KR20030077902A - Composition of anti-diabetes patient food using silkworm product - Google Patents

Abstract

The present invention relates to a composition of anti-dementia patients using silkworm products, which is obtained by lyophilizing silk fibroin having a molecular weight of 500 and a silkworm of 5 days and 3 days obtained by enzymatic hydrolysis of silk protein, 2: 1 (w / After mixing with w), 0.2 parts by weight of tyrosine and tryptophan were respectively added to 5 parts by weight of the mixture as a function enhancer, and 0.02 parts by weight of licorice and cinnamon extract were added to purify the odor, and 0.05 parts by weight of citric acid as a flavor and flavor enhancer. , 0.05 parts by weight of vitamin C is added, 0.1 parts by weight of taurine as an anti-fatigue agent is added and mixed, and then 200 ml of can packaging is prepared.

Description

COMPOSITION OF ANTI-DIABETES PATIENT FOOD USING SILKWORM PRODUCT}

The present invention relates to an anti-dementia patient composition using a silkworm product.

Diabetes is classified by the WHO report (1985) into insulin dependent diabetes (type I), insulin independent diabetes (type II), and malnutrition diabetes (MRDM). Fortunately, 84% of diabetic patients in Korea are type II diabetes, independent of insulin (Lee et al., 1984). Type II diabetes is particularly common among adults in their 40s and older, who are 1) obese and 2) less active, 3) have complications of hypertension, 4) have liver dysfunction, and 5) have high levels of triglycerides in their blood. It has the characteristic to

Therefore, in particular, as a complication, the development of a patient formula having a prevention effect on diabetes and senile dementia, known as intractable disease, has been requested.

An object of the present invention is to provide an antidementia patient food based on silk fibroin and silkworm powder.

Silk fibroin is mainly used as a silkworm-related product to be eaten, and even if silkworm powder is added, it is a problem to determine how much silk fibroin to use. Thus, silk protein was pretreated in distilled water, such as salt removal from silkworm-related products, and flavouryme (Novo Co., Denmark) was added at a concentration of 1% as a protease, and molecular weight 250, Obtaining silk fibroins of 500 and 1,000 is the most difficult task and requires considerable skill. In vitro experiments showed that silk fibroin having a molecular weight of 500 was the most effective through the antioxidant activity of DPPH according to the molecular weight of silk fibroin (Table 1), based on silk fibroin (MW 500), and silkworm powder Was added 2: 1 (w / w) to develop an senile antichime patient.

As a result of measuring the dose (μg) required to reduce 50% in order to compare the antioxidant capacity by electron donating ability, silk fibroin (4,774 ± 385 μg: 100%) having a molecular weight of 250 as shown in Table 1 The silk fibroin having a molecular weight of 500 was 73.6%, which is the most effective.

Silk fibroin is positioned as a component having functional physiological activity as a silkworm to be eaten as a hydrolyzate of silkworm protein. Silk fibroin is a neurotransmitter (acetylcholinc (ACh)) and related enzymes that are closely related to the treatment and prevention of senile dementia. Acetylcholineesterase (AChE) and catecholamine-based neurotransmitter breakdown enzymes were evaluated for their effects on the inhibition of the activity of monoamine oxidase-B (MAO-B) and the inhibitory effects on the production of free radicals. The antiaging effect of the powder was analyzed to form a composition for preventing dementia.

The present invention is a 2: 1 (w / w) mixed with silk fibroin (silkworm powder) excellent in inhibiting aging and silk fibroin is recognized to prevent dementia by increasing the activity of neurotransmitters and related enzymes 1.0 to 5.0 parts by weight of this mixture was prepared by adding 0.5 parts by weight of tyrosine and tryptophan, 0.1 part by weight of taurine, vitamin C, and citric acid to 0.05 parts by weight of Greenbia mild care (MC), respectively. 200 ml of Anti-dementia Patient Food was prepared.

