1. Patent Search
  2. Details

COMPOSITION COMPRISING SEA CUCUMBER GENITAL GLAND EXTRACT, COMPOUND DERIVED FROM SEA CUCUMBER OVARY EXTRACT, AND USE THEREOF

20240058392 ยท 2024-02-22

    Inventors

    Cpc classification

    International classification

    Abstract

    The present invention provides a sea cucumber genital gland extract, preferably, a novel saponin compound separated from an ovary extract, and a novel use of a sea cucumber genital gland extract or a compound separated from a sea cucumber genital gland extract. The present invention has an effect of treating or preventing any one or more diseases selected from the group consisting of obesity, inflammation, diabetes and dyslipidemia, by comprising a sea cucumber genital gland, preferably, an ovary extract, or a compound separated from a sea cucumber genital gland, preferably, an ovary extract.

    Claims

    1.-13. (canceled)

    14. A composition comprising a therapeutically effective amount of a compound represented by the following chemical formula S1, S2, S3, S4 or S9 and a pharmaceutically acceptable carrier: ##STR00006## ##STR00007##

    15. The composition according to claim 14, wherein the compound is extracted or separated from an ovary of a sea cucumber.

    16. A method for treating or improving any one disease selected from the group consisting of obesity, inflammation, diabetes and dyslipidemia in a subject in need thereof, comprising: administering a therapeutically effective amount of a sea cucumber genital gland extract, or a saponin compound derived from a sea cucumber genital gland to the subject.

    17. The method according to claim 16, wherein the sea cucumber genital gland is an ovary of a sea cucumber.

    18. The method according to claim 16, wherein the sea cucumber genital gland extract is obtained by extracting a sea cucumber genital gland with a 45 to 85% (V/V) ethanol aqueous solution as a solvent.

    19. The method according to claim 18, wherein the sea cucumber genital gland extract is obtained by extracting a sea cucumber genital gland with a 48 to 55% (V/V) ethanol solvent.

    20. The method according to claim 16, wherein the sea cucumber genital gland extract is a butanol fraction obtained by fractionizing an extract obtained by extracting a sea cucumber genital gland with a 48 to 55% (V/V) ethanol aqueous solution with butanol as a solvent.

    21. The method according to claim 16, wherein the saponin compound comprises any one or more selected from the group consisting of the following chemical formulas S1, S2, S3, S4, S5, S6, S8 and S9: ##STR00008## ##STR00009## ##STR00010##

    22. The method according to claim 16, wherein the sea cucumber genital gland extract, or saponin compound derived from the sea cucumber genital gland inhibits expression of any one or more proteins selected from the group consisting of PPAR, FAS, aP2, and C/EBP.

    23. The method according to claim 16, wherein the sea cucumber genital gland extract, or saponin compound derived from the sea cucumber genital gland inhibits differentiation from a preadipocyte to an adipocyte, or inhibits intracellular fat accumulation.

    24. A method for improving an effect of prevention or treatment of any one or more diseases selected from the group consisting of obesity, inflammation, diabetes and dyslipidemia of a sea cucumber genital gland extract, which increases any one or more saponin compounds selected from the group consisting of the following chemical formulas S1, S2, S3, S4, S5, S6, S8 and S9: ##STR00011## ##STR00012## ##STR00013##

    25. The method for improving an effect of prevention or treatment of any one or more diseases selected from the group consisting of obesity, inflammation, diabetes and dyslipidemia of a sea cucumber genital gland extract according to claim 24, wherein the sea cucumber genital gland extract is a sea cucumber ovary extract.

    26. The method according to claim 24, wherein the method extracts a sea cucumber ovary with a 48 to 55% (V/V) ethanol aqueous solution, and obtains a butanol fraction from the extract.

    Description

    BRIEF DESCRIPTION OF THE DRAWINGS

    [0036] The following drawings attached in the present description illustrate preferable examples of the present invention, and play a role of understanding the technical spirit of the present invention with the contents of the invention described above, so the present invention should not be interpreted as limited only to the matters described in such drawings.

    [0037] FIG. 1 is the result of confirming the cell viability of cells after treating an extract obtained from a sea cucumber ovary extract and internal organs except for an ovary by varying ethanol concentrations to 3T3-L1 cells.

    [0038] FIG. 2a is a drawing which shows an effect of inhibiting differentiation of adipocytes and fat accumulation of a sea cucumber extract.

    [0039] FIG. 2b shows O.D. values confirmed after treating Samples 1-4.

    [0040] FIG. 3 shows the result of analyzing adipocyte differentiation-related genes and proteins by part and concentration of a sea cucumber extract. FIG. 3a is the result of showing the degree of protein expression, and the graphs of FIG. 3b are the result of confirming the effect on mRNA expression of a sea cucumber extract.

    [0041] FIG. 4a is photographs showing adipocytes differentiation of a sea cucumber extract,

    [0042] and FIG. 4b is a graph showing the effect of inhibiting fat accumulation (25, 50 g/ml).

    [0043] FIG. 5 is the result of confirming the effect on expression of proteins involved in a process of adipocyte differentiation, PPAR, FAS, aP2, and C/EBP of a sea cucumber extract. It was confirmed that the sea cucumber ovary extract inhibits these proteins.

    [0044] FIG. 6 shows a process of separating a compound through extraction and fractionization of a sea cucumber ovary extract as a diagram.

    [0045] FIG. 7 shows an effect on the 3T3-L1 cell viability of compounds separated from a sea cucumber ovary extract.

    [0046] FIG. 8a shows the result of analyzing the degree of fat accumulation by compounds separated from a sea cucumber ovary extract through Oil red O staining after completing adipose differentiation of 3T3-L1 cells.

    [0047] FIG. 8b shows differentiated 3T3-L1 cells, and

    [0048] FIG. 8c shows optical density (O.D.) of Oil Red O measured at 500 nm. They were treated at a concentration of 2.5, 5 g/ml, respectively.

    [0049] Data are presented meanstandard deviation (SD). *, **, and *** indicate p<0.05, 0.01, and 0.001, respectively, when compared to the differentiated control group (MDI).

    [0050] FIG. 9 shows an adipogenesis-related protein level of differentiated 3T3-L1 cells when S1-S6, S8-S9 compounds are treated, respectively.

    [0051] FIG. 10 is the result of confirming the glucose uptake of a sea cucumber ovary extract and an extract of internal organs except for an ovary of a sea cucumber.

    [0052] FIG. 11 is the result of confirming the glucose uptake by condition of extraction of a sea cucumber ovary extract.

