KR102333879B1 - A hyperlipemia improvement health functional food composition containing hot water extract of Ulmus macrocarpa Hance and Moringa as an active ingredient and its preparing method - Google Patents

Abstract

The present invention relates to a hyperlipidemia-improving health functional food composition comprising a hot water extract of a mixture of King Elm and Moringa as an active ingredient, and a method for manufacturing the same, and more particularly, enzymatically treating and freezing the hot water extracted King Elm and Moringa extract. By providing a health functional food composition containing an active ingredient and a method for producing the same, a mixed extract of King Elm and Moringa produced by drying and powdering, there is no cytotoxicity using a natural hot water extract, and the ability to inhibit fat accumulation in hepatocytes and It can control the expression of triglyceride synthesis and degradation-related genes, can be involved in the regulation of blood total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol concentration, and is a protein for the synthesis and degradation of total cholesterol and triglycerides in liver tissue It has the effect of regulating their expression.

Description

A health food composition for improving hyperlipidemia comprising a mixed extract of King Elm and Moringa as an active ingredient, and a method for preparing the same {A hyperlipemia improvement health functional food composition containing hot water extract of Ulmus macrocarpa Hance and Moringa as an active ingredient and its preparing method}

The present invention relates to a functional health food composition for improving hyperlipidemia, and more particularly, to a functional health food composition for improving hyperlipidemia comprising a mixed extract of king elm and moringa as an active ingredient, and a method for manufacturing the same.

Fats in the blood can be divided into four types: total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglycerides. Low-density lipoprotein cholesterol (LDL cholesterol) is the bad cholesterol that accumulates on the walls of blood vessels and causes arteriosclerosis, which causes cardiovascular disease and cerebrovascular disease. Cholesterol contained in high-density lipoprotein is known as good cholesterol because it has the effect of preventing arteriosclerosis. Total cholesterol is a name that combines low-density lipoprotein cholesterol and high-density lipoprotein cholesterol into one, and triglycerides are created to store excess energy ingested from food. fat used.

Hyperlipidemia is a condition in which the concentrations of cholesterol, triglycerides, phospholipids and free fatty acids in the blood are abnormally increased. (hypercholesterolemia) is known to cause atherosclerosis. Atherosclerosis is a clinically important problem because atherosclerosis is caused by ischemic heart disease, angina pectoris, and myocardial infarction by accumulating thick lipids along the blood vessel wall and reducing blood flow (Korean J. Food Sci. Technol. 2003. 35:720- 725). Therefore, as a part of treatment and prevention measures to lower the level of these lipids in the blood, interest in health functional foods is being gathered.

Ulmus macrocarpa Hance is a deciduous broad-leaved tree, and its origin is Korea, Russia, and Mongolia. It is about 20 m high and about 80 cm in diameter. The bark is black brown, shallowly cracked, and the small branches are yellowish green, drooping down and hairy but disappearing. Leaves are alternate, long oval, flowers bloom in April - May, and bear fruit in May - June.

Moringa refers to a plant of the genus Moringa belonging to the family Moringaceae, which is one of the angiosperms belonging to the order Cruciferous. The moringa includes Moringa oleifera and Moringa stenopetala. Moringa is a plant native to India and Africa with a height of 5 to 10 m and has white flowers. The seeds are like beans, and 7 to 20 seeds exist inside the pod. carotene), and in southern Asia, all parts of plants, such as roots, bark, gum, leaves, pods, flowers, and seeds, have various diseases including inflammation and infectious diseases. It has been used as a folk remedy to treat asthma, and among them, it was reported that asthma symptoms and lung function indicators improved when moringa seed powder was drunk with water for asthma patients, and guinea pigs It has also been reported to have anti-asthmatic effects in animal models.

