KR20140066846A - Composition for preventing, treating or improving of metabolic disease comprising morus alba leaf and cudrania tricuspidata leaf as an active ingredient - Google Patents

Abstract

The present invention relates to a composition comprising Morus alba leaf and Cudrania tricuspidata leaf as active ingredients for preventing, treating, or alleviating a metabolic disease, wherein the metabolic disease is diabetes or diabetic complications. According to the present invention, the present invention can be used to develop a health product for improving metabolism by using high-functional natural food materials.

Description

[0001] The present invention relates to a composition for preventing, treating or ameliorating a metabolic disease comprising mulberry leaf and cucumber leaf as an active ingredient. [0002] Composition for Preventing, Treating or Improving of Metabolic Disease comprising Morus alba Leaf and Cudrania tricuspidata Leaf as An Active Ingredient [

The present invention relates to a composition for preventing, treating or ameliorating a metabolic disease comprising mulberry leaf and cucumber leaf as an active ingredient.

With the recent economic growth and the increase in national income, the prevalence of diabetes mellitus is on the rise as people 's diet patterns are westernized. Diabetes is caused by an absolute or relative deficiency of insulin, and it causes various pathological symptoms, including metabolic disturbances of carbohydrates, fats, and proteins due to insulin action in various tissues (1,2). In general, diseases and metabolic syndromes due to diabetes include lipid metabolism abnormalities such as blindness, stroke, myocardial infarction, chronic renal failure, increase in blood triglyceride, decrease in HDL-cholesterol and increase in LDL-cholesterol (3- 5).

On the other hand, Morus Alba L.) is a plant resource belonging to Moraceae and Morus widely distributed in the tropics and temperate regions. Mulberry leaves have been used for thousands of years as a feeding source of silkworms. Since 1998, As a possible raw material, it is listed as a foodstuff and has been steadily used as a raw material for various processed foods and health foods (6-8). The flavonoid rutin, known as a useful ingredient of mulberry leaves, exhibits capillary strengthening action and contraction, and GABA (γ-aminobutyric acid), which is known as a blood pressure lowering substance, is contained about 10 times as much as green tea leaves . In addition, plant sterols such as β-sitosterol, campesterol, β-sitosterol glycoside, β-exydone and inosterone exist in large amounts in the mulberry leaves to decrease the cholesterol and neutrophil in the blood, increase the HDL-cholesterol and antioxidant (9-11). Cudrania tricuspidata ) is a deciduous subfamily belonging to the genus Chalcis. It is used to treat eczema, mumps, pulmonary tuberculosis, chronic urinary retention, bruises, acute arthritis, etc. , Antimicrobial activity and inhibition of trypsin enzyme activity (12,13), lipid elevation and oxidation inhibition in mice (14), and hepatotoxicity inhibitory action (15).

The effectiveness of mulberry leaves or mallow leaves on diabetes has been well known for a long time, but recently, 1-deoxynojirimycin (DNJ, 1-deoxynojirimycin), which has an inhibitory activity against? -Glucosidase in mulberry leaves and several kinds of alkaloids . These results indicate that the diabetic rats fed with a mixture of mulberry leaves and silkworm powder or mulberry have a marked improvement in antioxidative and diabetic complications including hypoglycemia (16-18). However, there is little research to compare the efficacy of mulberry leaves, cucumber leaves, and mixtures thereof.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have sought to develop natural products that can prevent, treat or ameliorate metabolic diseases. As a result, mulberry leaves ( Morus alba Leaf and Cudrania tricuspidata leaf), it is possible to prevent, treat or ameliorate metabolic diseases such as diabetes and diabetic complications, thereby completing the present invention.

Accordingly, an object of the present invention is to provide a composition for preventing, treating or improving metabolic diseases.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a composition for preventing, treating or ameliorating a metabolic disease comprising Morus alba Leaf and Cudrania tricuspidata Leaf as an active ingredient, wherein the metabolic disease is diabetes mellitus or diabetes mellitus Lt; RTI ID = 0.0 > a < / RTI > complication.

The present inventors have sought to develop natural products that can prevent, treat or ameliorate metabolic diseases. As a result, it was confirmed that when mixed with mulberry leaves and cucumber leaves, diabetes and diabetic complications can be prevented, treated or ameliorated.

Mulberry leaves and cucumber leaves used in the composition of the present invention can be used in powder form. When the mulberry leaves and cucumber leaves of the present invention are used in the form of powder, they may be prepared in powder form by an additional process such as vacuum distillation and freeze drying or spray drying.

According to a preferred embodiment of the present invention, the mulberry leaves and cilantro leaves are used in the form of powder by hot air drying and crushing.

Mulberry leaves and cucumber leaves used in the composition of the present invention can be used in the form of an extract. When extracting the mulberry leaves or cucumber leaves with an extraction solvent, various extraction solvents may be used. Preferably, a polar solvent or a non-polar solvent can be used. Suitable polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, Or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- But are not limited to, pentane, 1-chlorobutane, 1-chloropentane, o -xylene, diisopropyl ether, 2- chloropropane, toluene, 1- chloropropane, chlorobenzene, benzene, diethyl ether, diethylsulfide, Methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF.

More preferably, the extraction solvent used in the present invention is (a) water, (b) anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.) (E) ethyl acetate, (f) chloroform, (g) butyl acetate, (h) 1,3-butylene glycol, (i) hexane and (j) diethyl ether. . Most preferably, the extract of the present invention is obtained by treating water, ethanol or a combination thereof with a mulberry leaf or a cedar leaf.