    [0053] FIG. 12 is the result of confirming the glucose uptake of saponin compounds separated from an ovary extract of a sea cucumber.

    [0054] FIG. 13 is the result of observing a change in body weight.

    [0055] FIG. 14 shows the final body weight confirmed after treating an extract of a high calorie diet intake.

    [0056] FIG. 15 is the result of white adipocyte size change confirmed through H&E stain.

    [0057] FIG. 16 is the result of representing an HDL content compared to total cholesterol.

    [0058] FIG. 17 is the result of confirming an effect of reducing total cholesterol.

    [0059] FIG. 18 shows the result of confirming an effect of reducing triglyceride.

    [0060] FIG. 19 is the result of confirming an effect of reducing PPAR expression of an extract treatment group.

    MODE FOR INVENTION

    [0061] Hereinafter, in order to help understanding of the present invention, it will be described in detail by examples and the like. However, the examples according to the present invention may be modified into various other forms, and the scope of the present invention should not be interpreted as limited by the following examples. The examples of the present invention are provided to more completely describe the present invention to those skilled in the art to which the present invention pertains.

    I. Experimental Example 1Confirmation of Effect of Extract by Sea Cucumber Part

    [0062] 1. Sample Extraction

    [0063] (1) Extraction Part and Solvent Concentration

    TABLE-US-00001 TABLE 1 Part Solvent Sea cucumber Intestinal 1 50% EtOH (Stichopus japonicus) organ 2 80% EtOH Ovary 3 50% EtOH 4 80% EtOH

    [0064] A sea cucumbers was divided into an ovary and intestinal organ excluding ovaries, and these were extracted. 50% ethanol and 80% ethanol were added 20 times each to the sea cucumber ovary and intestinal organs in the dried state, and they were stirred at a room temperature for 24 hours to extract. After that, through a process of filtration and decompression concentration, sea cucumber ovary and intestinal organ extracts were obtained.

    [0065] 2. Evaluation of Inhibitory Ability of Adipocyte Differentiation of Sea Cucumber Extract

    [0066] (1) Experimental Materials and Methods

    [0067] Reagents and materials are as follows.

    [0068] 3T3-L1 cells (ATCC, CL-173), DMEM high glucose (Gibco), fetal bovine serum (Gibco), penicillin-streptomycin-glutamin (Gibco), phosphate buffered saline (Gibco), Radioimmunoprecipitation assay (RIPA) buffer (Thermo scientific), protease- and phosphatase-inhibitor cocktails (Thermo scientific), PVDF membrane (Bio-rad), Primary antibody (PPAR (Santa cruz), C/EBP (cell signaling), FAS (cell signaling), aP2 (cell signaling), -actin (Santa cruz), Oli red O powder (sigma), and MTT powder (sigma) were used.

    [0069] (2) Measurement of Cell Viability

    [0070] 3T3-L1 cells were aliquoted in a 96 well plate at a concentration of 110.sup.4 cells/well and cultured for a day, and then 6 kinds of extracts were treated at a concentration of 100-200 g/ml. After 24 hours, MTT powder was dissolved in PBS at a concentration of 5 mg/ml and then filtered, and 20 l of cells was aliquoted each and cultured for 2 hours. After skimming the medium, 200 l of DMSO was aliquoted, and then the absorbance was measured at 570 nm to measure the cell viability.

    [0071] (3) Culture of 3T3-L1 Preadipocytes and Differentiation of Adipocytes

    [0072] 3T3-L1 cells were cultured in a high glucose DMEM medium in which 10% calf serum and penicillin streptomycin glutamin were added and subcultured per 2-3 days. After aliquoting the 3T3-L1 cells in a 96 well plate at a concentration of 410.sup.5 cells/well and then culturing them for 2 days, it was replaced with an MDI medium that dexamethasone, insulin, and 3-isobuty-1-methylxanthine (IBMX) were added to a medium containing 10% FBS instead of calf serum. After 2 days, it was replaced with a medium containing insulin, and the medium was replaced again per 2 days and differentiation was induced. The sea cucumber extract was treated at a concentration of 200 g/ml in 30 minutes after replacing the medium.

    [0073] (4) Oil Red O Staining

    [0074] The 3T3-L1 cells in which differentiation was completed were fixed with 4% formaldehyde for 1 hour, and then fats were stained using a 0.5% Oil red O staining solution to confirm the degree of differentiation. For quantitative analysis of adipose differentiation, the dye stained in the cells was dissolved in 100% isopropyl alcohol, and the absorbance was measured at 500 nm.

    [0075] (5) Quantitative Real-Time PCR

    [0076] Primer sequences were shown in Table 2.

    TABLE-US-00002 TABLE 2 Gene Forward primer (5.fwdarw.3) Reverse primer (5.fwdarw.3') C/EBP SEQ ID NO: 1 SEQ ID NO: 2 PPAR SEQ ID NO: 3 SEQ ID NO: 4 FAS SEQ ID NO: 5 SEQ ID NO: 6 aP2 SEQ ID NO: 7 SEQ ID NO: 8 -Actin SEQ ID NO: 9 SEQ ID NO: 10

    [0077] In order to investigate the effect on expression of adipose differentiation-related genes of the sea cucumber extract, the degree of mRNA expression of the related genes was confirmed using qRT-PCR. The differentiation-completed 3T3-L1 cells were skimmed using PBS and then it was extracted using Rneasy Mini kit (Qiagen), and RT-PCR was carried out by an SYBR green method by synthesizing cDNA (ReverTra Ace qPCR RT Master Mix, FSQ-201, TOYOBO) with 1 g of the extracted RNA. It was proceeded with a PCR condition of 40 cycles of pre-denaturation (95 C., 1 minute), 95 C., 15 seconds, and 60 C., 15 seconds. The used primer sequences were as Table 2.

    [0078] (6) Western Blotting

    [0079] In order to investigate the effect on expression of adipocyte differentiation-related proteins of the sea cucumber extract, expression of the related proteins was confirmed through western blot. The differentiation-completed 3T3-L1 cells were washed with PBS, and then they were lysed with RIPA buffer, and then proteins were separated by molecular weight through SDS-PAGE. After that, they were transferred to a PVDF membrane and blocking was performed using 5% skim milk for 30 minutes. Then, after reacting with a primary antibody, a secondary antibody was attached and protein expression was confirmed using an ECL solution.