Recently, research for developing functional foods or pharmaceutical compositions using plant extracts is being actively conducted. For example, Korean Patent Application Laid-Open No. 2014-92944 discloses a drug or health food using an extract obtained by extracting Yugeun skin with ethyl alcohol. A technology for a pharmaceutical composition for the prevention or treatment of obesity comprising a natural complex as an active ingredient is disclosed, and domestic registration patent No. 678519 discloses sageulgi, sulfur duck, eugeunpi, pineflower powder, mountain cheongmok, bamboo salt, wormwood, bellflower, and black bean. Disclosed is a composition for preventing and treating fatty liver comprising a complex extract consisting of Korean Patent Publication No. 2018-50634 discloses a health functional food composition for enhancing immunity containing alcohol extract of elm bark as an active ingredient, and Korean Patent No. 1931556 discloses unripe strawberry extract and milkweed of king elm The invention relates to a pharmaceutical composition for the prevention or treatment of gastrointestinal diseases comprising a mixture of blood extracts as an active ingredient, and Domestic Patent No. 2026409 discloses an invention related to a composition for skin regeneration using milk white skin powder. have.

In addition, Korean Patent Publication No. 2015-0127442 discloses a pharmaceutical composition for preventing and improving obesity comprising an active ingredient of Moringa oleifera leaves, and Korean Patent Publication No. 2015-144969 discloses a mother It is disclosed that the linga leaf extract has an effect of improving or preventing cancer, and Korean Patent Publication No. 2019-1089492 discloses that the extract of moringa leaf is effective in relieving alcoholic fatty liver. However, it has not been reported so far that Moringa is effective in the prevention or treatment of hyperlipidemia or that a mixed extract of King Elm and Moringa is effective in hyperlipidemia.

As a result of many years of research on health food for the improvement of hyperlipidemia, the present inventors have found that the extract obtained by putting King elm in purified water and hot water extraction has an effect of regulating the expression of total cholesterol and triglyceride synthesis and degradation-related proteins in liver tissue. After confirming the facts, the invention of "Healthy functional food composition for improving hyperlipidemia comprising hot water extract of King elm tree as an active ingredient and a method for manufacturing the same" was applied for as Patent Application No. 2018-157935 (2018.12.10.), After several research tests based on the patent application of

1. Domestic Patent Publication No. 2014-92944 1. Domestic Patent Publication No. 2014-145666 1. Domestic Patent Publication No. 2015-0127442 1. Domestic Patent Publication No. 2018-50634 1. Domestic Patent Publication No. 2015-0144969 1. Domestic Patent Publication No. 2019-1089492 1. Domestic Registered Patent No. 0678519 1. Domestic Registered Patent No. 2026409 1. Domestic Registered Patent No. 1931556

An object of the present invention is to provide a hyperlipidemia-improving health functional food composition comprising a mixed extract of king elm and moringa as an active ingredient.

In addition, the present invention is a mixed extract obtained by adding 500 to 1000 parts by weight of distilled water to 100 parts by weight of the mixed and pulverized product and then pulverizing the king elm and Moringa in a ratio of 9: 1 or 8: 2 to extract hot water. An object of the present invention is to provide a functional food composition for improving hyperlipidemia comprising as an active ingredient.

In addition, an object of the present invention is to provide a health functional food composition effective in preventing or improving hyperlipidemia derived from natural products without side effects even when taken for a long time.

In order to achieve the above object, the present invention is 75 to 98 parts by weight of 500 to 1000 parts by weight of purified water with respect to 100 parts by weight of a mixed pulverized product pulverized by mixing king elm and moringa in a weight ratio of 9: 1 to 8: 2 In order to extract it at ℃ to prepare it as a health functional food composition,

Drying the king elm and moringa at room temperature (20 - 25°C) and then chopping;

mixing the chopped King Elm and Moringa in a weight ratio of 9: 1 to 8: 2;

adding 100 parts by weight of the chopped king elm tree and moringa mixture and 500 to 1000 parts by weight of purified water to an extractor;

generating a mixed extract by heating and extracting the chopped King Elm and Moringa in the extractor for 6-8 hours;

Filtering the mixed extract to produce an enzyme-treated solution by inactivation for 10 minutes after an enzyme reaction for 2 hours;

Filtering the enzyme-treated liquid and concentrating under vacuum conditions to produce a concentrated liquid;

sterilizing the concentrate for 30-40 minutes and then freeze-drying; and

pulverizing the freeze-dried mixed concentrate; It is characterized by going through