As used herein, the term " extract " means that it is used in the art as a crude extract as described above, but broadly includes fractions obtained by further fractionating the extract. That is, the extracts of mulberry leaves or cucumber leaves include not only those obtained by using the above-described extraction solvent but also those obtained by further applying a purification process thereto. For example, a fraction obtained by passing the above extract through an ultrafiltration membrane having a constant molecular weight cut-off value, and a separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) The fraction obtained by the purification method is also included in the mulberry leaf and cucumber leaf extract of the present invention.

As used herein, the term "comprising as an active ingredient" means an amount sufficient to achieve the efficacy or activity of the following mulberry leaves and cucumber leaves. The present invention is a composition extracted from mulberry leaves and cucurbitan leaves which are natural plant materials. Since the composition does not adversely affect the human body even when administered in an excessive amount, the quantitative upper limit of mulberry leaves and cucumber leaves included in the composition of the present invention can be selected by a person skilled in the art have.

Preferably, the composition of the present invention comprises 1-40% by weight of mulberry leaves and 1-40% by weight of culm leaves. More preferably, the composition comprises 1-30% by weight of mulberry leaves and 2-38% by weight of cilantro leaves, and even more preferably, the composition comprises 1-20% by weight of mulberry leaves and 3-35% And most preferably, the composition comprises 5-15% by weight of mulberry leaves and 5-32% by weight of curly leaves.

Preferably, the composition has a content ratio of the mulberry leaf and the cucumber leaf of 1: 1 to 1: 4.

According to a preferred embodiment of the present invention, the composition of the present invention includes 10% mulberry leaves and 10% cucumber leaves, or 10% mulberry leaves and 30% cucumber leaves.

The composition of the present invention is a composition for preventing, treating or ameliorating a metabolic disease, wherein the metabolic disease is diabetes.

According to a preferred embodiment of the present invention, the diabetic model of the composition as compared to the blood glucose increase rate (29.2%) of the diabetic control group maintains almost the initial blood glucose level with a blood glucose increase rate of 6.7% and 7.6%.

Preferably, the composition reduces the glycated hemoglobin content.

According to a preferred embodiment of the present invention, it is reduced by 15-35% compared to the glycated hemoglobin content of the diabetic control.

The composition of the present invention is a composition for preventing, treating or ameliorating a metabolic disease, wherein the metabolic disease is diabetic complication.

Preferably, the diabetic fatty liver, diabetic cirrhosis, diabetic anemia, coronary artery disease, diabetic ketoacidosis, diabetic ketoacidosis, Hyperglycemia hyperosmolar state, hypoglycemia, diabetic coma, respiratory infections, periodontal disease, diabetic cardiomyopathy, diabetic nephropathy, diabetic nephropathy, Diabetic neuropathy, Diabetic retinopathy, Diabetic myonecrosis, Peripheral vascular disease or Stroke, and more preferably, Diabetic retinopathy, Diabetic retinopathy, Diabetic neuropathy, Diabetic retinopathy, Diabetic myonecrosis, Peripheral vascular disease, , Diabetic cirrhosis, diabetic anemia or coronary artery disease.

According to a preferred embodiment of the present invention, 10% by weight of mulberry leaves and 10% by weight of cilantro leaves were added to 32% and 45% of diabetic diabetic animals, respectively, as compared to AST (aspartate aminotransferase) and ALT (alanine aminotransferase) . This shows the effect of improving liver function by lowering the AST and ALT activity which are increased by diabetes induction.

According to a preferred embodiment of the present invention, the leukocyte count is increased by 38% in the diabetic model as compared to the diabetic control. Leukocytes resistant to diabetes mellitus and cedar leaves stimulate the transport of iron to hematopoietic action, thus increasing the number of blood cells due to diabetes.

The composition of the present invention can be provided as a food composition.

When the composition for preventing, treating or ameliorating a metabolic disease comprising mulberry leaf and cucumber leaf as an active ingredient of the present invention is prepared from a food composition, it is preferable that not only mulberry leaf and cedarwood leaf as an active ingredient, And includes, for example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared as a drink, it additionally contains citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, etc., .

The compositions of the present invention may be prepared with pharmaceutical compositions.

According to a preferred embodiment of the present invention, the composition of the present invention comprises (a) a pharmaceutically effective amount of the mulberry leaf and cucumber leaf of the present invention as described above; And (b) a pharmaceutically acceptable carrier. As used herein, the term " pharmaceutically effective amount " means an amount sufficient to achieve the efficacy or activity of the mulberry leaves and cucumber leaves described above.

When the composition of the present invention is manufactured from a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably administered orally.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . Typical dosages of the pharmaceutical compositions of this invention are in the range of 0.001-100 mg / kg on an adult basis.

The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The features and advantages of the present invention are summarized as follows:

(a) The present invention mulberry (Morus alba Leaf and Cudrania tricuspidata Leaf as active ingredients, and a composition for accelerating bowel movement.

(b) The present invention can develop a health product for improving metabolism through utilization of a highly functional natural food material.

Fig. 1 shows (a) changes in blood glucose level and (b) curves of the mice in diabetic mice fed with mulberry leaf powder and cucumber leaf powder. The experimental conditions are the same as in Table 1, and the results are shown in (a) as the median and (b) in the mean ± standard deviation (n = 8). Values indicated by other letters according to the Duncan multiple-range test method have significance of p < 0.05.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Materials and methods

Experimental material

The mulberry leaves and cucumber leaves used in this experiment were supplied from DaeTong Agricultural Cooperative Corporation in May, 2011, and collected in Sancheong-gun, Kyungnam Province. The leaves were dried in hot air at 50 ℃ and crushed.

Empirical formula

(AIN-93G) (19) was used as a basal diet to evaluate the physiological effects of mulberry leaves and cucumber leaves on the physiological efficacy of rats. (A), 10% of mulberry leaves, 10% of mixed leaf of Cucumber leaves, 10% of mulberry leaves, 30% of mixed leaf of Cucumber leaves, 10% of cucumber leaf powder, (E), and each experimental diet was fed for 6 weeks. Table 2 shows the results of measurement of general components of the experimental diet according to the AOAC method (20).

The basic diet (AIN-93G) was prepared from corn starch 397.486 g / kg diet, sucrose 100.00 g / kg diet, dextrose 132.00 g / kg diet, casein 200.00 g / kg diet, soybean oil 70.00 g / kg diet, / Kg diet, 35.00 g / kg of mixed mineral, 10.00 g / kg of mixed vitamin, 2.50 g / kg of tartaric acid choline, 3.00 g / kg of L-cystine and 0.014 g / kg of t-butylhydroquinone . ML in Table 1 represents Mulberry leaves, and CTL represents Cudrania tricuspidata leaves.

Group (n = 8) process A Basic diet (90%) + ML powder (10%) B Basic diet (80%) + ML powder (10%) + CTL powder (10%) C Basic diet (60%) + ML powder (10%) + CTL powder (30%) D Basic diet (90%) + CTL powder (10%) E The basic diet (100%, DM control)

group moisture
(g / 100 g) Crude fat
(g / 100 g) Crude protein
(g / 100 g) Views min
(g / 100 g) carbohydrate
(g / 100 g) Dietary Fiber
(g / 100 g) energy
(㎉ / 100 g) calcium
(Mg / 100 g) sign
(Mg / 100 g) A
B
C
D
E 10.08
10.05
10.15
10.59
9.55 15.56
16.78
16.26
16.47
15.91 17.74
18.08
18.30
18.25
18.10 6.00
5.88
5.95
5.92
6.00 56.62
55.09
55.29
54.69
56.44 19.73
20.29
20.83
19.49
19.05 437.48
443.70
440.70
439.99
441.35 1029.84
1045.28
1036.49
1053.28
1087.48 776.18
776.92
747.34
754.59
759.39

Breeding of experimental animals

Sprague-Dawley (SD) male rats, 3 weeks old, were purchased from Hanlim Experimental Animal Co., Ltd. (Korea) for 3 weeks, and were subjected to an adaptation with a general solid feed for 1 week before the experiment It was used in this experiment. The environment of the breeding room was maintained at constant temperature (23 ± 1 ℃) and humidity (50 ± 5%), and illumination was constantly adjusted to 12 hours / cancer cycle (08: 00-20: 00). During the experimental period, diets and drinking water were ad libitum , and body weights were measured once a week at regular intervals using an animal scales (GF-2000, AND, Korea) The remaining diet was refrigerated at 4 ° C and measured once a week. The feed efficiency ratio (FER) was calculated by dividing the weight gain by the dietary intake during the same period, and the insulin dose was measured every two days.

Induction of diabetes

Diabetes was induced by injecting streptozotocin (STZ, Sigma Chemical Co. Ltd., USA) to create an experimental model similar to insulin-dependent diabetes mellitus. SD male rats weighing 320 g were weighed and injected intraperitoneally in 0.2 ml of 0.1 M citrate buffer (pH 4.0) at a concentration of 47.5 ㎎ per ㎏ of body weight before the start of the experiment. Rats with a blood glucose level of 350 ㎎ / ㎗ were selected on the 7th day after STZ injection and were randomly assigned to 8 rats in a random-ized block design for a total of 6 weeks.

Treatment of experimental animals and sample collection

After completion of the experiment, blood was collected from the orbital plexus and allowed to stand for 1 hour at room temperature. After centrifugation at 2,500 rpm for 10 minutes, serum was isolated from the orbital plexus . The separated serum was used for analysis while being kept frozen at -70 ° C. After the blood collection, each organs (liver, kidney, pancreas, spleen and testes) were removed and washed with 0.9% physiological saline, and the water was removed with a filter paper and weighed.

Blood glucose measurement and serum component analysis

Blood glucose levels were measured weekly using a blood glucose meter (Accu-Chek ^ⓡ Active 601, Roche, Germany) and the area under the curve (AUC) was calculated using the Sigma Plot 8.0 program (Jandel Scientific, San Rafael, ). Serum biochemical values are shown in Table 3 according to the method of Han et al. (21).

