KR20230172710A - Health functional food composition for improving menopausal men's health containing Perilla frutescens var. acuta Kudo extract - Google Patents

Abstract

As a result of providing a health functional food composition for improving menopausal men's health containing the chazugi thermal water extract of the present invention as an active ingredient and conducting experiments using the same, the results showed that the number and motility of sperm were improved, testosterone levels were improved, antioxidant capacity was increased, and testicular disorders were improved. This had a confirmed effect.

Description

Health functional food composition for improving menopausal men's health containing chazugi extract and health functional food using the same {Health functional food composition for improving menopausal men's health containing Perilla frutescens var. acuta Kudo extract}

The present invention relates to a health functional food composition that can improve menopausal men's health, and a health functional food composition for improving menopausal men's health containing a Chazugi extract capable of improving menopausal men's health through improving testicular function, and This is about health functional foods using this.

Male menopause is a phenomenon caused by age-related testosterone deficiency. Unlike women, this symptom does not appear in all men and is a disease that varies greatly from person to person. According to the results of a recent study, as you enter your 40s, your blood testosterone concentration decreases by 1.6% every year, in your 70s it decreases to 50% of that in your 30s, and in your 880s it decreases to 30%.

Representative symptoms include decreased libido, erectile dysfunction, lethargy, decreased concentration, insomnia, decreased muscle mass, and depression, and the number of patients visiting hospitals complaining of symptoms is increasing every year.

According to the results of the 'Perception and Attitude Survey on Male Menopause' conducted on 1,000 men aged 45 or older in five Asia-Pacific countries (Korea, Australia, Hong Kong, Taiwan, and Thailand), one in two men aged 45 or older (approx. 53%) experience symptoms related to andropause syndrome, and among these, 76% of Korean men experience andropause symptoms, the highest figure among the five countries.

In Korea, symptoms of decreased male fertility due to reproductive aging are steadily increasing, but there is no clear treatment method. Widely used oral medications and hormonal treatments aim to restore sexual function, improve osteoporosis or bone density, restore muscle strength, and improve intellectual power. However, most of them must be administered for a long period of time and are accompanied by large and small side effects, so the development of natural functional foods that help improve symptoms is required.

The direct cause of male menopause is a decrease in testosterone levels secreted by the testes. Testosterone is a hormone that has a variety of functions, including the development of genital organs and sexual desire during secondary sexual characteristics, as well as inducing sperm production, sperm activity, promoting protein synthesis and bone growth, and maintaining reproductive organ function.

Meanwhile, Perilla frutescens (L.) Britton var. acuta (Thunb.) Kudo) is an annual medicinal plant belonging to the angiosperm phylum, dicotyledon class, and Lamiaceae in biological classification and is distributed in Korea and China. do. The stem is erect and square, the entire stem is purple and fragrant, and the leaves are opposite, broadly ovate, pointed at the end, round or somewhat wedge-shaped at the base, serrated at the edges, hairy on both sides, especially with veins. It has long hairs on top and long petioles.

Flowers bloom in light purple from August to September and grow in racemes at the ends of stems and branches, and on the upper leaves. The calyx is split into two, the upper one has three rows, the lower one has two rows, and there are hairs on the inside and outside of the tube. The corolla is usually short and labial, the lower lip is slightly longer than the upper lip, the stamens are two-strong, and the fruit is round and contained within the calyx.

The main species is purple overall, and the divisions are circular and have a net pattern. The leaves and stems are used for medicinal purposes, and the young leaves and seeds are edible. The young leaves are so similar to perilla leaves that it is difficult to distinguish them, and the sugar made from peryl aldehyde contained in chazugi is a sweetener that is about 2,000 times stronger than normal sugar, so it is used in cigarettes, intestines, toothpaste, etc. Leaves that are green rather than purple are called viridis. The leaves have white flowers, have a stronger scent than chazugi, and are often used as medicine.

