KR101413584B1 - A composition for preventing and treating bone diseases containing desmodii herba extracts as a active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating, or treating bone diseases, wherein the composition uses a desmodii herba extract as an active ingredient so as to control the differentiation of osteoclasts and the expression of c-Fos and NFATc1 so that bone diseases including osteoporosis can be prevented, alleviated, or treated. The composition according to the present invention is extracted from a natural material so that side effects do not occur and the differentiation of osteoclasts is controlled. Moreover, the expression of c-Fos and NFATc1 which is a major mechanism of the differentiation of osteoclasts is strongly controlled. Furthermore, the expression of osteoclast induction factors including TRAP, OSCAR, cathepsin K, and DC-STAMP which are activated by means of M-CSF and RANKL is controlled, and the activation of MAPK paths including ERK, JNK, p38, etc. is controlled.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing, improving or treating bone diseases, which comprises an extract of Ganoderma lucidum as an active ingredient. [0002] The present invention relates to a composition for preventing,

The present invention relates to a composition for preventing, ameliorating or treating bone diseases, which can inhibit osteoclast differentiation, c-Fos and NFATc1 expression using an extract of Ganoderma lucidum as an active ingredient, thereby preventing, ameliorating or treating bone diseases including osteoporosis ≪ / RTI >

A bone is a solid body composed of cells surrounded by a rigid intercellular space. It acts as a structural framework for the body of a vertebrate, provides a place where the muscles stick, And protects the brain, spinal cord and soft internal organs.

The bone consists of organic matter, minerals, and water. Calcium in the minerals is responsible for muscle contraction, release of neurotransmitters, regulation of heart rate, and blood coagulation. When the supply of calcium into the blood stops, the osteocyte is destroyed because of the abnormality such as the contraction of the muscles, so that the supply of calcium is continued in the blood. When bone cells are destroyed or old bone cells die, new bone cells are formed.

The osteoclast is the proliferation and differentiation of osteoclast progenitor cells. The osteoclast is a specific phenomenon that is seen only in osteoclasts. The osteoclast cell is skeletonized and the osteocyte is unnecessary in the process of bone growth. Destructs or absorbs and decomposes. When bone cells are degraded, osteoblasts secrete bone cells out of the cells, and they are transformed into osteocytes by themselves to produce new bone, and the secreted bone matrix is gradually mineralized to form bones. Thus, bone is formed by osteoblast differentiation of bone marrow progenitor cells into osteoclasts and bone formation by osteoblasts, and the homeostasis is maintained by balancing bone resorption and bone formation.

At this time, osteoclasts are differentiated into TRAP-positive multinucleated cells by osteoclast differentiation factors such as M-CSF (macrophage colony stimulating factor) and RANKL (receptor activator of NF-κB ligand) Are mainly involved in cell proliferation, survival and cytoskeletonization, and RANKL plays an important role in cell differentiation and survival.

Here, the osteoclast differentiation is described in more detail as follows.

Osteoclasts are produced from myeloid precursors, such as mononuclear cells in bone marrow cells or blood. M-CSF acts on myeloid progenitor cells to induce the expression of RANK (receptor activator of NF-κ) receptors.

RANKL, which is expressed in osteoblast cells, binds to RANK receptors and activates transcription factors such as NF-κB, c-Fos and NFATc1 to induce osteoclast induction such as TRAP and OSCAR Promoting the expression of the factor.

In particular, c-Fos induction of NFATc1 expression of a transcription factor which increases the differentiation initially, NFATc1 is in the c-Fos dimer AP-1 and the Ca ^{2 +} / calmodulin activated kinases are activated by a (CaMKs) signaling pathway osteoclasts Induces the expression of TRAF (TNF receptor associated factor), a cell-specific genetic material.

In addition, RANKL promotes the translocation of TRAF6 (TNF receptor assocated factor 6) into the cytoplasm of RANK, leading to important MAPKs such as ERK (extracellular signal-regulated kinase), JNK (c-Jun-N-terminal kinase) mitogen activated protein kinases. In particular, p38 and JNK play an important role in osteoclast differentiation, and ERK is known to be involved in osteoclast survival.