Experimental materials, experimental methods and results of in vivo experiments for the development of the antidementia patient formula are as follows.

1. Animal testing and feed composition

(1) Animals and experiment

Crj / Bgi rats (male, 180 ± 10 g) purchased from Biogenomics (Seoul) were preliminarily bred for two weeks in an animal nursery, and then divided into four groups of seven animals. The control group was silk fibroin- silkworm powder. Greenbia mild care (MC) only without (2: 1, w / w) and silk fibroin- silkworm powder (2: 1, w / w) in Greenbia MC were 1.0%, respectively. The 200 ml of 3.0% and 5.0% added senile anti-dementia patients were administered for 4 weeks to evaluate the production of neurotransmitters and related enzymes. The animal breeding room is automatically controlled with a 12-hour cycle (06: 00-18: 00) under constant temperature and humidity (22 ± 2 ℃, 65 ± 2% RH).

(2) The furtherance of senile antidementia patient meal

The feed composition used in this experiment was mixed with 29 g maltodextrin as carbohydrate, 6.0 g sunflower oil and MCT oil as lipid, 7.5 g sodium caseinate and 7.5 g soy protein as protein. Vitamins A (77.78 μgRE), D ₃ (0.56 μg), E (1.12 mgα-TE), C (6.12 mg), Folic acid (27.78 μg), Niacin (1.89 mg NE), Biotin (7.78 μg) and minerals Ca (90 mg), P (90 mg), Fe (1.34 mg), Mg (38.89 mg), K (254 mg), Na (149 mg) were added. 200 ml of this can is prepared so that the total amount of energy is 200 kcal.

2. Experimental materials

(1) Materials for the composition of antidementia patients

Pretreatment of silk protein with distilled water, such as salt removal, and addition of flavouryme (Novo Co., Denmark) at a concentration of 1% as a protease, silk at molecular weights of 250, 500 and 1,000 depending on the time of hydrolysis at 55 ° C for 2 hours. Among the fibroins, silk fibroin having a molecular weight of 500 with the highest bioactivity in the experiment of Table 1 and silkworm powder obtained by lyophilization of silkworms at 5 weeks of age were mixed at a ratio of 2: 1 (w / w), This mixture was added to Greenbia DM to 1.0%, 3.0% and 5.0%, and mixed and prepared to 200 ml with water.

(2) Reagent

As reagents used in this experiment, reagents such as acetylcholine (ACh) and choline acetyl transferase (ChAT), and other analytical reagents, all used special reagents made by Sigma.

3. Experimental method

(1) Administration of antidementia patient meal

Crj / Bgi rats (male, 180 g) were treated with a mixture of silk fibroin (SF) and silkworm powder (SWP) on a substrate of Greenbia diabetes millitus (SF-SWP 2: 1, w / w). Addition of 1.0%, 3.0%, and 5.0%, respectively, into a bottle of senile anti-dementia patients made from 200 ml of cans, followed by free intake for 4 weeks, inhibition of free radicals and measurement of neurotransmitters and related enzymes. The antidementia effect was evaluated. However, since rats (200 g body weight) consume about 20 ml of liquid per day, the daily intake of a mixture of silk fibroin and silkworm powder in antidementia patients per day corresponds to 100, 300 and 500 mg, respectively.

(2) Determination of hydroxy radicals (OH) and oxidized proteins

The production of the most powerful hydroxyl radical (OH) among free radicals was quantified according to the method of Chan et al. (1976), and the production of oxidized protein (OP) by the attack of free radicals by oxidative stress is Levine et al. According to the method of 1990), the amount of carbonyl group was measured and quantified.

(3) Determination of activity of the eliminating enzyme

The activity of superoxide dismutase (SOD), the most important free radical scavenger enzymes present in all aerobic cells in vivo, was measured and quantified according to the method of Oyanagui et al. (1984), and catalase : CAT) activity was measured and quantified according to the method of Rigo et al. (1997).