Item Way Analytical instrument Glucose Enzymatic method ADVIA 1650 (Bayer, Shiga, Japan) C-peptide ECLIA ^{1 )} Modular analytics E170 (Roche, Germany) HbA1c TIA ^{3 )} Integra 400 (Roche, Switzerland) insulin ECLIA Modular analytics E170 (Roche, Germany) Fructosamine Colorimetric Modular analytics P (Roche, Germany) FFA ^{4 )} ACS-ACOD ^{5 )} Method Modular analytics P (Hitachi, Japan) TC ^{6 )} Enzymatic method ADVIA 1650 (Bayer, Japan) HDL-C ^{7 )} Enzymatic method ADVIA 1650 (Bayer, Japan) LDL-C ^{8 )} EIA ^{9 )} ADVIA 1650 (Bayer, Japan) TG ^{10 )} Lipase, GK ^{11 )} , GPO ^{12 )} colorimetric method ADVIA 1650 (Bayer, Japan) AST ^{13 )} Enzymatic method by IFCC ADVIA 1650 (Bayer, Japan) ALT ^{15 )} Enzymatic method by IFCC ADVIA 1650 (Bayer, Japan) ALP ^{16 )} Enzymatic method by IFCC Modular analytics PE (Roche, Germany) Creatinine Jaffe reaction ADVIA 1650 (Bayer, Japan) BUN ^{17 )} Urease with GLDH ^{18 )} ADVIA 1650 (Bayer, Japan) RBC ^{19 )} Electronic impedance SYSMEX XE 2100 D (Sysmex, Japan) WBC ^{20 )} Flow cell analysis SYSMEX XE 2100 D (Sysmex, Japan) Hematocrit Electronic impedance SYSMEX XE 2100 D (Sysmex, Japan) hemoglobin Non-cyanogen hemoglobin spectrophotometer SYSMEX XE 2100 D (Sysmex, Japan) Platelet Electronic impedance SYSMEX XE 2100 D (Sysmex, Japan)

In Table 3, ECLIA is Electrochemiluminescence Immunoassay, HbA1c is Hemoglobin A1c, TIA is Turbidimetric Immunoassay, FFA is Free Fatty Acid, and ACS-ACOD is acetyl CoA synthetase-acetyl CoA oxidase, TC is total cholesterol, HDL-C is high-density lipoprotein-cholesterol, LDL- C is a low-density lipoprotein-cholesterol, EIA is an enzyme immunoassay, TG is a triglyceride, GK is a glycerokinase, and GPO is an L- α-Glycerol Phosphate Oxidase, AST is an aspartate aminotransferase (SGOT), IFCC is an International Federation of Clinical ALD is alkaline phosphatase, BUN is blood Urea Nitrogen, GLDH is glutamate dehydrogenase, Glutamate dehydrogenase is an enzyme, , RBC is Red Blood Cell, and WBC is White Blood Cell.

Statistical processing

The results obtained in this study were statistically analyzed using the SPSS program (Windows version 17.0, SPSS Inc., USA), and the results were expressed as mean ± standard error. The results of each group were analyzed by one-way ANOVA (Mutation Analysis) followed by Duncan's multiple range test at p <0.05.

Results and Discussion

weight, Dietary intake , Dietary efficiency  And Quantity of water

Table 4 shows the effect of dietary mulberry leaf and cucumber leaf powder on body weight, body weight gain, dietary intake, dietary efficiency and food intake of STZ-induced diabetic rats. At the end of the experiment, the body weight, body weight gain and dietary efficiency did not show any significant difference between the experimental groups. However, in the diabetic control group (E), the body weight gain was somewhat lower than that of the other experimental groups because STZ selectively destroyed the pancreatic β- (22). This is due to the imbalance of glucose metabolism caused by this, the excessive degradation of body fat and the continuous loss of body protein (22). Dietary intake and water intake were significantly lower in mulberry leaf and cucumber leaf 1: 1 mixed group (B) (p <0.05). The results of this study suggest that supplemental studies on the inhibition of weight gain of mulberry leaves and cucumber leaves are necessary.

group Initial weight (g) Final weight (g) Weight gain
(g / day) Dietary intake (g / day) FER Water volume (ml / day) A 356.0 ± 17.4 ^b 388.9 ± 65.1 ^NS 1.69 ± 1.21 ^NS 45.97 ± 3.26 ^b 0.044 0.031 ^NS 210.3 ± 14.7 ^b B 387.7 ± 48.9 ^ab 442.2 + - 106.3 2.18 ± 1.28 38.71 + 1.89 ^c 0.070 0.046 133.6 + - 6.5 ^c C 406.9 ± 14.5 ^a 458.2 + - 52.4 2.14 ± 3.15 51.37 + 4.77 ^a 0.052 + 0.062 219.6 ± 18.6 ^b D 407.8 ± 19.7 ^a 435.2 ± 29.3 1.61 ± 1.34 53.35 ± 6.77 ^a 0.034 0.031 251.4 ± 47.6 ^a E 367.5 ± 32.6 ^ab 373.0 + - 54.2 0.94 + - 1.27 48.28 ± 3.07 ^ab 0.019 0.031 221.0 ± 15.4 ^b

The FER in Table 4 represents a feed efficiency ratio and is a value obtained by dividing the weight gain by the dietary intake. The population of each group is 8, and according to the Duncan Multi-Range Test, the other superscripts indicate confidence of p <0.05. 'NS' means 'Not Significant'.

Long-term weight

Table 5 shows the effect of dietary mulberry leaf and cucumber leaf powder on the organ weight of STZ-induced diabetic rats. There was no significant difference in the liver, kidney, pancreas and testis weights among the experimental groups. In the spleen weight, the mulberry leaf and cucumber leaf 1: 3 mixed group (C) Respectively. In diabetic rats with STZ, diabetes mellitus is unbalanced due to decreased secretion of insulin (23,24) compared to hypertrophy of liver, kidney and heart (23,24). In this study, It turned out not to give.

group Organ Weight (g / 100 g body weight) liver kidney Pancreas spleen Testimony A 4.09 ± 0.65 ^NS 0.55 0.08 ^NS 0.26 ± 0.07 ^NS 0.18 ± 0.05 ^b 0.48 0.01 ^NS B 3.92 ± 0.29 0.55 + - 0.05 0.27 ± 0.07 0.23 ± 0.08 ^ab 0.54 + 0.03 C 4.64 ± 0.62 0.58 + 0.14 0.29 + 0.04 0.29 + 0.01 ^a 0.53 + 0.06 D 4.65 ± 0.62 0.61 + 0.06 0.24 + 0.02 0.22 0.02 ^ab 0.51 ± 0.05 E 4.21 + - 0.47 0.57 + 0.06 0.23 + 0.03 0.20 ± 0.04 ^b 0.51 ± 0.06

Blood sugar change

The area under the curve (AUC) of blood glucose changes and blood glucose levels during the 6-week feeding period after diabetic induction with mulberry leaves and cucurbit leaf powder were shown in FIG. The blood glucose level of the diabetic control group (E) and the cucumber leafy group (D) were higher than those of the other treatments during the experimental period, and the mulberry leaf and cucumber leaf 1: 1 mixture group (B) Which is similar to the pre-standard blood glucose level. The blood glucose increase rate was 6.7% and 7.6% in group B and 6, respectively, compared with the reference blood glucose (reference blood glucose), and the blood glucose level was lower in the group of diabetic control (29.2% ) Of group B and C were significantly lower than those of group D and E (p <0.05). In the present study, Lee et al. (25) showed similar trends in blood glucose lowering effects of STZ-treated hyperglycemic mice compared to diabetic control group in the water and ethanol layer of fasting mulberry leaf extracts. Alloxan ), The results of Kim et al. (26), which reported a statistically significant lowering of blood glucose level compared to the diabetic group, were also similar when fed anaerobically treated mulberry leaf powder (20%). This hypoglycemic effect of mulberry leaves is attributed to the fact that the deoxynojirimycin (DNJ) component does not affect the insulin concentration but causes the intestinal carbohydrate degrading enzymes? -Glucosidase,? -Mannosidase and? -Galactosidase (27) reported that there is an effect of inhibiting the action of the enzyme. In the 6th week of experiment, the blood glucose of mixed group (B, C) of mulberry leaves and cucumber leaves was significantly decreased (p <0.05) compared to control group (E) and mulberry leaves The results showed that the addition of two samples improves blood glucose lowering efficacy. However, since there was a difference in the level of addition of samples to each experimental group, it is considered that a supplementary experiment is needed for this part.

Index component of blood sugar

Table 6 shows the effect of dietary mulberry leaf and cucumber leaf powder on blood glucose-related index components of STZ-induced diabetic rats. After the experiment, fasting blood glucose values were significantly lower in the 1: 1 mixed group (B) and 1: 3 mixed group (C) of mulberry leaves and cucumber leaves than in the leaf added group (D) and diabetic control group (E) &Lt; 0.05). These results were similar to those of Yoo et al. (28), which reported that diabetic rats had a glucose reduction rate of about 22-25% when fed with YK-209 mulberry leaf powder for 3 weeks, It is believed that this effect is due to the hypoglycemic effect of the mulberry leaves mentioned above. The C-peptide content of the insulin-secreting pancreatic β-cells was 190.7 pmol / L, which was higher than that of the other experimental groups, but there was no significant difference. On the other hand, the degree of hyperglycemia seen in diabetes can be judged by observing changes in the content of glycated hemoglobin (HbA1c) or serum fructosamine in the blood (29). The glycated hemoglobin content, which is an index of long - term average blood glucose control, was significantly (p <0.05) lower than that of the diabetic control group in the mulberry leaf group (A), mulberry leaf and cucumber leaf 1: And insulin and fructosamine contents were not significantly different among the experimental groups.

group Glucose (mg / dl) C-peptide
(pmol / L) HbA1c (%) Insulin (ng / ml) Fructosamine (μmol / L) A 268.3 ± 46.9 ^ab 142.5 + - 71.0 ^NS 6.13 ± 1.07 ^b 0.07 ± 0.05 ^NS 249.0 ± 53.4 ^NS B 195.0 ± 48.7 ^b 146.0 + - 96.0 5.68 ± 1.34 ^b 0.06 0.02 211.8 ± 56.4 C 227.3 ± 37.9 ^b 190.7 ± 121.6 6.77 ± 1.80 ^ab 0.11 + 0.06 235.8 ± 52.7 D 298.8 + - 55.0 ^a 136.5 ± 65.2 6.53 ± 0.16 ^ab 0.14 + 0.06 259.0 + - 13.4 E 323.8 ± 73.5 ^a 131.3 ± 38.4 8.30 ± 1.29 ^a 0.10 0.09 238.0 ± 12.0

