KR20110027856A - Composition for increasing body height - Google Patents

Abstract

PURPOSE: A composition containing horse bone for increasing height is provided to prevent and treat dwarfism. CONSTITUTION: A composition for increasing height contains horse bone extract. The horse bone extract is obtained by isolating with methanol, ethanol, acetone, ethyl acetate, saturated normal butanol, chloroform, methylene chloride, water, or mixture solvent thereof. The composition further contains deer antlet powder or extract, Cibotii Rhizoma powder or extract, and Acanthopanacis Cortex powder or extract. The composition is a food composition, beverage.

Description

Composition for increasing kidneys {COMPOSITION FOR INCREASING BODY HEIGHT}

The present invention relates to a composition for increasing elongation. In particular, the present invention relates to a composition for increasing kidneys using the horse bone extract as an active ingredient.

In recent years, as the Korean people's perception of the ideal kidney has become more westernized, the preference for the large height is increasing. Accordingly, interest in physique growth in adolescence, in which kidney development is concentrated, is increasing. As height preferences are established as social aesthetics, although not pathological, relatively small heights are causing stress and alienation in growing adolescents. In fact, adolescents tend to be satisfied with their appearance and to respect themselves when they are tall (1. Seo Jung-hee, Baek Jae-hwa, Kim Hyo-chul. Satisfaction, evaluation, and self-esteem of youth consumers' appearances and abilities. 5 (2): 53-68), dissatisfaction with their appearance due to low height develops into abnormal body perception, and in severe cases not only impairs physical health but also causes mental side effects such as depression and eating disorders; (Shim, Mi-kyung, Lee Jong-han, Park Eun-ah. Physical management behavior according to body perception and control belief of adolescents. Proceedings of the Korean Psychological Association Annual Conference, 2008: 660-1). Treatment for growth disorders currently being practiced in adolescent growth clinics focuses on mild symptoms, such as poor growth and poor growth, than the average, rather than cases of extreme kidney growth such as Homunculus. This can be seen from this point.

Horse bone is not a widely used drug, but it is a special product of Jeju Island, and recent research has been conducted based on folklore that it has been applied to bone-related symptoms. High School (2006) reported the effects of horse bone supplementation on physiological function (Moon Koh Seok, Lee Young Jae. The effect of horse bone supplementation on rat physiology. Nanji Agricultural Research Institute 2006), Lee et al. (2006) The mineral content was compared and the ratio of calcium and phosphorus was 2: 1, indicating that it was an ideal ratio that had a positive effect on bone metabolism (Lee Jong-un. Nutritional characteristics of horse meat and horse bone. Nanji Agricultural Research 2006; 15: 20-2) . On the other hand, research data verifying osteoporosis, anti-cancer, and anti-inflammatory effects using osteoblasts have been newly studied. Kim (2007) reported that osteoporosis of horse bone in a rat model that caused osteoporosis by ovarian extraction and calcium diet control. (Jim, Min., 1998). The effect of horse bone powder on osteoporosis in rats induced by calcium deficiency diet after ovarian extraction. A combination of cell experiments with MG-63 cells and animal experiments in rats revealed that horse bone extracts are effective in osteoporosis by inhibiting bone resorption during bone metabolism (Pak, Sun-Soon. Factors Affected, Master's Thesis, Cheju National University, 2009.

However, there was no study on the effect of horse bones on kidney growth.

The technical problem of the present invention is to provide a composition for increasing kidneys using a horse bone.

The specific technical problem of the present invention will be presented below.

In order to solve the above technical problem, the present inventors extract the horse bone with 50% acetone and confirm that the extract is administered to the experimental animal to increase the length of the iliac bone and increase the length (height) of the growth plate, as confirmed in the following examples. We examined whether it promotes the expression of IGF-1 (Insulin-like growth factor) and BMF-2 (Bone morphogenic protein 2) in growth plate, and horse bone extract promotes long bone growth and growth plate length. And it was confirmed that promotes the expression of IGF-1 and BMF-2.

