KR101451755B1 - A composition for preventing bone metabolism-related diseases and increasing bone function comprising mix extracts of Mori Fructus and Aralia Continentalis - Google Patents

Abstract

The present invention relates to a composition for preventing or alleviating osteoporosis containing a mixed extract of Mori fructus and Aralia continentalis. The composition containing a mixed extract of Mori fructus and Aralia continentalis according to the present invention promotes proliferation of cells which affect bone metabolism, activates differentiation of osteoblast, and improves bone density in an animal test with ovariectomized mice which is an animal model of osteoporosis, thereby having an effect of preventing and treating diseases related to bone formation. Accordingly, the composition containing a mixed extract of Mori fructus and Aralia continentalis of the present invention can be used as a pharmaceutical medicine or a functional food for preventing bone metabolism-related diseases such as osteoporosis, osteopenia, osteomalacia, and calcium dysregulation, in other words, hypercalcemia or hypocalcaemia, and alleviating bone function.

Description

[0001] The present invention relates to a composition for preventing or improving osteoporosis, which comprises a mixed extract of a mental retarder and a chewing gum. [0002] The present invention relates to a composition for preventing or improving osteoporosis comprising Mori Fructus and Aralia Continentalis,

The present invention relates to a composition for preventing or ameliorating osteoporosis, which comprises a mixed extract of a remedy and a chewing gum. More specifically, the present invention relates to a composition for promoting and promoting cell proliferation and differentiation of mesenchymal stem cells and osteoblasts, And a composition for prevention or improvement of osteoporosis, which comprises a mixed extract of a topical and a non-essential oil having an effect of improving bone mineral density.

Bone tissue is a dense connective tissue surrounded by osteocyte and rigid calcium tissue around the osteocyte. It has the mechanical function of support and myofascial. Bone is formed by this bone tissue to form the skeleton of human body. It also functions to protect the organism and bone marrow, and to preserve calcium and phosphorus ions in the homeostasis. These bone tissues are composed of various kinds of cells such as collagen and glycoprotein, cell substrate and osteoblast, osteoclast, and osteocyte. Osteoblasts derived from bone marrow stromal cells play a major role in osteogenesis, and osteoclasts derived from hematopoietic stem cells are destroyed and are responsible for aging bone resorption. These osteoblasts and osteoclasts And bone remodeling through a balanced action of osteogenesis and bone resorption.

Bone metabolic disease occurs when the balance between osteoblasts and osteoclasts plays a major role in bone remodeling. As a representative example of bone metabolic diseases, there is osteoporosis. Osteoporosis increases osteoclast bone resorption activity of osteoblast compared with osteoblast-induced osteoblast, resulting in a decrease in total bone mass, resulting in fracture in mild shock Disease. To date, there is no effective treatment for osteoporosis cure and prevention is emphasized.

The osteoblasts responsible for osteogenesis originate from Mesenchymal Stem Cells. Mineralization, including calcium, formed by osteoblast differentiation can not only maintain the strength of the bones, And also plays an important role in the homeostasis of hormone metabolism. Calcium formation by osteoblast differentiation is regulated by vitamin D and parathyroid hormone, and it is known that bone morphogenetic protein (BMP), Wnt, MAP kinase, calcineurin-calmodulin Alkaline phosphatase (ALP), which is involved in osteoblast differentiation by cross-talk of various signaling systems such as calcineurin-calmodulin kinase, NF-κB and AP-1, After synthesis, osteopontin related to mineralization, osteocalcin, and type I collagen are known to be involved in bone formation by osteoblast differentiation.

In recent years, efforts have been made to prevent osteoclasts from being produced in vitro, to strengthen osteoblastic functions, or to treat bone damage. It is known that osteogenic supplement and Transforming Growth Factor beta (TGF-beta) subfamily play an important role in the formation and maintenance of bone tissue. In particular, BMP2 or BMP4 is added to the culture medium It has been reported that induction of differentiation into osteoblasts is increased. It is important to understand the factors and mechanisms involved in the induction of osteoclast differentiation in order to obtain a substance capable of performing clinically superior functions. In addition, it is very important to utilize the substance that regulates the activity of these substances to induce differentiation.

Accordingly, the present inventors have made intensive efforts to find a substance that not only proliferates osteoblasts but also induces differentiation. As a result, it has been found that a mixed extract of mature and unattached individuals is effective in cell proliferation and differentiation activity of mesenchymal stem cells and osteoblasts, The inventors of the present invention completed the present invention by confirming that there is an effect of improving the bone density in an animal test for an ovariectomized mouse which is an animal model.