As described above, osteoclast differentiated by RANKL expressed in osteoblast is activated by various gene expression and signal transduction proteins which are activated by binding to RANK receptor expressed by M-CSF. If bone resorption is not performed normally by osteoclasts, osteoporosis is caused by osteoporosis, osteoporosis and osteoporosis are caused by osteoporosis.

Among them, osteoporosis is the most common bone disease, which is a metabolic disorder in which the amount of bone is reduced and the quality of the bone is weakened due to qualitative changes, thereby increasing the risk of fracture of the spine, femur, and radius.

Osteoporosis is classified into primary and secondary, depending on the cause. Primary osteoporosis is divided into postmenopausal osteoporosis (type 1 osteoporosis) and senile osteoporosis (type 2 osteoporosis). Postmenopausal osteoporosis is osteoporosis caused by estrogen deficiency, which stimulates osteoblast cells in postmenopausal women, and senile osteoporosis occurs in both men and women due to aging-related changes and progresses slowly. Secondary osteoporosis is secondary to diseases such as rheumatoid arthritis, kidney failure and Cushing's syndrome in which the osteoclast activity is decreased.

Currently approved therapies for osteoporosis include female hormone therapy, a hormone therapy regulating bone resorption, regulators of female hormone receptors, bisphosphonates, parathroids, and growth hormones, fluoride and parathyroid hormone, which promote calcitonin and bone formation.

Among these, bisphosphonate is the most widely prescribed primary treatment for osteoporosis treatment. It inhibits osteoclast differentiation, weakens the function, and induces apoptosis and inhibits bone resorption. Recently, however, side effects of esophageal and gastrointestinal ulcers, jaw necrosis and atypical femoral fractures due to long term use have been reported. In addition, hormone drugs such as parathroids are expensive and less effective than bisphosphonates.

Accordingly, much research has been conducted to improve the effects and side effects of osteoporosis treatment agents by inhibiting osteoclast differentiation, which is a major cause of bone diseases, using natural materials including herbal medicines, which are low in human side effects and easy to obtain and have high added value.

As a conventional technique for this, a composition for preventing and treating bone diseases is disclosed in Japanese Patent Application Laid-Open No. 10-2010-0098317. Specifically, the present invention relates to a prophylactic and therapeutic agent for inhibiting the differentiation and production of osteoclasts and increasing bone density by containing Youngran Scent Extract as an active ingredient.

In addition, researches are being conducted to prevent osteoporosis and other bone diseases by inhibiting osteoclast differentiation using natural products such as omija, aloe, rhinoceros, and corn.

Desmodii Herba is a pre-plant of the leguminosae plant Desmodium styracifolium (Osbeck Mer.). Its medicinal area is above the ground, its taste is sweet and sour, Do.

Half-shrub herbaceous herbaceous stem, lying on its side or lying diagonally, is covered with a soft hair. Leaves are alternate phyllotaxis, one leaf is lobed, and the middle lobes are large and circular. The small lobes are relatively small in round shape and have no hairs on the front side. Hair) grows. There are many flowers bloomed and two flowers are lined up side by side. The calyx is a belliform, the corolla is a butterfly shape, and it is magenta. The shape of a substrate is umbrella (寬 倒 卵 形) is. Fruits are 3 to 6 joints (pods) with cones (pods), one side is plain and the other side is shrunken as a wave.

Ingredients are Alkaloid, Flavonoid glycoside, etc., and are known to have efficacy in urinary infection, urinary stone, nephritis, cholecystitis, lacquer, pediatric marker, hematoma, jaundice.

Currently, there is no known research to prevent and treat osteoporosis and other bone diseases by using the fungus.

The object of the present invention is to provide a composition for prevention and treatment of bone diseases, which contains, as an active ingredient, a fungicide extract capable of preventing, ameliorating or treating osteopathies including osteoporosis using natural substances with low human side effects and high added value I have to.

The present invention aims at solving the technical problem by providing a composition for prevention, improvement or treatment of bone diseases which contains an extract of Ganoderma lucidum as an active ingredient.

In addition, we are trying to solve the technical problem by confirming the effect of Gwanggeumcheoncho extract on osteoclast differentiation which is the main cause of bone disease.

In addition, we investigated the effect of Gwanggeumchochae extract on the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation, to solve the technical problems.

In addition, we investigated the effect of Gwangmaechoch extract on the expression of various genes involved in osteoclast differentiation and the activity of signal transduction proteins.

The present invention also relates to a method for preventing, ameliorating or preventing bone diseases, which comprises an extract of Ganoderma lucidum as an active ingredient and further comprising a pharmaceutically acceptable carrier, adduct or diluent to form a pharmaceutical unit dosage form, By providing a therapeutic composition, the technical problem is solved.

In addition, when the extract is formulated, it can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like which are usually used.

In addition, the above pharmaceutical dosage forms can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, and sterilized injection solutions according to conventional methods.

The composition for preventing, ameliorating or treating bone diseases containing the extract of the present invention as an active ingredient has an effect of inhibiting osteoclast differentiation without causing side effects extracted from natural materials. In addition, there is an effect of strongly inhibiting the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation. In addition, it inhibits the expression of osteoclast-like factors including TRAP, OSCAR, cathepsin K and DC-STAMP activated by M-CSF and RANKL, and inhibits the activity of MAPK pathway including ERK, JNK, and p38 have.

FIG. 1 is a graph showing the results of cytotoxicity measurement of the extracts of various concentrations of the extract.
FIG. 2 is a graph showing the cytotoxicity measurement results of the extract of 25 .mu.g / ml.
Fig. 3 is a photograph showing osteoclasts observed with a microscope using TRAP staining method.
FIG. 4 is a graph showing the results of counting TRAP-positive polynuclear cells (MNCs).
FIG. 5 shows the effect of the extract on the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation.

The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms and the inventor may properly define the concept of the term in order to best describe its invention It should be construed as meaning and concept consistent with the technical idea of the present invention.

Therefore, the embodiments, experimental examples and reference examples described in the present specification are merely the most preferred embodiments of the present invention and do not represent all the technical ideas of the present invention. Therefore, various equivalents It should be understood that water and variations may be present.

The bone maintains homeostasis due to the balance of bone resorption and bone formation in osteoclasts and osteoblasts. When bone resorption occurs, it leads to a decrease or increase in bone density, resulting in bone diseases such as osteoporosis and bone atrophy.

In osteoclast differentiation, the binding of RANKL and RANK is essential, and RANK is expressed by M-CSF. The expression of cFos is increased by RANKL, and NFATc1 is activated by the c-Fos and Ca2 ⁺ / calmodulin activated kinases (CaMKs) signaling system. The osteoclasts are differentiated into TRAP-positive polynuclear cells by the activity of c-Fos and NFATc1.

However, NFATc1 is a key substance in osteoclast differentiation because the precursor cells overexpressing NFATc1 without RANKL stimulation are differentiated into TRAP - positive polynuclear cells. Therefore, inhibition of c-Fos and NFATc1 expression inhibits osteoclast differentiation and inhibits bone resorption, thereby preventing and treating bone diseases such as osteoporosis.

Therefore, the present invention has been carried out through experiments to examine the effect of Gwanggimcheon extract, which is a natural substance with low human side effects and high added-value, on the expression of c-Fos and NFATc1, which are the main mechanisms of osteoclast differentiation and osteoclast differentiation.

The extracts were extracted with 70% ethanol. Here, the extract of the extract can be extracted by water or an organic solvent, depending on the conditions.

To investigate the cytotoxicity of the extracts, the extracts of various extracts were treated with macrophages derived from myeloid progenitor cells, and the cell proliferation of macrophages was measured.

The osteoclast differentiation ability of osteoclasts was investigated by inducing osteoclast differentiation by treating M-CSF and RANKL with macrophage to examine osteoclast differentiation affecting bone resorption directly.

In addition, we investigated the effect of Kwanggeumchochae extract on the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation.

Example  1. Prevention, improvement or treatment of bone disease

The present invention provides a preventive, ameliorative or therapeutic agent for bone diseases which is formulated into a pharmaceutical unit dosage form by adding a pharmaceutically acceptable carrier, excipient or diluent to the extract as an active ingredient.