IGF-1 is a growth factor that is stimulated by growth hormone and secreted from the anterior pituitary gland and affects the proliferation of chondrocytes in the growth plate (Gerard JT, Sandra RG.Principles of anatomy & physiolosy ( 10th edition) .USA.Wley.2003: 174), BMP-2 regulates the substances responsible for the proliferation of chondrocytes called retinoids (Hallahan AR, Pritchard JI, Chandraratna RAS, Ellenbogen RG, Geyer JR, Overland RP). , Strand AD, Tapscott SJ, Olson JM.BMP-2 mediates retinoid-induced apoptosis in medulloblastoma cells through a paracrine effect.Nature Med. 2003; 9: 1033-8; Cancedda R, Descalzi Cancedda F, Castagnola P. Chondrocyte differentiation. Int Rev Cytol. 1995; 159: 265-358) It is a growth factor known to promote bone formation by inhibiting a bone formation inhibitor called noggin (Wang EA, Rosen V, D'Alessandro JS, Bauduy M, Cordes). P, Harada T, Israel DI, Hewick RM, Kerns KM, LaPan P, Luxen berg DP, McQuaid D, Moutsatsos IK, Nove J, Wozney J M. Recombinant human bone morphogenetic protein induces bone formation.Proc. Nat. Acad. Sci. 1990; 87: 2220-4).

The present invention has been completed based on the above-described experimental results, characterized in that the composition for increasing kidney of the present invention comprises a horse bone extract as an active ingredient.

In the present specification, "horse bone extract" refers to an extract obtained by extracting a horse bone to be extracted with methanol, ethanol, acetone, ethyl acetate, saturated normal butanol, chloroform, methylene chloride, water, or a mixed solvent thereof regardless of the extraction method. It is understood as meaning including the extract fractionated with the solvents listed above in the extract. Regardless of the extraction method, as long as the extraction target is extracted through the step of immersing the horse bone in the extraction solvent, it should be understood that the extraction method can be applied to any method such as cold soaking, refluxing, heating, and ultrasonic wave. In addition, although the horse bone extract used in the animal experiments in the following examples is an extract obtained by the mixed solvent of acetone and water, the meaning of the horse bone extract is not limited to the extract obtained by the mixed solvent of acetone and water. The present inventors performed in vitro experiments using osteoblasts and chondrocytes to screen extracts having activity of increasing kidney, although not shown in the following examples, and all extracts obtained using the respective extraction solvents through the experiments. It has been confirmed that there is an activity for promoting the proliferation of osteoblasts and chondrocytes. The reason for using the extract obtained by the mixed solvent of acetone and water in the following examples is that only the solid weight yield of the obtained extract was the highest.

In addition, the meaning of "extract" in the present specification includes a concentrated liquid extract or solid extract from which the extraction solvent is removed.

In addition, in the present specification, the "active ingredient" means a component that can exhibit the desired activity alone or in combination with a carrier which is itself inactive.

In addition, in this specification, "extension increase" means an increase in the length of the long bone and / or an increase in the length of the growth plate. "Long bone" refers to a long, thick cylindrical bone that has a bone marrow in it and is ball-shaped at both ends, and typically includes the femur, tibia, radius, and ulna.

On the other hand, the composition for increasing kidney of the present invention may include any amount (effective amount) as long as it can exhibit the effect of increasing the horse bone extract, the effective ingredient, the conventional effective amount is 0.001 based on the total weight of the composition Will be determined within the range of weight% to 99.990 weight%. The term "effective amount" refers to the amount of the active ingredient that can induce an increase in height. Such effective amounts can be determined experimentally within the range of ordinary skill in the art.

The kidney increasing composition of the present invention includes a known natural product or compound, which is known to have an increase in nephropathy (or iliac length growth promoting activity) so as to increase and enhance the kidney increasing effect in addition to the horse bone extract which is an active ingredient thereof. can do.

As such natural products or compounds, antler powders or extracts, green rust powder or extracts, gucchi powders or extracts, zhaoga powder or extracts, fast powders or extracts, agapi powders or extracts, Guanyl cyclase B (GC-B), and the like. The meaning of the extract is as described above with respect to the horse bone extract. Such extracts may be obtained by mixing the extraction objects.

The composition for increasing kidney of the present invention can be regarded as a food composition, in particular a functional beverage, in a specific embodiment.

The food composition of the present invention may include sweeteners, flavoring agents, bioactive ingredients, minerals, etc. in addition to the active ingredients.