In the present invention, the mixed extract pulverized after mixing King Elm and Moringa is added to purified water and extracted with hot water, so that the mixed extract has no cytotoxicity, and the ability to inhibit fat accumulation in hepatocytes and the expression of genes related to triglyceride synthesis and degradation can be involved in the regulation of blood total cholesterol, triglyceride, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol concentration, and can regulate the expression of total cholesterol and triglyceride synthesis and degradation-related proteins in liver tissue. have

1 is a graph showing the cell viability of a mixed extract according to Examples and Comparative Examples.
2 is a micrograph showing the degree of inhibition of fat accumulation in HepG2 hepatocytes according to Examples and Comparative Examples.
3A is a graph showing the comparison of the expression level of Acetyl-CoA Carboxylase1 (ACC1), a triglyceride synthesis-related gene, in HepG2 hepatocytes according to Examples and Comparative Examples.
3B is a graph showing the comparison of the expression level of SREBP1c, a triglyceride synthesis-related gene, in HepG2 hepatocytes according to Examples and Comparative Examples.
3c is a graph showing the comparison of the expression level of cholesterol biosynthesis-related gene HMG-reducatase (HMGCR) in HepG2 hepatocytes according to Examples and Comparative Examples.
3D is a graph showing the comparison of the expression level of AMPK, a triglyceride synthesis-related gene, in HepG2 hepatocytes according to Examples and Comparative Examples.
Figure 4a is a graph showing the comparison of the concentration of AST, a hepatic toxicity index in the blood of rats according to Examples and Comparative Examples
Figure 4b is a graph showing the comparison of the concentration of ALT, an indicator of hepatic toxicity in the blood of rats according to Examples and Comparative Examples.
5A to 5D are graphs showing comparison of the contents of total cholesterol, triglyceride, LDL-cholesterol, and HDL-cholesterol in the blood serum of rats according to Examples and Comparative Examples.

Some terms used in the present invention are defined as follows. "Active ingredient" means a substance or group of substances expected to directly or indirectly express the efficacy and effect of a health functional food, including the main ingredient, and "improvement" means taking the health functional food composition It means all actions that improve the symptoms of hyperlipidemia through In addition, "health functional food" means food manufactured and processed using raw materials or ingredients useful for the human body in accordance with Health Functional Food Act No. 6727, and is a nutrient that is easily deficient in daily meals. A food that helps maintain health by using raw materials or ingredients that have useful functions for the human body (hereinafter referred to as "functional raw materials"). The Ministry of Food and Drug Safety evaluates scientific evidence such as animal tests and human application tests to recognize functional raw materials, and refers to products made with these functional raw materials.

"Functional" means ingestion for the purpose of obtaining useful effects for health purposes, such as regulating nutrients for the structure and function of the human body or physiological action. Various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloids It may contain thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like, and may contain pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. The components may be used independently or in combination.

According to another aspect of the present invention, in extracting from the pulverized mixture of king elm and moringa, drying the king elm and moringa at room temperature to mince and powder; Putting the powdered King Elm and Moringa mixed powder and purified water into an extractor: heating and extracting the powdered King Elm and Moringa mixed powder in the extractor for 6 hours to produce an extract; filtration of the extract, enzymatic reaction for 2 hours, and inactivation for 10 minutes to produce an enzyme-treated solution; Filtering the enzyme-treated liquid and concentrating under vacuum conditions to produce a concentrated liquid; sterilizing the concentrate for 30 minutes and then freeze-drying; And it provides a hyperlipidemia improvement health functional food composition comprising a mixed extract of king elm and moringa as an active ingredient, characterized in that it comprises the step of powdering the freeze-dried concentrate.

The enzyme is at least one selected from the group consisting of a starch degrading enzyme and a pectinase. A typical starch degrading enzyme is an amylase, and preferably, it may be one or more selected from the group consisting of α-amylase, β-amylase, β-gluconase, isoamylase, glucoamylase, and oligosaccharide amylase. Pectinase is one of the enzymes that hydrolyze glycosidic bonds, and is distributed in microorganisms such as bacteria, mold, yeast, higher plants, and reptiles. Preferably, it may be one or more selected from the group consisting of pectinase, polygalacturonase, and pectinesterase, which are enzymes involved in the degradation of pectinyl.

In the case of freeze-drying after general hot water extraction, the drying process is carried out in a concentrated state as much as possible in consideration of manufacturing cost and workability, and this is mainly done in the vicinity of 25brix to 30brix. In the case of King Elm, it is difficult to concentrate the concentration of the concentrate more than 5brix because the concentrate is a viscous substance (combination of starch and pectin), and the manufacturing cost increases when freeze-drying after simple extraction and concentration process. It is sold in the form of simple powder or extract, not in the form of concentrating and processing the mixed extract of Moringa and Moringa. For this reason, by decomposing and concentrating the viscous material using an enzyme, it is possible to concentrate the concentration of the concentrate by 20 brix or more, so that it is possible to lower the drying cost by three times or more, as well as to increase the efficiency of concentration and drying.

Hereinafter, specific contents for carrying out the present invention will be described in more detail through examples and the like. All terms used herein, including technical or scientific terms described in the present invention, have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs, and are not explicitly defined in this application. unless otherwise construed in an idealistic or overly formal sense. In addition, the embodiments according to the present specification may be modified in various other forms, and the scope of the present specification is not to be construed as being limited to the embodiments described below. The embodiments of the present specification are provided to more completely explain the present specification to those of ordinary skill in the art.

<Example 1> Preparation of a mixed extract of King Elm and Moringa

After naturally drying Cortex Ulmus macrocarpa Hance and Moringa, finely mince it and powder it. Then, using an electronic scale (CAS), the mixed powder of Cortex Ulmus macrocarpa Hance (CAS) is quantified at a ratio of 9: 1 did After that, purified water was put into the extractor according to the mixing ratio (5 times the weight of the mixed powder of King Elm and Moringa), the extraction temperature was set to 95°C, and the purified water was circulated while heating/extracting for 6 hours. After filtering the extract and transferring it to the enzyme reactor, the enzyme (0.4 % Viscozyme L, Novozymes, Denmark) was added according to the mixing ratio, and after 2 hours of enzymatic reaction, it was inactivated at 100° C. for 10 minutes. The enzyme-treated solution was filtered using a 0.1 μm mesh filter, concentrated under vacuum and reduced pressure, and then sterilized at 90°C or higher for 30 minutes. got angry

<Example 2> Preparation of a mixed extract of King Elm and Moringa (8: 2)

A mixed extract of King Elm and Moringa was prepared under the same conditions as in Example 1, except that the ratio of the powder mixture of King Elm and Moringa was 8:2.

<Comparative Example 1> Preparation of extract of King elm tree

An elm extract was prepared in the same manner as in Example 1, except that in Examples 1 and 2, only King elm powder was used instead of the mixed powder of King elm and Moringa.

<Animal Experiment>

Five-week-old 120-150 g male rats [Central Laboratory Animals] were purchased and acclimatized for 7 days, followed by animal experiments. Rats were bred in a constant environment of 24 ± 1 °C constant temperature, 50% humidity and constant humidity and a 12-hour photoperiod, and each control substance was administered. was put in The normal control group was fed a basic diet, not a hyperlipidemia-inducing diet. The control group and the positive control group were designed as shown in Table 3 below, and feed and drinking water were freely ingested for 6 weeks during the experiment.

animal testing classification test substance treatment concentration
(mg/kg) Experiment
number of animals (n) diet control Normal (N) - - 10 basic diet Control (C) - - 10 hyperlipidemia induction diet experimental group UME (Umbrella Elm Extract) 100 10 hyperlipidemia induction diet UM1 King Elm Extract Moringa Mixed Extract (9:1) 100 10 hyperlipidemia induction diet UM2 King Elm Extract Moringa Mixed Extract (8:2) 100 10 hyperlipidemia induction diet

< Experimental example 1> Cytotoxicity evaluation

Example 1 (UM1), Example 2 (UM2), Comparative Example 1 and MTT Assay System to confirm the cytotoxicity when untreated, the degree of change in cell proliferation for each sample was calculated as a percentage was indicated.