Examples of the carrier, excipient and diluent include tosse, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Vaginal cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical dosage forms may also be used in the form of pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable set.

In addition, when the extract is formulated, it can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like which are usually used.

In addition, each of the above pharmaceutical dosage forms may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or the like, oral preparations, suppositories, .

In the solid preparation for oral administration, at least one excipient such as starch may be prepared by mixing calcium carbonate, sucrose, lactose, gelatin or the like in the extract. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

The preparation for parenteral administration may include a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a freeze-dried preparation, and a suppository.

Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like.

Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The preferred dosage of the extract of the present invention varies depending on the condition and the weight of the patient, the degree of disease, age, sex, drug form, route of administration and period of time, but can be appropriately selected by those skilled in the art.

The extract of the present invention can be administered to mammals such as rats, mice, livestock, humans and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, muscular, subcutaneous injection.

The present invention is to provide a health functional food composition for preventing, ameliorating or treating bone diseases by adding a food supplementary additive to the extract.

Examples of foods to which the extract can be added include various foods, beverages, gums, tea, vitamin complexes, and health functional foods.

It can also be added to foods or beverages for the purpose of preventing, ameliorating or treating bone diseases such as osteoporosis.

At this time, the amount of the extract in the food or drink may be 0.01 to 20% by weight of the total food, and the health beverage composition may be added in an amount of 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 ml .

The health functional beverage composition of the present invention is not particularly limited to the components other than those containing the above extract, and may contain various flavors or natural carbohydrates as additional components such as ordinary beverages.

Examples of the above-mentioned natural carbohydrates are monosaccharides such as glucose, fructose; Disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. In addition to the above, natural flavoring agents (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavorings.

The ratio of the natural carbohydrate 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 extract of the present invention can be used as a nutritional supplement, a vitamin, a mineral (electrolyte), a flavor such as a synthetic flavor and a natural flavor, a coloring agent and an aggravating agent (cheese, chocolate etc.), a pectic acid and its salt, Colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.

In addition, the extracts of the present invention may contain flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages.

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 extract of the present invention.

Experimental Example  One. Light fence  Effect of Extract on osteoclast differentiation

(One) Light fence  Extract preparation

Gwangmukchoncho was purchased from Iksan Oriental Hospital of Wonkwang University and was identified through morphological evaluation after purchase. Sample samples were stored in Wonkwang University Oriental medicine physiology laboratory and biochemical laboratory. The fungus can also be purchased from a traditional Chinese medicine pharmacy.

The fungus was cleaned after purchase, dried for 5 days at the shade and room temperature, and pulverized to powder. The prepared powders were put into an extraction vessel, and about 20 times of 70% ethanol by weight of the powders were added and extracted twice at a temperature of 60 ° C for 10 hours. The extracted solution was filtered through a filter paper, and then concentrated under reduced pressure at 45 ° C by a rotary vacuum evaporator to obtain an extract of Fusarium spp.

(2) Test cells

Bone marrow cells were isolated from the femur of 5-week-old ICR mice and 50 ng / ml of M-CSF was added to α-MEM essential medium containing 10% FBS (fetal bovine serum) Lt; 0 > C for 3 days. After incubation, suspended cells were removed from the cultured cells and attached cells were used as bone marrow-derived macrophages (BMMs).

(3) Cytotoxicity analysis

To investigate the cytotoxicity of the extracts, the extracts of various fungicides were treated with macrophages (BMMs) and cell proliferation was measured by MTT assay.

(3-1) Light fence  Cytotoxicity analysis by extract

Macrophages (BMMs) were cultured in 96-well plates at 3 × 10 ⁴ cells / well in a cell incubator containing 5% CO ₂ and maintained at 37 ° C for 24 hours. After culturing, the extracts were treated with 0, 25, 50, and 100 ㎍ / ㎖, respectively, and cultured for 1 to 4 days in an incubator under the same conditions. The cultured cells were washed once with phosphate buffer (PBS), treated with 0.5 mg / ml of MTT reagent, and cultured for 30 minutes in the same incubator. Then, each well was treated with 100 μL of DMSO (dimethyl sulfoxide) to dissolve the formazan crystals. The absorbance was measured at 570 nm using a 3,550 microplate reader (Bio-Rad, Richmond, CA, USA). The cytotoxicity results were expressed as a percentage of the absorbance of the sample (0 ㎍ / ml of the extract).

FIG. 1 is a graph showing the results of cytotoxicity measurement of the extracts of various concentrations of extracts of various concentrations. Referring to FIG. 1, when the results are analyzed, when cell extracts of 25 μg / ml and 50 μg / The cells treated with 100 ㎍ / ㎖ showed a slight decrease but more than 90% cell proliferation. Therefore, it was found that the cytotoxicity was not observed when the concentration of the extract was less than 100 ㎍ / ㎖ based on 3 × 10 ⁴ cells.

(3-2) Light fence  Cytotoxicity analysis of extracts at 25 μg / ml

In order to further analyze the cytotoxicity of the extracts of Fusarium oxysporum, the cell proliferation of Fusarium fusiforme extract, which exhibited cell proliferation most similar to that of the control group, was treated with 25.

Cell culture and measurement were performed in the same manner as in the above cytotoxicity test, and cytotoxicity was measured 4 times daily for 4 days from the day after the treatment of the extract. The cytotoxicity results were expressed as a percentage of the absorbance of the sample (0 ㎍ / ml of the extract).

FIG. 2 is a graph showing the results of cytotoxicity measurement of 25 μg / ml of the extract of Fusarium oxysporum; FIG. 2 is a graph showing the results. The proliferation was increased, and it began to decrease gradually from 3 days, and appeared to be similar to the control group after 4 days of culture. Therefore, it was found that the extract of Gwangmaegumcho was not cytotoxic regardless of the incubation time.

(4) osteoclast Ability to distinguish  analysis

To investigate the effect of the extract on osteoclast differentiation, osteoclast differentiation ability was measured using TRAP staining method by inducing osteoclast differentiation by treating M-CSF and RANKL in macrophages.

Macrophages (BMMs) is the 48-well plates into a 3.5 × 10 ⁴ cells per well M-CSF 50 ng / ㎖ and RANKL then processes the 100 ng / ㎖ light geumjeoncho extracts were 0, 5, 10, 25, 50 Mu] g / ml were each treated for 4 days. The cultured cells were fixed with formaldehyde and the TRAP staining solution was dispensed and observed under a microscope. The results are shown in FIG. Fig. 3 is a photograph showing osteoclasts observed with a microscope using TRAP staining method.

As shown in FIG. 3, in the control group treated with 50 ng / ml of M-CSF and 100 ng / ml of RANKL, TRAP-positive polynuclear osteoclasts (MNCs) Induced the formation of TRAP-positive polynuclear cells (MNCs) in a dose-dependent manner.

In addition, TRAP-positive multinucleated cells (MNCs) having three or more nuclei were regarded as osteoclasts and the number was counted, and the results are shown in FIG. FIG. 4 is a graph showing the results of counting TRAP-positive polynuclear cells (MNCs).

As can be seen from FIG. 4, the number of TRAP-positive polynuclear cells (MNCs) was also decreased in dependence on the concentration of the extract. Therefore, it was confirmed that Gwanggimcheon extract inhibits osteoclast differentiation.

(5) c- Fos  And NFATc1  Expression analysis

We investigated the effect of Gwanggeumchochae extract on the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation.

Macrophages (BMMs) is put into a 3.5 × 10 ⁴ cells per well in 48-well plates M-CSF 50 ng / ㎖ and RANKL then processes the 100 ng / ㎖ light geumjeoncho extract 0, the process 25 ㎍ / ㎖ each And cultured for 4 days.

The cultured cells were washed once with cold phosphate buffer (PBS) and resuspended in extraction buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA, 0.5% Nonidet P-40, 0.01% protease inhibitor mixture) And the protein was extracted. SDS-PAGE and Western blotting were then performed to determine the expression of NFATc1 and c-Fos (Santa Cruz) from the proteins extracted from the cells.

