WO2017069506A1 - Pharmaceutical composition for prevention and treatment of menopausal symptoms, containing, as active ingredient, mixture of curcuma longa and licorice extracts - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention and treatment of menopausal symptoms, which contains, as an active ingredient, a mixture of Curcuma longa and licorice extracts. Specifically, the licorice may be heat-roasted licorice, but is not limited thereto, and the mixture of Curcuma longa and licorice extracts increases estrogen activity in human breast cancer cells (MCF-7), inhibits lipid accumulation in human liver cancer cells (HepG-2), and has effects of inhibiting lipid accumulation, suppressing weight gain, and improving liver protective activity and blood lipid concentration in animal models with menopausal hyperlipidemia induced through a high-fat and high-cholesterol diet after ovariectomy, and therefore, the mixture of Curcuma longa and licorice extracts of the present invention can be advantageously used as a pharmaceutical composition for the prevention and treatment of menopausal symptoms.

Description

Pharmaceutical composition for preventing and treating menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient

The present invention relates to a pharmaceutical composition for preventing and treating menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient.

Menopause is a transition period in which the gradual decline in ovarian function occurs and the physiological and sexual function decreases or is lost. Menopause is a permanent cessation of physiology after ovarian function is stopped as part of the menopausal process. ) Comes. When menopause comes, various diseases occur due to changes in hormones, such as decreased estrogen production, elevated follicular stimulating hormone (FSH) and luteininzing hormone (LH).

The disease caused by the hormonal change is called a menopausal disease (climacteric disorder), postmenopausal syndrome (menopausal syndrome) or menopausal syndrome, mainly in women 40s to 50s. Symptoms of menopausal disease include redness of the face, weakened bones, and relatively high levels of testosterone, resulting in increased body fat and decreased activity of fat cells leading to obesity. In addition, estrogen prevents the accumulation of visceral fat, lowers cholesterol levels, and protects blood vessels. As a result, menopausal women can easily accumulate cholesterol in blood vessels. As estrogen secretion decreases, total cholesterol and low density lipoprotein Increasing concentrations increase the risk of atherosclerosis and increase cardiovascular and liver related diseases (Carr, 2003).

One of the methods used to prevent or treat these symptoms is hormonal therapy, using estrogen monotherapy or estrogen-progesterone combination therapy. However, long-term use of hormone replacement therapy is problematic for side effects such as breast cancer, physical symptoms may include nausea, headache, breast pain, bloating, swelling and uterine bleeding, and mental symptoms of anxiety, anxiety and depression. And the like may appear.

Menopause usually occurs between 45 and 55 years old (mean 51 years old), but some women may be early menopause due to surgery or premature ovarian dysfunction, and have entered menopause if they have not had menstruation continuously for the past 12 months. As life expectancy increases, women spend one third of their lives after menopause (Pavelka et al., 1991), so there is a need for treatment of women's menopausal symptoms to improve their quality of life.

If the ovary is removed from the animal, there are signs similar to female menopausal symptoms, such as estrogen deficiency, weight gain, cholesterol levels, and fatty liver. High fat and high cholesterol diets exacerbate these symptoms, resulting in obesity, dyslipidemia, steatohepatitis and visceral adiposity (Zhu et al., 2013; Rios-Lu et al., 2010).

Curcumae Radix (CR) is a perennial herbaceous plant native to the Ginger family, native to tropical Asia, belongs to the roots of Curcuma Longa L., and consists of root stems used as medicinal herbs. It contains yellow pigments, curcumin, turmerone and dehydroturmerone, and promotes bile secretion, diuretic and detoxification effects (Kim et al., 1991), anti-cancer effect (Shinde et. al., 2007), anti-inflammatory effects (Connell and sutherland, 1969) and antioxidant effects (Connell, 1970).

Licorice (Ghycyrrhizae Radix et Rhizoma, GR) is a perennial herbaceous plant belonging to the fabaceae. It mainly uses the root and has reddish brown or dark brown color and has a unique sweet taste. Gycyrrhizae Radix Praeparata is made by roasting liquorice until the surface color of licorice is purple (Ko et al., 2007). The main components of licorice include glycyrrhizin, saponin, flavonoids, polysaccharides and asparagine, and some homonataloin, resins and fibers fibers) (Lee and Jang, 2010). In particular, glycyrazine lowers cholesterol and blood pressure in patients with hypertension, and has anticancer, antioxidant and antifungal effects. In addition, licorice has the advantage of being easily mixed with other medicines can be used as a raw material for medicine, health food and cosmetics.

