COMPOSITION COMPRISING PLANT EXTRACT AND USE THEREOF

Abstract

Provided is a composition comprising a plant extract. The plant extract includes a citrus extract and a green tea extract. The extract combination can effectively increase expression of GLUT4 gene.

Claims

1. A composition comprising a plant extract, wherein the plant extract comprises a citrus extract and a green tea extract.

2. The composition according to claim 1, wherein a ratio of the citrus extract to the green tea extract is 0.8-1.2:1.2-0.8.

3. The composition according to claim 2, wherein the ratio of the citrus extract to the green tea extract is 1:1.

4. The composition according to claim 1, wherein each of the citrus extract and the green tea extract is in a range of 0.03125-0.25 mg/ml.

5. A pharmaceutical composition comprising the composition according to claim 1 and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, wherein the pharmaceutical composition is in a form of a solution, a capsule or a lozenge.

7. A food composition comprising the composition according to claim 1 and a food ingredient, wherein the food ingredient is a component of a general food, a health food, a dietary supplement or a drink.

8. A method for promoting expression of a GLUT4 gene, comprising administering to a subject in need thereof an effective amount of the composition according to claim 1.

9. The method according to claim 8, wherein the composition is a pharmaceutical composition for promoting expression of the GLUT4 gene.

10. The method according to claim 8, wherein the composition is a food product, a health food, a dietary supplement or a drink for promoting expression of the GLUT4 gene.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The following drawings form part of the present specification and are included here to further demonstrate some aspects of the present invention, which can be better understood by reference to one or more of these drawings, in combination with the detailed description of the embodiments presented herein.

[0019] FIG. 1 is a diagram comparing the effects of different plant extracts or combinations on GLUT4 gene expression in an embodiment of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0020] In the following detailed description of the embodiments of the present invention, reference is made to the accompanying drawings, which are shown to illustrate the specific embodiments in which the present disclosure may be practiced. These embodiments are provided to enable those skilled in the art to practice the present disclosure. It is understood that other embodiments may be used and that changes can be made to the embodiments without departing from the scope of the present invention. The following description is therefore not to be considered as limiting the scope of the present invention.

Definition

[0021] As used herein, the data provided represent experimental values that can vary within a range of 20%, preferably within 10%, and most preferably within 5%.

Example 1

Preparation of Green Tea Extract

[0022] First, the green tea leaves were washed and dried, and the green tea leaves were coarsely crushed by a pulverizer. Water was used as a solvent, and the solvent and the resultant crude green tea were mixed at a liquid-solid ratio of 5-20:1-5. After that, the crude green tea was extracted at 50 C. to 100 C. for 0.5 to 3 hours. The extraction temperature is preferably from 75 C. to 95 C.

[0023] After the green tea extract obtained by the above extraction step was cooled to room temperature, it was filtered through a strainer of 400 mesh to remove residual solids. The filtered green tea extract can be further concentrated under reduced pressure at 45 C. to 70 C. to obtain a concentrated product.

Example 2

Preparation or Source of Other Plant Extracts

[0024] The citrus extract of the examples of the present invention was obtained by extracting Citrus reticulata fruit, which was commercially available from LAWTON Trading Co., Ltd. The red wine extract was obtained by extracting red wine, which was commercially available from Shanghai Boyoutang Biotechnology Co., Ltd. The green coffee bean extract was obtained by extracting seeds of unroasted Coffea spp., which was commercially available from ARJUNA NATURAL EXTRACTS Ltd. (India). The blueberry extract was obtained by extracting the Vaccinium Cyanococcus fruit, which was commercially available from Biomed Herbal Research Co., Ltd. These extracts are formulated in water at appropriate concentrations for use.

Example 3

Detection of Blood Sugar Regulating Genes

[0025] HepG2 cells (purchased from ATCC HB8065) were prepared and cultured in a cell culture medium (Dulbecco's modified Eagle's medium (Gibco) containing 1% penicillin/streptomycin and 10% fetal bovine serum). 2 ml of the cell culture medium was added to each well of a 6-well plate to have 1.510.sup.5 HepG2 cells per well.