(4) measuring acetylcholine (ACh) and related enzymes

Acetylcholine (ACh) was measured based on the reaction of O-acyl derivatives with alkaline hydroxylamine by the method of Hestrin et al. (1949), and of choline acetyltransferase (ChAT) in brain cells. The measurement was performed by adding and mixing 0.2 ml of 0.2 mM accetyl-CoA, 10 mM choline chloride, 0.2 M KCl, and 10 mM potassium phosphate with 1 mM EDTA to the total volume of 0.8 ml according to the method of Ellman (1959), and then choline acetyltransferase was added. Add 1 μl, 100 μl of cytosol, 190 μl of 1M tris buffer, and 5 μl of DTB (5,5'-dithiobis-2-nitrobenzoate) in 5 μl and 250 μl of 20% TCA (trichloroacetic acid). Using a photometer, the activity of ChAT was measured at a wavelength of 412 nm at 2 minute intervals.

4. Evaluation of Experimental Results

(1) inhibitory effect on free radicals and oxidative stress

Crj / Bgi rats containing 200 ml canned anti-dementia patients prepared by adding 1.0%, 3.0%, and 5.0% of silk fibroin-silk powder mixture (2: 1, w / w) to Greenbia MC. The results of the analysis on the effects of free radicals and oxidative stress on administration for 4 weeks are shown in Table 2. It was found that not only can the production of hydroxy radicals (.OH), the most powerful active oxygen, be inhibited very effectively, but also the production of oxidized proteins due to the attack of active oxygen can be suppressed very effectively. Therefore, as well as preventing the damage of neurotransmitters is expected to be very effective in inhibiting adult diseases and aging.

SF. SWP-1.0, 3.0, 5.0 administration group: daily silk fibroin (SF)-silkworm powder (SWP) 100, 300, 500 mg intake;

^a mean ± SD; ^b % of control; Significance test: compared with control group ^* p <0.05; ^** p <0.01; ^*** p <0.001.

(2) Evaluation of activity of free radical scavenger

Developmental dementia patients were placed in a water bottle and administered to Crj / Bgi rats for 4 weeks to compare the activity of scavenging enzymes against free radicals.

The effect of superoxide dismutase (SOD) activity on the active oxygen scavenging enzyme, the most powerful active oxygen, was found to be 2.88 ± Cu / Zn-SOD activity of SF.SWP-3.0 and SF.SWP-5.O group. As the 0.23, 3.06 ± 0.09 U / mg protein, the effect of 12 and 18% of the active oxygen scavenging enzyme of 12 and 18%, respectively, compared to the Cu / Zn-SOD activity of the control group (2.59 ± 0.08 U / mg protein: 100%) was recognized.

In addition, the effects of catalase (CAT) activity on the activity of catalase (CAT) as active oxygen scavenging enzymes such as hydroxy radicals and hydrogen peroxide, etc. As the ± 0.08 and 1.12 ± 0.04 μmol / ml serum, 16 and 18% of the significant active oxygen scavenging enzyme activities were recognized, respectively, compared to the CAT activity of the control group (0.95 ± 0.01 μmol / ml serum: 100%). Therefore, the development of these anti-dementia patients will not only prevent adult disease and prevent aging by inhibiting free radicals and increasing the activity of scavenger enzyme, but also effectively prevent senile dementia by effectively preventing neurotransmitter damage. It is expected.

SF.SWP-1.0, 3.0, 5.0 administration group: 100, 300, 500 mg intake of silk fibroin (SF) -silk powder (SWP) per day;

^a mean ± SD; ^b % of control; Significance test: compared with control group ^* p <0.05; ^** p <0.01; ^*** p <0.001.

