KR101376052B1 - The foodstuff composition comprising the extract of Rubus coreanus Miquel or ethyl acetate fraction therefrom for bone growth- promoting - Google Patents

Abstract

The present invention relates to a food composition exhibiting a bone growth promoting effect, including bokbunja (( Rubus coreanus Miquel) extract or ethyl acetate fraction, more specifically water or organic solvent extract of bokbunja and ethyl acetate fraction obtained from the It relates to a food composition having a bone growth promoting effect as an ingredient.
Food having an effect on promoting bone growth according to an embodiment of the present invention has a tibial length growth promoting effect can be usefully used for the treatment and prevention of tibial length growth disorders such as small authentication, dwarfism, stature growth.

Description

Food composition comprising the extract of Rubus coreanus Miquel or ethyl acetate fraction therefrom for bone growth-promoting}

The present invention relates to a food composition which has an effect on promoting bone growth containing bokbunja extract and / or ethyl acetate fraction separated therefrom as an active ingredient.

The development of children and adolescents has greatly increased due to the economic growth of modern society, changes in eating habits and improved nutritional status. As these average heights have increased, most people have become interested in growth, and the inferiority of children with smaller heights has increased.

Growth is predominantly controlled by the mother's environment before birth, but after birth it is a continuous and complex interaction of environmental factors such as genetic factors and nutrition (Cho Young-gil et al., Endocrinology, Medical Publishers, pp 247-265, 1985). In Chinese medicine, growth disorders of children are called emergency deficiency and renal insufficiency due to their physiological characteristics, congenital origin is kidney and spleen is spleen. 재) is the most closely related relationship (Jeong Hwan Jung, Oriental Medicine Review on Children's Growth. The Korean Journal of Oriental Pediatrics, 10 (1); 1-6, 1996). In general, the genetic effect is considered to be the largest factor in the growth of humans, but in fact it is only about 23%. The remaining 77% is determined by acquired effects, with research findings of nutritional 31%, exercise 20%, environment 16%, and other 10% (Kawada Aiyoshi, how to grow taller, Jeil Media, Rev. 1, 1997).

In addition, if growth is defined as a broad concept, not only an increase in height but also an increase in size and function of each organ of the body, but if defined as a concept of negotiations can be seen to mean an increase in height. This increase in kidney is achieved through skeletal metabolism, including the synthesis of cartilage tissue by nutrients and growth hormone, growth of skeletal length, and extensive proliferation of skeletal tissue. Bone (bone) tissue is a specially differentiated connective tissue, which is composed of several types of cells, including mesenchymal tissue, cartilage tissue, osteoblast, osteoclast, and bone marrow tissue. There is a balance between the amount of bone uptake by osteoclasts and the amount of bone formation by osteoblasts. During the growth phase, bone growth is greater than the amount of bone uptake because osteoblasts reach their maximum. Will be done. In this process, many hormones including growth hormone and nutritional status are combined, and among them, growth hormone is known to be involved in physical growth, especially skeletal growth, and the most important hormone for postnatal kidney growth (Byung Jin Choi, Korea Medicine). , 73-108, 1998).

Meanwhile, Bokbunja is an unripe fruit of Rubus coreanus Miquel, which belongs to the Rosaceae family, and contains a large amount of polyphenols, tannins, saponins, vitamins A, C, and anthocyanin in the fruit. Efficacy in traditional Chinese medicine is to boost the function of the kidney (유 精), montjung (夢 精), urination (遺尿), etc. to suppress the weakening of vision, lighten the body and blacken the head. It also makes the skin soft and beautiful (Kanbangpyeon, Namsandang, 1988).
The pharmacological effects of bokbunja were cytotoxic (Lee, Mi-Kyung. 11 (1); 5-12, 2003), antioxidant activity (Lee, Jong-Won, 29 (5); 943-947, 2000), testosterone secretion promotion Effect (Bae, BK. Et. Al ., Korean J. Oriental Physiol. Pathol. 18; 1007-1013, 2004) and immunity enhancing effect (Kim, Min-Chul, Journal of Medicinal Crop Science, 15 (6): 398-404, 2007) It has been reported.
In addition, growth-related studies (including patents) using herbal medicines containing bokbunja were described in (1) Effects of growth-related herbal preparations on the long bone length of rats (Lim Kang-Hyun. The Korean Journal of Herbology 18 (3); 181-186. Increased cartilage cell count in 2003), ② comparison of growth of height in health supplement food composition for growth promotion (Korea Patent Registration No. 10-0278206), ③ showing growth promotion effect by mixing 23 kinds of raw materials such as seaweed calcium Investigation of Weight Changes of Experimental Animals in Food (Korean Patent Registration No. 10-0715653) and Nutritional Effects of Herbal Combination Administration on Growth and Secretion of Growth Hormone in Pigs (Ko Bon-Hong et al., Korean Journal of Oriental Pediatrics 12 (1); 277- 287, 1998), but these results describe the appearance (weight, height) of growth or the effect of increasing the number of cartilage cells in the growth plate, but the tibial length growth has not been studied. In addition, the information described in these documents is a result of a study using a compound containing several kinds of herbal medicines such as bokbunja, and it is unclear whether bokbunja directly influenced growth hormone secretion or increased cartilage cell count. .

