KR101559068B1 - Composition for inhibiting the differentiation of adipocyte and lowering levels of lipid droplet in adipocyte - Google Patents

Abstract

The present invention relates to compositions for inhibiting adipocyte differentiation or growth, comprising, as an active ingredient, a cytokine which regulates the expression of chimeric (Chemerin, TIG2, RARRES2) or chimeric inhibiting adipocyte differentiation and growth. More particularly, the present invention relates to a pharmaceutical composition, a health food composition and a cosmetic composition comprising the above-mentioned chimeric as an active ingredient, which is useful as a composition for preventing or treating obesity, lipoma, arteriosclerosis, diabetes, hypertension, hyperlipidemia, hypercholesterolemia, It is useful not only for preventing or treating heart disease, but also for reducing fat tissue for cosmetic or cosmetic purposes.

Chimerin, arteriosclerosis, diabetes, hypertension, hyperlipidemia, hypercholesterolemia, hyperphagia, circulatory diseases or ischemic heart disease

Description

TECHNICAL FIELD The present invention relates to a composition for inhibiting differentiation of adipocytes containing chymeric and for reducing lipid levels in adipocytes. [0002] The present invention relates to a composition for inhibiting adipocyte differentiation and lipid droplet in adipocyte,

The present invention relates to a pharmaceutical composition, a health food composition and a cosmetic composition comprising chimeric as an active ingredient which inhibits adipocyte differentiation and inhibits adipocyte growth.

Chimeric is a 16 kDa chemoattractant that is secreted in the form of a precursor in the defense process for survival of the host and cleaved at the carboxy terminal by a serine protease And is known to play an important role in innate immunity and adaptive immunity.

It has been reported that the above-mentioned chimeric is used as a therapeutic agent for psoriasis or acne, and its expression is increased by the treatment of psoriasis, a recently developed synthetic retinoid, and tazarotene, which is reported to be used for some acne It is also referred to as tazarotene induced gene 2 (TIG2) or retinoic acid receptor responder 2 (RARRES2).

In addition, the above-mentioned chimeric was first found in inflammatory fluid of ovarian cancer ascites and rheumatoid arthritis synovial fluids, and is highly expressed in adipose tissue, but its role is unclear.

The chemerin receptor (cmklr1, ChemR23) is an orphan G-protein-coupled receptor, which binds to Gi protein and regulates calcium secretion and extracellular signal-regulated kinase 1/2 (ERK1 / 2).

The expression of the above-mentioned chimerin is present in the 3T3-L1 pre-adipocyte derived from mice at a very low expression level, but its expression and secretion are remarkably increased in the lipid differentiation process. In addition, when expression of the chimeric or chimeric receptor was reduced in 3T3-L1 preadipocytes using a small hairpin RNA (shRNA) system, it was not differentiated into adipocytes, Decreased expression of related genes has been known (Kerry et al., Chemerin, a novel adipokine that regulates adipogenesis and adipocyte metabolism. JBC 282: 28175-28188, 2007).

The Marin Keys were not white adipose tissue and the genetic expression amount of the difference of the white adipose tissue and lean ob / ob mouse model of obese ob / ob mouse models, fat sand rat (P. obesus

However, these findings are not yet clear from a medically relevant point of view, as the results of studies of the chimeric and chimeric receptors in various animal models are contradictory (Ormond et al., The rapidly expanding family of adipokines. Cell Metabolism 6: 159-161, 2007).

The mechanisms controlling the production of chimeric and chimeric receptors in adipocytes have not yet been elucidated. The secretion of immune cells and cytokines involved in inflammation and immunity, among other cells that interact with adipocytes, Although there is a possibility that chimerin can be regulated by influencing, there is no specific study or report. (Ormond et al., The rapidly expanding family of adipokines. Cell Metabolism 6: 159-161, 2007).