Sweeteners may be used in amounts that give the food a suitable sweet taste, and may be natural or synthetic. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.

Flavoring agents can be used to enhance the taste or aroma, both natural and combined. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. The natural flavor may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or may be obtained from green tea leaves, round leaves, jujube leaves, cinnamon, chrysanthemum leaves, jasmine and the like. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. In some cases, synthetic flavoring agents may be used, and synthetic flavoring agents may be used in the form of esters, alcohols, aldehydes, terpenes and the like.

As the physiologically active substance, catechins such as catechin, epicatechin, gallocatechin, epigallocatechin, vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine, riboflavin, and the like can be used.

As minerals, calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium, zinc and the like can be used.

In addition, the food composition of the present invention may contain a preservative, an emulsifier, an acidulant, a thickener, and the like, in addition to the sweetener.

Such preservatives, emulsifiers and the like are preferably added and used in very small amounts as long as the use to which they are added can be achieved. "Ultra trace" means numerically expressed in the range from 0.0005% to about 0.5% by weight based on the total weight of the food composition.

Examples of preservatives that can be used include sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), and the like.

Emulsifiers that can be used include acacia gum, carboxymethylcellulose, xanthan gum, pectin and the like.

Examples of acidulants that may be used include lead acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like. Such acidulant may be added so that the food composition is at an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing taste.

Thickeners that can be used include suspending implements, sedimenters, gel formers, swelling agents and the like.

In addition, the herbal composition may be added to the food composition of the present invention to improve flavor and palatability and to add other functionalities (such as prevention of arthritis or osteoporosis), which may be added as a medicinal herb extract, soybean extract, antler extract, safflower Phosphorus extract, Tosa extract, Sukjihwang extract, Tortoiseshell extract, Cornus extract, Goji berry extract, Licorice extract, Angelica extract, Root extract, Gangjinhyang extract, Haphwanpi extract, Sandugeun extract, Cultivated extract, Red ginseng extract may be exemplified. .

The composition of the present invention can be used as a pharmaceutical composition in a specific embodiment.

The pharmaceutical composition of the present invention is formulated for oral or parenteral administration by mixing the horse bone extract as an active ingredient with a pharmaceutically acceptable carrier, excipient, additives and the like.

Examples of pharmaceutically acceptable carriers or excipients include lactose, magnesium stearate, starch, talc, gelatin, agar, pectin, gum arabic, olive oil, sesame oil, cacao butter, ethylene glycol and the like, or other commercially available ones.

Examples of solid compositions for oral administration include tablets, pills, capsules, powders, granules and the like. In such a solid composition, horse bone extract, which is an active ingredient, contains at least one inert diluent such as lactose, mannitol, glucose, hydroxy propyl cellulose, microcrystalline cellulose, starch, polyvinyl pyrrolidone, magnesium aluminometa silicate, and the like. Are mixed. The solid composition may contain additives other than inert diluents, for example, lubricants such as magnesium stearate, disintegrants such as calcium cellulose glycolate, dissolution aids such as glutamic acid or aspartic acid according to conventional methods. The tablets or pills may be coated with a film made of a material such as sucrose, gelatin, hydroxypropyl methylcellulose phthalate and the like, if desired, and may be coated with two or more layers.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, and the like, and may include generally used inert diluents such as purified water and ethanol. This composition may contain, in addition to inert diluents, wetting agents, auxiliaries such as suspending agents, sweetening agents, flavoring agents, fragrances, preservatives and the like.

Parenteral injections include sterile aqueous or nonaqueous solutions, suspensions, and emulsifiers. Aqueous solutions and suspensions include, for example, water for injection and physiological saline for injection. Non-aqueous solutions and suspending agents include, for example, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, polysorbate 80® and the like. Such a composition is, in addition, a preservative, a humectant, an emulsifier, a dispersant, a stabilizer (e.g. lactose), a dissolution aid (e.g. glutamic acid, aspartic acid).

Adjuvants, such as lactic acid). These can be sterilized by conventional sterilization methods such as, for example, filter sterilization by microfiltration membranes, heat sterilization such as autoclaving, or a sterilizing agent. Injectables can be solution solutions or freeze-dried preparations which can be dissolved and used in use. As an excipient for freeze-drying, for example, sugar alcohols or sugars such as mannitol and glucose can be used.