The present invention seeks to provide a health functional food composition containing perilla extract, a natural raw material, to reduce side effects occurring in existing health functional foods for improving menopausal health.

Domestic Patent Publication No. 10-2020-0072148 relates to a composition for improving male menopause using complex herbal medicine materials and a health food using the same. It uses herbal medicine ingredients that act not only on individual symptoms of andropause but also on various symptoms. A composition for improving male genitalia using complex herbal medicine materials that can improve depression, reproductive function, sexual function, osteoporosis, muscular atrophy, and ultimately improve male menopause, and a health food using the same are disclosed. Domestic registered patent number 10-1870940 discloses a health food composition useful for health by improving male menopausal syndrome by enhancing the secretion of testosterone, a male hormone, by using night gate and red ginseng as ingredients for health food. Domestic Patent Publication No. 10-2018-0000136 discloses a health functional food for improving male menopause and a pharmaceutical composition for preventing or treating male menopause containing Shinchae, which has the effect of inhibiting the death of TM3 cells, as an active ingredient. . Domestic Patent Publication No. 10-2015-0038906 includes extracts of herbal medicines such as red ginseng, wild herb, goji berry, cornelian cherry, and turmeric, and contains extracts of herbal medicines that are effective in preventing or treating various symptoms of male menopausal syndrome. Disclosed is a composition for preventing or treating male hypogonadism or male menopausal syndrome and a health functional food.

In order to solve the problem of side effects occurring in existing health foods for improving male menopause, the present invention confirmed the male health improvement effect of chazugi (perilla leaf), which is of natural origin and used as a medicinal plant in East Asia, and included extracts using it. The aim is to provide health functional food compositions and health functional foods thereof.

The present invention provides a health functional food composition for improving menopausal health containing Chazugi thermal water extract as an active ingredient. The health functional food composition for improving menopausal men's health containing the Chazugi thermal water extract as an active ingredient further includes a pharmaceutically acceptable carrier excipient or a mixture thereof, and the formulation of the composition is tablet, capsule, or soft capsule. It may be any one selected from the group including tablets, pills, granules, and drinks.

In addition, another embodiment of the present invention may be to provide a health functional food for improving menopausal health, preferably manufactured by the above method.

In the present invention, an animal model in which testicular dysfunction was induced by busulfan and andropause symptoms appeared was used. As a result of administering various concentrations (100, 200 mg/kg) of Chazugi extract to animal models showing menopausal symptoms, it was confirmed that symptoms were improved, including improvement in sperm count and motility, improvement in testosterone levels, increase in antioxidant capacity, and improvement in testicular disorders.

Figure 1 shows the results of measuring body weight, testes, and relative testicular weight according to Experimental Example 2.
Figure 2 shows the results of testicular histological analysis according to Experimental Example 3.
Figure 3 shows the results of measuring sperm count and motility according to Experimental Example 4.
Figure 4 shows the results of measuring blood testosterone and FSH standards according to Experimental Example 5.
Figure 5 shows the results of antioxidant enzyme activity and gene expression levels encoding the enzyme according to Experimental Examples 6 and 7.

Hereinafter, the health functional food composition for improving menopausal men's health containing the Chazugi extract of the present invention and the health functional food using the same will be described in detail with an experimental example as follows.

<Manufacture of Chazugi hot water extract>

100 g of dried chazugi leaves were washed with distilled water, then 500 ml of distilled water was added, and extracted by heating in a reflux extractor at 100°C for 3 hours. After filtration using filter paper, the filtrate was concentrated under reduced pressure.

The residue remaining after filtration was extracted twice more using the same amount of distilled water, filtered, and concentrated under reduced pressure. The concentrated hot water extract was freeze-dried for 48 hours using a freeze dryer. By the above method, the hot water extract of Chazugi leaf was obtained according to the extraction solvent and used as a sample in the following experimental example.