Therefore, the present inventors are Korean specialty Bokbunja ( Rubus) that grows in Korea The present invention was completed by identifying the fact that the tibial length has a growth promoting function in a specific fraction of the coreanus Miquel) extract and can be usefully used for the growth and development of children.

The present invention has been made to solve the problems of the prior art as described above, the object is to provide a food composition that has an effect on promoting bone growth containing the bokbunja extract or an active fraction obtained therefrom as an active ingredient. .

In order to achieve the above object,
The present invention comprises the steps of extracting the bokbunja with alcohol to obtain a primary extract; And extracting the primary extract with ethyl acetate to obtain an ethyl acetate-soluble heavy component extract.

In preparing the bokbunja extract, which is an active ingredient of the food of the present invention, the solvent may be limited to water and organic solvents such as alcohol, hexane, acetone, and the like. The alcohol is preferably a lower alcohol such as methanol, ethanol, propanol, butanol, and among these, ethanol is preferably used, but is not necessarily limited thereto. In addition, the concentration of the organic solvent is preferably 10 to 100% by volume, more preferably 70% by volume.

The present invention also provides a food composition exhibiting an effect on promoting bone growth containing water or an organic solvent extract of Bokbun, ethyl acetate fraction of the extract, or an active ingredient of the extract as an active ingredient.

The ethyl acetate fraction of the bokbunja extract may be prepared in the form of tablets for the purpose of promoting children's growth, in addition, it may be prepared in the form of powders, granules, capsules, powders, or extracts in accordance with conventional manufacturing methods. In addition, it may be added to a variety of health foods for the purpose of promoting children's growth, when used as a food additive, it may be added as it is, or used together with other foods or food ingredients, may be appropriately used according to conventional methods.

There is no particular limitation on the kind of the food. Examples of foods to which the substance may be added include candy, confectionery, noodles, gum, ice cream, beverages, drinks, vitamin complexes, and the like, and in general sense, children's food, including health foods.

Bokbunja extract of the present invention or the ethyl acetate fractions isolated therefrom are effective in bone growth and can be used as a health food to be applied in the prevention and treatment of bone length growth disorders such as subauthentication, dwarfism and stature growth.

Figure 1 is a graph measuring the interval between the growth plate and the light emitting line 48 hours after the tetracycline (tetracycline) injection in the rat.
Control group: saline administration group, experimental group 1: ethanol extract (100 mg / kg) administration group, experimental group 2: ethanol extract (500 mg / kg) administration group, experimental group 3: ethanol extract ethyl acetate fraction (100 mg / kg) administration group, experimental group 5 : Water extract (100mg / kg) administration group, ^** : P <0.01

The present invention provides a food composition effective for promoting bone growth in children consisting of bokbunja extract or ethyl acetate fraction isolated therefrom.

Example 1 Preparation of Bokbunja Extract

1-1 Preparation of Bokbunja Extract Using Water

Purchasing Bokbunja (Gyeongdong Market, Seoul) to remove impurities, washing and then extracting twice at 80 ° C for 4 hours with distilled water of 10 times the weight of the sample (w / v). , England), and the filtrate was concentrated in a rotary vacuum evaporator (Buchi) and then lyophilized to prepare a water extract of bokbunja.

1-2 Preparation of Bokbunja Extract Using 70% Ethanol

The ethanol extract of Bokbunja was prepared by extracting and concentrating using 70% ethanol in the same manner as in 1-1.

Preparation of Ethyl Acetate Fraction of 1-3 Bokbunja Extracts

50 g of the crude extract of Bokbunja prepared using 70% ethanol was suspended in 1 L of 70% ethanol, and then fractionated three times with soluble fraction of each solvent while gradually increasing the polarity of the solvent with hexane, ethyl acetate and unsaturated butanol. The ethyl acetate fractionated layers were collected, concentrated completely with a reduced pressure concentrator, and lyophilized. As a result, 16.5 g of ethyl acetate fraction powder was obtained.

Experimental Example 1 Determination of tibial bone growth by bokbunja extract and ethyl acetate fractions isolated therefrom

1-1 Animal Materials

For the measurement of bone length growth by bokbunja extract, the animals used in the experiment were Sprague-Dawley male rats (3 weeks old) purchased from Santaco (Gyeonggi-do, South Korea). Adapted to the experimental environment for one week while supplying enough, divided into the control group and each sample administration group was used in the experiment. The external conditions of the animal breeding room were adjusted to a temperature of 20 ± 2 ° C., a relative humidity of 50 ± 10%, and an illumination time of 12 hours (07:00 lit, 19:00 off). Experimental animals were housed in 5 polycarbonate cages (10.25W x 18.75D x 8H inch) during the entire breeding period. Feed was freely fed solid animal feed (Samyang Feed, Korea) for experimental animals, and water was freely ingested using water bottles.