The pharmaceutical composition of the present invention is administered by oral administration or parenteral administration.

Preferably, parenteral administration method, for example, administration by injection (subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection, etc.), transdermal administration, transmucosal administration (rectal administration, etc.), transpulmonary administration, etc. to be. Oral administration methods may also be used.

The dosage of the active ingredient of the pharmaceutical composition of the present invention can be determined depending on the severity of the disease, the age of the patient, etc., but is generally in the range of 0.005 µg / kg to l00 mg / kg, preferably 0.02 µg / kg to 5 mg / kg. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as the route of administration, the age, sex, weight of the patient, and the severity of the patient, the dosage limit the scope of the present invention in any aspect. It should not be understood.

As described above, according to the present invention can provide a composition for increasing elongation. The composition for increasing kidney of the present invention may be commercialized into a food composition or a pharmaceutical composition.

The composition for increasing kidney of the present invention can be usefully used not only for the treatment or prevention of small authentication or dwarf development, but also for the purpose of increasing the height of children and adolescents during the growing season.

Hereinafter, the present invention will be described with reference to Examples. However, the scope of the present invention is not limited to these examples.

≪ Example 1 > Materials and Methods of Experiment of Elongation Effect of Horse Bone Extract

<Example 1-1> Experimental material

 The animals used for the experiment were purchased from 80 ± 10g Sprague-Dawley male rats (3 weeks old) from the experimental animals, and adapted to the experimental environment for 1 week with sufficient feed and water, and then divided into control and experimental groups. (n = 10).

Used in the experiments malppyeo the (main) was used femur of jejuma industry the Jeju 36 month old male horse (Equus caballus) purchased from, Jeju Island Acanthopanax (Acanthopanax koreanum Nakai) is provided in Jeju Biodiversity Research Institute And it was verified and used in Semyung University main school.

<Example 1-2> Experiment method

<1> Preparation of Sample

Horse bone powder extract and sesame seed extract used in the experiment were used in this test through the following process.

The femur bones of 36-month-old males from Jeju were washed with water and crushed through a milling machine (NIRO Atomizer, Denmark) to obtain 12 kg of horse bone powder, which was first dried in a hot air dryer, and then frozen and stored. Add 5 times 50% (v / v) acetone to horse bone powder, extract at 60 ℃ for 2 hours, filter with Whatman NO.1, concentrate the filtrate under reduced pressure, and freeze-drier (FD 8512, Ilsin Lab, Korea) was used by lyophilization.

Isogapi was added to 10 times 70% EtOH and extracted for 2 hours at 60 ℃, filtered with Whatman paper (Whatman NO. 1) and the filtrate was concentrated under reduced pressure to prepare a powder. The horse bone powder extract was subjected to the sample A as a sample A, and the mixture of horse bone powder extract and sesame stem extract in a 1: 1 weight ratio was used as sample B.

<2> administration of sample and setting of experimental group

In order to observe the efficacy of Samples A and B on bone growth in rats, Hansson's first method was used to determine bone growth (Hansson LI, Menander-Sellman K, Stenstrom A, Thorngren KG.Rate of normal longitudinal bone growth). in the rat.Calcif Tissue Res. 1972; 10: 238-51). This is a method for observing normal bone metabolism by using the phenomenon that the fluorescent factor is deposited on the bone through calcium chelate bonds. The bone growth was most active during the growth phase is deposited on the bone angiogenesis at the bottom of the bone growth plate to form a line, which was administered over a period of time using the fluorescence microscope to measure the length between each line was measured. The administered drug takes at least 12 hours to reach and act on the growth plate (De Luca F, Barnes KM, Uyeda JA, De-Levi S, Abad V, Palese T, Mericq V, Baron J. Regulation of growth plate chondrogenesis by bone morphogenetic protein-2.Endocrinology.2001; 142 (1): 430-6) In this experiment, we decided to administer the sample once a day from 4 days before autopsy for sufficient drug expression.

 The control group (distilled water administration group) was orally administered with distilled water, and the sample A administration group had three concentrations of high, medium and low concentrations (250, 50 and 10 mg / kg, po), and the sample B administration group had one high concentration (250 mg). / kg, po) was administered once daily for 5 days from the start of the experiment (day 1). All four experimental groups were administered calcein (calcein: 10 mg / kg, i.p .: Sigma, U.S.A.) 4 days after the start of the experiment.