According to an experimental example of the present invention, in order to confirm the effect of Chazugi extract on improving andropause, Chazugi extract was administered to experimental animals and the presence and extent of symptom improvement were analyzed, and a representative testicular disorder-induced animal model was used. Testosterone levels, the causes of andropause, number and movement of pauses, histological analysis of testes, and antioxidant capacity were evaluated.

<Experimental Example 1> Animal model and induction of menopausal symptoms

The animal model used according to Experimental Example 1 of the present invention was C57B6/L mice, and after a one-week adaptation period, they were divided into four groups of 10 mice each. Group1 is the control group; Group 2 is the group that was not treated with any substances, Group 2 is the menopause induction group, Group 3 is the menopause induction + 100mg/kg intake of chazugi extract, and Group 4 is the menopause induction + 200mg/kg intake of chazugi extract.

Menopausal symptoms were induced by administration of Busulfan (1,4-butanediol dimethanesulfonate), and 40 mg/kg of Busulfan was administered twice at 3-week intervals. Changes in symptoms were examined after oral administration of Chazugi extract at each concentration for 35 days to an animal model in which symptoms were induced.

<Experimental Example 2> Measurement of body weight and testicular weight

After completion of Experimental Example 1, the weight of the experimental animals was measured, the testes were removed, and the weight was measured. Figure 1 shows the results of measuring body weight, testes, and relative testicular weight according to Experimental Example 2.

Compared to the control group, body weight, testicular weight, and testicular weight relative to body weight were all decreased in the andropause induced model ( p <0.05). However, in the chazugi extract administration group, the decreased values tended to recover, and absolute and relative testicular weight recovered significantly ( p <0.05) in both the low and high concentration administration groups.

<Experimental Example 3> Histological analysis of testes

Histological analysis was performed to confirm the condition of the seminiferous tubules where sperm are produced in the testes extracted according to Experimental Example 2 and the germinal epithelium that constitutes them.

After fixing the extracted testis, a paraffin block was made according to the usual tissue preparation process, and then a 5-μm-thick section was obtained and hematoxylin and eosin (H&E) staining was performed. Twenty round or near-circular tubes within the section were randomly selected and the diameters of the seminiferous ducts and seminiferous genital epithelium were measured.

Figure 2 shows the results of testicular histological analysis according to Experimental Example 3. Micrographs of the seminiferous tubules in the control group showed normal spermatogenic activity in the seminiferous tubules, but the menopausal induction group showed seminiferous tubule atrophy with reproductive epithelial dysplasia.

The diameters of spermatozoa (arrows), cellular germ epithelium (black bars), and Sertoli cells (asterisks) shown in Figure 2 can be observed (A) and the cross-sectional values of seminiferous tubule total diameter (TD) and germ cell diameter (CD). is expressed graphically (B). The histological changes induced by Busulfan were significantly improved by administration of Chazugi extract, and when administering 100 and 200 mg/kg of extract, the diameters of the seminiferous ducts and the genital epithelium were significantly ( p <0.05) compared to the menopausal induction group. increased. (* p < 0.001 vs control, # p < 0.001 vs menopausal induction group, PFA: Chazugi extract, P. frutescens var . acuta).

<Experimental Example 4> Measurement of sperm count and motility

The epididymis of an experimental animal cut with scissors was placed in phosphate-buffered saline and incubated for 10 minutes to collect sperm. The number of sperm collected was measured through microscopic observation, and the ratio of motile sperm to the number of sperm was calculated.

Figure 3 shows the results of measuring sperm count and motility according to Experimental Example 4. The number and motility of sperm were significantly reduced in the menopausal induction model by busulfan, and when administered with Chazugi extract, the number of sperm was significantly increased at both concentrations. However, a significant increase in sperm motility was observed only in the group administered high-dose chazugi extract.