Serum free fatty acid and lipid concentration

Table 7 shows the effect of dietary mulberry leaf and cucumber leaf powder on serum free fatty acid (FFA) and lipid concentration in STZ-induced diabetic rats. Serum free fatty acid levels were 773.25 μEq / L, 882.00 μEq (L), respectively, in the mulberry leaf group (A), mulberry leaf and cucumber leaf 1: 1 mixed group (B) / L and 814.00 μEq / L, respectively, but there was no significant difference. The total cholesterol (TC) concentration was decreased by 14% on average in the mulberry leaf and cedar leaf application groups (A, B, C and D) compared to the diabetic control group (E), but there was no statistically significant difference. The HDL- There was no significant difference between the experimental group and the experimental group. LDL - cholesterol levels were significantly different between group C (8.25 ㎎ / ㎗) and group D (17.50 ㎎ / ㎗) (p <0.05) and triglyceride (TG) A, B and D (129.50, 117.25 and 93.75 mg / dl), respectively (p <0.05). Hyperlipidemia due to high blood cholesterol and triglyceride content is a complication accompanied by diabetes, and arteriosclerosis resulting from this is the main cause of death of diabetic patients. In particular, hypercholesterolemia is a major cause of ischemic heart disease and atherosclerosis, and is known to occur in patients with non-insulin dependent diabetes mellitus (30). In this regard, Oh et al. (31) reported a decrease in serum TG levels in db / db mice fed with mulberry leaf extract for 5 weeks. Kim et al. (32) reported that experimental cholesterol- Induced TG concentration in rats fed the mulberry leaf methanol extract. These results suggest that dietary mulberry leaves and cucumber leafy diets are effective in improving the serum lipid composition of STZ-induced diabetic rats. Quercetin, kaempferol, chlorogenic acid, (33), such as chlorogenic acid, arthocarpesin, and caffeic acid.

group FFA (μEq / L) Lipid (㎎ / ㎗) TC HDL-C LDL-C TG A 773.25 + 267.22 ^NS 76.75 ± 18.34 ^NS 65.75 ± 19.40 ^NS 12.00 ± 6.06 ^ab6 129.50 ± 35.89 ^b B 882.00 ± 212.75 77.75 ± 17.33 67.00 ± 19.61 14.25 ± 5.12 ^ab 117.25 + - 31.85 ^b C 1038.25 ± 222.48 80.50 ± 19.30 61.25 + - 22.02 8.25 ± 2.63 ^b 256.50 ± 45.66 ^a D 814.00 ± 198.95 78.00 ± 17.94 66.25 + - 16.76 17.50 ± 5.45 ^a 93.75 ± 23.20 ^b E 1042.00 ± 404.34 90.50 ± 47.66 51.25 + 16.74 9.75 ± 4.50 ^ab 249.00 ± 30.35 ^a

Liver function test and kidney function test

Table 8 shows the effect of dietary mulberry leaf and cucumber leaf powder on liver and kidney function in STZ-induced diabetic rats. AST and ALT activities were significantly decreased in mulberry leaf group (A), mulberry leaf and cucumber leaf 1: 1 mixture group (B) and cucumber leafy group (D) compared to diabetic control group (p < 0.05). Alkaline phosphatase (ALP) activity was slightly decreased in A, B, and D groups compared with DM control group, and was significantly different from mulberry leaves and Cryptomeria japonica 1: 3 mixed group (C) (P <0.05). Kim et al. (34) reported that elevated serum AST and ALT are hepatocyte necrosis and are useful indicators for the diagnosis and treatment of liver disease. The diabetic rats in this study were presumed to have liver damage by STZ. Dietary supplementation of mulberry leaf and cucumber leaf decreased the AST and ALT activity, which were induced by diabetes mellitus. Creatinine and blood urea nitrogen (BUN) concentrations (35), which are important indicators in the diagnosis of kidney disease, were not significantly different between the experimental groups, and the effects on renal function of mulberry leaves and cucumber leaves were not clearly revealed.

group Liver Function Value (U / L) Kidney function value (mg / dl) AST ALT ALP Creatinine BUN ³⁾ A 250.25 + - 72.68 _b 132.00 ± 39.95 ^b 259.25 + - 38.87 ^b 0.40 + 0.08 ^NS 33.98 ± 16.08 ^NS B 283.00 ± 49.37 ^b 137.75 ± 32.00 ^b 233.25 + - 57.36 ^b 0.40 + 0.08 31.13 ± 18.19 C 373.75 ± 82.92 ^ab 193.75 ± 40.60 ^ab 411.50 ± 43.11 ^a 0.40 + 0.08 33.58 ± 12.17 D 287.75 ± 49.73 ^b 145.50 ± 45.01 ^b 203.75 ± 68.59 ^b 0.30 ± 0.00 30.43 + - 4.76 E 413.25 ± 59.31 ^a 250.00 ± 42.46 ^a 321.00 ± 77.51 ^ab 0.30 ± 0.00 27.45 ± 9.92

Whole blood cells  inspection

The effects of dietary mulberry leaves and cucumber leaf powder on the total blood count (CBC) of STZ-induced diabetic rats are shown in Table 9. [ The number of red blood cells (RBC) was significantly higher in group B than in group A (mulberry leaf) and in group C (mulberry leaf, mixed culm leaf and cucumber leaf 1: (P <0.05) in group B and C compared to control group (P <0.05). The hematocrit and hemoglobin concentrations were not significantly different between the experimental groups. Platelet counts were significantly higher in the mulberry leaves and cucumber leaves (A, B, C, D) than in the E (P <0.05). In this regard, Woo et al. (36) reported leukocyte depletion due to oxidative stress in diabetes mellitus in STZ-treated diabetic rats. In this study, mulberry leaf and cedarwood leaf stimulated iron transport, It is thought that it slightly increases the decreased blood count. The number of individuals in Table 8 is 8.