    [0080] (7) Statistics

    [0081] Statistical analysis of the result was performed using GraphPad Prism 7 software (San Diego, CA, USA), and a significant difference between each group was analyzed through One-way ANOVA. (p<0.05)

    [0082] 3. Experimental Result

    [0083] (1) Cell Viability

    [0084] As a result of investigating the effect on the viability of the 3T3-L1 cells of the sea cucumber extract, according to each condition, all of the 6 extracted sea cucumber extracts did not show a significant difference from a non-treatment group up to 200 g/ml.

    [0085] After treating the sea cucumber extract, the viability of the 3T3-L1 cells was shown in FIG. 1.

    [0086] (2) Oil Red O Staining Result

    [0087] In order to investigate the effect on adipose differentiation of the 3T3-L1 cells by the sea cucumber extract, as a result of analyzing the degree of fat accumulation through Oil red O staining, in the cells treated with the sea cucumber ovary 50% and 80% ethanol extracts, the degree of staining was reduced, and an effect of reducing fat accumulation was shown. In addition, in order to quantitively represent this effect, as the result of measuring the absorbance by dissolving Oil red O stained in the cells with 100% isopropyl alcohol, in the cells treated with the sea cucumber ovary 50% and 80% ethanol extracts, it was confirmed that the absorbance was significantly reduced and fat accumulation was reduced.

    [0088] In other words, it was confirmed that the sea cucumber ovary extract had an effect of inhibiting fat accumulation compared to other organs of the sea cucumber.

    [0089] In FIG. 2, the effect of inhibiting adipocyte differentiation and fat accumulation of the sea cucumber extract was shown. As could be confirmed in FIG. 2, in the treatment groups 3 and 4 (ovary extracts), fats were less accumulated compared to the treatment groups 1 and 2, so the O.D. value was almost half of that of the treatment groups 1 and 2.

    [0090] (3) Analysis of Adipocyte Differentiation-Related Genes and Proteins

    [0091] As a result of confirming the effect of the sea cucumber extract affecting expression of proteins and mRNA of PPAR, FAS, aP2, C/EBP known to play an important role in the adipocyte differentiation process, it could be confirmed that the protein expression was reduced in the cells treated with the sea cucumber ovary 50% and 80% ethanol extracts, similarly to the previous experiment. In addition, it could be confirmed that the mRNA expression was also significantly reduced in the cells treated with the sea cucumber ovary 50% and 80% ethanol extracts compared to other intestinal organ parts of the sea cucumber. Therefore, taking these results together, the sea cucumber extract seems to inhibit adipose differentiation by regulating expression of the genes and proteins related to adipose differentiation. In FIG. 3, the result of the effect of inhibiting expression of the adipose differentiation-related genes and proteins of the sea cucumber extract was shown.

    [0092] When comparing the effect of inhibiting adipose differentiation of the sea cucumber extract extracted according to each condition, the most effective extraction condition was confirmed as the sea cucumber ovary 50% (Sample name: SJ-O-D1-50) or 80% ethanol extract (Sample name: SJ-O-D1-80). In other words, the treatment groups 3 and 4 had the excellent effect of inhibiting expression of the adipose differentiation-related genes.

    II. Experimental Example 2Verification of Effect of Extract and Effect of Fraction According to Solvent Condition

    [0093] 1. Experimental Materials and Methods

    [0094] (1) Preparation of Sea Cucumber Samples

    TABLE-US-00003 TABLE 3 Sample name Extract information 1 SJ-O-D1-30 Dried ovary 30% EtOH, 20-fold, 24 h stirring, primary extraction 2 SJ-O-D1-50 Dried ovary 50% EtOH, 20-fold, 24 h stirring, primary extraction 3 SJ-O-D1-80 Dried ovary 80% EtOH, 20-fold, 24 h stirring, primary extraction 4 SJ-O-D1-H Hexane fraction of Sample 2 5 SJ-O-D1-B BuOH fraction of Sample 2

    [0095] (2) Reagents and Materials

    [0096] They were prepared as the items of I-2-(1).

    [0097] (3) Culture of 3T3-L1 Preadipocytes and Differentiation of Adipocytes

    [0098] The 3T3-L1 cells were cultured in a high glucose DMEM medium in which 10% calf serum and penicillin streptomycin glutamine were added, and subcultured per 2-3 days. The 3T3-L1 cells were aliquoted in a 96 well plate at a concentration of 410.sup.5 cells/well and then it was replated with an MDI medium that dexamethasone, insulin, and 3-isobuty-1-methylxanthine (IBMX) were added to a medium containing 10% FBS instead of calf serum, and the sea cucumber extract was treated in 30 minutes after replacing with the MDI medium. After 2 days, it was replaced with a medium containing insulin, and it was replaced with a new medium again per 3 days to induce differentiation.

    [0099] (4) Oil Red O Staining

    [0100] The differentiation-completed 3T3-L1 cells were fixed with 4% formaldehyde for 1 hour, and then fats were stained using a 0.5% Oil red O staining solution to confirm the degree of differentiation. For quantitative analysis of adipose differentiation, the dye stained in the cells was dissolved with 100% isopropyl alcohol and the absorbance was measured at 500 nm.

    [0101] (5) Western Blotting

    [0102] In order to investigate the effect on expression of the adipocytes differentiation-related proteins by the sea cucumber extract, expression of the related proteins was confirmed through western blot. The differentiation-completed 3T3-L1 cells were washed with PBS, and then lysis was conducted with RIPA buffer, and then the proteins were separated by molecular weight through SDS-PAGE. After that, it was transferred to a PVDF membrane and it was blocked using 5% skim milk for 30 minutes. Then, it was reacted with a primary antibody, and then a secondary antibody was attached to confirm protein expression using an ECL solution.

    [0103] (6) Statistics

    [0104] Statistical analysis of the result was conducted using GraphPad Prism 7 software (San Diego, CA, USA), and the significant difference between each group was analyzed through One-way ANOVA. (p<0.05)

    [0105] 2. Experimental Result

    [0106] (1) Process of Preparing Extracts and Fractions

    [0107] Ethanol extracts were obtained by extracting intestinal organs except for an ovary and an ovary of a sea cucumber as follows, respectively. After 30% EtOH, 50% EtOH or 80% EtOH was added to the dried sea cucumber ovary 20 times, they were stirred for 24 hours, and then filtered and concentrated in the same manner.

    [0108] Fractions of the sea cucumber ovary extract were obtained by the following method.

    [0109] After suspending the concentrated sea cucumber ovary extract by adding distilled water, the same amount of n-hexane was added and shaken and then stood. After completing layer separation, the n-hexane layer was separated and concentrated. Water saturated butanol was added to the remaining suspension and shaken, and then stood sufficiently, and after the layer separation was completed, the water saturated butanol layer was separated and concentrated.