<Experimental Example 5> Measurement of blood testosterone and FSH (follicle stimulating hormone) levels

Testosterone is an important hormone that induces secondary sexual characteristics and acts on the seminiferous duct to produce sperm and induce sexual desire. FSH is a hormone that stimulates the gonad gland, inducing sperm differentiation and maturation and promoting sex hormone secretion. do. To check the levels of the two hormones, a blood sample was obtained from the heart of the animal model, the serum was separated by centrifugation, and the hormone levels were confirmed using enzyme-linked immunosorbent assay kits.

Figure 4 shows the results of measuring blood testosterone and FSH standards according to Experimental Example 5. Serum testosterone and FSH levels were significantly decreased in the menopausal induction group compared to the control group ( p < 0.001). However, while administration of Chazugi extract significantly ( p < 0.01) improved busulfan-induced abnormal serum testosterone hormone levels, it had no significant effect on serum FSH levels.

<Experimental Example 6> Antioxidant activity test

To test antioxidant capacity, experiments were conducted by homogenizing freeze-dried testicular tissue. Malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activity measurement kits were used.

Figure 5 shows the results of the antioxidant capacity test according to Experimental Example 6. SOD and GPx enzyme activities in testicular tissue were significantly decreased compared to the control group in the busulfan-induced menopausal model, but administration of Chazugi extract significantly improved the decreased activity. In contrast, MDA levels, which are a result of lipid peroxidation, significantly increased in the menopausal induction model and were significantly improved after administration of Chazugi extract (Figure 5 A, B, C).

<Experimental Example 7> Comparative analysis of mRNA through real-time qPCR

TRIzol reagent was added to the freeze-dried testicular tissue to extract total RNA, which was then converted to cDNA and subjected to qPCR analysis. Genes confirmed to be active were Sod1 and Gpx1, and beta-actin was used as an internal control to display relative expression levels.

Figure 5 shows the results of gene expression levels encoding antioxidant enzymes according to Experimental Example 7. Similar to the previous results, the expression level of genes Sod1 and Gpx1 encoding SOD and GPx in the menopausal induction model was significantly decreased compared to the control group, and significant improvement was confirmed in the group administered with chazugi extract ( p < 0.01), (Figure 5 D, E).

Experimental Examples 1 to 7 above confirmed the effect of Chazugi extract on a menopausal model in which testicular disorders were induced by busulfan. Compared to the control group, a decrease in testicular weight, a decrease in relative weight (weight-to-body weight), a decrease in the number and motility of sperm, a decrease in testosterone level, and histological changes were observed in the menopausal induction model. However, when chazugi extract was administered, the menopausal symptoms significantly worsened. It was improved, and antioxidant enzyme activity increased and MDA concentration in testicular tissue was significantly reduced.

Considering these results, it is believed that Chazugi extract will help improve men's health by relieving menopausal symptoms that increase with decreased testosterone levels due to aging and activating antioxidant enzymes.

<Manufacture of health foods using chazugi extract>

A health functional food composition for improving menopausal men's health containing chazugi leaf extract as an active ingredient can be manufactured in the form of tablets, capsules, soft capsules, granules, and liquid.

According to another embodiment of the present invention, the composition can be prepared as a beverage additive. The health functional food for improving menopausal men's health is powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, transdermal agent to contain 0.01 to 99.9% by weight of the composition containing the Chazugi leaf extract as an active ingredient. , can be manufactured by formulating a suppository or sterile injectable solution.

As interest in menopausal men's health increases, a lot of demand is expected by providing naturally derived compositions for improving menopausal men's health without side effects and health foods containing them, which will help increase profits for those involved in the business and have industrial applicability. There is.

Claims (3)

Health functional food composition for improving menopausal health containing Chazugi thermal water extract as an active ingredient
The composition of claim 1 is a health functional food for improving menopausal health containing the composition as an active ingredient.
The method of claim 2, wherein the health functional food further comprises a pharmaceutically acceptable carrier excipient or a mixture thereof, and the dosage form of the composition is selected from the group including tablets, capsules, soft capsules, pills, granules, and drinks. Health functional food for improving menopausal health, characterized by being manufactured from one of the following: