KR100859986B1 - Fomitopsis pinicola Extract Having Diabetes Complication Preventive Effect and The Use Thereof - Google Patents

Abstract

The present invention relates to pine needles mushroom extract and its use. Pine needles fruiting body extract and culture mycelia extract of the present invention inhibits the activity of aldose reductase of the lens and kidney, and reduces the concentration of neutral lipids, total cholesterol and LDL cholesterol, such as cataracts or kidney disease The present invention may be widely used for the development of functional foods that exhibit a therapeutic or prophylactic effect of diseases caused by increased activity of aldose reductase, such as diabetic complications.

Pineapple Butterfly, Diabetic Complications, Aldose Reductase, β-1,3-Glucano-β-1,6-heterogalactomannan-protein Complex

Description

Fomitopsis pinicola extract having diabetes complication preventive effect and the use thereof}

1 is a chart of the mechanism of development of diabetes mellitus and diabetic complications. The aldose reductase in the polyol pathway is involved in metabolizing glucose into sorbitol and inhibits the activity of the former aldose reductase to block progression to diabetic complications.

Figure 2 is a graph showing the effect of aldose reductase activity of the rat lens by the addition of pine needles fruiting body hot water extract in vitro.

Figure 3 is a graph showing the effect of pine needles mushroom fruiting hot water extract on the kinetics of the lens aldose reduction enzyme in rats.

The present invention relates to pine needles mushroom extract and its use. More specifically, the present invention relates to pine needles and fruit mycelia extracts exhibiting the preventive effect of diseases caused by increased activity of aldose reductase, such as diabetic complications such as cataracts, kidney disease.

As the eating habits are westernized, the incidence of lifestyle diseases such as obesity, hyperlipidemia, and diabetes is increasing, which is a serious social problem. In particular, the incidence of diabetes among Koreans has risen sharply in recent decades, making it the fourth largest cause of death among Koreans (see Annual Report on Cause of Death Statistics, 1999, 2002).

Diabetes is classified into insulin-dependent type 1 diabetes and insulin-independent type 2 diabetes. About 90 to 95% of diabetes is known as type 2 diabetes. When diabetes develops due to hormonal imbalance, metabolism of the three major nutrients including carbohydrates is abnormally progressed, resulting in high blood sugar, increased levels of triglycerides, total cholesterol and LDL cholesterol in the blood, and decreased content of HDL cholesterol. It is known that severe changes cause atherosclerosis, which causes cardiovascular disease, complications including neurological disorders and kidney damage, leading to death.

On the other hand, when diabetes occurs and hyperglycemia persists, hexokinase that is involved in anaerobic glycolysis, a normal carbohydrate metabolism, is saturated, so that excess glucose is an aldose reductase enzyme of the polyol pathway, a metabolic process of glucose metabolism. It is metabolized to sorbitol by aldose reductase and to fructose by sorbitol dehydrogenase. At this time, the sorbitol produced has a low cell membrane permeability, causing osmotic stress, which causes tissue cell damage. In addition, excess glucose or excess fructose is combined with protein to produce final glycated product (AGEs). It is known to cause diabetic complications by causing damage to tissues by causing damage to or generating reactive oxygen species (ROS).

Diabetes treatment is generally combined with diet, exercise therapy and drug therapy, all of which aim to prevent or delay diabetic complications by continuously maintaining normal blood sugar. However, many researches on diabetes complications have been conducted so far, but there is still a need for continuous research on functional foods and medicines related to the treatment or prevention of diabetes complications.

Fomitopsis pinicola ( Fomitopsis pinicola ) is known to be extinct as a kind of native mushroom in Korea, but recently discovered by a farmer and registered as a 'regenerated mushroom' at the National Seed Control Center (application number: 2003-498) . Electric pine grass butterfly is a semi-circular large mushroom that grows attached to a tree trunk in a shelf shape. Its top surface is covered with thick cuticles, and its surface is flat and black or reddish brown. Recently, many studies have reported that mushrooms not only contain various nutrients but also contain various kinds of bioactive substances involved in the prevention and treatment of metabolic diseases such as diabetes, hyperlipidemia and atherosclerosis, and anticancer and immune enhancement. have.