Accordingly, a technical problem to be solved by the present invention is to provide a composition for preventing or improving osteoporosis.

In order to accomplish the above object, the present invention provides a composition for preventing or improving osteoporosis, which comprises a topical extract and a fragrant extract as an active ingredient.

The composition comprising the mixed extract of the extract of the present invention according to the present invention is effective for the osteoblast differentiation activity while promoting cell proliferation affecting the bone metabolism and is useful for the ovariectomized animal model, It has been confirmed that there is an effect of improving bone density, and it has a preventive and therapeutic effect on diseases related to bone formation. Therefore, the composition containing the mixed extract of the present invention as an active ingredient can be used as a bone metabolism-related disease such as osteoporosis, osteopenia, osteomalacia, calcium control abnormalities such as hypercalcemia, prevention of hypocalcemia, As a pharmaceutical preparation or a functional food.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments of the invention and, together with the description of the invention, It should not be construed as limited.
FIG. 1 shows the results of cell proliferation of C3H10T1 / 2 cells treated with mesenchymal stem cells.
FIG. 2 is a result of observing cell proliferation by treating MC3T3-E1 cells, an osteoblast, with a mixed extract of MB3 and MB3.
FIG. 3 shows the result of ALP (alkaline phosphatase) activity of the cells treated with C3H10T1 / 2 cells, which is a mesenchymal stem cell mixture, with a mixture of a mixture of a remedy and a chewing gum.
FIG. 4 shows the results of ALP activity of cells treated with MC3T3-E1 cells, osteoblasts, mixed extracts of the left and right cheeks.
FIG. 5 shows the results of real-time PCR for the gene expression levels of bone-associated gene markers Alp, Runx2, and Bglap (Osteocalcin, Ocn ) in C3H10T1 / 2 cells.
FIG. 6 shows the results of real-time PCR for the gene expression levels of bone-associated gene markers Alp, Runx2, and Bglap (Osteocalcin, Ocn ) on the mixed extract of MB3T3 -E1 cells derived from differentiation-induced MC3T3-E1 cells .

The present invention relates to a composition for preventing or ameliorating osteoporosis, which comprises a topical extract and a fragrant extract as an active ingredient.

Mori Fructus used in the present invention is a black berries mulberry fruit. It has the effect of a nourishing lotion that it is said that the head is blackened when a lot of mulberry is eaten. It gradually turns from blue to purple to black when fully ripe. It is known as an ageless medicine that cleanses the blood and helps the kidneys and kidneys to regulate petechiae (diabetes), which is caused by depletion of the essence by showing the blood (阴血), dry mouth, tongue dryness and dizziness due to lack of adulthood , It has an effect on insomnia. It is also known as a medicinal product that is effective for constipation that softens the intestines and comes from lack of blood.

Aralia Continentalis is a perennial herb that belongs to Araliaceae, and is also called Aralia Continentalis , Aralia Continentalis , Kangchung, and Ganghwang. The juice is used for food and roots are used for medicinal purposes. Especially, it is effective for fever, tension, headache, toothache, neuralgia, arthralgia. The roots contain a large amount of diterpene pimaric acid derivatives as fat-soluble components and oleanolic acid or araloside as hydrazine saponin as a water-soluble component .

The composition for prevention or improvement of osteoporosis according to the present invention is characterized in that a mixed extract of a fresh mint and a pure mint is contained in a concentration of 1 ug / ml to 100 ug / ml in the whole composition.

If the concentration of the mixed extract of the above-mentioned drowsiness and obesity is less than 1 ug / ml in the total composition, it is difficult to expect a preventive effect and a bone function improvement effect of the desired bone metabolism-related diseases. If the concentration exceeds 100 ug / ml, Increasing the concentration is meaningless because it is not excellent.

In the present invention, the left and right cheeks can be mixed and extracted or may be separately extracted and mixed. Preferably, the extract is used as a mixture of 2: 1 and 1: 2 by weight, preferably 1: 1 by weight. do.

The bulk extract or fragrant extract of the present invention can be obtained by a process of drying and crushing the bulkhead or chewing gum and then extracting, followed by primary concentration and filtration, and then secondary concentration. For example, according to the method described in Korean Patent Registration No. 10-0169290, it is possible to obtain a mixed extract of fresh or dried fruit.