Accordingly, the present inventors have studied to find a substance that is effective in preventing and treating menopausal symptoms among natural products and has a low side effect. As a result, a mixture of turmeric and persimmon extract increases estrogen activity in human breast cancer cells (MCF-7), and human liver cancer. Inhibition of lipid accumulation in cells (HepG-2) was confirmed. In addition, when the mixture of turmeric and persimmon extract was administered to an animal model inducing menopausal hyperlipidemia through high fat and high cholesterol diet after ovarian extraction, lipid accumulation inhibition, weight gain inhibition, hepatoprotective activity and blood lipid concentration were improved. Confirmed that the effect was completed the present invention.

An object of the present invention to provide a pharmaceutical composition for preventing or treating menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient.

Another object of the present invention to provide a health functional food for preventing or improving menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient.

Another object of the present invention provides a use of the composition containing a mixture of turmeric and licorice extract as an active ingredient for preventing or treating menopausal symptoms.

The mixture of turmeric and Jacorice extract of the present invention increases estrogen activity in human breast cancer cells (MCF-7), inhibits lipid accumulation in human liver cancer cells (HepG-2), and suppresses high fat and high cholesterol diet after ovarian extraction. In animal models inducing menopausal hyperlipidemia through lipid accumulation inhibition, weight gain inhibition, hepatoprotective activity and blood lipid concentrations have the effect of improving it can be useful as a composition for preventing and treating menopausal symptoms.

1 is a diagram showing the change in estrogen activity by treatment of the mixture of turmeric and persimmon extract in human breast cancer cells.

Figure 2 is a diagram showing the change in lipid accumulation by the treatment of turmeric and persimmon extract in human liver cancer cells.

Figure 3a to 3d is a diagram showing the weight change and weight gain by administration of the mixture of turmeric and persimmon licorice extract in the menopausal hyperlipidemia animal model.

Figure 4 is a diagram showing the morphological changes of liver tissue by administration of the mixture of turmeric and persimmon licorice extract in the menopausal hyperlipidemia animal model.

Figures 5a and 5b is a diagram showing the change of adipose tissue by administration of a mixture of turmeric and persimmon extract in menopausal hyperlipidemia animal model.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient.

In addition, the present invention provides a dietary supplement for preventing or improving menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient.

The present invention also provides the use of a pharmaceutical composition containing a mixture of turmeric and licorice extract as an active ingredient for preventing or treating menopausal symptoms.

Licorice in the mixture of the extract may be specifically, but not limited to Licorice. Licorice is a roasted licorice with heat Licorice is mentioned in the present invention may be different in the manufacturing method, but includes all those that are marketed and classified as a persimmon in Chinese medicine.

The mixture of turmeric and licorice extract may be prepared by a manufacturing method including the following steps, but is not limited thereto.

1) drying each of turmeric and licorice to remove moisture and pulverizing;

2) extracting at room temperature by adding an extraction solvent to each of the dried turmeric and licorice of step 1); And

3) mixing the extract of step 2) by filtration and concentration under reduced pressure.

In the above method, turmeric and licorice may be used without limitation, such as grown or commercially available.

In the above method, the drying of step 1) may be, but not limited to, drying under reduced pressure, vacuum drying, boiling drying, spray drying or freeze drying.

In the above method, the extraction method of step 2) may be used a conventional method in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction, extracted once to 5 times by hot water extraction method It may be, and more specifically, may be extracted three times, but is not limited thereto. The extraction solvent may be added 0.1 to 10 times to the dried sample, more specifically 10 times. Extraction temperature may be 20 to 40 ℃ but is not limited thereto. In addition, the extraction time may be 12 to 48 hours, but is not limited thereto.

In the above method, the extraction solvent of step 2) may be water, a lower alcohol of C ₁ to C ₂ or a mixture thereof, but is not limited thereto.

In the above method, the decompression concentration in step 3) may be to use a vacuum decompression concentrator or a vacuum rotary evaporator, but is not limited thereto.

In the composition of the present invention, the mixture of turmeric and licorice extract was prepared by adding water or alcohol to each dried product of turmeric and licorice, and prepared by mixing the single extract by weight ratio, specifically, turmeric extract and licorice extract Can be mixed in a weight ratio of 5: 1 to 1: 5. In addition, the mixture of turmeric and licorice extract includes but is not limited to those extracted in a composite composition by mixing in a weight ratio.

The menopausal symptoms are any one selected from the group consisting of estrogen deficiency, obesity, cholesterol increase and fatty liver, but are not limited thereto.

The inventors tested whether the mixture of turmeric and persimmon extract increased the estrogen activity in human uterine cancer cells (MCF-7). It was confirmed to exhibit high estrogen activity (FIG. 1).

In addition, as a result of experiments on whether the mixture of turmeric and persimmon extract inhibited lipid accumulation in human liver cancer cells (HepG-2), methyl-beta-cyclodextrin-palmitic acid (MβCD, Sigma) ) The treatment group was increased lipid accumulation compared to the control group, the mixture treatment group of turmeric and persimmon extract was confirmed that the lipid accumulation was inhibited compared to the control group (FIG. 2).