<3> weight measurement

The same visual weight was measured every day from the start date to the end date of the experiment.

<4> tissue sampling

One day after fluorescein administration, rats were anesthetized with chloral hydrate (Japan, 35.0 mg / kg, ip), deformed, incisions were made in the right ventricle, and the needle (No. 18) was injected into the left ventricle. 5% sodium nitrite (Sigma, USA) saline was perfused to the heart and then perfused with 4.0% formalin fixative at pH 7.4 dissolved in 0.1 M phosphate buffer. The left and right tibia of the rats were then detached, fixed in 0.1 M phosphate buffered formalin fixative for 2 hours, left in 50 mM EDTA for 2-3 days, demineralized, and then 30% protected against freezing. Soak in sucrose was maintained at 4 ℃ overnight. After freezing the fixed bone tissue, a sagittal section of the proximal part of the tibia was collected every 40 μm using a sliding microtome (HM440E, Zeiss, Germany).

<5> bone growth measurement and growth plate observation

The proximal tibial sagittal section collected at every 40 μm was placed on a slide glass and dried, and then, the fluorescence microscope was used to measure the length of the growth plate and the line formed by the deposition of fluorescent factors in bone tissue.

After measuring the length, the cresyl violet was stained, and the chondrocytes of the resting part, the proliferating part and the hypertrophic part of the growth plate were observed, and the total height was measured.

<6> immunohistochemical staining

Immunohistochemical staining was used to observe the growth plate expression of IGF-1 and BMP-2. Tissue sections prepared at 40 μm as described above were used. The primary antibodies used for immunostaining were IGF-1 antibody (Santa Cruz Biotech., U.S.A.) and BMP-2 antibody (Santa Cruz Biotech., U.S.A.), respectively. Anti-rabbit antibody (Vector Lab, U.S.A.) was used as a secondary antibody of IGF-1, and anti-goat antibody (Vector Lab, U.S.A.) was used as a secondary antibody of BMP-2. In addition, a Vectastatin elite ABC kit (Vector Lab, U.S.A.) was used. Experimental method was soaked tissue sections in 0.1 M PBS (phosphate-buffered saline, pH 7.2) for 5 minutes, twice in 15 minutes in Triton-X 100 solution (Sigma, USA), 0.1 M PBS + 0.5% BSA (Bovine Serum) Albumin, Sigma, USA) was washed twice for 15 minutes, and then reacted with the primary antibody overnight at room temperature. The next day, rinse in 0.1 M PBS + 0.5% BSA twice for 15 minutes, then react the section with secondary antibody for 60 minutes, then rinse twice for 15 minutes with 0.1 M PBS + 0.5% BSA for 60 minutes, then ABC (avidin- The biotin-peroxidase complex was added at a concentration of 1:50 and reacted at room temperature. Each tissue section was washed twice with 0.1 M PB for 15 minutes and finally reacted with 0.05% DAB (3,3-diaminobenzidine, Sigma, U.S.A.) and 0.1 M PB with 0.03% hydrogen peroxide. After color development, each tissue section was placed in a new 0.1 M PB to stop the reaction, and then produced a slide sample for observation.

<7> Statistical method

One way ANOVA test was used to determine the effect of the drug, and Dunnett's post hoc test was used to compare with the control group. Statistical significance was assumed to be p <0.05 or less.

<Example 2> Experiment result

<1> Effect on weight gain

The weight measured daily for 5 days is shown graphically in <Fig. 1> by MEAN ± SE, and in FIG. 2 for the weight gain on the 4th and 5th days.

Referring to Figure 2, compared with the control group (Nomal), high concentration of horse bone extract group (A-high), medium concentration of horse bone extract group (A-middle) and low concentration of horse bone extract group (A-low) ) Showed statistically significant difference. However, no statistically significant difference was found in the mixed group (B-high) of horse bone extract and organ extract (* p <0.005, ** p <0.01).

<2> Effect on iliac length growth

Melia ± SE by measuring the length of iliac length for each experimental group for one day, that is, the calcin (10mg / kg), one time, and the length between the growth plate and the growth plate formed by the infiltration of the phosphor in bone tissue 24 hours later. 3 is shown.