    [0110] (2) Oil Red O Staining Result

    [0111] In order to investigate the effect on adipose differentiation of the 3T3-L1 cells of the sea cucumber extract, each sea cucumber extract was treated with a differentiation medium at a concentration of 25 and 50 g/ml (Sample No. 6, 10, 25 g/ml) for 48 hours to proceed an experiment. As a result of analyzing the degree of fat accumulation through Oil red O staining, all the samples except for Sample No. 1 showed a significant effect of inhibiting fat accumulation. (FIG. 4)

    [0112] (3) Analysis of Adipocyte Differentiation, Lipids in Blood, and Inflammation-Related Proteins

    [0113] Various proteins are involved in the process of adipocyte differentiation, and when these proteins are inhibited, adipose differentiation and accumulation are inhibited. As a result of confirming the effect of the sea cucumber extract on expression of major related proteins, PPAR, FAS, aP2, and C/EBP, similarly to the previous experiment, it could be confirmed that expression of adipose differentiation-related proteins was reduced in five samples except for No. 1 sample in two independent experiments, and accordingly, taking these results together, it can be seen that the sea cucumber extract inhibits expression of proteins related to adipose differentiation and this inhibitory effect inhibits adipose differentiation and accumulation.

    [0114] Through the above experiment, as a result of experimenting an effect of inhibiting adipocytes differentiation and fat accumulation of the sea cucumber extract extracted under each condition, a concentration-dependent effect of inhibiting fat accumulation was confirmed at a concentration of 25 and 50 g/ml. It seems that this effect inhibits adipocyte differentiation and expression of fat accumulation-related proteins such as PPAR, FAS, aP2, and C/EBP and regulates a process of differentiation into adipocytes, and it seems that the sea cucumber extract has a significant effect of inhibiting adipocyte differentiation. In case of PPAR, it is a factor having a correlation with inflammatory disease, and in case of FAS, it is an enzyme involved in fatty acid biosynthesis, and it has been known as an enzyme which regulates a concentration of triglyceride in blood and SREBP-1 (transcription factor regulating synthesis of cholesterol and fatty acids in fats and liver tissue). Therefore, through inhibition of expression of these proteins, an effect of improving dyslipidemia and inflammation can be obtained.

    III. Experimental Example 3Effect of Compound Separated from Sea Cucumber Ovary Extract

    [0115] 1. Separation, Structure Identification of 5 Kinds of Novel Saponins and 3 Kinds of Conventionally Reported Saponin Compounds

    [0116] Compounds corresponding to Chemical formulas S1, S2, S3, S4, S5, S6, S8 and S9 were separated from butanol fractions of the sea cucumber ovary extract by the following method. In order to isolate active components from the extract, separation and purification were performed according to activity guided fractionation. In order to separate active components from water saturated butanol fractions of the sea cucumber ovary, 8 kinds of saponin components were separated/purified by conducting various kinds of column chromatography.

    [0117] The separation/purification method was shown in FIG. 6.

    [0118] The specific separation/purification method is as follows. [0119] 1) The sea cucumber ovary butanol fractions obtained from the above were divided into small fractions through reversed-phase column chromatography. [0120] 2) polysaccharides were removed by erupting with 50% methanol in 100% distilled water, and after that, 60% methanol and 70% methanol fractions 1 and 2 were obtained, respectively. [0121] 3) The 60% methanol fraction was erupted with 30% ACN through preparative LC to obtain Saponin 1. [0122] 4) The 60% methanol fraction was erupted with 30% ACN through preparative LC to obtain Saponin 1. [0123] 5) The 70% methanol fraction 1 was divided into 4 small fractions through reversed-phase column chromatography again, and among them, No. 1 small fraction was erupted with 30% ACN through preparative LC to obtain Saponin 2. [0124] 6) The above No. 2 small fraction was erupted with 32% ACN through preparative LC to obtain Saponins 3 and 9, respectively. [0125] 7) The above No. 3 small fraction was divided into MC:MeOH=3:1, 2:1, 1:1 fractions through normal phase column chromatography again, and among then, the MC:MeOH=2:1 fraction was erupted with 35% ACN through preparative LC to obtain Saponins 4 and 5. [0126] 8) The 70% methanol fraction 2 was divided into MC:MeOH=7:1, 5:1. 3:1 fractions through normal phase column chromatography again, and among them, the MC:MeOH=3:1 fraction was erupted with 35%-40% ACN through preparative LC to obtain Saponins 6 and 8, respectively.

    [0127] Among them, the compounds corresponding to Chemical formulas S1, S2, S3, S4 and S9 corresponding to saponin-1, 2, 3, 4, and 9, respectively, were confirmed as novel compounds, and Saponin-5, 6, and 8 were confirmed as a chemical structure of holotoxin D1, holotoxin B, and holotoxin A, respectively.

    [0128] 2. Experimental Result

    [0129] (1) Cell Viability

    [0130] In order to investigate the effect on the 3T3-L1 cell viability of a total 8 kinds of the single compounds, as a result of confirming the cell viability by treating each of the single compounds at a concentration of 2.5-5 g/ml, it could be confirmed that a significant change was not shown in a majority of samples. In the samples of S3, S4, and S5, a significant decrease was shown at a concentration of 2.5 g/ml, and the cell viability of about 80% or more was shown.

    [0131] (2) Oil Red O Staining Result

    [0132] In order to investigate the effect on adipose differentiation of the 3T3-L1 cells of the single compound in the sea cucumber ovary extract, a total 8 of compounds were treated with a differentiation medium at a concentration of 2.5 and 5 g/ml for 48 hours. After completing differentiation, it could be confirmed that a significant difference was not shown at all the concentrations, but when S3-S9 were treated, fat accumulation was significantly reduced at a concentration of 2.5, 5 g/ml. It could be confirmed that the sample of S3 inhibited fat accumulation of about 16, 40, 80% from 1 g/ml to 5 g/ml and showed a tendency to have adipose differentiation inhibitory efficacy in a concentration dependent manner, and samples of S4-S8 inhibited fat accumulation at a level of non-differentiated cells at 2.5 g/ml.