Since various diseases are presumed to be related to abnormal adipocyte differentiation or lipid body content in adipocyte, studies on adipocyte differentiation and lipid body content in adipocyte have been actively conducted, and in this study, human mesenchymal stem cells are used as well as human adipocytes and mesenchymal stem cells. In particular, the mesenchymal stem cells of a human are similar to the method of differentiating the adipose precursor cells collected from the subcutaneous tissues into complete adipocytes (Xu et al., The use of small interfering RNAs to inhibit adipocyte differentiation in human preadipocytes and fetal-derived mesenchymal cells, Cell Res. 312: 1856-1864, 2006) and is widely used in adipocyte differentiation studies (Pittenger et al., Multilineage potential of adult human mesenchymal stem cells. S cience 284: 143-146, 1999).

It is known that the mesenchymal stem cells of a human capable of differentiating into adipocytes are distributed in most tissues of the body and can be differentiated into other types of cells such as chondrocytes and bone cells as well as adipocytes Mesirelles et al., Mesenchymal stem cells reside in virtually all post-natal organs and tissues. J. Cell Sci. 119: 2204-2213, 2006).

The human mesenchymal stem cells are generally known to have similar properties or characteristics, although there are slight differences in their properties or characteristics depending on which tissue they are derived from (Sakaguchi et al., Comparison of human stem cells derived from various mesenchymal tissues, Arthritis Rheumat., 52: 2521-2529, 2005).

Despite the steady progress of studies using adipocytes or human mesenchymal stem cells to control lipid differentiation or lipid levels in adipocytes, which are presumed to be associated with the onset of the various diseases, There is still a need in the art for factors or substances that can effectively control lipid levels in differentiation or adipocytes.

Accordingly, the present inventors have found a characteristic that is different from the variation of the chimeric gene in the existing mouse-derived sex cells as a result of confirming the variation of the chimerin in the process of differentiation of human mesenchymal stem cells into adipocytes. In the process of differentiation of human mesenchymal stem cells into adipocytes, the expression of chimeric and chimeric receptors decreased. Furthermore, when the chimeric protein itself was treated with mesenchymal stem cells, lipidation was inhibited in a concentration - dependent manner. In addition, the present inventors have completed the present invention by confirming that the amount of chimerin is regulated by various cytokines secreted by immune cells, and the degree of lipid differentiation can be controlled thereby.

Accordingly, an object of the present invention is to provide a composition for inhibiting adipocyte differentiation or growth, which comprises, as an active ingredient, a cytokine that regulates expression of chimeric or chimeric.

It is still another object of the present invention to provide a method for the treatment or prophylaxis of obesity, lipoma, arteriosclerosis, diabetes, hypertension, hyperlipidemia, hypercholesterolemia, hyperphagia, A pharmaceutical composition for treating or preventing ischemic heart disease, a health food composition and a cosmetic composition.

The present invention relates to a composition for inhibiting adipocyte differentiation or growth using chimeric as an active ingredient.

More particularly, the present invention relates to a method for the treatment and prophylaxis of adipocyte differentiation or adipocyte differentiation which is characterized in that the expression of chimeric and chimeric receptors is reduced in the differentiation process of human mesenchymal stem cells, To a composition for inhibiting growth.

The present inventors confirmed that expression of a marker marker gene associated with adipocyte differentiation is reduced when adipocyte differentiation is inhibited by chimeric. That is, the transcription level of fatty acid binding protein 4 (FABP4), adiponectin, and γ type peroxisome proliferators-activated receptor gamma (PPARγ) is decreased. In addition, IL-4 (Interleukin-4), IL-6 (Interleukin-6), IL-17 (Interleukin-17) and IFN-r (Interferon-gamma) It was confirmed that lipidation can be controlled by controlling the expression of chimeric and chimeric receptors.

The present invention will be described in more detail.