The measurement was performed once before the treatment of M-CSF and RANKL, and four times daily for 4 days from the day after the culture, and the result is shown in FIG. FIG. 5 shows the effect of the extract on the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation.

As shown in FIG. 5, in the control group treated with 50 ng / ml of M-CSF and 100 ng / ml of RANKL from day 3 after the incubation, the expression of c-Fos was significantly increased from the third day after the incubation and the expression of NFATc1 Which was significantly increased from the next day. However, the expression of NFATc1 and c-Fos was significantly inhibited in the experimental group treated with 25 ㎍ / ㎖ of Gwanggimcheon extract.

Thus, it was found that the extracts of Gwangmaechochae strongly inhibited the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation.

As a result of osteoclast differentiation by inducing differentiation into osteoclasts by treating M-CSF and RANKL with macrophage cells isolated from bone marrow, .

In addition, we investigated the effects on the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation. As a result, it was found that Kwanggeumchoncho extract strongly inhibited the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation.

Reference example  One. Light fence  Effect of Extract on Gene Expression and Signal Transduction Protein Activity

Osteoclasts are produced in bone marrow cells or in myeloid progenitor cells such as monocytes in the blood. M-CSF acts on myeloid progenitor cells to induce the expression of RANK receptors. RANKL expressed in hematopoietic cells binds to the RANK receptor and activates NF-κB, PU1, MITF (Mi transcription factor), c-Fos, NFATc1 and the like to induce osteoclast induction such as TRAP, OSCAR, cathepsin K, DC- ≪ / RTI > In particular, NFATc1 is activated by AP-1 and CaMKs signal transduction system, which is a dimer of c-Fos, and induces the expression of TRAF, an osteoclast-specific genetic material.

In addition, RANKL binds to RANK receptor and promotes TRAF 6 migration into the cytoplasm of RANK, activating MAPKs (mitogen activated protein kinases) such as ERK, JNK, and p38. p38 and JNK play an important role in osteoclast differentiation, and cells showing overexpression of p38 are known to suppress NFATc1 expression.

In other words, the combination of RANK and RANKL induced by M-CSF induces the expression of osteoclast inducers including TRAP, OSCAR, cathepsin K, DC-STAMP, and the like to form a set of TRAF family proteins such as TRAF6 And activation of MAPK pathways including ERK, JNK, p38, etc., thereby differentiating osteoclasts.

The present invention provides a method for inhibiting osteoclast differentiation induced by M-CSF and RANKL in macrophages and strongly inhibiting the expression of c-Fos and NFATc1, the main mechanism of osteoclast differentiation, Could know.

From these results, it is possible to confirm that the extracts of Ganoderma lucidum were activated by M-CSF and RANKL and inhibited various gene expression and signal transduction protein activities involved in osteoclast differentiation. In addition, it can be shown that the extract of Gwanggimcheon itself can be used as a preventive and therapeutic agent for bone diseases including osteoporosis.

It should be understood that the foregoing description of the embodiments, examples and reference examples are for illustrative purposes only and that the present invention is not limited thereto as long as various designs can be made within the technical scope of the present invention.

Claims (5)

A composition for preventing, ameliorating or treating a bone disease including osteoporosis and bone atrophy containing an extract of Desmodii Herba as an active ingredient,
A composition for preventing, ameliorating, or treating bone diseases, which is characterized by inhibiting osteoclast differentiation and inhibiting expression of c-Fos and NFATc1 (nuclear factor of activated T-cells, cytoplasmic 1).
delete delete The method according to claim 1,
The composition for preventing, ameliorating, or treating bone diseases, wherein the concentration of the extract is in the range of 5 μg / ml to 100 μg / ml based on 3.5 × 10 ⁴ cells.
A health functional food composition for preventing or ameliorating a bone disease including osteoporosis and bone atrophy containing an extract of Desmodii Herba as an active ingredient,
The composition for preventing or ameliorating a bone disease according to claim 1, wherein the extract of the present invention inhibits osteoclast differentiation and inhibits the expression of c-Fos and NFATc1 (nuclear factor of activated T-cells, cytoplasmic 1).

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