    [0133] (3) Analysis of Adipocyte Differentiation-Related Proteins

    [0134] In order to figure out the mechanism of the fat accumulation inhibitory efficacy of the single compound examined above, the degree of expression of proteins of FAS, PPAR, C/EBP, and FABP4, which were adipogenesis-related proteins was confirmed. As a result, it could be confirmed that in the 3T3-L1 cells treated with S2, S2, compared to the control in which anything was not treated, a significant difference in protein expression was not shown, but in the cells treated with S3, S4, S5, from the cell treated with 2.5 g/ml, expression of the related proteins was definitely reduced, and it could be confirmed that it was definitely reduced in the cells treated with 1 g/ml compared to the control group. Next, as a result of confirming protein expression in the cells treated with S6, S8, S9, compared to the control group, a definite decrease in protein expression could be confirmed in S6 and S8. Therefore, taking these results together, it seems that the single compounds, S3-6, S8 inhibits adipocyte differentiation and fat accumulation through inhibition of expression of adipogenesis-related proteins.

    [0135] Through the experiment at this time, an effect of inhibiting adipocyte differentiation and fat accumulation of the single compound in the sea cucumber ovary extract was confirmed. As the experimental result of the previous extract, it seemed that expression of FAS, PPAR, C/EBP, and FABP4, which are adipogenesis-related proteins, was significantly reduced, and it inhibited adipocyte differentiation through such protein inhibition.

    [0136] <Nmr Data>

    [0137] Tables 4 to 9 below show the result of .sup.13C and .sup.1H NMR chemical shifts of S1-S4 and S9 among the compounds separated from the sea cucumber ovary extract. Through this, it was confirmed that S1-S4 and S9 were newly separated saponin compounds.

    TABLE-US-00004 TABLE 4 .sup.13C NMR (200 MHz) data for aglycones of compounds 1-3, 9 in C.sub.5D.sub.5N/D.sub.2O( in ppm) Position 1 2 3 9 1 36.0 36.0 36.0 36.0 2 27.1 27.2 27.2 27.2 3 89.0 89.1 89.1 89.1 4 39.5 39.6 39.6 39.6 5 47.7 47.8 47.8 47.8 6 23.4 23.5 23.5 23.4 7 122.6 122.6 122.6 122.6 8 147.9 148.0 148.0 148.0 9 46.2 46.2 46.2 46.2 10 35.7 35.7 35.7 35.7 11 22.1 22.1 22.1 22.0 12 20.6 20.6 20.6 20.6 13 54.8 54.9 54.9 54.9 14 46.0 46.0 46.1 46.1 15 44.7 44.7 44.7 44.7 16 79.8 79.9 79.9 79.9 17 62.5 62.5 62.5 62.5 18 182.1 182.2 182.2 182.3 19 24.1 24.1 24.2 24.1 20 71.3 71.3 71.3 71.3 21 26.7 26.7 26.7 26.6 22 42.6 42.5 42.5 42.5 23 22.1 22.1 22.1 22.1 24 38.5 38.5 38.5 38.5 25 146.0 146.0 146.1 146.0 26 110.5 110.5 110.5 110.6 27 22.4 22.4 22.4 22.5 30 17.4 17.3 17.3 17.3 31 28.8. 28.7 28.7 28.7 32 34.6 34.6 34.6 34.6

    TABLE-US-00005 TABLE 5 .sup.1H NMR (800 MHz) data for aglycones of compounds 1-3, 9 in C.sub.5D.sub.5N/D.sub.2O( in ppm) Position 1 2 3 9 1 1.47 (2H) 1.47 (2H) 1.47 (2H) 1.47 (2H) 2 2.09, 1.88 2.09, 1.88 2.10, 1.87 2.10, 1.87 3 3.25 3.26 3.25 3.25 4 5 0.98 0.97 0.99 0.98 6 2.05, 1.97 2.04, 1.96 2.05, 1.97 2.04, 1.96 7 5.67 5.66 5.67 5.67 8 9 3.25 3.25 3.23 3.24 10 11 2.05, 1.84 2.05, 1.83 2.05, 1.84 2.05, 1.87 12 2.67, 2.18 2.66, 2.17 2.66, 2.18 2.66, 2.19 13 14 15 2.17, 2.03 2.18, 2.01 2.19, 2.03 2.19, 2.02 16 5.11 5.10 5.11 5.12 17 2.69 2.67 2.69 2.70 18 19 1.06 (3H) 1.05 (3H) 1.06 (3H) 1.05 (3H) 20 21 1.52 (3H) 1.51 (3H) 1.52 (3H) 1.52 (3H) 22 1.82, 1.78 1.82, 1.77 1.82, 1.78 1.82, 1.78 23 1.72, 1.53 1.72, 1.53 1.71, 1.53 1.72, 1.53 24 2.08 (2H) 2.07 (2H) 2.08 (2H) 2.08 (2H) 25 26 4.83 (2H) 4.83 (2H) 4.83 (2H) 4.83 (2H) 27 1.73 (3H) 1.73 (3H) 1.73 (3H) 1.73 (3H) 30 1.11 (3H) 1.11 (3H) 1.11 (3H) 1.10 (3H) 31 1.28 (3H) 1.28 (3H) 1.28 (3H) 1.27 (3H) 32 1.44 (3H) 1.43 (3H) 1.45 (3H) 1.45 (3H)

    TABLE-US-00006 TABLE 6 .sup.13C NMR (200 MHz) data for glycoside moiety of compounds 1-3, 9 in C.sub.5D.sub.5N/D.sub.2O( in ppm) Position 1 2 3 9 Xy1.sup.1(1.fwdarw.C3) Xy1.sup.1(1.fwdarw.C3) Xy1.sup.1(1.fwdarw.C3) Xy1.sup.1(1.fwdarw.C3) 1 105.3 105.2 105.2 105.2 2 82.6 83.4 83.3 83.3 3 75.7 75.8 75.7 75.8 4 77.6 77.4 77.4 77.4 5 64.1 64.1 64.1 64.1 Glc.sup.1(1.fwdarw.4Xyl.sup.1) Glc.sup.1(1.fwdarw.4Xyl.sup.1) Glc.sup.1(1.fwdarw.4Xyl.sup.1) Glc.sup.1(1.fwdarw.4Xyl.sup.1) 1 103.4 103.3 103.3 103.3 2 74.4 74.3 74.4 74.4 3 78.0 78.0 78.0 77.9 4 71.5 71.5 71.5 71.4 5 78.7 78.6 78.3 78.6 6 62.4 62.4 62.4 62.3 Glc.sup.2(1.fwdarw.2Xyl.sup.1) Quin.sup.1(1.fwdarw.2Xyl.sup.1) Quin.sup.1(1.fwdarw.2Xyl.sup.1) Quin.sup.1(1.fwdarw.2Xyl.sup.1) 1 105.4 105.4 105.4 105.4 2 76.2 76.2 76.2 76.2 3 75.9 75.7 75.8 75.8 4 81.5 87.3 87.2 87.1 5 76.6 71.7 71.7 71.7 6(or Me) 61.5 18.2 18.2 18.3 Glc.sup.3(1.fwdarw.4Glc.sup.2) Glc.sup.2(1.fwdarw.4Quin.sup.1) Glc.sup.2(1.fwdarw.4Quin.sup.1) Glc.sup.2(1.fwdarw.4Quin.sup.1) 1 104.4 104.8 104.8 105.3 2 73.7 73.8 73.8 74.8 3 88,0 88.0 87.7 78.0 4 69.7 69.8 69.8 71.5 5 77.9 77.9 77.9 78.4 6 62.1 62.2 62.1 62.4 Glc.sup.4(1.fwdarw.3Glc.sup.2) Glc.sup.3(1.fwdarw.3Glc.sup.2) MeGlc.sup.1(1.fwdarw.3Glc.sup.2) 1 105.6 105.6 105.3 2 75.4 75.4 75.0 3 78.0 78.0 87.8 4 71.5 71.5 70.6 5 78.6 78.5 78.6 6 62.4 62.4 62.1 Me 60.8