Specifically, it was found that the cell viability was high when the mixed extract of King Elm and Moringa was treated at a ratio of 8: 2, but when the mixing ratio of King Elm and Moringa was 9: 1, It was confirmed that extracts of elm only also did not have a significant effect on cell viability, and none of them had significant cytotoxicity, and the results are shown in FIG. 1 .

★ In Fig. 1, UME: King elm extract, UM1: Mixed extract of 9:1 ratio of King elm and Moringa, UM2: Mixed extract of 8:2 ratio of King elm and Moringa, 0 is radish It means the treated state (purified water).

<Experimental Example 2> Fat accumulation inhibitory ability analysis

Oil Red O staining was used to measure and compare the fat accumulation inhibitory ability of king elm extract, a mixed extract of king elm and moringa (9: 1, 8: 2), and the results are shown in FIG. 2 . The staining state of the cells was confirmed under a microscope, and as fat accumulation increased, the degree of Oil Red O staining increased.

As a result, fat accumulation did not occur in hepatocytes without any treatment (normal control group), and significant fat accumulation occurred in the positive control group without any treatment after fat accumulation. In addition, the fat accumulation inhibitory effect was the highest in cells treated with a mixed extract of King Elm and Moringa (8:2).

<Experimental Example 3> Expression regulation of triglyceride synthesis and degradation related genes

Acetyl-CoA Carboxylase1 (ACC1), SREBP1c, HMG-reducatase (HMGCR), which are genes related to triglyceride synthesis in hepatocytes of mixed extracts of King Elm and Moringa prepared according to Examples and Comparative Examples, and triglycerides involved in decomposition In order to measure the expression regulation of the AMPK gene, it was measured using a real time PCR analysis method, and the results are shown in FIGS. 3A to 3D .

ACC1 is an enzyme involved in the synthesis of malonyl-CoA from acetyl-CoA and is known as an enzyme that regulates the rate of fatty acid synthesis. When the expression of ACC1 gene decreases, fatty acid synthesis is reduced. SREBP-1c is a transcription factor that plays an important role in the synthesis of triglycerides in liver tissue. When the expression of the SREBP-1c gene is suppressed, fatty acid synthesis is reduced, and as a result, triglyceride accumulation in liver tissue is suppressed.

In hepatocytes, HMG-reducatase is involved in the cholesterol biosynthesis process, and when the expression of HMGCR gene is reduced, it has the effect of lowering the concentration of blood cholesterol. It has the effect of inhibiting the synthesis of triglycerides by reducing the phosphorus SREBP-1 expression.

It is also known to inhibit the expression of acetyl-CoA carboxylase (ACC), a target gene of AMPK and a rate-regulating enzyme in the fatty acid synthesis process.

As shown in Figures 3a to 3d, the expression of each of the ACC1, SREBP1c, and HMGCR genes was significantly increased in the positive control group compared to the normal control group, and the mixed extract of King Elm and Moringa (9:1, 8) :2) was treated with OA, and the group treated with a mixed extract of King Elm and Moringa (8:2) was treated with a King Elm extract and a mixed extract of King Elm and Moringa (9:1). It showed the effect of suppressing the expression of ACC1, SREBP1c, HMGCR gene and increasing the expression of AMPK gene than one group.

Therefore, when the mixing ratio of King Elm and Moringa was 8: 2, the expression of ACC1, SREBP1c, and HMGCR genes was reduced compared to the case of King Elm extract alone or the mixing ratio of King Elm tree and Moringa was 9: 1. It was confirmed that the effect of suppressing and increasing the expression of the AMPK gene was excellent.

<Experimental Example 4> Rat blood analysis: hepatic toxicity index AST analysis

ALT, which is an indicator of hepatic toxicity in the blood of rats, was analyzed and the concentration thereof is shown in FIG. 4 .