Preferably, the inventive extract or fragrance extract of the present invention can be obtained by a method comprising the following steps:

(S1) hot-air drying at 50 to 60 DEG C and then pulverizing to a size of 60 mesh or less;

(S2) adding an extracting solvent having a weight ratio of 6 to 10 to the pulverized bulkhead or chewing gum and extracting it at 70 to 80 ° C for 2 to 6 hours to produce an extract by repeating the process twice or three times;

(S3) centrifuging the obtained fresh or dried extract at 12,000 to 15,000 rpm for 10 to 15 minutes, and then concentrating the dried extract at a first reduced pressure at 60 to 80 ° C; And

(S4) filtration of the concentrated solution or concentrated extract, followed by concentration under reduced pressure, to obtain a fresh or concentrated extract.

In step (S2), the extraction solvent may be water, an aqueous solution of ethanol, or a mixture thereof. Particularly, it is preferable to mix water and an aqueous ethanol solution in a volume ratio of 1: 2 to 1: 3. The concentration of the aqueous ethanol solution is preferably 20 to 40%.

In the present invention, the bone metabolism activity of the mixed extracts of the above-mentioned MB and the non-MB was measured. Specifically, the mesenchymal stem cell C3H10T1 / 2 cell line and the osteoblast MC3T3-E1 cell line, The in vitro experiment was carried out by treating mixed extract of virulence.

Mesenchymal stem cells are cells that have the ability to differentiate into osteoblasts or adipocytes, and osteoblasts are cells before the differentiation into osteoblasts. In order to confirm the cell proliferation effect of the mixed extracts of the two types of cell lines, the MTT test was performed. To confirm the differentiation activity, the degree of Alkaline phosphatase (ALP) activity and osteoblast differentiation activity We investigated the changes of osteoblast differentiation gene markers at the molecular level.

In vivo experiments were performed to observe bone mineral density improvement after short - term administration (8 weeks) of mixed extracts of old and newborn rats to ovariectomized mice.

Since most osteoporosis patients are associated with estrogen deprivation induced by menopause in women, an ovariectomized animal model is the most suitable animal model for the development of health food for the prevention of osteoporosis.

According to the general embodiment of the present invention, it was confirmed that a composition containing a mixture of a fresh and a mixed extract promotes cell proliferation and differentiation of mesenchymal stem cells and osteoblasts. The MTT assay and the ALP activity And to increase the expression level of gene markers related to bone metabolism. In the animal experiment of ovariectomized mice, it was confirmed that bone mineral density was improved by short - term administration (8 weeks)

The composition containing the active ingredient of the present invention as an active ingredient may further comprise a suitable carrier, excipient and diluent commonly used in the production of a pharmaceutical composition.

Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition according to the present invention can be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like oral preparation, external preparation, suppository and sterilized injection solution according to a conventional method Can be used.

When the pharmaceutical composition according to the present invention is formulated into an oral formulation, an additional food coloring agent may be added to impart color. The amount thereof may be added in an amount of 0.1 to 1% by weight based on the total weight of the composition of the present invention.

The amount of the pharmaceutical composition of the present invention may vary depending on the age, sex, and body weight of the patient. However, in order to be administered in an amount of 1 to 300 mg / kg per day, Lt; / RTI > The dosage of the composition may also be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

In the present invention, bone disease refers to, for example, osteoporosis, osteopenia, osteomalacia, calcium-controlling abnormalities, that is, hypercalcemia and hypocalcaemia.

The present invention also provides a functional food composition comprising a composition effective for prevention of bone metabolic diseases and improving bone function and a pharmaceutically acceptable food-aid additive.

Since the composition of the present invention has little toxicity and side effects, it can be safely used for prolonged consumption even for prophylactic purposes.

The composition of the present invention may be added to foods or beverages for the purpose of preventing bone metabolic diseases and improving bone function. In this case, the functional food composition of the present invention in the food or beverage may be added at 0.01 to 15% by weight of the total food, and the health beverage composition may be added at 0.02 to 10 g, preferably 0.3 to 1 g, It can be added in a ratio.

The functional food composition of the present invention is not limited to any particular ingredient, except that it contains the composition of the present invention as an essential ingredient at the specified ratio, and may contain various flavors or natural carbohydrates or calcium as an additional ingredient ≪ / RTI > Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of the flavor other than those mentioned above include natural flavors (tautatin, stevia extract such as rebaudioside A and glycyrrhizin) and synthetic flavors (such as saccharin and aspartame), and calcium , Coral powder (weathered calcium carbonate) may be added. The proportion of the additive is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents such as, for example, , Alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. As the coloring agent, an edible pigment may be further added. The amount thereof may be added in an amount of 0.1 to 1% by weight based on the total weight of the food composition of the present invention. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The composition comprising the mixed extract of the recovered extract according to the present invention promotes the proliferation of cells affecting bone metabolism and induces osteogenic differentiation activity, thereby preventing and treating bone related diseases. Therefore, the composition containing the mixed extract of the present invention as an active ingredient can be used as a bone metabolism-related disease such as osteoporosis, osteopenia, osteomalacia, calcium control abnormalities such as hypercalcemia, prevention of hypocalcemia, As a pharmaceutical preparation or a functional food.

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

≪ Example 1 > Preparation of bulk extract

Prior to the purchase of a newborn, the drug product quality team of Dong-dang Pharmaceutical Co., Ltd. confirmed its suitability as a test substance through close examination based on pharmacopoeia, residual sulfur dioxide analysis, residual heavy metal measurement, residual pesticide analysis and so on.

1.0 kg of the spermatozoa were cleaned and dried by hot air at 60 ° C and pulverized to 60 mesh or less. Then, 1.8 l of water and 4.2 l of ethanol were added thereto, and the mixture was extracted at 80 ° C for 4 hours.

Subsequently, the supernatant extract was centrifuged at 15,000 rpm for 15 minutes, and then concentrated under reduced pressure until the water content became about 30% or less at about 70 ° C., filtered and then concentrated again to obtain a final concentration of 190.1 g < / RTI > At this time, the concentration of sickle cell extract was 85.3 brix.

≪ Example 2 > Preparation of fragrant extract

Prior to the purchase of a newborn, the drug product quality team of Dong-dang Pharmaceutical Co., Ltd. confirmed its suitability as a test substance through close examination based on pharmacopoeia, residual sulfur dioxide analysis, residual heavy metal measurement, residual pesticide analysis and so on.

0.95 kg of pure water was cleaned and dried at 60 ° C with hot air and pulverized to 60 mesh or less. Then, 1.995 L of water and 4.655 L of ethanol were added and extracted at 80 ° C for 4 hours to prepare a polyunsaturated extract.

Then, the viscous extract solution was centrifuged at 15,000 rpm for 15 minutes, and then concentrated under reduced pressure until the water content became about 30% or less at about 70 ° C., then filtered and then concentrated again to obtain a final concentration of 60 g Lt; / RTI > extract. At this time, the concentration of the extract was 60 brix.

Test Example 1 Cell culture

The mouse mesenchymal stem cell C3H10T1 / 2 cells and the osteoblast MC3T3-E1 cells were cultured in Basal Medium Eagle (Sigma) supplemented with 10% fetal bovine serum, 1 mM sodium pyruvate, 100 units / L penicillin and 100 mg / L streptomycin BME) and α-minimal essential medium (α-MEM) medium at 37 ° C and 5% CO ₂ .

In the case of MC3T3-E1 cells, osteogenic differentiation inducers such as ascorbic acid (50 ug / ml) and β-glycerophosphate (10 mM) were added to induce differentiation for 3 days.

The cells were treated with C3H10T1 / 2 cells and MC3T3-E1 cells, respectively, by mixing 10 ug / ml of the extract from the above extracts obtained in Examples 1 and 2 and 10 ug / ml of the extract with the extract, and further cultured for 48 hours. As a control group, cells cultured in the absence of a mixed extract of the left kidney and the right kidney were used.

≪ Test Example 2 > Cell proliferation assay [

Cell proliferation assays were performed using the EZ-Cytox Enhanced Cell Viability Assay Kit with water-soluble Tetrazolium salt.

Specifically, C3H10T1 / 2 cells and MC3T3-E1 cells ( ³ × 10 ³ cells / well) were inoculated on a 96-well plate and cultured in a growth medium for 24 hours at 37 ° C. in which 5% CO ₂ was supplied. 10 and 10 ug / ml of the extracts of the extracts were added to the cells and cultured for 24 hours. WSTs (Tetrazolium salt) were treated with each of the cultured cells and samples and cultured at 37 ° C for 2 hours. The absorbance was measured at 450 nm, and the reference wavelength was 655 nm, and the results are shown in FIGS. 1 and 2, respectively.