Human mesenchymal stem cells (hBM-MSCs) were cultured in the recommended manner and differentiated into adipocytes. In the experimental group, After 15 days after human mesenchymal stem cells began to differentiate into adipocytes, chimerin was added to the medium to remove the cell culture medium, fix the cells, and then easily separate the lipid bodies and nuclei in the adipocytes through the staining and then microscopically As a result of observing adipocytes, it was confirmed that the chimerin inhibited the differentiation of adipocytes without inducing apoptosis of human mesenchymal stem cells. The results are shown in Fig. A: human mesenchymal stem cells (control group), B: mesenchymal stem cells that induced adipocyte differentiation without addition of chimerin, C: human mesenchymal stem cells that induced adipocyte differentiation after addition of chimeric.

To quantitatively determine the inhibitory effect of the chimerin on the adipocyte differentiation, the adipocyte differentiated by the same induction method was treated with different concentrations of chimeric to differentiate adipocytes. As a result, adipocyte differentiation of human mesenchymal stem cells It was also confirmed that chimerin inhibited adipocyte differentiation of human mesenchymal stem cells in a dose-dependent manner. This result indicates that chimerin inhibits adipocyte differentiation in experimental groups with high concentration of chimerin.

In order to induce adipocyte differentiation of human mesenchymal stem cells in the same manner as above, chimeric RNA was isolated from the cells, and RNAs isolated from the cells were isolated using Qiagen RNA Kit (Qiagen, Valencia, CA) , CDNA was synthesized from the RNA using a reverse transcription kit (Superscript Reverse Transcriptase (RT) II kit, Invitrogen, Carlsbad, Calif.), And the resultant was subjected to real time reverse transcription The expression of adipocyte-associated marker genes by chimerin was investigated by polymerase chain reaction (Q-RT-PCR). As a result, the inhibition of adipocyte differentiation of human mesenchymal stem cells and the decrease of lipid levels in differentiated adipocytes, Adiponectin, FABP4, PPARgamma, and the marker marker gene of the chimeric receptor were decreased, and the results are shown in FIG. A: Human mesenchymal stem cells (control), B: Human mesenchymal stem cells that induced adipocyte differentiation without addition of chimerin, C: Human mesenchymal stem cells that induced adipocyte differentiation after addition of 10 ng / D: Human mesenchymal stem cells that induced adipocyte differentiation after addition of 100ng / ml of chimeric.

In order to investigate the expression of the above-mentioned chimeric and chimeric receptors in the lipid differentiation process of human mesenchymal stem cells, human mesenchymal stem cells were treated with chimeric at the same treatment conditions to induce differentiation into adipocytes, After RNA was isolated using the same method as above, cDNA was synthesized and its expression was detected by real-time reverse transcription polymerase chain reaction. As a result, it was confirmed that the expression level of the chimeric receptor as well as the chimeric receptor was decreased And the results are shown in Fig. 4 below.

In order to confirm that the expression of the lipoprotein-related protein can also be regulated by controlling the amount of expression of chimeric and chimeric by cytokine IL-17 secreted by immune cells interacting with adipocytes, In the present invention, human mesenchymal stem cells are treated with chimerin to induce differentiation into adipocytes, RNA is isolated using the same method as described above, and then cDNA is synthesized and then ligated with chimeric, Receptor expression and expression of FABP4 and adiponectin, the expression of chimeric receptor was increased when treated with IL-17 among the chimerin and chimeric receptors decreased with lipid differentiation, And it was confirmed that the expression of FABP4 and adiponectin, which are proteins involved in lipid differentiation, is decreased, and the results are shown in Fig. 5 Respectively.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating obesity, lipoma, arteriosclerosis, diabetes, hypertension, hyperlipidemia, hypercholesterolemia, hyperphagia, circulatory system disease or ischemic heart disease A pharmaceutical composition for the treatment or prevention of cancer, and a health food composition.

The present invention also provides a method for screening candidate substances for controlling obesity by measuring the amount of expression of chimerin and the degree of modulation of expression level, and also provides an obesity-controlling candidate substance containing cytokine IL-17 from adipocyte stem cells to adipocytes Treating at the induction of differentiation; And measuring the expression of chimerin expressed in the adipocytes differentiated from the mesenchymal stem cells to select an obesity-controlling candidate substance that increases the expression level of chimerin. , And a method for diagnosing local obesity and obesity by measuring the expression of the above-mentioned keymerin.