Referring to Figure 3, the control group (Nomal) showed a length of 404.0 ± 9.5 ㎛ growth, high concentration of horse bone extract group (A-high) and medium concentration of horse bone extract group (A-middle) 482.2 respectively Length growth of ± 12.2 μm and 518.6 ± 21.7 μm showed statistically significant difference (*** p <0.001). However, the low concentration of horse bone extract group (A-low) and the mixed group of horse bone extract and organ extract (B-high) showed length growth of 405.5 ± 11.9㎛ and 428.6 ± 11.9㎛, respectively. It was presumed that the statistically significant difference did not appear in the mixed administration group of horse bone extract and organ extract.

For reference, the fluorescence micrograph of the horse bone extract administration group of medium concentration is shown in <4>. In FIG. 4, A is a control group, the upper arrow is a boundary line between the growth plate and bone, and the lower arrow is a line formed by absorption of calcein.

<3> Effect on Growth Metabolism of Chondrocytes

The total length of the growth plate was measured and shown in FIG. 5 by MEAN ± SE. The control group (Nomal) showed a growth of 561.3 ± 1.1 ㎛, the high concentration of horse bone extract group (A-high), medium concentration of horse bone extract group (A-middle) and low concentration of horse bone extract group (A-low), respectively 606.6 ± 9.1 μm, 615.6 ± 11.4 μm, and 615.1 ± 8.9 μm, respectively, and the growth rate of 642.4 ± 16.9 μm was observed in the mixed group (B-high) of horse bone extract and Ogapi extract. There was statistically significant difference in all experimental groups (* p <0.05, *** p <0.001).

On the other hand, the increase in long bone length did not show statistically significant difference in the low concentration of horse bone extract group and the mixed group of horse bone extract and organ extract, but there was a statistically significant difference in all the experimental groups including the above groups in the growth plate length. It is assumed that this is because the results appear faster. In other words, the growth of bone length is likely to accelerate after the growth plate length is increased. Therefore, the bone length growth is not promoted, but the growth plate length is very high in many cases, the growth plate height is not the case that the growth plate height is not high.

For reference, the fluorescence micrograph of the horse bone extract administration group of medium concentration is shown in Figure 6, where A is the control and the length between the arrows is the length of the growth plate.

<4> Effect on the expression of IGF-1 in the growth plate

The fluorescence microscopy image after immunohistochemical staining of the administration group of the medium concentration horse bone extract is shown in FIG. 7. (E, f, g and h in Fig. 7) is that of the administration group of the horse bone extract of medium concentration.

IGF-1 expression was weakly expressed in the resting part (a and e of FIG. 7) and the proliferating part (b and f of FIG. 7) in both the control group and the administered group of the horse bone extract of medium concentration, but the hypertrophy (c and g of FIG. 7). ) And ossification part (d and h of Figure 7) was observed that the IGF-1 expression was increased by the increase of immunostaining in the group of administration of horse bone extract of medium concentration.

In addition, when observing the details of the proliferative chondrocytes, IGF-1 expression of the marker (*) chondrocyte cytoplasm by the proliferating region (Fig. 7b) of the control group was observed very weakly, but the proliferation of the administration group of the medium concentration of horse bone extract It was observed that IGF-1 expression in the minor (*) star marker (*) chondrocyte cytoplasm was slightly stronger.

In addition, when observed in detail for hypertrophic chondrocytes, IGF-1 expression of marker (*) chondrocyte cytoplasm of the control hypertrophy (Fig. 7c) was observed at normal levels, but the hypertrophy of the administration group of medium horse bone extract was observed. (G of FIG. 7) IGF-1 expression of the star marker (*) chondrocyte cytoplasm was stronger and it could be observed that it was expressed in a greater number of chondrocytes.

In the case of osteochondral chondrocytes, the hypertrophic chondrocytes were similar to the hypertrophic chondrocytes. 7 h) star marker (*) chondrocytes IGF-1 expression was stronger and more chondrocytes expressed was observed.

<4> Effect on the expression of BMP-2 in the growth plate

Fluorescence microscopy images after immunohistochemical staining of the group of horse bone extracts of medium concentration are shown in FIG. 8. The photographs on the left column (a, b, c and d in FIG. 8) are those of the control, and the photographs on the right column. (E, f, g and h in Fig. 8) is that of the administration group of the horse bone extract of medium concentration.