On the other hand, there have been reports of hypoglycemic activity against pine xanabi mushrooms, which have only been known to have anti-tumor inhibitory effects (see Korean Patent Publication No. 2005-60726). The research is not yet actively conducted, and it is not clear exactly what polysaccharide or protein polysaccharide acts.

Therefore, the necessity of verifying various physiological activities of pine needles butterfly, development of a method for preparing fruiting body and culture mycelium extract of pine needles butterfly and analyzing the components in the extract has emerged.

Thus, the inventors of the present invention to investigate whether the physiological activity of pine zanabi mushrooms can prevent or delay diabetic complications, while dieting and observing the extract of pine zanabi mushrooms in rats that induced diabetes, Inhibition of DOS reductase activity and reduction of neutral lipid, total cholesterol and LDL cholesterol concentrations were confirmed, and the present invention was completed. In addition, by analyzing the structure of the extract of the pine zanbi mushroom, it was confirmed that the main component is 'β-1,3-glucano-β-1,6-heterogalactomannan-protein complex'.

After all, the main object of the present invention is to provide a pine fruit or mushroom mycelium extract exhibiting a therapeutic or prophylactic effect of a disease caused by an increase in the activity of aldose reduction enzymes such as diabetic complications such as cataracts or kidney disease.

Another object of the present invention is a composition comprising the pine berry butterfly fruit extract or culture mycelium extract as a precaution, and exhibits the treatment or prevention effect of a disease caused by the activity of aldose reduction enzymes such as diabetic complications such as cataracts or kidney disease To provide.

Still another object of the present invention is to increase the activity of aldose-reducing enzymes, including the β-1,3-glucano-β-1,6-heterogalactomannan-protein complex, such as diabetic complications such as cataracts or kidney disease It is to provide a composition exhibiting a therapeutic or prophylactic effect of a disease caused by the.

The present invention is a pine grass butterfly ( Fomitopsis) pinicola ) provides a composition for the treatment or prevention of diseases caused by increased activity of aldose reductase, including fruiting body extract or culture mycelia extract, for example, diabetic complications.

The fruiting body extract of pine needles butterfly is a hot water extract or an alkali extract, and the hot water extract is extracted by cutting, crushing, watering and heating the pine fruit butterfly, which is concentrated and precipitated with ethanol, and the precipitate is obtained by freeze drying. do. In this case, the valence is not particularly limited thereto, but it is preferable to add purified water corresponding to about 30 times the weight of the crushed fruiting body, and the extraction is not particularly limited, but it is preferable to extract at about 24 hours at 100 ° C. However, the concentration is not particularly limited, but it is preferable to concentrate at 40 ° C. until it is about 1/10 of the initial volume.

In addition, the alkaline extracts are chopped and crushed pine needles mushroom fruit body and swelled with 1N KOH for 1 hour, homogenized using a crusher, filtered through a 100mesh sieve, neutralized with concentrated hydrochloric acid and washed with distilled water, 60 ℃ Obtained by drying in the. At this time, the swelling is not particularly limited, but it is preferable to swell by adding 1N KOH at about 1: 1 (w / v). The alkali extraction method is a drastically improved yield compared to the conventional alkali extraction method, also called a HAS (homogenization after alkali swelling) method.

On the other hand, the culture mycelium extract of pine zanbi mushroom is cultured by subculture and seed culture of the mycelia of pine zanbi mushroom, and then inoculated in sterilized and cooled potato medium, and cultured by neutralization and ethanol precipitation, The precipitate is obtained by dialysis with distilled water. At this time, the neutralization of the culture solution is not particularly limited thereto, but neutralizing to pH 6.5 with a sodium bicarbonate (NaHCO ₃ ).

In the present invention, the disease caused by the increased activity of aldose reductase means a disease including diabetic complications such as cataract or kidney disease. Glucose is metabolized by aldose-reductase to sorbitol, and the resulting sorbitol has low cell membrane permeability, causing osmotic stress, resulting in damage to organs such as glomeruli of the lens, retina or kidney (see FIG. 1).

The disease caused by the increase of aldose reductase activity of the present invention pine zanbi mushroom extract obtained from the above method, for example, the prevention effect of diabetic complications or diabetic induced hypercholesterolemia improvement effect was investigated as follows. After the oral administration of the extract to rats induced diabetes with STZ (streptozotocin), the lens and kidney were extracted from the mice, homogenized under ice-cooling, and the activity of aldose reductase from the homogenized lens and kidney was investigated. The diabetic complications were prevented, and blood was obtained from the mice, and the blood was analyzed to examine triglyceride, total cholesterol, HDL cholesterol, and LDL cholesterol content to confirm the effect of improving diabetes-induced hypercholesterolemia.

Furthermore, in order to find out the active source of the above-mentioned effect of pine needles mushroom extract of the present invention, using a DEAE-cellulose ion exchange resin and Sepharose CL-4B gel filtration column sequentially fractionated, purified, GC Methylation analysis was carried out to determine the composition and molecular weight of the pine needles mushroom extract.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example

Example 1 Preparation of Fruiting Body Extract and Cultured Mycelia Extract of Pine Sprout Butterfly

Example 1-1 : Preparation of fruiting body hot water extract of pine tree butterfly

90 g of pine berry butterfly fruit body provided from regenerated agricultural products in Pohang was cut into 5 × 5 mm, crushed, 2.5L of purified water was added thereto, and extracted at 100 ° C. for 24 hours. Thereafter, the extract was concentrated to 40 mL at 40 ° C, and precipitated by addition of ethanol to obtain an alcohol insoluble substance. The precipitated alcohol-insoluble substance was lyophilized to obtain pine needles mushroom fruiting hot water extract.

Example 1-2 Preparation of Fruiting Body Alkali Extracts of Pine Turtle Butterfly

90 g of pine berry butterfly fruit body provided from regenerated agricultural products in Pohang was cut into 5 × 5 mm and crushed and swelled with 1N KOH (1: 1, w / v) for 1 hour, and then homogenized using a crusher. Thereafter, the resultant was filtered through a 100mesh sieve, neutralized with concentrated hydrochloric acid, washed with distilled water, and dried at 60 ° C. to obtain a fruiting body alkali extract of pine needles butterfly.

Example 1-3 Preparation of Cultured Mycelium Extract of Pine Turtle Butterfly

The mycelium spawns of pine beetle mushrooms from Pohang-si regenerated farms were collected in YM agar medium (yeast extract: 0.5% (w / v), peptone: 0.5% (w / v), malt extract: 0.2% (w / v), Glucose: 1.0% (v / v), agar: 2.0% (w / v), pH 6.5), and subcultured every 15 days at 30 ℃, the culture medium using a rotary shaker (YM medium) Spawn at 150 rpm).

After 10 days of incubation, the culture solution was sterilized (120 ° C., 30 minutes) and cooled to 2% (v / v) in potato medium (water: 16L, potato powder: 300g, sugar: 150g, peptone: 0.1g). Inoculated, incubated with mycelium liquid for 10 days while injecting sterilization air (10cm ³ / min) in a rotary shaker at 150rpm, 30 ℃, neutralized to pH 6.5 with sodium bicarbonate (NaHCO ₃ ) and precipitated by adding 10-fold ethanol I was.

The precipitate obtained above was dialyzed with distilled water for 48 hours using a dialysis membrane to remove molecular weight 3,000 or less to obtain a pine myrtle mushroom culture mycelium extract.

Example 2 : Prevention of Diabetic Complications of Pine Barberry Butterfly Extract

Example 2-1 : Experimental group and test method

SD male rats with a body weight of 200 ± 5g were fed the normal control group (NC), the diabetic control group (DM), 1% supplemented hot water extract after diabetic induction (DM-WE), and 1% added alkaline extract after diabetic induction ( DM-AE) and after the induction of diabetes were classified into 5 groups, such as dietary mycelial extract 1% diet group (DM-CM), each group was grown to four weeks according to the dietary composition of Table 1. Diabetes induced STZ (streptozotocin, 55mg / kg) was injected intramuscularly, and blood glucose was measured 48 hours after STZ administration, and animals over 300mg / dL were regarded as diabetes-induced model experimental animals and used in the experiment. Blood glucose was measured using Bio-sensor and kit reagent.