group RBC (× 10 ⁶ / μl) WBC (占 10 ³ / 占 퐇) Hct (%) Hb ^{5 )} (g / dl) Platelets (占 10 ³ / 占 퐇) A 8.78 ± 0.69 ^b 11.66 ± 1.75 ^ab 44.75 ± 1.85 ^NS 16.55 + 0.81 ^NS 1023.8 ± 210.21 ^a B 9.21 ± 0.57 ^a 12.73 + 1.50 ^a 44.60 + - 2.48 16.60 ± 1.07 1115.6 ± 109.60 ^a C 9.54 + 0.29 ^a 13.78 ± 1.23 ^a 45.40 +/- 0.79 16.68 ± 0.57 1222.1 + - 115.00 ^a D 9.06 ± 0.63 ^ab 11.63 ± 1.95 ^ab 44.76 ± 1.64 16.90 + - 0.41 912.25 ± 133.90 ^a E 8.97 ± 0.62 ^ab 9.98 ± 1.90 ^b 44.30 ± 6.09 16.45 + - 2.34 651.00 ± 109.28 ^b

These results suggest that STZ-induced diabetic rats are effective in reducing blood glucose levels and improving blood glucose-related indicators, serum lipid levels, and hepatic function when fed with a 1: 1 mixture of mulberry leaf and cucumber leaf powder . In order to compare the efficacy of mulberry leaf and cucumber leaf in individual and mixed diets, it is considered that supplementary studies such as diversification of sample addition level will be needed. In the future, development of diverse products for improving diabetes using these functional food materials Is expected to be possible.

references

1. Kannel WB, Megee DL. 1979. Diabetes and cardiovascular disease. JAHA 241: 2035-2038.

2. Abrams JJ, Ginsberg H, Grundy SM. 1982. Metabolism of cholesterol and plasma triglycerides in non-ketotic diabetes mellitus. Diabetes 31: 903-910.

3. Steinberg HO, Chaker H, Leaming R, Johnson A, Brechtel G, Baron AD. 1996. Obesity / insulin resistance associated with endothelial dysfunction. Implications for the syndrome of insulin resistance. J Clin Invest 97: 2601-2610.

4. Hayden JM, Reaven PD. 2000. Cardiovascular disease in diabetes mellitus type 2: a potential role for novel cardiovascular risk factors. Curr Opin Lipidol 11: 519-528.

5. Treadway JL, Mendys P, Hoover DJ. 2001. Glycogen phosphorylase inhibitors for the treatment of type 2 diabetes mellitus. Expert Opin Investig Drugs 10: 439-454.

6. Onogia, Osawa K, Yasuda H, Sakaia, Morita H, Tokawa H. 1993. Flavonol glycosides from the leaf of Morus alba . Shoyakugaku Zasshi 47: 423-425.

7. Shin KH, Young HS, Lee TW, Choi JS. 1995. Studies on the chemical component and antioxidative effects of Sola - num lyratum . Korean J Pharm 26: 130-138.

8. Kim MW, Ahn MS, Lim YH. 2003. The antioxidative activities of mulberry leaves extracts on edible soybean oil. Korean J Food Culture 18: 1-8.

9. Kodama T, Ishida H, Kokubo T, Yamakawa T, Noguchi H. 1990. Glucosylation of quercetin by a cell suspension culture of Vitis sp. Agric Biol Chem 54: 3283-3288.

10. Shin DH. 1998. Antioxidation substances in mulberry leaf. J Korean Oil Chemists Soc 16: 27-31.

11. Chae JY, Lee JY, Hoang IS, Whangbo D, Choi PW, Lee WC, Kim JW, Kim SY, Choi SW, Rhee SJ. 2003. Analysis of functional components of leaf mulberry cultivars. J Korean Soc Food Sci Nutr 32: 15-21.

12. Kim SH, Kim NJ, Choi JS, Park JC. 1993. Determination of flavonoid by HPLC and biological activities from the leaves of Cudrania tricuspidata Bureau. J Korean Soc Food Sci Nutr 22: 68-72.

13. Ottersen T, Vance B, Doorenbos NJ, Chang BL, EI-Feraly FS. 1977. The crystal structure of cudranone, 2,6,3'-trihydroxy-4-methoxy-2 '- (3-methyl-2-butenyl) -benzophenone: a new antimicrobial agent from Cudrania cochinchi - nensis . Acta Chem Scand 31: 434-436.

14. Chang CH, Lin CC, Hattori M, Namba T. 1994. Effects of anti-lipid peroxidation of Cudrania cochinchinensis var. gerontoge . J Ethnopharomacol 44: 179-185.

15. Joo HY, Lim KT. 2009. Protective effect of glycoprotein isolated from Cudrania tricuspidata on liver in CCl ₄ -treat-ed A / J mice. Korean J Food Sci Technol 41: 93-99.

16. Asano N, Oseki K, Tomioka E, Kizu H, Matsui K. 1994. N-containing sugars from Morus alba and their glycosidase inhibitory activities. Carbohyr Res 259: 243-255.

17. Jang MJ, Rhee SJ. 2004. Hypoglycemic effects of pills made of mulberry leaves and silkworm powder in streptozotocin-induced diabetic rats. J Korean Soc Food Sci Nutr 33: 1611-1617.

18. Kwon EH, Jung MA, Rhee SJ, Choi SW, Cho SH. 2006. Antioxidant effects of improvement of lipid metabolism of mulberry fruit, mulberry leaves and silkworm powder with different mixing ratios in streptozotocin-induced diabetic rats. Korean J Nutr 39: 91-99.