1-2 Experimental Design

The rats were divided into 6 groups of 10 rats, saline solution for the control group, 100 mg / kg bokbunja 70% ethanol extract (experimental group 1), 500 mg / kg bokbunja 70% ethanol extract (experimental group 2), 100 mg / kg Ethyl acetate fraction of 70% ethanol extract (Experimental group 3), 100 mg bokbunja water extract (Experimental group 4) and 500 mg bokbunja water extract (Experimental group 5) were all once a day for 6 days from the start of the experiment to the end of the experiment. Physiological saline and samples were administered orally. Both groups were treated with tetracycline (10 mg / kg, i.p., Sigma, U.S.A.) two days before and one day before the end of the experiment and maintained until the end of the experiment. On day 7 of the experiment, the ankles were sacrificed by anesthesia and sacrificed, and the tibia was separated from the rats, fixed in 4% phosphate-buffered paraformaldehyde for 48 hours, and then immersed in 30% Sucrose solution. . After the frozen bone tissue was frozen, the sagittal section of the proximal part of the tibia was sectioned every 40 μm using cryocut and collected and used as a tissue sample. Sections were then used for length growth analysis. In order to measure the growth, the distance between the growth plate and the emission line developed by tetracycline was measured by fluorescence microscopy (Hansson et al., Calcified Tissue Research, 10 (3), 238, 1972, FIG. 1), The average value for each section was calculated.

Tibial Growth and Growth Rate by Experimental Group division Growth (μm / day) Growth rate (%) Control (saline) 278.5 ± 19.2 100 Experiment 1 285.2 ± 21.3 102.4 Experiment 2 295.8 ± 18.6 106.2 Experiment group 3 308.8 ± 15.9 110.9 ** Experiment group 4 279.9 ± 18.3 100.5 Experimental group 5 286.7 ± 23.6 103.0
As a result, the administration of bokbunja extract or ethyl acetate fractions had a statistically significant tibial growth compared to the control group, and the bokbunja ethanol extract administration group tended to have an effect on the tibial growth in a concentration-dependent manner, especially in the bokbunja ethyl acetate fraction administration group. Tibial growth was shown (Table 1). ^** indicates P <0.01.

In addition, when the growth rate of the control group administered with saline was 100%, the growth rate of the bokbunja extract (experimental group 1, experimental group 2), acetate fraction administration group was calculated. As a result, the bokbunja extract administration group and ethyl acetate fraction administration group was significantly higher than the control group (Fig. 1).

Fig.

Control group: saline, experimental group 1: ethanol extract (100 mg / kg), experimental group 2: ethanol extract (500 mg / kg), experimental group 3: ethyl acetate fraction (100 mg / kg), experimental group 5: water extract (100 mg / kg) kg)

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<Experimental Example 2> Rubus coreanus extract and the acute toxicity test of the ethyl acetate fraction

In order to determine the acute toxicity for the bokbunja extract or ethyl acetate fraction, the following experiment was performed.

2-1 Animal Materials

SPF (specific pathogens free) ICR mice were divided into 4 groups (3 females and 3 females / experimental group) at 4 weeks of age, temperature 22 ± 3 ℃, humidity 55 ± 10%, illumination 12L / 12D It was bred in a condition animal room. Mice were purified for a week before being used in the experiment. Feed for experimental animals (Samyang Feed, Korea) and drinking water were sterilized and fed free.

2-2 Experimental Design

Bokbunja 70% ethanol extract and ethyl acetate fractions prepared in Example were prepared at a concentration of 50 mg / ml in 0.5% tween 80, and then 0.04 ml (100 mg / kg) and 0.2 ml (500) per 20 g of mouse body weight. Mg / kg) and 0.4 ml (1,000 mg / kg). Samples were administered orally once and observed for side effects or lethality for 7 days after administration. That is, the change of general symptoms and the presence or absence of dead animals were observed at least once every morning and every afternoon from the day of administration to the 7th day. In addition, on the 7th day of administration, animals were killed and dissected, and the internal organs were visually examined. The change in body weight was measured at the interval of 1 day from the day of administration, and the weight loss phenomenon of the animals by the bokbunja extract was observed.

As a result, no significant clinical symptoms were observed in all the mice to which the sample was administered, no mice died, and no toxicity change was observed in weight change, blood test, blood biochemical test, autopsy findings, and the like. Therefore, bokbunja 70% ethanol extract and ethyl acetate fraction were judged to be safe substances over 1,000 mg / kg in all mice. Therefore, 50% lethal dose (LD50) by oral toxicity test is considered to be a safe substance of at least 1,000 mg / kg or more.

Claims (4)

Bonebun ( Rubus coreanus Miquel) 70% (v / v) is added to 70% (v / v) food composition for promoting bone growth containing bokbunja extract obtained by heating and reflux heating for 8 hours. delete delete The food composition according to claim 1, which is a pizza, a wire, a noodles, a bread, a confectionery, a drink.

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