Anti-obesity composition comprising extract of rosa rugosa flower buds as active ingredient

Abstract

The present invention relates to an anti-obesity composition comprising an extract of Rosa rugosa flower buds as an active ingredient, and by virtue of comprising the extract, aims to provide a composition having excellent anti-obesity efficacy by exhibiting the effects of inhibiting fat accumulation, reducing triglyceride content, increasing the expression of adipose triglyceride lipase (ATGL) and AMP-activated protein kinase (AMPK) proteins, and decreasing the expression of G0S2 (G0 / G1 switch 2) and perilipin proteins.

Description

Anti-obesity composition containing extract of dandelion flower buds as an active ingredient

[0001] The present invention relates to an anti-obesity composition comprising a safflower bud extract as an active ingredient, and more specifically, to an anti-obesity composition comprising a safflower bud extract having an excellent anti-obesity effect by confirming the anti-obesity functionality of a safflower bud extract and establishing optimal extraction conditions.

[0002] Obesity, defined as excessive accumulation of body fat, is recognized as one of the most important diseases that humanity must overcome. Obesity is a chronic condition characterized by insufficient energy expenditure compared to nutrient intake, resulting in the accumulation of excess energy as body fat. The World Health Organization (WHO) defines obesity as abnormal or excessive fat accumulation that poses a health risk. A body mass index (BMI) of 25 or higher is considered overweight, and a BMI of 30 or higher is considered obese.

[0003] Meanwhile, global obesity has nearly tripled since 1975, with more than 1.9 billion adults aged 18 and older being overweight in 2016, and more than 650 million of them obese. Furthermore, as of 2020, 39 million children under the age of five were overweight or obese, indicating a sharp increase in the number of obese patients.

[0004] Obesity is caused by a complex genetic and phenomenological relationship between various neuroendocrine substances and abnormalities in various elements related to energy metabolism. Its causes are very diverse, including irregular eating habits, excessive food intake, lack of exercise, endocrine diseases, genetic factors, psychological factors, and medications. If obesity occurs and persists, it causes various complications such as hypertension, diabetes, hyperlipidemia, fatty liver, obstructive sleep apnea, and degenerative arthritis, and it also increases the risk of various cancers such as colon cancer, pancreatic cancer, prostate cancer, and breast cancer.

[0005] Accordingly, various treatments are being studied to treat obesity, but the available obesity treatments are very limited due to various side effects. As of 2020, obesity treatments available in Korea include short-term approved medications such as phentermine, diethylpropion, phendimetrazine, and mazindol, and long-term approved medications such as orlistat, naltrexone / bupropion sustained-release tablets, phentermine / topiramate capsules, and liraglutide. The obesity treatment market increased from $1.2 billion in 2016 to $1.75 billion in 2020, and various obesity treatments are being studied both domestically and internationally (Ministry of Trade, Industry and Energy “Bioindustry Technology Development Project” Information Provision Report, July 2021, Korea Pharmaceutical and Bio-Pharma Manufacturers Association).

[0006] Accordingly, the present invention aimed to manufacture a composition that can effectively improve obesity by using Rosa rugosa THUNB., a natural product, and in particular, by including a Rosa rugosa flower bud extract, it was aimed to develop a composition that exhibits excellent anti-obesity effects by inhibiting fat accumulation, reducing triglyceride content, increasing ATGL (Adipose triglyceride lipase) and AMPK (AMP-activated protein kinase) protein expression, and reducing G0S2 (G0 / G1 switch 2) and Perilipin protein expression.

[0007] The present invention aims to develop and provide an anti-obesity composition that suppresses fat accumulation, reduces triglyceride content, and regulates the expression of proteins related to adipogenesis and lipogenesis in 3T3-L1 adipocytes by containing an extract of the flower buds of the common dandelion as an effective ingredient.

[0008] The present invention provides an anti-obesity composition comprising a flower bud extract of Rosa rugosa THUNB. as an active ingredient.

[0009] In the anti-obesity composition of the present invention, the extract may be characterized by being obtained by hot water extraction of the flower buds of the dandelion.

[0010] In the anti-obesity composition of the present invention, the composition may be characterized as being a food composition.