In the pharmaceutical composition or health food composition of the present invention, the chimeric is derived from a mammal, preferably a human, and may be recombinant chimeric or purified natural chimeric produced by recombinant techniques.

The above-mentioned chimeric may be used in the form of full-length chimeric or fragment thereof, and is preferably used in the form of full-length chimeric form.

The pharmaceutical composition of the present invention contains chimeric as an active ingredient in an amount of 0.00001 to 15% by weight of the total pharmaceutical composition. The pharmaceutical composition thus prepared is used at an effective content of 0.00001 mg / kg to 400 mg / kg per dose, It should be understood that the actual dosage of the active ingredient should be determined in light of various relevant factors such as the severity of the symptoms, the route of administration chosen, the age, sex, weight and health of the subject, and so on, And are not intended to limit the scope of the invention.

The pharmaceutical composition of the present invention may further contain a pharmaceutical adjuvant such as a preservative, a stabilizer, a wetting agent or an emulsifying accelerator, a salt for controlling osmotic pressure, a buffer, and other therapeutically useful substances, Or parenterally administered forms.

Examples of formulations for oral administration include tablets, pills, hard and soft capsules, liquids, suspensions, emulsions, syrups, granules and the like. These formulations may contain, in addition to the active ingredient, a diluent such as lactose, dextrose, Sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (such as silica, talc, stearic acid and magnesium or calcium salts thereof and polyethylene glycols). Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, optionally mixed with starch, agar, alginic acid or its sodium salt Such as disintegrants, absorbents, coloring agents, flavoring agents, and sweetening agents. Tablets can be prepared by conventional mixing, granulating or coating methods. Also typical of parenteral administration formulations are isotonic aqueous solutions or suspensions which are suitable for injectable use.

A health food composition for preventing or ameliorating obesity, lipoma, arteriosclerosis, diabetes, hypertension, hyperlipidemia, hypercholesterolemia, hyperphagia, circulatory system disease or ischemic heart disease containing the above-mentioned chimeric according to the present invention as an active ingredient to provide.

The health food composition of the present invention contains chimeric as an active ingredient in an amount of 0.00001 to 10% by weight in the whole health food composition. Examples of the food form include glucose, citric acid, liquid oligosaccharide, corn syrup, Caramel, chocolate, diet bar, or confectionery, by using commonly used ingredients such as vegetable oils, vegetable hardening oils, skim milk, sugar, margarine, salt, starch, wheat flour, starch syrup, maltose, ≪ / RTI >

There is provided a cosmetic composition for improving obesity containing the above-mentioned chimeric according to the present invention as an active ingredient.

The cosmetic composition of the present invention contains chimeric as an active ingredient in an amount of 0.00001 to 10 wt% of the total cosmetic composition. The formulation of the cosmetic composition is not particularly limited and may be appropriately selected according to the purpose. For example, the cosmetic composition of the present invention can be prepared into any one form selected from the group consisting of massage cream, body lotion, body cream, body oil and body essence.

As described above, the composition for inhibiting adipocyte differentiation or growth, which contains chimeric as an active ingredient, is useful as a therapeutic agent for obesity, lipoma, arteriosclerosis, , Diabetes, hypertension, hyperlipidemia, hypercholesterolemia, hyperphagia, circulatory system diseases or ischemic heart diseases, health food compositions and cosmetic compositions.

In addition, cytokine can regulate the expression of chimeric and chimeric receptors to inhibit lipid differentiation. By measuring the expression level of chimeric and receptor, it is possible to efficiently detect local obesity and obesity, Research and diagnostic methods.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, it should be understood that the present invention is not limited by the following examples, and that various modifications or changes may be made therein without departing from the spirit and scope of the present invention.