Statistical analysis was performed using the Student's t-test for the control group without extract treatment and the mixed extract treatment group for the sickle-tailed and unexposed group. The significance level was ^* p <0.05, ^** p <0.01 and ^*** p < 0.001.

As shown in FIGS. 1 and 2, no statistically significant difference was observed in the C3H10T1 / 2 cell line and the MC3T3-E1 cell line in all the groups treated with the mixed extract of the old and newborn.

<Test Example 3> ALP activity assay

Cell ALP activity was measured after culturing for 48 hours in an experimental group and a control group in which 10 ug / ml of the extract of the fresh and concentrated extracts obtained in Examples 1 and 2 was mixed with 10 ug / ml of the non-toxic extract. ALP activity was measured using TRACP and ALP Assay Kit.

Specifically, after cells were washed with physiological saline, the cells dissolved in the cells were treated with p-nitrophenylphosphate, which is an ALP substrate, and cultured at 37 ° C for 1 hour. The absorbance was then measured at a wavelength of 405 nm after addition of 0.5N NaOH as a reaction stop solution.

The effect of ALP activity on the cell extracts of C3H10T1 / 2 cells and MC3T3-E1 cells induced by differentiation was analyzed. The results are shown in FIG. 3 and FIG. Statistical analysis was performed with Student's t-test for the control group without extract treatment and with each extract treatment group. ^* P <0.05, ^** p <0.01 and ^*** p <0.001 depending on significance level .

As shown in FIG. 3, the C3H10T1 / 2 cell line showed a statistically significant difference in the group treated with the mixture of MB and extract, and it was confirmed that the ALP activity was increased.

As shown in FIG. 4, the results of the same experiment conducted on the MC3T3-E1 cell line showed that ALP activity was statistically increased in the mixed extract treatment group of the old and the new.

As described above, in both cell lines, it was confirmed that ALP activity was increased with a statistically significant difference in the experimental group treated with the mixed extract of MB and non-DM.

Test Example 4 Analysis of Osteoblast Differentiation Gene Marker Expression Level

Real-time PCR was used to confirm the gene expression level of three osteoblast-differentiated gene markers, Alp, Runx2 and Bglap (Osteocalcin, Ocn ) among the bone metabolism-related gene markers showing the change in the expression level upon osteoblast differentiation.

Total RNAs were isolated using Trizol after harvesting the control cells treated with the mixed extracts of control cells, dendritic cells and virulence. To remove genomic DNA, DNase I was reacted at room temperature for 15 minutes, treated with EDTA, I was inactivated.

Next, reverse transcription was induced using oligo dT primer as a primer to synthesize cDNA, and real-time PCR was performed using sequence specific primers of each gene and SYBR Green. For the analysis of the expression level, relative quantification of the gene of interest using the Gapdh gene was used. Gene-specific primer sequence information for confirming the expression level of the genetic marker used to confirm the degree of osteoblast differentiation is shown in Table 1 below.

The expression levels of three gene markers related to bone metabolism , Alp, Runx2 and Ocn , on the C3H10T1 / 2 cell and differentiated MC3T3-E1 cell extracts were compared, 5 and Fig. Statistical analysis was performed with Student's t-test for the control group without extract treatment and with each extract treatment group. ^* P <0.05, ^** p <0.01 and ^*** p <0.001 depending on significance level .

As shown in FIG. 5, in the C3H10T1 / 2 cell line, the expression levels of all three gene differentiation markers ( Alp, Runx2, Ocn ) were significantly increased.

As shown in Fig. 6, only the amount of Alp expression was statistically increased in the differentiated MC3T3-E1 cell line.

&Lt; Test Example 5 > Bone mineral density improvement of osteoporosis animal model

Four 5-week-old sham-operated ddY female mice and 16 ovariectomized ddY female mice were supplied from the Central Laboratory Animal Co., and received a purification period of 1 week in the quarantine room of Ajou University, Respectively. The weight of each mouse was measured and group separation was performed so that there was no statistically significant weight difference between the experimental groups. The stock solution was prepared at a concentration of 100 mg / ml in a mixture of 1: 1 mixture of diluted and untreated mixed extracts to be used in the experiment. To carry out the laboratory animal center, Soya Green Tech Co., And sterilization was carried out through gamma irradiation. Initial bone mineral density was measured with PIXImus bone densitometer prior to administration of mixed extracts. Bone mineral density (BMD) was measured by anesthetizing with 50 μl of Zoletil and Rompun 1: 2 mixed solution (diluted 2: 3 with physiological saline) , And Bone Mineral Density) were measured. From the next day after the bone mineral density measurement, the mixture of low-dose and high-dose mixed extracts was started to be free-flowing in the negative water so that the intake of 50 mg / kg, 150 mg / kg and 300 mg / kg, respectively. Negative bottles containing the extracts were replaced every 3 days, weighed once a week, and the cages were changed once a week. The final body weight was measured 8 weeks after the administration of the mixture of the left and right mixed rats. The results are shown in Table 2 below. BMD (Bone Mineral Density) was measured after injection anesthesia and the results are shown in Table 3 below.