(3) Evaluation of the activity of neurotransmitters and related enzymes

Developmental dementia patients were placed in water bottles and administered to Crj / Bgi rats for 4 weeks to synthesize acetylcholine (ACh), a major neurotransmitter, and choline acetyltransferase (CHAT) as a synthase of ACh. ) And the activity of acetylcholinesterase (ACHE) directly related to neurotransmission.

The amount of ACh synthesized in the SF, SWP-3.0 and SF.SWP-5.0 administration groups as antidementia patients was 39.35 ± 1.95 and 40.12 ± 2.90 U / mg protein, respectively, and the amount of ACh in the control group (36.04 ± 2.19 ng / mg protein). : Significantly increased the amount of ACh synthesis of 9.2% and 12.3%, respectively. In addition, the activity of ChAT as a synthase of ACh was increased in dose-dependent manner, but the significance was significantly increased by 12.5% of ACh synthase ChAT activity compared to the control group only in SF.SWP-5.0 administration group. On the other hand, the effect of AChE directly related to neurotransmission in the antidementia patient SF / SWP group also increased AChE activity in a dose-dependent manner of the SF / SWP group, especially the activity of AChE in the SF / SWP-5.0 group. The activity of AChE was significantly increased by about 10% compared to the control group.

The development of these anti-dementia patients can effectively prevent senile dementia through the effective expression of neurotransmitter through the synthesis of neurotransmitters as well as the effect of inhibiting free radicals and oxidative stress and increasing the activity of scavenger enzyme. In addition, it has been demonstrated that it can promote neurotransmission. Therefore, it is expected that the anti-dementia patients may not only prevent adult disease and aging, but also effectively prevent senile dementia.

Effect of Antidementia Patients on the Neurotransmitter ACh and Related Enzymes

SF.SWP-1.0, 3.0, 5.0 administration group: 100, 300, 500 mg intake of silk fibroin (SF) -silk powder (SWP) per day;

^a mean ± SD; ^b % of control group; Significance test: compared with control group ^* p <0.05; ^** p <0.01.

Example 1

Silk fibroin of molecular weight 500 obtained by enzymatic hydrolysis of silk protein and silkworm of 5 days and 3 days were lyophilized and mixed at 2: 1 (w / w), and then, 5 parts by weight of the mixture was used as a function enhancer. 0.2 parts by weight of tyrosine and tryptophan were respectively added, and 0.02 parts by weight of licorice and cinnamon extract were added to purify the odor, and 0.05 parts by weight of citric acid and 0.05 parts by weight of vitamin C were added as a flavor and flavor enhancer, and taurine as an anti-fatigue agent was added thereto. After 0.1 parts by weight was added and mixed, a functional anti-dementia patient was prepared in 200 ml of can packaging.

The present invention is the most important acetylcholine (acetylcholine (ACh) among the neurotransmitters of silk fibroin (silk fibroin) in order to create an anti-dementia patient that can prevent and prevent the dementia of the elderly, the most important cause of poor quality of life As a related enzyme, choline acetyltransferase (CAhT), a synthetase, and acetylcholinesterase (AChE), which are directly involved in neurotransmission, are mainly used for silkworm powder. Since the anti-dementia patient formula was prepared by combining geriatric disease and the inhibitory effect of aging, the beverage of the present invention is effective for the health of dementia patients and diabetics.

Claims (1)

A freeze-dried silk fibroin of molecular weight 500 obtained by enzymatic hydrolysis of silk protein and silkworms of 5 days and 3 days were mixed at 2: 1 (w / w), and then, 5 parts by weight of the mixture was used as a function enhancer. 0.2 parts by weight of tyrosine and tryptophan were respectively added, and 0.02 parts by weight of licorice and cinnamon extract were added to purify the odor, and 0.05 parts by weight of citric acid and 0.05 parts by weight of vitamin C were added as a flavor and flavor enhancer, and taurine as an anti-fatigue agent was added thereto. An anti-dementia patient composition using a silkworm product, which is prepared by adding and mixing 0.1 parts by weight, and then, in a 200 ml can packaging.