In addition, an animal model of menopausal hyperlipidemia was prepared in order to confirm whether the mixture of turmeric and persimmon licorice extracts had a therapeutic effect on menopausal symptoms, and the mixture of the extract was administered at 200 and 450 mg / kg / day concentrations. As a result of confirming the weight, it was confirmed that the body weight was reduced by 9 and 8% in the group administered with the mixture of turmeric and persimmon extract, respectively, as compared to the weight of the high fat and high cholesterol diet group after ovarian extraction (FIG. 3).

In addition, as a result of visual observation of the changes in the liver, the liver color of the high fat and high cholesterol diet group and the high fat and high cholesterol diet group after ovarian extraction turned yellow and the liver was swollen. The liver of the mixture administration group was yellowish brown, and the degree of swelling was also confirmed. In the liver tissues, high fat and high cholesterol diet group and ovarian extraction, high fat and high cholesterol diet group showed large and small fear-like fat cells in hepatocytes resulting in liver damage and fatty liver findings. In the mixture administration group of the liver damage was significantly reduced and it was confirmed that the large and small fear-shaped fat globules in the hepatocytes was significantly reduced (Fig. 4).

In addition, in the high fat and high cholesterol diet group and ovarian extraction, the high fat and high cholesterol diet group induced obesity and increased the cell size of the surrounding ovary compared to the normal diet group, while the ovarian fat in the mixture administration group of turmeric and persimmon extract It was confirmed that the cell size of significantly decreased (FIG. 5).

In addition, in the high fat and high cholesterol diet group and the ovarian extraction group, the high fat and high cholesterol diet group increased the weight of the peripheral and ovarian fats compared with the normal diet group, and obesity with weight gain was induced. In the mixture administration group of licorice extract, it was confirmed that the fat weight was reduced (Table 2).

In addition, total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) concentrations, and atherosclerosis (Atherogenic index) were compared with those of normal diets. The high fat and high cholesterol diet group and the high fat and high cholesterol diet group after ovarian extraction were increased, but decreased in the administration group of turmeric and persimmon extract (Table 3).

In addition, ALT and AST activity was significantly increased in the high fat and high cholesterol diet group and the high fat and high cholesterol diet group after ovarian extraction compared with the normal diet group, resulting in hepatotoxicity. And it was confirmed that the AST enzyme activity was reduced (Table 4).

Therefore, the mixture of turmeric and licorice extract of the present invention can be used as an active ingredient of the pharmaceutical composition for preventing and treating menopausal symptoms.

The composition of the present invention contains from 0.1 to 99.9% by weight of the mixture of turmeric and licorice extract of the present invention with respect to the total weight of the composition as an active ingredient, and may include a pharmaceutically acceptable carrier, excipient or diluent.

The compositions of the present invention may be in various oral or parenteral formulations. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or lactose (at least one compound). lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, and syrups, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.

As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The composition of the present invention may be administered orally or parenterally, and parenteral administration may select external or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injection methods. It can be used for external skin.

The dosage of the composition of the present invention varies depending on the weight, age, sex, health condition, diet, time of administration, administration method, excretion rate and severity of the disease of the patient, the daily dosage is a mixture of turmeric and licorice extract 0.01 to 1000 mg, specifically 30 to 500 mg / kg, more specifically 50 to 300 mg / kg, based on the amount, may be administered 1 to 6 times per day.

The compositions of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.

In addition, the present invention provides a dietary supplement for preventing or improving menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient.

The mixture of turmeric and Jacorice extract of the present invention increases estrogen activity in human breast cancer cells (MCF-7), inhibits lipid accumulation in human liver cancer cells (HepG-2), and suppresses high fat and high cholesterol diet after ovarian extraction. In the animal model induced by menopausal hyperlipidemia, lipid accumulation inhibition, weight gain inhibition, hepatoprotective activity and blood lipid concentrations are effective in improving the health functional food for preventing and improving menopausal symptoms.

The health functional food of the present invention may contain various flavors or natural carbohydrates as additional ingredients. The above-mentioned natural carbohydrates are sugars such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose and polysaccharides such as dextrin and cyclodextrin, xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin, aspartame, and the like can be used. The ratio of the natural carbohydrate can be selected from 0.01 to 0.04 parts by weight, specifically about 0.02 to 0.03 parts by weight per 100 parts by weight of the health functional food of the present invention.

In addition to the above, the health functional food of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, And a carbonation agent used for the carbonated beverage. These components can be used independently or in combination. The ratio of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health food of the present invention.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention in detail, and the content of the present invention is not limited by the Examples and Experimental Examples.