As a result, the group administered with the king elm and moringa mixed extract (8:2) had a lower AST concentration than that of the king elm extract, which is used as an indicator of hepatic toxicity in the blood. It was also measured lower than the group fed the mixed extract of linga (9:1).

Therefore, it was possible to confirm the effect of improving the fatty liver of the mixture of King elm and Moringa extracts of Example through a significant decrease in the blood of the rats administered the mixed extract of King elm and Moringa.

<Experimental Example 5> Rats blood analysis: hepatic toxicity index ALT analysis

ALT, which is an indicator of hepatic toxicity in the blood of rats, was analyzed and the concentration thereof is shown in FIG. 5 .

As a result, in the group administered with the mixed extract of King Elm and Moringa (8:2), the concentration of ALT, which is used as an indicator of hepatic toxicity in the blood, was higher in the concentration of King Elm extract and the mixed extract of King Elm and Moringa (9: 1) showed a significant reduction effect compared to the group fed.

<Experimental Example 6> Analysis of cholesterol content in rat blood

The content of total cholesterol, triglyceride, LDL-cholesterol, and HDL-cholesterol in the blood serum of rats was analyzed and shown in FIG. 6 .

As a result, the group administered with a high cholesterol diet and a mixed extract of King Elm and Moringa (8:2) was administered with a King Elm extract or a mixed extract of King Elm and Moringa (9:1). Compared to that, the serum triglyceride and LDL-cholesterol content was decreased, and the HDL-cholesterol content was increased.

According to these results, it was found that the mixed extract of king elm and moringa (8:2) was more effective in inhibiting fatty liver progression than the extract of king elm or the mixed extract of elm and moringa (9:1).

From the above experimental results, the mixed extract of King elm and Moringa of the present invention (8: 2) has neutral lipids and It was confirmed that it was effective in preventing and treating fatty liver and hyperlipidemia by reducing the LDL-cholesterol content, increasing the HDL-cholesterol content, and controlling the synthesis and decomposition mechanism of triglycerides, fatty acids and cholesterol.

Claims (9)

The extract obtained by extracting the mixed powder of 80% by weight of King Elm and 20% by weight of Moringa with hot water was filtered, followed by an enzyme reaction for 2 hours, and then inactivated for 10 minutes. , A health functional food composition for improving hyperlipidemia comprising, as an active ingredient, a mixed extract of King Elm and Moringa, which is powdered by sterilizing the concentrate for 30-40 minutes and then freeze-drying. delete delete According to claim 1,
A health functional food composition for improving hyperlipidemia comprising a mixed extract of king elm and moringa as an active ingredient, characterized in that the enzyme is any one selected from the group consisting of starch-degrading enzymes or pectin-degrading enzymes. Drying king elm and moringa at room temperature, then shredding and powdering;
mixing the chopped King Elm and Moringa;
adding 100 parts by weight of the powdered king elm and moringa mixture and 500 to 1000 parts by weight of purified water to an extractor;
Heating the chopped King Elm and Moringa in the extractor at 75 to 98° C. for 6 - 8 hours and extracting the extract;
Filtering the extract, enzymatic reaction for 2 hours, and then inactivating for 10 minutes to produce an enzyme-treated solution;
Filtering the enzyme-treated liquid and concentrating under vacuum conditions to produce a concentrated liquid;
sterilizing the concentrate for 30-40 minutes and then freeze-drying;
pulverizing the freeze-dried concentrate; as
A method for producing a health food composition for improving hyperlipidemia comprising a mixed extract of king elm and moringa as an active ingredient, characterized in that it consists of. delete delete 6. The method of claim 5,
The mixed extract is for improving hyperlipidemia comprising a mixed extract of King Elm and Moringa as an active ingredient, characterized in that 500 to 1000 parts by weight of purified water are mixed and extracted with respect to 100 parts by weight of the mixed pulverized product of King Elm and Moringa A method for producing a health food composition. 6. The method of claim 5,
The enzyme is one or more selected from the group consisting of starch-degrading enzymes and pectin-degrading enzymes. Method for producing a health food composition for improving hyperlipidemia comprising a mixed extract of king elm and moringa as an active ingredient.

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