    TABLE-US-00007 TABLE 7 .sup.1H NMR (800 MHz) data for glycoside moiety of compounds 1-3, 9 in C.sub.5D.sub.5N/D.sub.2O( in ppm) Position 1 2 3 9 Xy1.sup.1(1.fwdarw.C3) Xy1.sup.1(1.fwdarw.C3) Xy1.sup.1(1.fwdarw.C3) Xy1.sup.1(1.fwdarw.C3) 1 4.73 4.72 4.73 4.73 2 4.14 4.04 4.04 4.04 3 4.26 4.24 4.25 4.25 4 4.29 4.33 4.33 4.33 5 4.43, 3.66 4.43, 3.66 4.44, 3.67 4.44, 3.67 Glc.sup.1(1.fwdarw.4Xyl.sup.1) Glc.sup.1(1.fwdarw.4Xyl.sup.1) Glc.sup.1(1.fwdarw.4Xyl.sup.1) Glc.sup.1(1.fwdarw.4Xyl.sup.1) 1 5.02 5.02 5.02 5.02 2 4.03 4.03 4.02 4.03 3 4.25 4.25 4.24 4.26 4 4.18 4.16 4.15 4.15 5 4.01 4.00 4.00 4.01 6 4.55, 4.31 4.55, 4.29 4.54, 4.27 4.55, 4.28 Glc.sup.2(1.fwdarw.2Xyl.sup.1) Quin.sup.1(1.fwdarw.2Xyl.sup.1) Quin.sup.1(1.fwdarw.2Xyl.sup.1) Quin.sup.1(1.fwdarw.2Xyl.sup.1) 1 5.26 5.13 5.13 5.13 2 4.05 4.03 4.03 4.04 3 4.22 4.10 4.10 4.12 4 4.33 3.66 3.66 3.67 5 3.86 3.77 3.76 3.77 6(or Me) 4.55, 4,43 1.74 1.74 1.74 Glc.sup.3(1.fwdarw.4Glc.sup.2) Glc.sup.2(1.fwdarw.4Quin.sup.1) Glc.sup.2(1.fwdarw.4Quin.sup.1) Glc.sup.2(1.fwdarw.4Quin.sup.1) 1 5.16 4.97 4.97 4.98 2 4.09 4.07 4.05 4.04 3 4.21 4.27 4.29 4.27 4 4.05 4.02 4.03 4.13 5 3.96 4.02 4.03 4.09 6 4.45, 4.17 4.50, 4.17 4.50, 4.17 4.61, 4.25 Glc.sup.4(1.fwdarw.3Glc.sup.2) Glc.sup.3(1.fwdarw.3Glc.sup.2) MeGlc.sup.1(1.fwdarw.3Glc.sup.2) 1 5.28 5.33 5.33 2 4.07 4.08 4.00 3 4.25 4.25 3.75 4 4.15 4.14 4.08 5 4.03 4.04 4.01 6 4.55, 4.27 4.55, 4.26 4.51, 4.23 Me 3.89

    TABLE-US-00008 TABLE 8 .sup.13C and .sup.1H NMR data for aglycone moiety of compound 4 in C.sub.5D.sub.5N/D.sub.2O( in ppm) C.sup.a) H.sup.b) 1 36.3 1.84, 1.42 2 27.0 2.20, 2.01 3 88.9 3.24 4 39.9 5 52.8 0.87 6 21.1 1.62, 1.46 7 28.5 1,62, 1.21 8 38.8 3.24 9 151.2 10 39.7 11 111.2 12 32.1 2.65, 2.55 13 55.9 14 42.1 15 52.1 2.45, 2.22 16 214.0 17 61.3 2.94 18 176.5 19 22.1 1.37 (3H) 20 83.5 21 26.9 1.49 (3H) 22 38.4 1.82, 1.66 23 22.3 1.81, 1.54 24 38.0 2.00, 1.97 25 145.6 26 110.6 4.79 (2H) 27 22.3 1.71 (3H) 30 16.7 1.11 (3H) 31 28.1 1.23 (3H) 32 20.7 0.95 (3H)

    TABLE-US-00009 TABLE 9 .sup.13C and .sup.1H NMR data for glycoside moiety of compound 4 in C.sub.5D.sub.5N/D.sub.2O( in ppm) 8 C.sup.a) 8 H.sup.b) Xy1.sup.1(1.fwdarw.C3) Xy1.sup.1(1.fwdarw.C3) 1 105.2 4.78 2 81.9 4.20 3 75.5 4.29 4 77.7 4.29 5 63.9 63.9 Glc.sup.1(1.fwdarw.2Xyl.sup.1) Glc.sup.1(1.fwdarw.2Xyl.sup.1) 1 104.9 5.30 2 75.9 4.05 3 75.8 4.20 4 81.4 81.4 5 76.5 76.5 6 (or Me) 61.5 61.5 Glc.sup.2(1.fwdarw.4Glc.sup.1) Glc.sup.2(1.fwdarw.4Glc.sup.1) 1 104.1 5.14 2 73.8 4.08 3 87.6 4.25 4 69.5 3.99 5 77.6 3.98 6 61.9 4.44, 4.15 Glc.sup.3(1.fwdarw.3Glc.sup.2) Glc.sup.3(1.fwdarw.3Glc.sup.2) 1 105.2 5.28 2 75.3 4.07 3 77.7 4.25 4 71.4 4.09 5 78.3 4.04 6 62.3 4.55, 4.22 Me Glc.sup.4(1.fwdarw.4Xyl.sup.1) Glc.sup.4(1.fwdarw.4Xyl.sup.1) 1 102.6 5.01 2 73.5 4.03 3 87.4 4.31 4 69.6 69.6 5 77.8 77.8 6 61.9 61.9 MeGlc.sup.1(1.fwdarw.3Glc.sup.4) MeGlc.sup.1(1.fwdarw.3Glc.sup.4) 1 105.1 5.33 2 74.9 3.99 3 87.5 3.77 4 70.6 1.04 5 78.1 4.04 6 62.1 4.52. 4.20 Me 60.9 3.90