As a result of comparing the control group and the administration group of the horse bone extract of the concentration in the sample, the expression in the growth plate tissue of BMP-2 is as follows. In the control group and the administration group of the horsetail extract of medium concentration, BMP-2 expression was weakly expressed in the resting part (a and e of FIG. 8) and the proliferating part (FIG. 8 b and f), but the hypertrophy (c and g of FIG. 8). As a result of observing the ossification part (d and h of FIG. 8), it was observed that BMP-2 expression was increased due to the increase of immunostaining in the administration group of medium concentration of horse bone extract.

In addition, when observed in detail for the proliferation of chondrocytes BMP-2 expression of control chondrocytes (Fig. 8b) of the chondrocyte cytoplasm is very weak and was hardly observed, but the growth group of the administration of horse bone extract of medium concentration (f of Fig. 8f). BMP-2 expression was weaker in the chondrocytes (see asterisk *) of the maturation stage below, but stronger than in the control group.

In addition, when observed in detail for hypertrophic chondrocytes, BMP-2 expression of the control hypertrophy (Fig. 8c) marker (*) chondrocyte cytoplasm was observed at normal levels, but the hypertrophy of the administration group of medium horse bone extract (G) of the star marker (*) chondrocytes It was observed that the BMP-2 expression of the cytoplasm was stronger and the number of expressed chondrocytes was expressed in a greater number of cells.

In the case of osteochondral chondrocytes, the hypertrophy of the administration group of the horse bone extract of medium concentration than the BMP-2 expression of the control hypertrophy (*) chondrocytes (*) cartilage cytoplasm similar to the hypertrophic chondrocytes (Fig. 8d) 8 h) star marker (*) chondrocytes BMP-2 expression was found to be stronger.

1 is a graph showing the weight of the experimental animals measured daily for 5 days.

Figure 2 is a graph showing the weight gain on the 4th and 5th day of the experimental animal.

Figure 3 is a graph showing the degree of growth of iliac bone for 1 day.

Figure 4 is a fluorescence micrograph of the longitudinal section of the proximal tibia of the control group and the medium concentration (50mg / kg) horse bone extract administration group of distilled water administration group.

5 is a graph showing the degree of increase of the growth plate length.

Figure 6 is a fluorescence microscope picture of the growth plate of the control group and the medium concentration (50mg / kg) horse bone extract administration group which is a distilled water administration group.

Figure 7 is a fluorescence micrograph showing the expression level of IGF-1 in the growth plate tissue of the control group and medium concentration (50mg / kg) horse bone extract administration group of distilled water administration group.

8 is a fluorescence micrograph showing the expression level of BMP-2 in the growth plate tissue of the control group and the medium concentration (50mg / kg) horse bone extract administration group of distilled water administration group.

Claims (9)

Elongation composition, characterized in that it comprises a horse bone extract. The method of claim 1, The horse bone extract is an extract for increasing kidneys, characterized in that the extract obtained by extracting the horse bone with methanol, ethanol, acetone, ethyl acetate, saturated normal butanol, chloroform, methylene chloride, water, or a mixed solvent thereof. The method of claim 1, The horse bone extract is a composition for increasing kidneys, characterized in that the extract obtained by extracting the horse bone with water, acetone or a mixed solvent thereof. The method of claim 1, The increase in height is a composition for increasing kidneys, characterized in that the increase in the length of the long bone (長 骨) or growth plate length. The method of claim 4, wherein The long bone is a composition for increasing kidneys, characterized in that one of the femur, tibia, radial and ulna. The method of claim 1, The composition is one selected from antler powder or extract, green rust powder or extract, Guchu powder or extract, Zhaoga powder or extract, Sokdan powder or extract and Ogapi powder or extract Elongation composition, characterized in that it further comprises above. The method according to any one of claims 1 to 6, The composition for increasing the kidney, characterized in that the food composition. The method of claim 7, wherein The food composition is a composition for increasing kidneys, characterized in that the beverage. The method according to any one of claims 1 to 6, The composition for increasing kidneys, characterized in that the pharmaceutical composition.

Images