC group, but plasma cholesterol concentration was decreased compared to DM group by the addition of the extract of Example 2. The increase in total cholesterol in the diabetic group is presumed to be due to carbohydrates not being used as an energy source in metabolism of diabetic rats, but because free fatty acids are used as energy to synthesize cholesterol.

In addition, the plasma triglyceride concentration was increased in the DM group compared with the NC group, but the neutral lipid content was decreased in the DM-Water group. Hyperlipidemia, one of the main complications of diabetes, is known to increase plasma triglyceride levels by increasing the rate of fatty acid conversion to triglycerides than normal. In this experiment, the concentration of triglycerides in diabetic was consistently increased, and the concentration of triglycerides in plasma was decreased when the extract of Example 2 was added.

Group ALT (g / L) AST (g / L) TC (g / L) TG (g / L) NC 34.87 ± 0.98 35.87 ± 0.34 78.32 ± 1.90 118.09 ± 1.65 DM 93.05 ± 0.76 71.22 ± 1.87 95.81 ± 1.04 132.87 ± 1.02 DM-Water 62.23 ± 0.99 62.43 ± 1.23 83.65 ± 1.47 120.32 ± 1.80

Experimental Example  4: Indicator Material Analysis

LC / MS / MS was analyzed using Agilent Technologies Agilent 6410 with ion souce negative and fragmentor 150. The gas temperature was 320 ℃ and the gas flow was analyzed at a flow rate of 35 mL / min. The capillary volt was analyzed at 4000. The column of HPLC used Epic C18 and the column temperature was kept at 40 degreeC. Distilled water (A) containing 0.1% formic acid and acetonitrile (B) containing 0.1% formic acid were used as the mobile phase. Gradient conditions of the mobile phase are shown in Table 5 below. 300 mg of the sample was injected with 2.0 mL of 80 wt% methanol, sonicated for 10 minutes, and 20 uL of the solution passed through a 0.45 um membrane filter was injected.

min A (%) B (%) 0 ~ 0.1 85 15 0.1-15 0 100 15-20 0 100 20-21 85 15

Three representative indicators known to have antidiabetic effects (protocatechualdehyde), 2,5-dihydroxybenzoic acid, and 2,5-dihydroxyacetophenone (2, 5-dihydroxyacetophenone)) was quantified using LC / MS / MS as shown in Table 6 below. In particular, protocatechualdehyde, which is known to have an excellent effect on the treatment of diabetes, was found to contain 18.79 μg per gram of the extract of Example 2.

sample Protocatechualdehyde (μg / g) 2,5-dihydroxybenzoic acid (μg / g) 2,5-dihydroxyacetophenone (μg / g) Example 2 18.79 ± 0.87 37.65 ± 1.32 20.76 ± 1.09

As it discussed above, according to the present invention and the prototype catheter weight aldehydes known to be an excellent effect on the treatment of diabetes can provide three Syrian lipoic lock Sera other culture extract containing at least 10㎍ / g or more, specifically, the category estimation protocol and aged between 10 and aldehyde in an amount of 25㎍ / g lipoic Ria is possible to provide a lock Sera other culture extract.

Claims (13)

(S1) three other repositories Ria Rock Serra (Ceriporia the mycelium of lacerata) preparing a three reports Ria lock Sera other mycelium culture liquid by liquid culture;
(S2) comprising: a powdered freeze-dried three reports Ria lock Sera other mycelium culture liquid; And
(S3) the three reports Ria lock sera extract the other mycelium broth powder with water, ethanol, methanol, acetone, butanol and ethyl acetate, one solvent or two or mixture of two or more solvents selected from the group consisting of the steps of filtered and concentrated under reduced pressure the method of the three repositories Ria Rock Serra another culture medium containing the extract;
The method of claim 1,
(S1) phase liquid culture method of manufacturing a three reports Ria lock Sera other broth extract is performed 5-16 days at 20 ℃ ~ 30 ℃ in.
The method of claim 1,
(S1) phase liquid culture is 1 ~ 3 vvm of injecting air (air volume / volume of medium / minute) and the method of manufacturing a three reports Ria lock Sera other broth extract is performed in.
The method of claim 1,
Medium composition for liquid culture in step (S1) is 0.2 to 3% by weight of glucose, 0.2 to 3% by weight of sugar, 0.2 to 4% by weight of starch, 0.2 to 3% by weight of soy flour, magnesium sulfate (MgSO ₄ ) 0.05 ~ 0.1 wt.%, potassium dihydrogen phosphate _{(KH 2 PO 4) 0.05 ~} 0.1 the method of producing a three reports Ria lock Sera other culture extract comprises wt% and water 90 to 99% by weight.
The method of claim 1,
(S2) phase is freeze-dried Method of manufacturing a three reports Ria lock Sera other broth extract is performed for from 20 ℃ ~ 30 ℃ 48 sigan ~ 72 hours at.
The method of claim 1,
Solvent in (S3) step is the production of three reports Ria lock Sera other culture extract, characterized in that the aqueous ethanol solution in water or 50% (w / w) ~ 80% (w / w).
The method of claim 1,
Extraction in (S3) is phase process for producing a three reports Ria lock sera of three other reports RIA is performed by adding a solvent of 20 ~ 50mL / g with respect to the mycelium powder lock Sera other culture extract.
The method of claim 1,
(S3) is phase extraction method for producing a three reports Ria lock Sera other broth extract is performed for 5 to 7 hours at 20 ℃ ~ 30 ℃ in.
Any one of claims 1 to 8 prepared in the three reports Ria any one of the production method of the anti-lock Sera other culture extract.
10. The method of claim 9,
Three lipoic Ria lock Sera other culture extract comprising the protocol category weight aldehydes (Protocatechualdehyde) 10 ~ 25㎍ / g .
10. The method of claim 9,
Three lipoic Ria lock Sera other culture extract comprising the protocol category weight aldehydes (Protocatechualdehyde) 17 ~ 25㎍ / g .
Section 9-year-old Ria Repo Rock Serra another culture extracts pharmaceutical composition for the treatment and prevention of diseases, including diabetes.
The three repositories paragraph 9 Leah supplements containing the extract of Rock Serra another culture.

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