    [0138] a) Measured at 200 MHz in C.sub.5D.sub.5N/D.sub.2O, b) Measured at 800 MHz in C.sub.5D.sub.5N/D.sub.2O.

    [0139] Through the NMR data, it was confirmed that S1-S4 and S9 were novel saponin compounds.

    IV. Experimental Example 4Confirmation of Anti-Diabetic Effect of Sea Cucumber Ovary Extract

    [0140] 1. Experimental Materials and Methods

    [0141] (1) Reagents and Materials

    [0142] 3T3-L1 cells (ATCC, CL-173), DMEM high glucose (Welgene), Fetal bovine serum (MPBio), penicillin-streptomycin (Welgene), Dulbecco's phosphate-buffered saline (Welgene), Trypsin-EDTA (Welgene), Bovine calf serum (Welgene), Glucose Uptake-Glo Assay Kit (Promega)

    [0143] (2) Culture of 3T3-L1 Preadipocytes and Differentiation of Adipocytes

    [0144] The 3T3-L1 cells were cultured in a high glucose DMEM medium in which 10% Calf serum and penicillin-streptomycin were added, and subcultured per 2-3 days. The 3T3-L1 cells were aliquoted in a 96 well plate at a concentration of 210.sup.4 cells/well and then cultured for 3 days, and then it was replaced with an MDI medium that dexamethasone, insulin and 3-isobuty-1-methylxanthine (IBMX) were added to a medium containing 10% PBS instead of calf serum. After 2 days, it was replaced with a medium containing insulin, and the medium was replaced again per 2 days and differentiation was induced.

    [0145] (3) Glucose Uptake-Glo Assay

    [0146] The differentiation-completed 3T3-L1 cells were under starvation for 16 hours, and then the sea cucumber extract was treated in a serum-free medium at a concentration of 25, 50 ug/mL for 6 hours. The cultured supernatant was recovered and in the 3T3-L1 cells, using Glucose Uptake-Glo Assay Kit, a glucose transport activity degree was measured using a luminometer.

    [0147] 2. Experimental Result

    [0148] (1) Result of Confirming Degree of Glucose Uptake into Cells (Muscle Glucose Uptake) of Sea Cucumber Ovary Extract

    [0149] As can be confirmed in FIG. 10, the intracellular glucose uptake of the sea cucumber ovary extract is significantly excellent. FIG. 10 is the result of confirming the glucose uptake of the sea cucumber ovary extract and extract of intestinal organs except for an ovary of a sea cucumber. FIG. 11 is the result of confirming the glucose uptake by extraction condition of the sea cucumber ovary extract. At all the concentrations, the glucose uptake was excellent compared to the control group. In particular, it could be seen that the sea cucumber ovary extract had an excellent anti-diabetic effect, as it had glucose uptake efficiency increased more than Rosiglitazone conventionally used as a therapeutic agent for diabetes.

    [0150] As can be confirmed in Table 10 below, the sea cucumber ovary extract had the glucose uptake increased compared to the untreated group, and in particular, the sea cucumber ovary extract showed the uptake at a much higher level (1.29-fold, 1.43-fold, 1.35-fold) at the entire concentrations compared to the 1.24-fold increase of Rosiglitazone.

    TABLE-US-00010 TABLE 10 Sample Fold change Rosiglitazone (Ref.) 1.24 30% EtOH ~1.29 50% EtOH ~1.43 80% EtOH ~1.35

    [0151] Through the result, it was confirmed that it had increased glucose uptake by treatment of the sea cucumber ovary extract.

    V. Experimental Example 5Confirmation of Anti-Diabetic Effect of Saponin Compounds Separated from Sea Cucumber Ovary Extract

    [0152] 1. Experimental Method

    [0153] (1) Reagents and Materials

    [0154] They were prepared as the items of IV-1-(1).

    [0155] (2) Culture of 3T3-L1 Preadipocytes and Differentiation of Adipocytes

    [0156] The 3T3-L1 cells were cultured in a high glucose DMEM medium in which 10% Calf serum and penicillin-streptomycin were added, and subcultured per 2-3 days. The 3T3-L1 cells were aliquoted in a 96 well plate at a concentration of 210.sup.4 cells/well and cultured for 3 days, and then it was replaced with an MDI medium that dexamethasone, insulin and 3-isobuty-1-methylxanthine (IBMX) were added to a medium containing 10% FBS instead of calf serum. After 2 days, it was replaced with a medium containing insulin, and the medium was replaced again per 2 days to induce differentiation.

    [0157] (3) Glucose Uptake-Glo Assay

    [0158] For the differentiation-completed 3T3-L1 cells, starvation was conducted for 16 hours, and then a sea cucumber-derived single compound was treated in a serum-free medium at a concentration of 2.5 ug/mL for 6 hours. The cultured supernatant was recovered and for the 3T3-L1 cells, using Glucose Uptake-Glo Assay Kit, the glucose transport activity degree was measured using a luminometer.

    [0159] For the saponin compounds obtained previously, the anti-diabetic effect was confirmed.

    [0160] 2. Experimental Result

    [0161] FIG. 12 is the result of confirming the glucose uptake of the saponin compounds separated from the sea cucumber ovary extract. As could be confirmed in FIG. 12, when the glucose uptake of the untreated group was considered as 100%, all the compounds showed the uptake of 100% or more. Through this result, it could be seen that the separated compounds increased the glucose uptake compared to the untreated group, and it could be seen that each had an anti-diabetic effect.

    [0162] Table 11 below is fold change values, showing the degree of the glucose uptake compared to the control group. A glucose uptake at a similar level to Rosiglitazone was also shown.