According to the above Table 2, there was no statistically significant difference between the ovariectomized group and the old-fashioned and untreated mixed extract group. There was no difference between the groups in the initial body weight measurement and in the final body weight measurement after 8 weeks Student's t-test showed no significant difference. However, weight change was statistically significantly increased in the high dose group. As shown in Table 3, the BMD of the Sham group was significantly higher between the ovariectomy group and the Sham group than the initial BMD value. This tendency shows a larger difference between the final BMD and BMD changes. This fact indicates that the induction into an animal model of osteoporosis is good. Based on these facts, early BMD improvement was not significantly different from ovariectomized group, but the final BMD of low-dose group and mid-dose group was statistically significantly higher than that of ovariectomized group (P <0.05), but there was a significant trend (p = 0.097, p = 0.055) in the BMD variation.

As a result of the above experimental results, it was confirmed that the effect on the bone marrow stem cell line and the osteoblastic cell line affected the osteoblast differentiation activity while promoting cell proliferation, The results of the experiment with mice showed that bone density was improved. Therefore, it is considered that the mixed extract of the old and new dogs shows the effect of prevention and treatment of diseases related to bone formation through promotion of cell proliferation and induction of bone cell differentiation.

The composition comprising the mixed extract of the extract of the present invention according to the present invention is effective for the osteoblast differentiation activity while promoting cell proliferation affecting the bone metabolism and is useful for the ovariectomized animal model, It has been confirmed that there is an effect of improving bone density, and it has a preventive and therapeutic effect on diseases related to bone formation. Therefore, the composition containing the mixed extract of the present invention as an active ingredient can be used as a bone metabolism-related disease such as osteoporosis, osteopenia, osteomalacia, calcium control abnormalities such as hypercalcemia, prevention of hypocalcemia, As a pharmaceutical preparation or a functional food.

<110> DONG WOO DANG CO., LTD          AJOU UNIVERSITY INDUSTRY-ACADEMIC COOPERATION FOUNDATION <120> A composition for preventing bone metabolism-related diseases and          increasing bone function comprising mix extracts of Mori Fructus          and Aralia Continentalis <130> A00327 <160> 8 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Gapdh-F <400> 1 tgaccacagt ccatgccatc 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Gapdh-R <400> 2 gacggacaca ttgggggtag 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Alp-F <400> 3 tcccacgttt tcacattcgg 20 <210> 4 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Alp-R <400> 4 cccgttacca tataggatgg cc 22 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Runx2-F <400> 5 taaagtgaca gtggacggtc cc 22 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Runx2-R <400> 6 tgcgccctaa atcactgagg 20 <210> 7 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Bglap-F <400> 7 tagtgaacag actccggcgc ta 22 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Bglap-R <400> 8 tgtaggcggt cttcaagcca t 21

Claims (5)

A composition for prevention or improvement of osteoporosis characterized by comprising a mixed extract obtained by mixing a heder extract and a combed extract at a weight ratio of 2: 1 to 1: 2 as an active ingredient,
The composition for prevention or improvement of osteoporosis according to claim 1, wherein the topical or visceral extract is prepared by a method comprising the steps of:
(S1) hot-air drying at 50 to 60 DEG C and then pulverizing to a size of 60 mesh or less;
(S2) The process of adding 6 to 10 times of water, an aqueous solution of ethanol or a mixture thereof to the pulverized bulkhead or chewing gum at a weight ratio of 2 to 6 hours at 70 to 80 ° C is repeated 2-3 times Producing an extract;
(S3) centrifuging the obtained fresh or dried extract at 12,000 to 15,000 rpm for 10 to 15 minutes, and then concentrating the dried extract at a first reduced pressure at 60 to 80 ° C; And
(S4) filtration of the concentrated solution or concentrated extract, followed by concentration under reduced pressure, to obtain a fresh or concentrated extract.
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