Table 1 : Basic Dietary Composition for Animal Experiments (g / kg)

Diet NC (normal group) WE (Hot Water Extract Group) AE (alkali extract group) CM (mycelium culture group) Casein 200 200 200 200 cornstarch 150 150 150 150 Sugar 500 500 500 500 Cellulose 50 40 40 40 Corn oil 50 50 50 50 AIN-mineral mix ^{1 )} 35 35 35 35 AIN-Vitamin Mixture ^{2 )} 10 10 10 10 DL-Methionine 3 3 3 3 Cholinevitatarate 2 2 2 2 Fruiting water extract - 10 - - Fruiting body alkali extract - - 10 - Mycelial Culture Extract - - - 10     total 1,000 1,000 1,000 1,000

¹⁾ AIN-mineral mixture (g / kg): calcium lactate 620.0, sodium chloride 74.0, potassium diphosphate 220.0, potassium sulfate 52.0, magnesium oxide 23.0, manganese carbonate 3.3, iron citrate 6.0, zinc carbonate 1.0, copper carbonate 0.2 , 1,000 g of potassium iodide 0.01, sodium selenite 0.01 and potassium chromium sulfate 0.5 as fine powder

²⁾ AIN-vitamin mixture (mg / kg): Thiamine hydrochloride 600, Riboflavin 600, Pyridoxine hydrochloride 700, Nicotinic acid 3,000, D-calcium pantothenate 1,600, Folic acid 200, D-biotin 20, Vitamin B12 2.5, Vitamin A 400,000 IU, 1,000 g of Vitamin D3 100,000 IU, Vitamin E 7,500 IU and Vitamin K 75 as fine powder

In addition, the weight gain, dietary intake and dietary efficiency of the experimental animals were observed during four weeks of growth by varying the diet (see Table 2). The weight gain and dietary efficiency of the DM group, the diabetic control group, were significantly reduced compared to the NC group. However, in the diet group added 1% of pine needles fruiting body alkali extract, it decreased compared to NC group, but showed a significant increase compared to DM group. The dietary intake was significantly increased in the DM group, the hot water extract, and the cultured mycelial extract group compared with the NC group, but was restored to the control level in the alkaline extract group. Dietary efficiency was significantly decreased in all experimental diet groups compared to NC group, but increased in all pine needles extracts compared to diabetic control group.

Table 2 : Feeding Effect of Pine Bark Butterfly Extracts on the Weight Gain, Dietary Intake and Dietary Efficiency of STZ-induced Diabetic Rats for 4 Weeks

Group ^{1 )} Final weight (g) Weight gain (g / week) Dietary Intake (g / week) Dietary Efficiency ^{2 )}  NC 315.8 ± 7.08 28.96 ± 5.88 175.1 ± 5.4 0.165 ± 0.006  DM 172.8 ± 22.6 -6.79 ± 2.54 206.5 ± 11.2 -0.033 ± 0.034  DM-WE 203.8 ± 32.3 0.95 ± 7.21 199.8 ± 10.3 0.005 ± 0.028  DM-AE 260.2 ± 21.4 15.06 ± 6.11 179.5 ± 6.2 0.084 ± 0.019  DM-CM 193.9 ± 17.1 -1.51 ± 7.03 198.6 ± 8.7 0.008 ± 0.031

¹⁾ NC: normal control group, DM: diabetic control group, DM-WE: dietary supplementation of 1% pine needles mushroom fruiting hot water extract after STZ treatment, DM-AE: dietary supplementation of 1% pine needles fruiting body alkali extract after STZ treatment, DM-CM: Addition of 1% of pine myrtle mushroom culture mycelium extract after STZ treatment

²⁾ feed efficiency ratio: weight gain / food intake

After four weeks of animal experiments, the rats were fasted with water for 16 hours, and then opened through the abdominal midline under ether anesthesia, blood was collected from the abdominal aorta, blood glucose was analyzed, and kidney and eye lenses were extracted. By homogenizing with a homogenizer under ice-cooling to obtain a postmitochondria fraction, it was used as an enzyme solution to determine the activity of aldose reductase (AR).

Example 2-2 : Prevention of Diabetic Complications of Pine Needle Butterfly Extract

Complications such as cataracts, peripheral neuritis, and kidney damage caused by diabetes are hemokinase, the first enzyme in anaerobic glycolysis, the main metabolic process of normal sugars, and is metabolized by the polyol pathway, which is the metabolic process of sugars. It is known to occur by. Sorbitol, produced by reducing glucose through aldose-reductase, the first stage of the polyol pathway, is a substance with low cell membrane permeability. It has been reported to cause peripheral nerve and kidney damage, and substances that inhibit the activity of aldose reductase in such organs are known to prevent or delay diabetic complications.

Therefore, by measuring the activity of the aldose reductase in the enzyme solution of each of the lens and kidney prepared in Example 2-1, it was to investigate the diabetic complication prevention effect of pine needles mushroom extract of the present invention.

The aldose-reducing enzyme activity was reduced by the addition of lithium sulfate, NADPH, DL-α-glycerol and enzyme solution to 50mM phosphate buffer (pH 6.2) according to the method of Hayman and Kinoshita (1965). Was measured at 340 nm.

Example 2-2-1 : Inhibitory Effect of the Lens on Aldose Reductase Activity

The results of observing the activity change of aldose-reducing enzyme were added to the test tube by adding the hot-water extract from pine fruit butterfly, fruiting body, as a enzyme source, using the postmitochondria fraction obtained by centrifugation of the pulverized lens homogenate of rats in FIGS. It was. The activity of aldose-reducing enzyme was decreased in proportion to the concentration of hot water extract of fruiting body, and the activity of enzyme was significantly inhibited by about 50% compared to the control at the concentration of 125 ㎍ / mL. On the other hand, when the kinetics of the action of activating the aldose-reducing enzyme activity by the addition of pine needles fruiting body extract was observed, the Vmax value was not significantly changed, but the km value increased significantly.

Example 2-2-2 : effect of reducing aldose reductase activity of kidney

The activity of aldose-reducing enzyme (AR) was investigated in each case using the postmitochondria fraction obtained by centrifugation of the kidney ground homogenate of rats as an enzyme source. In the diabetic control group (DM) by STZ administration, the activity of aldose-reducing enzyme was increased by about 47% compared to the normal control group (NC). Compared with the decrease of about 25%, dietary group containing the hot water extract (WE) and culture mycelia extract (CM) of the fruiting body showed a tendency to decrease (see Table 3).

Table 3 : Feeding Effects of Four Pine Butterflies Extracts on the Activity Changes of Renal Tissue Aldose Reductase (AR) in STZ-Induced Diabetic Rats

Group ^{1 )} AR activity (oxidized NADPH nmoles / mg protein / min)  NC 4.89 ± 0.71  DM 7.17 ± 0.87  DM-WE 6.27 ± 0.93  DM-AE 5.36 ± 0.86  DM-CM 6.39 ± 0.80

¹⁾ NC: Normal control group, DM: Diabetic control group, DM-WE: Dietary supplementation of pine needle butterfly fruit fruit hot water extract after diabetes induction, DM-AE: Dietary addition of pine needle butterfly fruit fruit alkali extract after diabetes induction, DM-CM: Addition of 1% of pine myrtle mushroom culture mycelium extract after diabetes induction

Example 3 Diabetes Induction Hypercholesterolemia Improvement Effect of Pine Bark Butterfly Extract

Both the experimental group and the dietary administration method were the same as in Example 2, and the five experimental groups were fed for four weeks to measure serum triglyceride, total cholesterol, LDL cholesterol and HDL cholesterol content. Serum triglycerides, total cholesterol and HDL cholesterol content were measured using commercially available kit reagents (AM 157S-K, AM 202-K, AM 203-K, Asanpharm Co., Korea), and LDL cholesterol content was Fridwell. (1972).

Serum triglyceride content was significantly increased in the diabetic control group treated with STZ compared to the normal control group, but the dietary group containing 1% (w / w) pine needles mushroom fruit alkali extract returned to the normal control level. However, in the diet group added 1% (w / w) of pine needles fruiting body fruit hot water extract and the culture group added 1% (w / w) of mycelial extract were similar to the diabetic control (see Table 4).

Table 4 : Feeding Effect of Pine Bark Butterfly Extract for 4 Weeks on Serum Neutral Lipid Content of STZ-induced Diabetic Rats

Group ^{1 )} Serum triglyceride (mg / dL)  NC 98.5 ± 15.2  DM 128.7 ± 13.0  DM-WE 126.8 ± 15.6  DM-AE 112.9 ± 14.1  DM-CM 123.6 ± 14.4

¹⁾ NC: Normal control group, DM: Diabetic control group, DM-WE: Dietary supplementation of pine needle butterfly fruit fruit hot water extract after diabetes induction, DM-AE: Dietary addition of pine needle butterfly fruit fruit alkali extract after diabetes induction, DM-CM: Addition of 1% of pine myrtle mushroom culture mycelium extract after diabetes induction

In addition, the serum total cholesterol content was decreased by 18% in the diet group containing 1% (w / w) pine needles fruit fruit alkali extract compared to the diabetic control group, but it was not significantly decreased in the diet group containing the hot water extract and culture mycelia extract. (Table 5).

Table 5 : Dietary Effect of Pine Stem Butterfly Mushroom Extracts on Serum Total Cholesterol Content in Diabetic Rats Induced by STZ for 4 Weeks

Group ^{1 )} Serum Total Cholesterol (mg / dL)  NC 107.0 ± 11.2  DM 142.8 ± 16.3  DM-WE 134.5 ± 22.1  DM-AE 117.8 ± 12.5  DM-CM 125.1 ± 20.4

¹⁾ NC: Normal control group, DM: Diabetic control group, DM-WE: Dietary supplementation of pine needle butterfly fruit fruit hot water extract after diabetes induction, DM-AE: Dietary addition of pine needle butterfly fruit fruit alkali extract after diabetes induction, DM-CM: Addition of 1% of pine myrtle mushroom culture mycelium extract after diabetes induction

On the other hand, LDL cholesterol content was significantly increased by 95% in the diabetic control group compared to the normal control group by STZ administration, and significantly decreased compared to the diabetic control group in the diet group containing 1% (w / w) pine needles and fruit body alkali extracts. The control group was restored to control level and significantly decreased in the diet group containing 1% (w / w) cultured mycelia extract compared to the diabetic control group, but not significantly decreased in the diet group with 1% (w / w) of hot water extract. (Table 6).

Table 6 : Supplemental Effect of Pine Bark Butterfly Extract for 4 Weeks on Serum LDL Cholesterol Content in STZ-Induced Diabetic Rats

Group ^{1 )} Serum LDL Cholesterol (mg / dL)  NC 34.9 ± 5.7  DM 68.2 ± 12.3  DM-WE 57.5 ± 11.8  DM-AE 39.7 ± 9.7  DM-CM 47.1 ± 11.2

¹⁾ NC: Normal control group, DM: Diabetic control group, DM-WE: Dietary supplementation of pine needle butterfly fruit fruit hot water extract after diabetes induction, DM-AE: Dietary addition of pine needle butterfly fruit fruit alkali extract after diabetes induction, DM-CM: Addition of 1% of pine myrtle mushroom culture mycelium extract after diabetes induction

However, serum HDL cholesterol content was increased in all experimental groups compared to the diabetic control group, but was not significant (Table 7).

Table 7 : Feeding Effect of Pine Bark Butterfly Extract for 4 Weeks on Serum HDL Cholesterol Content in STZ-Induced Diabetic Rats

Group ^{1 )} Serum HDL Cholesterol (mg / dL)  NC 52.4 ± 5.8  DM 48.9 ± 10.2  DM-WE 51.6 ± 10.4  DM-AE 55.5 ± 9.6  DM-CM 56.1 ± 8.9

¹⁾ NC: Normal control group, DM: Diabetic control group, DM-WE: Dietary supplementation of pine needle butterfly fruit fruit hot water extract after diabetes induction, DM-AE: Dietary addition of pine needle butterfly fruit fruit alkali extract after diabetes induction, DM-CM: Addition of 1% of pine myrtle mushroom culture mycelium extract after diabetes induction

Taken together, the results of four weeks of dietary administration of pine beetle mushroom extract to the diabetic rats, it was confirmed that can reduce the content of triglycerides, total cholesterol and LDL cholesterol induced by diabetes, the effect is pine It was found that the fruit extract of the fruit of Janavier mushroom was the most excellent.

Example 4 Component Analysis of the Pine Pine Butterfly Extract of the Present Invention

As described in Examples 2 to 3, the pine needles mushroom extract of the present invention has an effect of preventing diabetic complications or improving diabetes-induced hypercholesterolemia in diabetic rats. Therefore, in order to find out the active source of the pine berry butterfly extract was analyzed its components including the molecular weight.

The obtained pine leaf butterfly fruit fruit hot water extract, pine leaf butterfly fruit fruit alkali extract and pine leaf butterfly mushroom culture mycelium extract is a known method (see: Structure analysis of extracellular anticancer polysaccharides by liquid culture of Lee Shin-young, Kang Tae-soo, Ganoderma lucidum mycelium, According to The Korean J. Mycology, 27: 76-81, (1999)), fractionated and purified sequentially using DEAE-cellulose (Cl-) ion exchange resin and Sepharose CL-4B gel, and then methylated using GC. The analysis was performed. In addition, the molecular weight of 2 million, 500,000, and 300,000 dextran was used as a standard (product made by Sigma) for molecular weight measurement. The absorbance at 280 nm was measured to know the protein composition of the fraction, and hexasaccharide was analyzed by anthrone method (Spiro RG, Analysis of sugars found in glycoprotein in Method in Enzymology, Academic Press, New York 8). : 4-10, (1966)).

As a result of affinity chromatography to determine the steric coordination of the polysaccharides extracted from the fruit water extract, the alkaline extract and the culture mycelium extract of pine needles, the polysaccharides were β-structured because no sugar was detected in the adsorption part. And it was found. As a result of performing methylation analysis to specifically identify this, as shown in Table 8, 2,3,4,6-tetramethyl glucose and 2,4,6-trimethyl glucose were detected and β-glucose 1,3 bound. β-1,3-glucan was contained, and β-1,6-heterogalactomannan, which has not been identified until now, was mainly composed of β-1,3-glucan and β-1. It was confirmed that β-1,3-glucano-β-1,6-heterogalactomannan to which 6-heterogalactomannan is bound, and the complex polysaccharide had a molecular weight of 30 to 500,000 as a result of gel filtration. It was identified as a protein polysaccharide present in combination with.

Table 8 : Structural Analysis of Polysaccharides Isolated from the Fruiting Body Extract of Pineapple Butterfly and Cultured Mycelia

 Methylated sugar Hydrothermal Extract Polysaccharide (Content) Alkali Extract Polysaccharide (Content) Mycelium broth (content ratio)  Coupling method 2,3,4,6-tetramethyl-glucose  1.0  1.0  1.0    [Glc] 1 → 2,4,6-trimethyl glucose 4.2 3.8 4.0 → 3 [Glc] 1 → 2,3,6-trimethyl glucose 0.8 0.5 0.5 → 4 [Glc] 1 → 2,4-dimethylglucose 0.5 0.6 0.5 → 3, → 6 [Glc] 1 → 2,3,4-trimethyl-mannose  0.7  0.4  0.4 → 6 [Man] 1 → 2,3,4-trimethyl-galactose  1.2  0.9  0.8 → 6 [Gal] 1 →

The extract of the fruiting body extract and culture mycelia of pine needles of the present invention inhibits the activity of aldose reductase of the lens and kidney, and reduces the concentration of triglyceride, total cholesterol and LDL cholesterol, thereby treating diabetic complications or It can be widely used in the development of functional foods with a protective effect.

Claims (11)

Fomitopsis pinicola fruiting body ( Fomitopsis pinicola ) is cut and crushed, swelled with 1N KOH for 1 hour, homogenized using a crusher, filtered through a 100 mesh sieve, neutralized with hydrochloric acid and washed with distilled water, dried pine at 60 ℃ It contains the fruiting body alkali extract of Xavier mushroom, The fruiting body alkali extract of the pine needles butterfly has a molecular weight of 30 to 500,000 as a result of gel filtration, 2,3,4,6-tetramethyl glucose, 2,4,6-trimethyl glucose, 2,3,6-trimethyl glucose , Β-1,3-glucano, 2,4-dimethyl glucose, 2,3,4-trimethyl mannose and 2,3,4-trimethyl galactose bound in a weight ratio of 1.0: 3.8: 0.5: 0.6: 0.4: 0.9 a -β-1,6-heterogalactomannan-protein complex as an active ingredient, A composition for the treatment or prevention of diabetic complications of cataracts or kidney disease caused by an increase in the activity of aldose reductase. delete delete delete delete delete delete delete delete delete delete

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