    TABLE-US-00011 TABLE 11 Sample Fold Relative to Control Rosiglitazone (Ref.) 1.49 S1 1.25 S2 1.43 S3 1.1 S4 1.6 S5 1.38 S6 1.24 S8 1.43 S9 1.03

    VI. Experimental Example 6In Vivo Experiment of Sea Cucumber Ovary Extract

    [0163] [Effect of Reducing Body Weight (Anti-Obesity Use)]

    [0164] 1. Experimental Method

    [0165] (1) Experimental Animal Group Information

    [0166] An experiment was performed for C57BL/6 male mice (5-week-old), for a total of 50 animals, 10 in each group. In all other groups except for the normal diet group, a high-fat diet was fed. [0167] 1) Normal (Normal diet group) [0168] 2) HFD (High fat diet group) [0169] 3) HFD+0.05% sea cucumber ovary extract (SCOE) [0170] 4) HFD+0.1% sea cucumber ovary extract (SCOE) [0171] 5) HFD+0.2% sea cucumber ovary extract (SCOE)

    [0172] (2) Breeding Condition

    [0173] The extract was fed through a diet by preparing a feed, and an experiment was carried out for 8 weeks. The body weight was measured once a week, and the diet intake was measured 3 times a week.

    [0174] 2. Experimental Result

    [0175] FIGS. 13 and 14 are results which observe the change in body weight and the finally confirmed body weight of the high calorie diet intake mice. Through FIG. 13, it could be seen that in the group treated with the sea cucumber ovary extract (50% ethanol), the body weight increase rate was significantly reduced as a result of confirming up to 8 weeks. As confirmed through FIG. 14, it could be seen that the final body weight was reduced, when compared to the untreated group with the sea cucumber ovary extract indicated by HFD.

    [0176] [White Adipose Tissue Reducing Effect (Anti-Obesity Use)]

    [0177] 1. Experimental Method

    [0178] (1) H&E Stain of Adipose Tissue

    [0179] Hematoxilin is a basic/cationic substance, which stains the nucleus (anion) in dark blue or purple, and eosin is an acidic/anionic substance, which stains the cytoplasm (cation) in red or pink.

    [0180] The process of H&E staining is as follows.

    [0181] Dewaxing.fwdarw.dehydration.fwdarw.hematoxylin.fwdarw.differentiation.fwdarw.blueing.fwdarw.eosin.fwdarw.dehydration.fwdarw.clearing.fwdarw.cover-slipping

    [0182] (2) White fats among adipose tissue are related to obesity and overweight, and in the present experiment, lipid accumulation of epididymis white fats was histologically analyzed by H&E staining to confirm the tissue size of the white adipose tissue.

    [0183] 2. Result

    [0184] FIG. 15 is the result of the change in the white adipocyte confirmed through H&E stain.

    [0185] It was confirmed that when a high calorie diet was administered (HFD), the size of the white adipocyte related to obesity became significantly larger, and when the sea cucumber ovary extract was administered herein (HFD+SOCE 0.05%, HFD+SOCE 0.1%, HFD+SOCE 0.2%), the adipocyte size was significantly reduced. Through this, it was confirmed that an adipocyte size decrease, an anti-obesity effect, and the like could be obtained by treatment of the sea cucumber ovary extract.

    [0186] [Confirmation of Decrease in Total Cholesterol and Triglyceride in Blood (Improvement of Hyperlipidemia, Metabolic Syndrome)]

    [0187] 1. Experimental Method

    [0188] (1) High-Density Lipoprotein Content Compared to Total Cholesterol in Blood (HDL/TC)

    [0189] A sinking reagent of 200 l was added to a blood sample of 200 l, and then they were mixed and left at a room temperature for 5 minutes. After 5 minutes, centrifugation was performed at 3000 rpm for 10 minutes, and an enzyme solution of 3 ml was added to the centrifuged supernatant of 100 l each, and mixed. They were reacted in a 37 C. constant-temperature water bath for 5 minutes. Using a blind test as a control, the absorbance was measured at a wavelength of 500 nm.


    HDL-C(mg/dl)=(absorbance of specimen/absorbance of standard)100(mg/dl)

    [0190] (2) TC (Total Cholesterol) in Blood

    [0191] Test tubes for a blind test/standard/specimen were prepared, respectively. An enzyme solution of 3 ml was added to a blood specimen of 20 l, and they were mixed well. They were reacted in a 37 C. water bath for 5 minutes. Using a blind test as a control, the absorbance of the standard and specimen was measured at a wavelength of 505 nm.


    Total cholesterol (mg/dl)=(absorbance of specimen/absorbance of standard)concentration of standard solution

    [0192] (3) TG (Total Glyceride) in Blood

    [0193] Test tubes for a blind test/standard/specimen were prepared, respectively. An enzyme solution of 3 ml was added to a blood specimen of 20 l, and they were mixed well. They were reacted in a 37 C. water bath for 5 minutes. Using a blind test as a control, the absorbance of the standard and specimen was measured at a wavelength of 550 nm.


    Amount of triglyceride (mg/dl)=(absorbance of specimen/absorbance of standard)300(mg/dl)

    [0194] (Amount of triglyceride of standard solution: 300 mg/dl)

    [0195] 2. Result

    [0196] FIGS. 16 to 18 show the result of representing the HDL content compared to total cholesterol, the result of confirming the effect of reducing total cholesterol, and the result of confirming the effect of reducing triglyceride. Through these results, it was confirmed that the sea cucumber ovary extract treatment group showed an effect of reducing levels of cholesterol and triglyceride in blood. In particular, it should be noted here that although the total cholesterol numerical value was reduced, the HDL numerical value was increased. Through this result, it could be seen that it showed an effect for treatment of dyslipidemia.

    [0197] In addition, as a result of confirming expression of PPAR known to be involved in inflammation regulation, in particular, inflammatory bowel disease and/or rheumatoid arthritis using the extract, an effect of reducing PPAR compared to the high calorie diet could be confirmed. This result can be confirmed in FIG. 19. Through this, it was confirmed that the sea cucumber ovary extract could obtain the effect of inhibiting inflammation.

    INDUSTRIAL APPLICABILITY

    [0198] The present invention can provide a composition for reducing body weight, a composition for inhibiting obesity, a composition for anti-diabetes, a composition for anti-inflammation, and a composition for improving dyslipidemia, which comprises a sea cucumber ovary extract. The composition of the present invention can provide a method for reducing body weight, a method for inhibiting obesity, a method for treating or improving diabetes, a method for treating or improving inflammation, and a method for treating or improving dyslipidemia.