Composition comprising <i>Hordeum vulgare </i>extract for preventing or treating short stature

Abstract

Disclosed is a composition, which comprises Hordeum vulgare extract as an active ingredient and is effective at promoting longitudinal bone growth.

Claims

1. A method for promoting longitudinal bone growth of an animal in need thereof, comprising administering a feed composition comprising an effective amount of a Hordeum vulgare extract to the animal.

2. A method of promoting longitudinal bone growth of a subject in need thereof, comprising administering a pharmaceutical composition comprising an effective amount of a Hordeum vulgare extract to the subject.

3. The method of claim 2, wherein the Hordeum vulgare extract is extracted in water, a C1-C4 alcohol, or a mixture thereof.

4. The method of claim 2, wherein the pharmaceutical composition is orally administered.

5. The method of claim 1, wherein the Hordeum vulgare extract is extracted in water, a C1-C4 alcohol, or a mixture thereof.

6. The method of claim 1, wherein the feed composition further comprises an additional active ingredient having an effect of treating a growth disorder.

7. The method of claim 2, wherein the pharmaceutical composition further comprises an additional active ingredient having an effect of treating a growth disorder.

Description

DESCRIPTION OF DRAWINGS

(1) FIG. 1 illustrates the fluorescence microscope images of luminescence generated from the deposited tibial epiphyseal growth plate after injection of tetracycline to rats; and

(2) FIG. 2 is a graph illustrating the extent of longitudinal bone growth in rats.

MODE FOR INVENTION

(3) A better understanding of the present invention may be obtained through the following example and experimental example, which are set forth to illustrate, but are not to be construed as limiting the scope of the present invention.

Example 1

Preparation of Hordeum vulgare (H. vulgare) Extract

(4) Hordeum vulgare was purchased from Yaksudang, and used after having been validated by the Dept. of Herbal Pharmacology, College of Oriental Medicine, Kyung Hee University. In order to prepare the Hordeum vulgare extract, 1 L of 70% ethanol was added to 100 g of Hordeum vulgare, reflux extracted at 80° C. for 3 hours, and then filtered with filter paper. The resulting extract liquid was concentrated under reduced pressure and then lyophilized, thus obtaining 13.47 g (yield 13.5%) of the Hordeum vulgare extract.

Experimental Example 1

Evaluation of Longitudinal Bone Growth Effect of Hordeum vulgare Extract—Tetracycline Staining Method

(5) <1-1> Preparation of Experimental Animals

(6) As experimental animals, four-week-old Sprague-Dawley female rats (Samtako, Korea) weighing about 100 g were purchased and cared for so as to be acclimatized to the experimental environment while supplying sufficient feed and water. After about 1 week for the acclimatization period, animal testing was performed.

(7) <1-2> Evaluation of Longitudinal Bone Growth Effect

(8) Two days after administration, 20 mg/kg of tetracycline hydrochloride (Sigma T7660) was injected intraperitoneally to all test groups. Two days thereafter, the rats were sacrificed via cervical dislocation, after which the left and right tibiae were removed, fixed in a fixing solution at 4° C. for 3 days, allowed to stand in 50 mM EDTA for 1 day, demineralized, and immersed in 30% sucrose overnight to protect freezing. The bone tissue thus dehydrated was frozen, and cut into 40 μm sections using a sliding microtome (HM440E, Zeiss, Germany), thus collecting sagittal sections of the proximal part of the tibia. To measure the bone growth, each sagittal section of the proximal part of the tibia, 40 μm thick, was placed on a slide glass, dried, and then observed under UV light at a wavelength of 400 nm using a fluorescence microscope (FIG. 1). Using image analysis software ImageJ (NIH, USA), the length between the fluorescence bands formed by the deposition of tetracycline in the growth plate and the bone tissue was measured, and Student's t-test and ANOVA test for comparison with a control group were performed in order to determine the growth effect.

(9) <1-3> Administration of Hordeum vulgare Extract

(10) The Hordeum vulgare extract prepared in Example 1 was administered after having been dissolved in a concentration of 30 mg/kg in type II distilled water in a volume of 1.0 ml per 100 g of rat weight, and as a control group, the same volume of type II distilled water was administered. The Hordeum vulgare extract was orally administered two times a day for a total of four days, starting from 2 days before the administration of tetracycline until the test termination day.

(11) <1-4> Administration of Positive Control Group

(12) Based on the reports of increasing the bone growth using recombinant human growth hormone (Isgaard J, Nilsson A, Lindahl A, Jansson J O, and Isaksson O G: Effects of local administration of GH and IGF-1 on longitudinal bone growth in rats. Am J Physiol. 250:E367-72, 1986), as a positive control group, 20 μg/kg of a recombinant human growth hormone (rhGH; LG Lifescience, Eutropin®) was administered at a volume of 0.1 ml per 100 g of rat weight. For the positive control group, the recombinant human growth hormone was subcutaneously injected once a day for a total of 4 days, starting from 2 days before the administration of tetracycline until the test termination day, and for the other group, distilled water was subcutaneously injected.

(13) <1-5> Evaluation of Longitudinal Bone Growth Effect of Hordeum vulgare Extract

(14) Based on the results of direct measurement of the longitudinal bone growth through the tetracycline staining method, the longitudinal bone growth rate of the positive control group was found to be 360.1±35.8 μm/day (n=5), which is 6.6% higher than 337.8±30.0 μm/day (n=6) in the normal control group, but the difference was not significant.

(15) In the group administered with 30 mg/kg of Hordeum vulgare extract (p.o.), the longitudinal bone growth rate was 394.5±41.38 μm/day (n=6) and thus a significant bone growth effect was exhibited compared to the normal control group (16.8%, p<0.001) (FIG. 2). Thus, when the Hordeum vulgare extract was administered, the longitudinal bone growth effect could be confirmed to remarkably increase.

(16) The test results are summarized in Table 1 below.

(17) TABLE-US-00001 TABLE l Dose Longitudinal bone (Administration route) growth rate (%) Control group — — Positive control group 20 μg/kg (s.c.) 6.6% Hordeum vulgare extract 30 mg/kg (p.o.) 16.8%

INDUSTRIAL APPLICABILITY

(18) According to the present invention, a composition comprising Hordeum vulgare extract as an active ingredient can prevent, ameliorate or treat growth disorder.