[ Example  ] Human bone marrow Intermediate lobe  For stem cell differentiation of adipocytes Chimeric  effect

Human bone marrow mesenchymal stem cells (hBM-MSCs) were purchased from Lonza, Inc. (Walkersville, MD, USA) and cultured according to Lonza's guidelines. Adipocyte differentiation was induced by using indomethacin in accordance with the method recommended by Ronza Co., Ltd. Specifically, in the untreated control medium (A) in which mesenchymal stem cells were cultured, 1 μg / ml of insulin was added to the medium of mesenchymal stem cells , Stem cells were differentiated into adipocytes (B) in the presence of 1 μM dexamethasone (DEXA) and 0.5 mM isobutylmethylxanthine (IBMX). In the experimental group, 100 ng chimerin / ml was added to the medium (C). Chimeric was purchased from R & D Systems (Minneapolis, Minn., USA), prepared as a solution according to the manufacturer's instructions, and sterilized.

After 15 days from the start of adipocyte differentiation of mesenchymal stem cells, the cell culture medium was removed and the cells were washed with 10 ml of phosphate buffered saline (PBS), and then 0.2 ml / cm2 of 10% formalin / PBS solution And the cells were fixed for 10 minutes. The fixed cells were washed once with a 60% isopropanol solution 0.5 ml / cm 2, and the lipids in the adipocyte were stained with 0.2 ml / cm 2 of a dye solution obtained by dissolving Oil Red O in 60% isopropanol. Subsequently, re-staining with hematoxylin was performed to make it easy to distinguish the cell nuclei. Then, the cells were observed under a microscope. As a result of observation, as shown in Fig. 1, when chimeric did not induce apoptosis of mesenchymal stem cells And inhibited the differentiation of adipocytes.

[ Example  2] Chimeric  human's Of mesenchymal stem cells  Quantitative measurement of inhibitory effect of adipocyte differentiation

As shown in Example 1, the differentiation of adipocytes was induced from mesenchymal stem cells, and divided into four groups. In Group A, no induction factors were treated (control group). In Group B, insulin 1 μg / ml, DEXA 1 μM and IBMX (IDX). Group C was treated with IBMX 0.5 mM (IDX) and chimeric 10 nM. Group D was treated with IBMX 0.5 mM (IDX) and 100 mg / ml chimeric.

In order to measure the degree of adipocyte differentiation, the fat cells were stained with Oil Red Oto as in Example 1, the stained oil red oe was eluted with a 100% isopropanol solution, and the absorbance was measured at a wavelength of 500 nm As shown in Fig.

As can be seen from FIG. 2, when differentiation into adipocytes was induced by IBMX (IDX), adipocyte differentiation of mesenchymal stem cells was observed at 100 ng / ml compared to the control without treatment of chimeric (Fig. 2B) ml of chimerin treated group (D) and 31% of experimental group (C) treated with 10ng / ml of chimeric. From these results, it was confirmed that chimeric inhibits adipocyte differentiation of human bone marrow mesenchymal stem cells in a dose-dependent manner.

[ Example  3] associated with fat cells Marker marker  The level of expression of genes Chimeric  effect

As shown in Example 1, the following experiment was conducted to investigate the change in the expression pattern of adipocyte-related genes when lipidation of human bone marrow mesenchymal stem cells was inhibited by chimeric

Specifically, the bone marrow mesenchymal stem cells of human cells induced differentiation into adipocytes were treated with chimeric at a concentration of 10 ng / ml or 100 ng / ml for 15 days in the same manner as described in Example 1, RNA was isolated from cells using a sol solution (Trizol reagent, Invitrogen, Carlsbad, Calif., USA). The separated RNA was further purified with an RNA kit (Qiagen RNeasy kit, Qiagen, Valencia, Calif.) From Agilent Technologies, Inc. (Agilent 2100 BioAnalyzer, Agilent Technologies, Santa Clara, Calif. Were used to confirm the quality of the RNA. CDNA was synthesized from the RNA using a reverse transcription kit (Superscript Reverse Transcriptase (RT) II kit, Invitrogen, Carlsbad, Calif.), And the cDNA was synthesized using real time reverse transcription polymerase chain reaction Q-RT-PCR). Adiponectin (Hs00605917_m1), FABP4 (Hs00609791_m1), PPARγ (Hs00233423_m1), and the chimeric receptor (Hs03063327_m1), which are the following adipose cell-associated mark marker genes, were identified at the time of inhibition of adipocyte differentiation and differentiation of lipid levels in differentiated adipocytes. Were evaluated using a TaqMan gene expression assay kit (Applied Biosystems, Foster City, Calif.) Of Applied Biosystems.