[0011] The present invention provides a health functional food containing a flower bud extract of Rosa rugosa THUNB. as an effective ingredient.

[0012] The present invention relates to an anti-obesity composition comprising a daisy flower bud extract, and by containing a daisy flower bud extract as an active ingredient, it is possible to provide a composition having excellent anti-obesity efficacy by exhibiting effects of inhibiting fat accumulation, reducing triglyceride content, increasing ATGL (Adipose triglyceride lipase) and AMPK (AMP-activated protein kinase) protein expression, and reducing G0S2 (G0 / G1 switch 2) and Perilipin protein expression.

[0013] Figure 1 is a graph showing the effect of different concentrations of the extract of the common chrysanthemum on the cell viability of 3T3-L1 preadipocytes.

[0014] Figure 2 is a graph showing the effect of the extract of the Chinese chrysanthemum on fat accumulation.

[0015] Figure 3 is a graph showing the effect of the extract of the common perilla flower on triglyceride content.

[0016] Figure 4 shows the results of confirming the intracellular lipid reduction efficacy of extracts from each part of the corresponding flower.

[0017] Figure 5 shows the results of confirming the effect of the extract of the flower buds of the common chrysanthemum on the expression of ATGL (Adipose triglyceride lipase), G0S2 (G0 / G1 switch 2), Perilipin, and AMPK (AMP-activated protein kinase) proteins.

[0018] Figure 6 shows the results of measuring body weight changes according to administration of extract of the flower buds of the common chrysanthemum.

[0019] Figure 7 shows the results of DEXA (Dual Energy X-ray Absorption Meter, InAlyzer, Medikors, KOREA) body composition analysis.

[0020] Figures 8 to 13 show the results of serum biochemical analysis of the extract of the flower buds of the common chrysanthemum.

[0021] Figure 14 shows the results of tissue weight measurement of the extract of the flower buds of the common chrysanthemum.

[0022] Figure 15 shows the results of measuring the fat globules area of ​​retroperitoneal fat and liver of the extract of the flower buds of the common chrysanthemum.

[0023]

[0024] The present invention provides an anti-obesity composition comprising a flower bud extract of Rosa rugosa THUNB. as an effective ingredient.

[0025] Rosa rugosa (THUNB.) is a deciduous broadleaf shrub in the rose family, also known as "maegwae." Its young shoots are eaten as greens, and its roots are known to be beneficial for diabetes, toothache, and arthritis. The flowers are used to relieve pain and stop bleeding, and are also used as an ingredient in perfume. Rosa rugosa grows well in a variety of soils, but thrives in moist, fertile sandy loam soils, primarily found on sandy beaches. It can also overwinter outdoors and can be cultivated anywhere in the country. It is drought-tolerant and salt-resistant.

[0026] Meanwhile, in the anti-obesity composition of the present invention, the extract may be characterized by being a hot water extract of the flower buds of the common daisy. More preferably, the flower buds of the common daisy are hot water extracted at 80 to 120°C for 5 to 7 hours.

[0027] In the present invention, a composition capable of effectively improving obesity was prepared using Rosa rugosa THUNB., a natural product, and by including Rosa rugosa THUNB. flower bud extract, a composition exhibiting excellent anti-obesity effects was developed by exhibiting effects of inhibiting fat accumulation, reducing triglyceride content, increasing ATGL (Adipose triglyceride lipase) and AMPK (AMP-activated protein kinase) protein expression, and reducing G0S2 (G0 / G1 switch 2) and Perilipin protein expression.

[0028] Meanwhile, in the anti-obesity composition of the present invention, the composition may be characterized as being a food composition.

[0029] In addition, the present invention can provide a health functional food containing a flower bud extract of Rosa rugosa THUNB. as an effective ingredient.

[0030]

[0031] Hereinafter, the present invention will be described in more detail through the following examples and experimental examples. However, the scope of the present invention is not limited to the following examples and experimental examples, but includes all variations of technical concepts equivalent thereto.

[0032]

[0033] [Example 1: Preparation of extract from the flower buds of the common daisy]

[0034] In this example, the effects of solvents were compared by extracting safflower buds with different ethanol contents. Table 1 below shows the extraction conditions for safflower buds.