Example 1 Preparation of Licorice

Licorice is purchased from Gyeongdong Market (Seoul, South Korea), evenly and thinly cut and sprinkled with water, and when the surface is wet enough, it is baked on fire until licorice becomes yellowish violet (re-edited by Dong-Eui Library, Dong Pharmaceutical Co., Yeogang Publishing Co., Seoul, 61-64, 1993).

Example 2 Preparation of Mixture of Turmeric and Licorice Extracts

<2-1> Genetic analysis of turmeric sample

Nuclear ribosomal DNA (nuclear ribosomal DNA) was analyzed by using biological samples (leaves) of turmeric and blast searched by NCBI National Center for Biotechnology Information, which can be found in Table 1 below. As shown, 99% base homology with Curcuma longa (NCBI Genbank number-KF304502) was confirmed (Table 1).

Table 1

name	Length	Base homology (KF304502)	name	Length	Base homology (KF304502)
CL1	669 bp	99%	CL6	99%	676 bp
CL2	675 bp	99%	CL7	99%	677 bp
CL3	676 bp	99%	CL8	99%	675 bp
CL4	677 bp	99%	CL9	99%	669 bp
CL5	675 bp	99%	CL10	99%	677 bp

<2-2> Preparation of Turmeric and Licorice Extracts

Ulum was grown in Jeju Island and purchased by Jeju Yellow Ulum Co., Ltd., and verified by Dr. Gil Gi-jung of Jungbu University, and 10 times the amount of water or ethanol in the turmeric and persimmon plants to produce extracts of turmeric and persimmon. After the addition was extracted for 12 hours at 70 ℃. The extract was filtered through a 0.4 ㎛ filter, concentrated in a rotary evaporator (rotary evaporator), and lyophilized to prepare an extract. The mixture of turmeric and persimmon extract was prepared by mixing the turmeric extract and the persimmon extract in a weight ratio of 5: 1 to 1: 5.

<Example 3> Curcuminoid content measurement in turmeric extract

High performance liquid chromatography was performed to determine the curcuminoid content in turmeric water or ethanol extract. As a result, as can be seen in Table 2, the peak of the curcuminoids could be confirmed, the content was found to be high in the order of curcumin (curcumin), methoxycurcumin (methoxycurcumin) and bismethoxycurcumin (bismethoxycurcumin) Table 2).

TABLE 2

Curcuminoids	Content (mg / g)
	Ethanol extract	Water extract
Curcumin	0.34 ± 0.12	0.032 ± 0.009
Methoxycurcumin	1.05 ± 0.45	0.025 ± 0.007
Bismethoxycurcumin	2.59 ± 0.39	0.024 ± 0.005
all	3.98 ± 0.56	0.081

Experimental Example 1 Confirmation of Increased Estrogen Activity in Human Breast Cancer Cells (MCF-7) by Administration of Turmeric and Licorice Extracts

In order to determine the change in estrogen activity by the administration of the mixture of turmeric and persimmon extract.

Specifically, human breast cancer cells MCF-7 (ATCC, USA) is 10% fetal bovine serum (Hyclone, USA), 100 U / mL penicillin, 100 mg / mL streptomycin (streptomycin, Hyclone) , USA) was added and cultured in RPMI medium, where dextran-coated charcoal (dextran-coated charcoal, Sigma-Aldrich, USA) was used to remove steroid hormones from fetal bovine serum. The MCF-7 cell line was cultured at 39 ° C. and 5% CO ₂ , and the cells were divided into 96 well plates at 1 × 10 ⁴ cells / mL. The dispensed cells were replaced with RPMI medium without phenol red and incubated for 24 hours, followed by 10 ^-9 M estrogen (17β-estradiol, Sigma-Aldrich, USA), turmeric, and licorice water or ethanol. Extracts (10 μg / ml) were treated respectively. The degree of cell differentiation by estrogen activation was determined by sulforhodamine-B assay.

As a result, as shown in FIG. 1, in the turmeric extract treatment group and the licorice extract treatment group, the estrogen activity was 93.1% and 103.4%, respectively, while the mixture treatment group of the turmeric and licorice extract showed 111.8%. It was confirmed that the mixture treatment group of licorice extract significantly increased the estrogen activity (Fig. 1a). In addition, the mixture treatment group of turmeric and licorice extract also showed a high estrogen activity (Fig. 1b). These results indicate that the mixture of turmeric and persimmon licorice extracts have a strong binding affinity for estrogen receptors in MCF-7 cells, which may be useful for treating estrogen-related symptoms such as menopause.

Experimental Example 2 Confirmation of Lipid Accumulation Inhibitory Effect by Administration of Turmeric and Licorice Extracts in Hepatocellular Carcinoma Cells (HepG-2)

Lipid accumulation by treatment of methyl-beta-cyclodextrin-palmitic acid (MβCD, Sigma-Aldrich, USA) by administration of a mixture of turmeric and persimmon licorice extract in hepatic cancer cells (HepG-2) In order to evaluate whether this is inhibited, the experiment was as follows.

Specifically, HepG2 cells were cultured in DMEM medium containing 10% fetal bovine serum (Hyclone, USA), 100 U / mL penicillin, 100 mg / mL streptomycin (Hreplone, USA). When cells were about 70% full, the cells were replaced with 0.2% BSA-DMEM medium containing MβCD (20 μg / ml), a mixture of turmeric and persimmon extract (10 μg / ml) or simvastatin (simvastatin, 30 μm), respectively. It was. After 8 hours of incubation, the cells were fixed with 10% formalin for 10 minutes and washed three times with PBS. Cells were then rinsed with 60% isopropanol and stained with diluted oil red o solution (stock solution, 3 mg / ml in isopropanol; working solution, 60% Oil Red O stock solution diluted in water) for 1 hour. The stained cells were rinsed with 60% isopropanol and PBS and treated with 100% isopropanol to dissolve intracellular lipids and the absorbance was measured at 500 nm with a spectrophotometer.

As a result, as shown in Figure 2 compared with the control (100%) MβCD treated group was increased 1.62 times (162%) lipid accumulation, simvastatin treatment group was confirmed that lipid accumulation was significantly inhibited. In addition, it was confirmed that the mixture treatment group of turmeric and Jasimcho extract compared with the control group inhibited lipid accumulation in both the water extract and the ethanol extract (FIG. 2).

Experimental Example 3 Effects of Mixtures of Turmeric and Licorice Extracts in Animal Models Inducing Menopausal Hyperlipidemia

The following experiment was performed to evaluate the change by the administration of mixture of turmeric and persimmon extract in ovarian hyperlipidemia induced ovarian extraction and high fat and high cholesterol feeding.

<3-1> Preparing Menopausal Guidance Animal Models

 Seven week-old female rats (Sprague Dawley rat, SD, 200-220 g) were supplied by Orient Bio, South Korea. The mice were fed a week (Ralston-Purina, St. Louis, USA) with plenty of water and water, and adapted for 1 week in a light-dark cycle with a temperature of 22 ± 1 ° C, 50 ± 5% humidity, and 12 hours. . After one week, the rats were randomly divided into two groups, all of which were identical to the ovarian resection group except that the ovariectomy operation group removed the ovary and the sham operation group removed the ovary. Was performed.

<3-2> Preparation of Animal Model Inducing Menopausal Hyperlipidemia

To induce hyperlipidemia in menopausal animal models, the following experiments were performed.

Specifically, the rats prepared in Experimental Example <3-1> were further divided into seven small groups and reared for 8 weeks: (A) Siamese group (ordinary diet group) orally administered normal feed and saline, (B) high fat and Sham group orally administered high cholesterol feed and saline (high fat and high cholesterol diet group), (C) Ovariectomy group orally administered high fat and high cholesterol feed and saline (high fat and high cholesterol diet after ovarian extraction), ( D) Ovariectomy group (simvastatin 20) orally administered high fat and high cholesterol feed and simvastatin (20 mg / kg / day), (E) High fat and high cholesterol feed and a mixture of turmeric and persimmon extract (50 mg / kg) ovarian resection group (or mixture of turmeric and persimmon extract 50) orally administered (day) oral administration of a mixture of high fat and high cholesterol feed and turmeric and persimmon extract (200 mg / kg / day) Temperance Group (Sult and Ruler) Mixture of licorice extract 200) and (G) Ovariectomy group (mixture of turmeric and licorice extract 450) orally administered a mixture of high fat and high cholesterol feed with turmeric and licorice extract (450 mg / kg / day). Normal feed was supplied by Ralston-Furrina (St. Louis, USA), and high fat and high cholesterol feeds (45% / w / w fat and 0.15% w / w cholesterol) were supplied by Feedlab (Korea). received.

<3-3> Inhibition of weight gain by administering a mixture of turmeric and persimmon licorice extract in animal models inducing menopausal hyperlipidemia

The body weight of the test animals was measured once a week during the 8-week test period, and the weight gain was calculated by subtracting the initial weight from the final weight. Dietary intake was measured three times a week.

As a result, as shown in Figure 3a, the weight of the entire experimental group showed a tendency to increase during the experimental period, especially in the high-fat and high-cholesterol diet group after ovarian extraction from 2 weeks after administration, diet and high fat and high cholesterol diet The group and the degree of weight gain were significantly different (Fig. 3a).

As a result of examining the body weight at the end of the experiment, the high fat and high cholesterol diet group was increased by 30% compared with the normal diet group and 12% compared with the high fat and high cholesterol diet group after ovarian extraction. In addition, weight loss was reduced by 10% in simvastatin 20, 9% in 200% mixture of turmeric and persimmon extract, and 450% in 450% mixture of turmeric and persimmon extract in comparison with the body weight of high fat and high cholesterol diet after ovarian extraction. It was confirmed (FIG. 3A).

In addition, the high fat and high cholesterol diet group after ovarian extraction showed a statistically significant weight gain compared to the normal diet or the high fat and high cholesterol diet group, and the mixture 200 and 450 of simvastatin 20, turmeric and Licorice extract were ovaries Menopausal induction due to extraction and weight gain due to high fat and high cholesterol diet was significantly reduced (Fig. 3b).

The dietary intake of the experimental animals showed a slightly higher tendency than the other experimental groups in the administration of the mixture of turmeric and Jasimcho extract, and the change in dietary intake efficiency showed the same trend as the weight gain (FIGS. 3C and 3D).

On the basis of these results, it was confirmed that the mixture of turmeric and persimmon extract suppressed weight gain by menopause.

Experimental Example 4 Pathological Morphological Changes of Liver Tissue by Administration of Turmeric and Persimmon Licorice Extracts in an Animal Model Inducing Menopausal Hyperlipidemia

In order to examine the hepatoprotective effect by administration of the mixture of turmeric and persimmon extract in the menopausal hyperlipidemia animal model of Experimental Example <3-2>, the following experiment was performed.

<4-1> Confirmation of external shape change

After the experiment, blood was collected from the vena cava of the experimental animal, and liver and adipose tissue (peripheral ovary and peritoneal fat) were immediately separated and soaked in 10% formalin overnight.

When visually examined, the liver of the normal diet group showed chocolate color, and the liver color of the high fat and high cholesterol diet group and the high fat and high cholesterol diet group changed to yellow and the liver was swollen after ovarian extraction. On the other hand, the liver of the mixture administration group simbastatin 20 and turmeric and jasmine herb extract was yellowish brown, it was confirmed that the degree of swelling is small.

<4-2> Pathological examination of liver tissue

The liver tissue of Experimental Example <4-1> was stained with hematoxylin & eosin (hematoxylin & eosin) by a method well known in the art, and the shape of liver tissue was confirmed by an optical microscope.

As a result, as shown in Figure 4, in the high fat and high cholesterol diet group (B) and ovarian extraction after the high fat and high cholesterol diet group (C), large and small fear-shaped fat globules are deposited in the liver cells, liver damage and fatty liver findings Confirmed. On the other hand, simvastatin 20 (D) has a decreased degree of liver damage, a significant decrease in large and small terror-like fat globules in hepatocytes, and turmeric and persimmon extracts compared to high fat and high cholesterol diet group (C) after ovarian extraction. Mixture administration group (E, F, G) also showed a similar pattern to simvastatin 20 (Fig. 4). On the basis of these results, it was confirmed that the mixture of turmeric and persimmon extract has the effect of inhibiting fat accumulation in liver tissue.

Experimental Example 5 Pathological Morphology Changes of Adipose Tissue by Administration of Turmeric and Licorice Extracts in Animal Model Inducing Menopausal Hyperlipidemia

In order to examine the effect of inhibiting lipid accumulation by administration of a mixture of turmeric and Jacocci extract in the menopausal hyperlipidemic animal model of Experimental Example <3-2>, the following experiment was performed.

Specifically, the diameter, cell number, and size change of adipocytes in the adipose tissue of Experimental Example <4-1> were measured using Image J software. Indicated.

As a result, as shown in Figure 5, in the high fat and high cholesterol diet group (B) and ovarian extraction after high fat and high cholesterol diet group (C), the number and size of fat cells markedly increased while obesity was induced, normal diet group Compared with (A), the cell size of the peripheral ovary fat was increased by 105% and 115%, respectively. On the other hand, compared with the high-fat and high-cholesterol diet group (C), the cell size of peripheral ovary decreased by 37% compared to the high-fat and high-cholesterol diet groups (C), and the mixture of turmeric and persimmon extracts (E, F, G) ), It was confirmed that the cell size of the ovarian peripheral fat was reduced (FIG. 5).

In addition, as can be seen in Table 3, in the high fat and high cholesterol diet group and the high fat and high cholesterol diet group after the ovarian extraction compared to the normal diet group, the weight of the periphery fat, peripheral ovary fat increased, but simvastatin 20 and In the group administered with the mixture of turmeric and persimmon extract, fat weight was decreased after ovarian extraction compared with the high fat and high cholesterol diet groups (Table 3). On the basis of these results, it was confirmed that the mixture of turmeric and persimmon extract has an effect of suppressing ovarian fat accumulation by menopause.

TABLE 3

Metric (UI / L)	Normal diet.	High fat and high cholesterol diet	High fat and high cholesterol diet after ovarian extraction	Simvastatin	20	Mixture of turmeric and persimmon extract 50	Mixture of Turmeric and Licorice Extract 200	Mixture of Turmeric and Licorice Extract 450
Kidney fat	5.44 ± 0.87	10.42 ± 1.04	11.49 ± 1.56	10.07 ± 0.49	10.99 ± 1.71	9,75 ± 1.00	9.73 ± 1.16
Ovary	5.57 ± 1.85	7.01 ± 1.11	10.77 ± 1.40	9.06 ± 1.30	9.55 ± 1.34	8.93 ± 1.85	8.21 ± 1.79

Experimental Example 6 Changes in Serum Lipid Compositions According to the Administration of Turmeric and Licorice Extracts in Animal Model Induced by Climacteric Hyperlipidemia

In order to examine the antihyperlipidemic effect of the administration of the mixture of turmeric and persimmon extract in the menopausal hyperlipidemia animal model of Experimental Example <3-2>, the following experiment was performed.

Specifically, the blood obtained in Experimental Example <4-1> was collected by centrifugation (3000 rpm, 10 minutes), and serum was collected using a Roche Modular P Autoanalyzer (Roche diagnotics, USA). Lipid concentrations were analyzed according to the manufacturer's instructions.

As a result, as shown in Table 4, the total cholesterol (TC) concentration increased by 47% and 54% in the high fat and high cholesterol diet group and the high fat and high cholesterol diet group after ovarian extraction, respectively, compared to the normal diet group. Simovastatin 20 decreased ovary by 11% compared to the high fat and high cholesterol diet group, and decreased 22% and 17% in 200 and 450 mixtures of turmeric and persimmon extracts, respectively.

In addition, triglyceride (TG) concentrations were increased by 58% and 80% in the high fat and high cholesterol diet group and the high fat and high cholesterol diet group after ovarian extraction, respectively, compared to the normal diet group, especially high fat and high cholesterol after ovarian extraction. The TG concentration in the diet group was increased by 14% compared to the high fat and high cholesterol diet groups, indicating a significant change in TG concentration due to ovarian extraction. However, simovastatin 20 decreased TG concentrations by 19% compared to the high fat and high cholesterol diets after ovarian extraction, and 14% and 12% decreases in 200 and 450 mixtures of turmeric and persimmon extracts, respectively.

In addition, low density lipoprotein cholesterol (LDL-C) concentrations were increased by 173% and 220% in the high fat and high cholesterol diet group and the high fat and high cholesterol diet group after ovarian extraction, respectively, compared to the normal diet group. In ovary 20, ovary was reduced by 25% compared to the high fat and high cholesterol diet groups, and in the mixtures 50, 200, and 450 of turmeric and persimmon extract, 16%, 86% and 67%, respectively. High density lipoprotein cholesterol (HDL-C) concentration did not show statistical significance in the experimental group.

Atherosclerotic index (AI) was higher in the high-fat and high-cholesterol group and the higher-fat and high-cholesterol diet group by 1.56 and 2.03, respectively, by 143% and 217%, compared to 0.64 in the normal diet group. Simvastatin 20 Was 1.53, which was 33% lower than the high fat and high cholesterol diets after ovarian extraction. At 50, 200, and 450, the mixtures of turmeric and Jasimcho extracts showed that the arteriosclerosis index was 1.61, 1.08, and 1.17, respectively, which decreased by 26%, 88%, and 74% after ovarian extraction. Table 4). On the basis of these results, it was confirmed that the mixture of turmeric and persimmon extract had an effect of improving hyperlipidemia caused by menopause.

Table 4

Indicator (mg / dL)	Normal diet.	High fat and high cholesterol diet	High fat and high cholesterol diet after ovarian extraction	Simvastatin	20	Mixture of turmeric and persimmon extract 50	Mixture of Turmeric and Licorice Extract 200	Mixture of Turmeric and Licorice Extract 450
TC	67.40 ± 8.02	99.20 ± 2.77	103.50 ± 3.56	93.00 ± 4.80	100.17 ± 7.57	85.17 ± 7.94	88.33 ± 4.37
TG	54.80 ± 6.80	86.40 ± 4.83	98.50 ± 8.76	82.80 ± 5.63	93.17 ± 7.36	86.33 ± 2.58	88.33 ± 2.07
HDL-C	41.00 ± 2.45	39.80 ± 6.87	34.33 ± 2.80	37.00 ± 2.74	38.83 ± 4.26	41.33 ± 3.72	41.00 ± 3.63
LDL-C	15.44 ± 4.96	42.12 ± 7.52	49.47 ± 6.64	39.44 ± 5.00	42.70 ± 9.31	26.57 ± 10.47	29.67 ± 4.65
AI	0.64 ± 0.12	1.56 ± 0 .46	2.03 ± 0.29	1.53 ± 0.26	1.61 ± 0.38	1.08 ± 0.32	1.17 ± 0.19

TC, Total Cholesterol; TG, Triglyceride; HDL-C, high density lipoprotein -cholesterol; LDL-C, low density lipoprotein cholesterol; AI, Atheroscrelosis Index

Experimental Example 7 Changes of Enzyme Activities in Blood by Administration of Turmeric and Licorice Extracts in Animal Model Induced by Climacteric Hyperlipidemia

Aspartate aminotransferase (AST) and alanine amino to determine the presence of liver disease in order to determine the enzyme activity by administration of the mixture of turmeric and persimmon licorice extract in the menopausal hyperlipidemic animal model of Experimental Example <3-2> The activity of the transferase (alanine aminotransferase, ALT) was measured.

Specifically, the blood obtained in Experimental Example <4-1> was collected by centrifugation (3000 rpm, 10 minutes), and serum was collected using a Roche Modular P Autoanalyzer (Roche diagnotics, USA). Enzyme activity was analyzed according to the manufacturer's instructions.

As a result, as shown in Table 5, the high fat and high cholesterol diet group and the high fat and high cholesterol diet group after ovarian extraction compared to the normal diet group was found to increase ALT and AST activity resulting in hepatotoxicity, simvastatin In 20, the ALT and AST enzyme activities were reduced by 30% and 38%, respectively, compared to the high fat and high cholesterol diet groups after ovarian extraction. In addition, ALT enzyme activity was reduced by 34%, 43% and 35%, respectively, in the mixtures 50, 200 and 450 of turmeric and persimmon extracts compared to the high fat and high cholesterol diet groups after ovarian extraction, and AST enzyme activity was 9%, respectively. , 19% and 22% reduction (Table 5). The mixtures of turmeric and Jacorice extracts significantly reduced liver disease-related enzyme activities such as ALT and AST, thus protecting the liver by inhibiting damage to liver tissue caused by ovarian extraction and high-fat diets. there was.

Table 5

Metric (UI / L)	Normal diet.	High fat and high cholesterol diet	High fat and high cholesterol diet after ovarian extraction	Simvastatin	20	Mixture of turmeric and persimmon extract 50	Mixture of Turmeric and Licorice Extract 200	Mixture of Turmeric and Licorice Extract 450
ALT	30.20 ± 5.72	51.60 ± 4.51	57.83 ± 4.26	44.40 ± 6.80	43.17 ± 5.88	40.50 ± 1.87	42.83 ± 2.64
AST	61.80 ± 12.85	124.00 ± 16.05	131.20 ± 11.86	95.00 ± 2.16	120.83 ± 10.07	110.17 ± 5.53	107.67 ± 6.56

ALT, alanine aminotransferase; AST, aspartate aminotransferase

Claims (10)

1. A pharmaceutical composition for preventing or treating menopausal symptoms, which contains a mixture of turmeric and licorice extract as an active ingredient.
2. The pharmaceutical composition for preventing or treating menopausal symptoms according to claim 1, wherein the licorice is a persimmon licorice.
3. The pharmaceutical composition for preventing or treating menopausal symptoms according to claim 1, wherein the mixture of the extract is extracted using water, C ₁ to C ₂ lower alcohols, or a mixture thereof as a solvent.
4. The pharmaceutical composition for preventing or treating menopausal symptoms according to claim 1, wherein the extract is a mixture of turmeric extract and licorice extract in a weight ratio of 5: 1 to 1: 5.
5. The pharmaceutical composition for preventing or treating menopausal symptoms according to claim 1, wherein the menopausal symptoms are any one selected from the group consisting of estrogen deficiency, weight gain, cholesterol increase, and fatty liver.
6. Health functional food for preventing or improving menopausal symptoms containing a mixture of turmeric and licorice extract as an active ingredient.
7. 7. The dietary supplement for preventing or improving menopausal symptoms according to claim 6, wherein the licorice is persimmon licorice.
8. The dietary supplement for preventing or improving menopausal symptoms according to claim 1, wherein the mixture of the extract is extracted using water, C ₁ to C ₂ lower alcohols, or a mixture thereof as a solvent.
9. The dietary supplement for preventing or improving menopausal symptoms according to claim 1, wherein the extract is a mixture of turmeric extract and licorice extract in a weight ratio of 5: 1 to 1: 5.
10. The dietary supplement for preventing or improving menopausal symptoms according to claim 1, wherein the menopausal symptoms are any one selected from the group consisting of estrogen deficiency, weight gain, cholesterol increase, and fatty liver.

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