When treated with 10ng / ml or 100ng / ml of chimeric, adiponectin, FABP4 and PPARgamma transcript levels, which are adipocyte related genes, decreased dependent on the amount of chimeric, and the results are shown in FIG. From the results shown in FIG. 3, it was confirmed that chimeric regulates the intracellular specific mechanism involved in adipocyte-related genes, thereby inhibiting adipocyte differentiation and reducing lipid levels in differentiated adipocytes.

[ Example 4] Adipose tissue differentiation process Chimeric  And Chimeric  Changes in receptor expression

The following experiment was conducted to investigate changes in the expression pattern of the chimeric and chimeric receptors in the lipid differentiation process of human bone marrow mesenchymal stem cells.

Specifically, human bone marrow mesenchymal stem cells were induced to differentiate into adipocytes for 15 days in the same manner as described in the step of Example 1 (lipid differentiation without chimerin and change in the amount of chimerin ) And Trizol reagent (Invitrogen, Carlsbad, Calif., USA). The separated RNA was further purified with an RNA kit (Qiagen RNeasy kit, Qiagen, Valencia, Calif.) From Agilent Technologies, Inc. (Agilent 2100 BioAnalyzer, Agilent Technologies, Santa Clara, Calif. Were used to confirm the quality of the RNA. CDNA was synthesized from the RNA using a reverse transcription kit (Superscript Reverse Transcriptase (RT) II kit, Invitrogen, Carlsbad, Calif.), And the cDNA was synthesized using real time reverse transcription polymerase chain reaction Q-RT-PCR). The expression patterns of the chimeric and chimeric receptor genes in the adipocyte differentiation of mesenchymal stem cells were measured using a TaqMan gene expression assay kit (Applied Biosystems, Foster City, Calif.) From Applied Biosystems Hs00161209_g1), and the expression level of the chimeric receptor (Hs03063327_m1).

When human bone marrow mesenchymal stem cells were differentiated into adipocytes by IBMX (IDX) for 15 days, the transcription level of the chimeric receptor as well as the chimeric receptor decreased, and the result is shown in FIG. From the results shown in Fig. 4, it can be confirmed that the regulation of the expression level of the chimeric and chimeric receptors in human bone marrow mesenchymal stem cells is an important phenomenon in differentiation into adipocytes.

[ Example  5] in the process of adipose differentiation Chimeric  And Chimeric  Effect of cytokines on the expression of receptor

The following experiments were carried out to investigate the changes in expression patterns of the chimeric and chimeric receptors by cytokines in the lipid differentiation process of human bone marrow mesenchymal stem cells.

Specifically, 1 μM Troglitazone was added to the cells in the same manner as described in Example 1, and 50 ng of IL-4, IL-6, and IL-17 were added to human bone marrow mesenchymal stem cells that induced differentiation into adipocytes (Trizol reagent, Invitrogen, Carlsbad, Calif., USA) for 15 days at a concentration of 200 units / ml or IFN-r. The separated RNA was further purified with an RNA kit (Qiagen RNeasy kit, Qiagen, Valencia, Calif.) From Agilent Technologies, Inc. (Agilent 2100 BioAnalyzer, Agilent Technologies, Santa Clara, Calif. Were used to confirm the quality of the RNA. CDNA was synthesized from the RNA using a reverse transcription kit (Superscript Reverse Transcriptase (RT) II kit, Invitrogen, Carlsbad, Calif.), And the cDNA was synthesized using real time reverse transcription polymerase chain reaction Q-RT-PCR). The expression patterns of the chimeric and chimeric receptor genes in the adipocyte differentiation of mesenchymal stem cells were measured using a TaqMan gene expression assay kit (Applied Biosystems, Foster City, Calif.) From Applied Biosystems Hs00161209_g1), and the expression level of the chimeric receptor (Hs03063327_m1).