[0035] T1 (0%)T2 (30%)T3 (50%)T4 (70%)Extraction temperature100℃60℃60℃60℃Extraction time6 hours6 hours6 hours6 hours

[0036]

[0037] [Experimental Example 1: Confirmation of cell viability of extract from the flower buds of the common daisy]

[0038] In this experiment, an MTT assay (MTT experiment) was performed to confirm the effect of the extract of the safflower buds prepared in Example 1 on the cell viability of 3T3-L1 preadipocytes.

[0039] 3T3-L1 preadipocytes were cultured in DMEM (10% FBS, 1% PEST) medium and subcultured until sufficient cell numbers were secured. After securing sufficient cell numbers, 3 × 10 5Cells were seeded and the medium was replaced at 48-hour intervals. After the cells filled the culture dish, the medium was replaced and cultured for 48 hours. After 48 hours, the medium was replaced with a culture medium containing 0.5 uM dexanethasone, 500 uM IBMX, and 10 ng / mL insulin, and cultured for 48 hours to induce differentiation. After 48 hours, the medium was replaced with a culture medium containing 10 ng / mL insulin, and cultured for 48 hours to maintain differentiation induction. After 48 hours, the general medium (10% FBS, 1% PEST in DMEM) was replaced twice at 48-hour intervals to complete differentiation induction.

[0040] Figure 1 is a graph showing the effect of various concentrations of the extract of the daisy flower bud on the cell viability of 3T3-L1 preadipocytes. As shown in Figure 1, the experimental results confirmed that the extract of the daisy flower bud did not exhibit toxicity to 3T3-L1 cells up to a concentration of 0.5 mg / mL under all extraction conditions.

[0041]

[0042] [Experimental Example 2: Confirmation of the Body Fat Reduction Effect of Rose of Sharon Flower Bud Extract]

[0043] In this experiment, the effect of the extract of the flower buds of the common daisy prepared in Example 1 on lipid accumulation and triglyceride contents was investigated.

[0044] First, the fat accumulation experiment was conducted as follows. 3T3-L1 preadipocytes were cultured using DMEM (10% FBS, 1% PEST) medium and subcultured until sufficient cell numbers were secured. (-2 days) After securing sufficient cell numbers, 3 × 10 5Cells were seeded and the medium was replaced at 48-hour intervals. After the cells filled the culture dish, the medium was replaced and cultured for 48 hours. (Day 0) After 48 hours, the medium was replaced with a culture medium containing 0.5 uM dexanethasone, 500 uM IBMX, and 10 ng / mL insulin, and cultured for 48 hours to induce differentiation. When the medium was replaced, the materials were diluted by concentration and treated. (Day 2) After 48 hours, the medium was replaced with a culture medium containing 10 ng / mL insulin, and cultured for 48 hours to maintain differentiation induction. When the medium was replaced, the materials were diluted by concentration and treated. (Day 4-6) After 48 hours, the general medium (10% FBS, 1% PEST in DMEM) was replaced twice at 48-hour intervals to complete differentiation induction. When the medium was replaced, the materials were diluted by concentration and treated. (8 days) After 8 days, cells were washed with PBS, treated with 10% formalin, and fixed for 1 hour at 4°C. After fixation, cells were washed with PBS, treated with 60% oil red O solution, and stained for 1 hour at room temperature. After fixation, cells were washed with PBS, and oil red O dye was extracted with isopropanol. The absorbance was measured at 520 nm and compared with the control group (adipocytes).

[0045] Figure 2 is a graph showing the effect of the extract of the daisy flower bud on fat accumulation. As shown in Figure 2, it was confirmed that the effect of inhibiting fat accumulation increased as the treatment concentration of the extract of the daisy flower bud increased in all samples.

[0046] Meanwhile, the experiment to measure triglyceride content was conducted as follows. 3T3-L1 preadipocytes were cultured using DMEM (10% FBS, 1% PEST) medium and subcultured until sufficient cell numbers were secured. (-2 days) After securing sufficient cell numbers, 3 × 105 cells were seeded into 6 wells and the medium was replaced at 48-hour intervals. After the cells filled the culture dish, the medium was replaced and cultured for 48 hours. (0 day) After 48 hours, the medium was replaced with a culture medium containing 0.5 uM dexanethasone, 500 uM IBMX, and 10 ng / mL insulin, and differentiation was induced by culturing for 48 hours. When the medium was replaced, natural and composite materials were diluted and treated according to concentration. (2 days) After 48 hours, the medium was replaced with a culture medium containing 10 ng / mL insulin, and differentiation was maintained by culturing for 48 hours. When exchanging the medium, natural and composite materials were diluted and treated according to concentration. (4~6 days) After 48 hours, the general medium (10% FBS, 1% PEST in DMEM) was exchanged twice at 48-hour intervals to complete differentiation induction. When exchanging the medium, natural and composite materials were diluted and treated according to concentration. (8 days) After 8 days, the cells were washed with PBS, 200 ㎕ of PBS was added to each well, the cells were harvested, and triglyceride (TG) was analyzed using the EZ-Triglyceride Quantification Assay Kit.

[0047] Figure 3 is a graph showing the effect of the extract of the daisy flower bud on the triglyceride content. As shown in Figure 3, it was confirmed that the effect of suppressing the intracellular triglyceride content increased as the treatment concentration of the extract of the daisy flower bud increased in all samples.

[0048] That is, it was found that the extract of the safflower buds of the present invention has an excellent effect of suppressing fat accumulation and triglyceride content, thereby having a body fat reduction effect. Meanwhile, the present invention conducted the following experiment using the water extract of the safflower buds (0% ethanol), which is safer and more economical when applied to health functional food products while also having an excellent effect of suppressing fat accumulation and triglyceride content.

[0049]

[0050] [Experimental Example 3: Confirmation of the Lipid-Reducing Effect of the Rose of Sharon Flower Bud Extract]

[0051] In this experiment, extracts of each part of the rose (flower bud, stem, and root) were prepared using the water selected as the final extraction solvent in Experimental Example 2 and under the same extraction conditions, and the intracellular lipid reduction efficacy was confirmed. Meanwhile, the lipid accumulation inhibitory effect of the extracts of each part of the rose was confirmed using the same method as Experimental Example 2.

[0052] Figure 4 shows the results of confirming the intracellular lipid reduction efficacy of extracts from each part of the common daisy. As shown in Figure 4, among the extracts from each part of the common daisy extracted under T1 extraction conditions (0% ethanol), it was confirmed that the intracellular lipid reduction efficacy of the common daisy flower bud extract was the best.

[0053]

[0054] [Experimental Example 4: Confirming the Effect of Rose of Sharon Flower Bud Extract on Protein Expression]

[0055] In this experiment, the extract of the flower buds of the safflower selected in Experimental Example 2 was treated on 3T3-L1 adipocytes, and the effect on the expression of biomarker proteins that regulate fatty acid synthesis (lipogenesis) was confirmed through a western blot experiment.

[0056] 3T3-L1 preadipocytes were cultured in DMEM (10% FBS, 1% PEST) medium and passaged until sufficient cell numbers were secured. After securing sufficient cell numbers, 3 × 10 5Cells were seeded and the medium was replaced at 48-hour intervals. After the cells filled the culture dish, the medium was replaced and cultured for 48 hours. After 48 hours, the medium was replaced with a culture medium containing 0.5 uM dexanethasone, 500 uM IBMX, and 10 ng / mL insulin, and cultured for 48 hours to induce differentiation. After 48 hours, the medium was replaced with a culture medium containing 10 ng / mL insulin, and cultured for 48 hours to maintain differentiation induction. After 48 hours, the general medium (10% FBS, 1% PEST in DMEM) was replaced twice at 48-hour intervals to complete differentiation induction. The differentiated cells were replaced with a serum-free medium (containing only 1% PEST) and cultured for 18 hours. (Serum starvation) After starvation, the materials were diluted by concentration, treated, and cultured for 24 hours. After 24 hours, the culture medium was removed, the cells were washed with PBS, lysed with RIPA buffer, and centrifuged at 8,000 rpm for 10 minutes. After collecting the supernatant, the protein content was quantified at 20 μg using the Bradford method, mixed with sample buffer at a 3:1 ratio, heated at 100°C for 10 minutes, and then cooled. The protein sample was electrophoresed and transferred to a PVDF membrane, and then blocked with 8% skim milk (in TBST) for 1 hour. After washing three times with TBST, the membrane was treated with each antibody and reacted at 4°C for 18 hours. After washing three times with TBST, the membrane was treated with each secondary antibody and reacted at 4°C for 1 hour. After washing three times with TBST, the membrane was treated with ECL solution, film developed, and the bands were quantified using the ImageJ program and compared with the control group (adipocytes).

[0057] Figure 5 shows the results of confirming the effect of the water extract of the flower buds of the common lily of the valley on the expression of ATGL (Adipose triglyceride lipase), G0S2 (G0 / G1 switch 2), Perilipin, and AMPK (AMP-activated protein kinase) proteins in 3T3-L1 cells at a concentration of 75 ㎍ / ml.

[0058] As shown in Fig. 5, it was confirmed that the extract of the flower buds of the present invention showed an effect of increasing the expression of ATGL (Adipose triglyceride lipase) and AMPK (AMP-activated protein kinase) proteins, while decreasing the expression of G0S2 (G0 / G1 switch 2) and Perilipin proteins.

[0059] In this way, it was confirmed that the extract of the safflower bud of the present invention exhibits a body fat reduction effect by increasing ATGL (Adipose triglyceride lipase) and AMPK (AMP-activated protein kinase) protein expression and decreasing G0S2 (G0 / G1 switch 2) and Perilipin expression.

[0060]

[0061] [Experimental Example 5: In vivo anti-obesity efficacy evaluation of the extract from the flower buds of the common daisy]

[0062] In this experiment, the anti-obesity efficacy of the extract of the daisy flower buds selected in Experimental Example 2 above was evaluated in vivo. The anti-obesity efficacy of the extract of the daisy flower buds was evaluated in an obesity model induced by a 12-week high-fat diet using male SD rats.

[0063] A total of 48 6-week-old male SD rats without specific pathogens were purchased and used as experimental animals. After a one-week quarantine and adaptation period, healthy animals without weight loss were selected for use in the experiment. The experimental animals were housed in an SPF breeding facility under the following conditions: temperature 23 ± 3℃, relative humidity 50 ± 20%, ventilation rate 10 to 15 times / hour, lighting time 12 hours, and light intensity 150 to 200 Lux. During the acclimatization and testing periods, they were housed three animals each in cages made of polysulfone with PG3 beta chips (lavendi PG-3, LASvendi, Germanry) in an SPF breeding facility. During the adaptation period, the experimental animals were fed a dedicated feed (Teklad global 18% protein, Cat No. 2018C, Enviog) sterilized by UV irradiation, and drinking water was provided freely from tap water sterilized by an autoclave. Meanwhile, Table 2 below is a table summarizing the contents of this experiment.

[0064] Test groupTest substanceAdministration dose(mg / kg)Administration amount(ml / kg)Administration cycleAdministration routeNumber of animalsG1_ND(normal feed group)0.1%MC-5QD / 12WPO8G2_HFD(high-fat feed group)0.1%MC-8G3_HFD_HCA(positive control group)HCA1508G4_HFD_Test substance low doseNG-RRF(rose hips flower bud extract)508G5_HFD_Test substance medium doseNG-RRF(rose hips flower bud extract)758G6_HFD_Test substance high doseNG-RRF(rose hips flower bud extract)1008

[0065]

[0066] As shown in Table 2 above, the test substance administration to the test group was carried out by selecting healthy animals after a one-week adaptation period and classifying them into G1_ND (normal feed group), G2_HFD (high-fat feed group), G3_HFD_HCA (positive control group), G4_HFD_low-dose test substance (50 mg / kg), G5_HFD_medium-dose test substance (75 mg / kg), and G6_HFD_high-dose test substance (100 mg / kg) groups based on the egg block method.

[0067]

[0068] Meanwhile, the experimental method and results are as follows.

[0069] ① Inducing obesity

[0070] Obesity was induced using a 60% kcal high fat diet (D12492, Rodent Diet With 60kcal%, Research Diets, USA) during a 12-week trial period.

[0071]

[0072] ② Weight measurement

[0073] Body weights of test animals were measured upon separation from each group and twice a week (Monday and Thursday). Fasting body weights were measured after 10 hours of fasting before necropsy.

[0074] Figure 6 shows the results of body weight measurement, and as seen in Figure 6, significant body weight loss was observed in the control group administered with 150 mg / kg of G3_HFD_HCA and in the group administered with 50 mg / kg of G4_NG-RRF and 75 mg / kg of G5_NG-RRD compared to the high-fat feed group.

[0075]

[0076] ③ DEAX measurement

[0077] Body composition, including body fat, body fat mass, and BMC, was measured using DEXA (Dual Energy X-ray Absorption Meter) 2-3 days before autopsy.

[0078] Figure 7 shows the results of DEXA (Dual Energy X-ray Absorption Meter, InAlyzer, Medikors, KOREA) body composition analysis. As shown in Figure 7, the results of body composition analysis using DEXA before autopsy showed that the fat mass was statistically significantly reduced in the G3_HCA 150 mg / kg, G4_NG-RRF 50 mg / kg, G5_NG-RRF 75 mg / kg, and G6_NG-RRF 100 mg / kg administration groups compared to the high-fat feed feeding group.

[0079]

[0080] ④ Autopsy

[0081] At autopsy, after anesthesia with 2% isoflurane, the animals were opened and blood was collected from the abdominal aorta. Blood was collected into SST tubes (367957, BD STT II vacutainer, BD, USA) and centrifuged at 5,000 rpm for 10 minutes to obtain serum. The blood was placed in tubes labeled with the animal number and stored in an ultra-low temperature freezer (-80°C). After blood collection, the abdominal aorta and vein were incised to exsanguineate the animals, and the weights of the liver, retroperitoneal fat, epididymal fat, and visceral fat were measured and fixed in 10% neutral formalin.

[0082]

[0083] ⑤ Serum biochemistry test

[0084] At autopsy, serum was collected and analyzed using an automatic serum biochemical analyzer (AU680, BECKMAN COULTER) to measure FFA (Free fatty acids), ALP (Alkaline phosphatase), AST (Aspartate aminotransferase), ALT (Alanine aminotransferase), CHOL (Total cholesterol), LDL (Low density lipoprotein), HDL (High density lipoprotein), TG (Triglyceride), CREA (Creatinine), BUN (Blood urea nitrogen), ALB (Albumin), A / G ratio (Albumin / Globulin ratio), GLO (Globulin), GGT (Gamma glutamyl transferase), GLU (Glucose), LDH (Lactate dehydrogenase), Total protein, TBIL (Total bilirubin), and CK (Creatine kinase).

[0085] Figures 8 to 13 show the results of serum biochemical analysis. As seen in Figures 8 to 13, CHOL and LDL were significantly reduced in the G4_NG-RRF 50 mg / kg administration group, TG was significantly reduced in the G3_HCA 150 mg / kg and G6_NG-RRF 100 mg / kg administration groups, and FFA was significantly reduced in the G6_NG-RRF 100 mg / kg administration group.

[0086] The levels of ALT and ALP were significantly reduced in the G4_NG-RRF 50 mg / kg, G5_NG-RRF 75 mg / kg, and G6_NG-RRF 100 mg / kg administration groups, but GGT showed no difference from the high-fat feed group.

[0087] BUN levels showed a significant decrease in the G3_HCA 150 mg / kg administration group and the G4_NG-RRF 50 mg / kg, G5_NG-RRF 75 mg / kg, and G6_NG-RRF 100 mg / kg administration groups compared to the high-fat feed group, but no statistical significance was observed in the CREA levels.

[0088] For CK and LDH, there was a significant decrease in the G3_HCA 150 mg / kg administration group and the G4_NG-RRF 50 mg / kg, G5_NG-RRF 75 mg / kg, and G6_NG-RRF 100 mg / kg administration groups compared to the high-fat feed group.

[0089] GLU levels were significantly reduced in the G4_NG-RRF 50 mg / kg and G5_NG-RRF 75 mg / kg administration groups.

[0090] In summary of the above results, it was determined that the test substance NG-RRF was effective in reducing body fat in an obesity model induced by high-fat diet intake.

[0091]

[0092] ⑥ Production and interpretation of histopathology

[0093] At autopsy, fixed liver and retroperitoneal tissues were dehydrated, transparent, and infiltrated using a tissue processor (Tissue-Tek VIP 5 Jr, SAKURA Fineteck, Japan). Then, paraffin-embedded tissues were prepared in an embedding center (LEICA Eg1150H, LEICA, Germany) using a rotary microtome (HM 340E, Thermo scientific, USA) and 5 μm-thick sections were prepared. After deparaffinization and rehydration, they were stained with hematoxyline and eosin (H&E). Image analysis was performed under a microscope using the DIXI eXcope image analysis program to measure the lipid droplet area in the stained liver tissues to evaluate steatosis (% area). At least 10 adipocytes were counted in the retroperitoneum to determine the average size (μm).

[0094] Figure 14 shows the results of tissue weight measurement. As shown in Figure 14, the absolute weights of liver, epididymal fat, retroperitoneal fat, and visceral fat showed a decreasing trend in the G3_HCA 150 mg / kg, G4_NG-RRF 50 mg / kg, G5_NG-RRF 75 mg / kg, and G6_NG-RRF 100 mg / kg administration groups, and statistically significantly decreased in all tissues except epididymal fat. In visceral fat, both the control and NG-RRF administration groups showed a statistically significant decrease compared to the high-fat feed feeding group, which means that fat accumulation was suppressed by NG-RRF administration.

[0095] Meanwhile, Fig. 15 shows the results of measuring the area of ​​fat globules in the retroperitoneal fat and liver. As shown in Fig. 15, the histopathological evaluation of the liver showed that the area of ​​fat globules accumulated in the liver was significantly reduced in the high-fat feed group at 150 mg / kg of G3_HCA, 75 mg / kg of G5_NG-RRF, and 100 mg / kg of G6_NG-RRF compared to the high-fat feed group.

[0096] In addition, histopathological evaluation of retroperitoneal fat showed that the size of fat cells was significantly reduced in the G3_HCA 150 mg / kg administration group and the G4_NG-RRF 50 mg / kg, G5_NG-RRF 75 mg / kg, and G6_NG-RRF 100 mg / kg administration groups, which could be judged to be due to the inhibition of fat accumulation by NG-RRF administration.

[0097]

[0098] ⑦ Statistical analysis

[0099] Test data are expressed as mean±SD. If an individual's data falls outside the mean±SD, it can be excluded as an outlier. Statistical significance was assessed using ANOVA using the acquired data. Post hoc tests were performed using the Dunnett or Newman-Keuls method. If variances were not equal, ANOVA on ranks was used. The statistical method used was GraphPad Prism 8.4.3, a widely used commercial statistical package.

[0100]

[0101] As such, it was found that the extract of the safflower buds of the present invention exhibits effects of inhibiting fat accumulation, reducing triglyceride content, increasing ATGL (Adipose triglyceride lipase) and AMPK (AMP-activated protein kinase) protein expression, and reducing G0S2 (G0 / G1 switch 2) and Perilipin protein expression, and reduces body weight and body fat mass, CHOL (Total cholesterol), LDL (Low density lipoprotein), TG (Triglyceride), and FFA (Free fatty acids) in experimental animals, and exhibits effects of reducing liver, epididymal fat, retroperitoneal fat, and visceral fat, thereby producing a composition with excellent anti-obesity effects.

Claims

1. An anti-obesity composition comprising a flower bud extract of Rosa rugosa THUNB. as an active ingredient.

2. In paragraph 1, The above extract is, An anti-obesity composition characterized by hot water extraction of the flower buds of the rose of Sharon.

3. In paragraph 1, The above composition, An anti-obesity composition characterized by being a food composition.

4. A health functional food containing Rosa rugosa THUNB. flower bud extract as an active ingredient.

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