19. Reeves PG, Nielsen FH, Fahey GC. 1993. AIN-93 purified diets for laboratory rodent: final report of the American Institute Nutrition Ad Hoc Writing Committee in the reformulation of the AIN-76A rodent diets. J Nutr 123: 1939-1951 .

20. AOAC. 1980. Official Methods of Analysis . 15th ed. Associate of Official Analytical Chemists, Washington, DC, USA. p 211-260.

21. Han CK, Kim SS, Choi SY, Park JH, Lee BH. 2009. Effects of rice added with mulberry leaves and fruit on blood glucose, body fat and serum lipid levels in rats. J Korean Soc Food Sci Nutr 38: 1336-1341.

22. Odaka H, Matsuo T. 1992. Ameliorating effects of an intestinal disaccharidase inhibitor, AO-128, in streptozotocin-diabetic rats. J Jpn Soc Nutr Food Sci 45: 33-38.

23. Mogensen CE, Anderson MJF. 1973. Increased kidney size and glomerular filtration rate of early jurvenile diabetes. Diabetes 22: 706-712.

24. Heaton KW. 1973. Food fiber as an obstacle to energy intake. Lancet 2: 1418-1421.

25. Lee WC, Kim SE, Han SJ, Yun SC. 1993. Effect of number of days after urea foliar spray on mulberry leaf components and silkworm. Korea J Seric Sci 35: 86-88.

26. Kim SY, Ryu KS, Lee WC, Ku HO, Lee HS, Lee KR. 1999. Hypoglycemic effect of mulberry leaves with anaerobic treatment in alloxan-induced diabetic mice. Korean J Pharmacogn 30: 123-129.

27. Chung SH, Yu JH, Kim EJ, Ryu KS. 1996. Blood glucose lowering effect of silkworm. Bull KH Pharma Sci 24: 95-100.

28. Yoo SK, Kim MJ, Kim JW, Rhee SJ. 2002. Effects of YK-209 mulberry leaves on disaccharidase activities of small intestine and blood glucose-lowering in streptozotocin-induced diabetic rats. J Korean Soc Food Sci Nutr 31: 1071-1077.

29. Pinnell AE, Northam BE. 1978. New automated dye-binding method for serum albumin determination with bromo-cresol purple. Clin Chem 24: 80-86.

30. Levy RI. 1991. Cholesterol, lipoproteins, apolipoproteins and heart disease; present status and future prospects. Clin Chem 27: 653-662.

31. Oh UJ, Kim GP, Cho YW, Chung SH, Gu SJ. 1999. Effect of extracts from mulberry leaves on serum glucose and lipid levels in db / db mouse. Annual Meet-ing of Korean Soc Food Sci Technol, Seoul, Korea. p 430.

32. Kim SY, Lee WC, Kim HB, Kim AJ, Kim SK. 1998. Antihy-perlipidemic effects of methanol extracts from mulberry leaves in cholesterol induced hyperlipidemia in rats. J Korean Soc Food Sci Nutr 27: 1217-1222.

33. Kim MS, Choue RW, Chung SH, Koo SJ. 1998. Blood glucose lowering effects of mulberry leaves and silkworm extracts on mice fed high carbohydrate diet. Korean J Nutr 31: 117-121.

34. Kim YG, Cho GB, Kim MS, Ko YH, Park MI, Woo IK, Ahn SY, Park BC. 1993. The diagnostic significance of AST / ALT ratio in patients with hepatocellular carcinoma. Korean J Gastroenterology 25: 958-963.

35. Seoul Medical Science Institute. 2005. SCL examination guidebook .

36. Woo HJ, Kim JH, Kim JK, Kim HJ, Park KB. 2006. Effect of gamma irradiation and Cichorium products on oxidative damage and lipid metabolism in streptozotocin-induced diabetic rats. English J Environ Biol 24: 102-111.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (8)

Morus alba Leaf and Cudrania tricuspidata Leaf as active ingredients, wherein the metabolic disease is diabetes or diabetic complication.
The method of claim 1, wherein the diabetic complication is selected from the group consisting of diabetic fatty liver, diabetic cirrhosis, diabetic anemia, coronary artery disease, diabetic ketoacidosis, Diabetic ketoacidosis, hyperglycemia hyperosmolar state, hypoglycemia, diabetic coma, respiratory infections, periodontal disease, diabetic cardiomyopathy, diabetic ketoacidosis, A diabetic neuropathy, a diabetic neuropathy, a diabetic retinopathy, a diabetic myonecrosis, a peripheral vascular disease, or a stroke. Composition.
The composition according to claim 2, wherein the diabetic complication is diabetic fatty liver, diabetic cirrhosis, diabetic anemia or coronary artery disease.
4. The composition according to any one of claims 1 to 3, wherein the composition reduces the glycated hemoglobin content.
4. The composition according to any one of claims 1 to 3, wherein the mulberry leaf and the cucumber leaf are in the form of a powder or an extract.
4. The composition according to any one of claims 1 to 3, wherein the composition comprises 1-40% by weight of mulberry leaves and 1-40% by weight of cilantro leaves.
The composition according to any one of claims 1 to 3, wherein the composition has a content ratio of the mulberry leaf and the cucumber leaf of 1: 1 to 1: 4.
The composition of claim 1, wherein the composition is a food composition or a pharmaceutical composition.

Images