Similar to the results shown in FIG. 4, when the human bone marrow mesenchymal stem cells were differentiated into adipocytes by IBMX (IDX) for 15 days, the level of transcription of the chimeric receptor as well as the chimeric receptor decreased, Was treated with cytokine and differentiated into adipocytes by IBMX (IDX) for 15 days, the chimerin was greatly increased again in the treatment with IL-17, and the chimeric receptor was further reduced, and the results are shown in FIG. 5 Respectively. From the results shown in FIG. 5, it was confirmed that the expression of chimeric and chimeric receptors is regulated by various cytokines in human bone marrow mesenchymal stem cells, which is an important phenomenon in differentiation into adipocytes. In addition, when IL-17 and IFN-r were treated, the transcription level of adiponectin, FABP4, which is an adipocyte related gene, was decreased, and the result is shown in Fig. From the results shown in Fig. 5, it was confirmed that cytokines such as IL-17 regulate the expression of chimerin and inhibit adipocyte differentiation by using a specific mechanism in cells involved in adipocyte-related genes.

[Pharmaceutical composition Formulation Example 1] Tablets

80 mg of chimerin, 200 mg of galactooligosaccharide, 60 mg of lactose and 140 mg of maltose were mixed and granulated by using a fluidized bed dryer, followed by addition of 6 mg of sugar ester and tableting. The final weight of the contents is 600 mg.

[Health food composition Formulation Example  2] Drinks

80 mg of chimerin, 10 mg of glucose, 0.6 g of citric acid and 25 g of liquid oligosaccharide were mixed, and 300 ml of purified water was added, and each bottle was filled with 200 ml each. And then sterilized at 130 DEG C for 4 to 5 seconds to prepare a beverage.

[ Cosmetics  Composition Formulation Example  3] Massage cream

A massaging cream formulation containing chimeric as an active ingredient was prepared according to a conventional method (weight%).

FIG. 1 shows the results of observing the inhibitory effect of chimeric adipocyte differentiation on the adipocyte of the experimental group treated with human mesenchymal stem cells (control) and chimerin at a concentration of 100 ng / ml.

FIG. 2 shows the results of measurement of the adipocyte differentiation of the experimental group treated with the control and 10 mg / ml of chimeric and 100 ng / ml of adipocyte differentiation of human mesenchymal stem cells.

FIG. 3 shows the results of RT-PCR for marker marker genes of the marker markers adiponectin, FABP4, PPARγ and the chimeric receptor after treating human mesenchymal stem cells with a control and chimeric at a concentration of 100 ng / ml.

FIG. 4 shows the results of reverse transcription polymerase chain reaction for the chimeric and chimeric receptors in the process of differentiating human mesenchymal stem cells into adipocytes.

FIG. 5 is a graph showing the effect of differentiation of human mesenchymal stem cells on the differentiation of adipocytes into chimeric and chimeric receptors treated with 50 ng / ml IFNr and 200 nits / ml IL-4, 6 and 17, adiponectin, This is the result of the reverse transcription polymerase chain reaction for FABP4.

Claims (18)

delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete Treating the adipocyte differentiation-controlling candidate substance comprising cytokine IL-17 from the mesenchymal stem cells at the induction of adipocyte differentiation; And measuring the expression of chimerin expressed in the adipocytes differentiated from the mesenchymal stem cells to select an adipocyte differentiation-controlling candidate substance that increases the expression level of chimerin. 17. The method of claim 16, Wherein said substance reduces the level of transcription of adiponectin or FABP4. A method of inhibiting the differentiation or growth of adipocytes from mesenchymal stem cells by treating chemerin with isolated mesenchymal stem cells.

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