[0026] The samples were then divided into two groups, of which group A was the blank control group, and group B was the test group. In group B, according to the plant extracts and component types listed in Table 1, at doses of 0.03125 mg/ml to 0.5 mg/ml, single extract prepared in Example 1 or Example 2 or combination of two extracts prepared in Example 1 or Example 2 in a ratio of 1:1 were added, and then reacted at 37 C. for 6 and 24 hours. After that, the expression of the GLUT4 gene was analyzed. The doses shown in the tables are the doses of the single extract, or the components of the combination extract.

[0027] Cells in each of the above groups were recovered, RNA was extracted with an RNA extraction kit (Genemark), and the RNA (2000 ng) was reverse-transcribed into cDNA by the reverse transcriptase (Superscript III Reverse Transcripatase, Invitrogen). Then, qPCR (KAPA CYBR FAST qPCR Kits, KAPA Biosystems) was performed using the primer pairs listed in Table 2 and the ABI Step One Plus Real-Time PCR system to quantify the expression of the GLUT4 gene (the expression of the ACTB (beta-actin) gene was used as an internal control group). During the real-time polymerase chain reaction, the melting curve was analyzed, and the SCORE method was used. The cycle threshold (Ct) of the aforementioned ACTB gene was used as the internal control group, and the blank control group was used as the control gene for relative quantitative analysis of gene expression. When analyzed by the SCORE method, the relative expression of mRNA of the target gene (GLUT4) was derived from the equation 2.sup.Ct, wherein Ct=C.sub.target geneCt.sub.ACTB, and then the difference between the expression level of GLUT4 gene in each group and the blank control group was calculated. The sum of the differences was used as the blood sugar gene index. The blood sugar gene index corresponding to gene expression is shown in Table 1 and FIG. 1, and FIG. 1 is a histogram graphically shown in Table 1. The higher the index value, the better the promotion of GLUT4 gene expression.

TABLE-US-00001 TABLE 1 Plant extract/ blood sugar gene composition dose/ratio index blank control group 0 0 citrus 0.5 mg/ml 0.13 citrus 0.25 mg/ml 1.10 citrus 0.03125 mg/ml 0 green tea 0.25 mg/ml 0 blueberry 0.125 mg/ml 0.96 red wine 0.03125 mg/ml 0.33 citrus + green tea 0.25 mg/ml (1:1) 2.84 citrus + blueberry 0.5 mg/ml (1:1) 0 citrus + red wine 0.03125 mg/ml (1:1) 0.61

TABLE-US-00002 TABLE 2 Primer length Product length Gene Primer SEQ ID NO (ntds) (ntds) GLUT4 GLUT4-F SEQ ID NO: 1 18 200 GLUT4 -R SEQ ID NO: 2 24

[0028] From the results of Table 1 and FIG. 1, the index of the promoting effect of the citrus extract alone (0.25 mg/ml) on GLUT4 gene expression was 1.10, and the index of promoting effect of the green tea extract alone (0.25 mg/ml) was 0. However, when the two were combined, the index of the promoting effect can be as high as 2.84, which was greatly increased to 258% compared with that of the citrus extract alone. In contrast, if the citrus extract was combined with the blueberry extract, the index of the promoting effect was reduced to 0. Therefore, the effect of the combination between the extracts is not necessarily the synergistic effect. In the examples of the present invention, the promoting effect obtained by combining the citrus extract with the green tea extract is preferred.

[0029] It can be seen from the above test that the composition for promoting GLUT4 gene expression in the embodiment of the present invention, when combined with the two extracts of the citrus extract and the green tea extract, can produce an unexpected synergistic effect, so that the expression of GLUT4 gene is increased significantly. Accordingly, these compositions can be utilized to prepare related pharmaceutical or food compositions that allow the applicator or consumer to achieve an adjustment in blood sugar concentration, thereby improving hyperglycemia.

[0030] Further, the composition having the function of promoting GLUT4 gene expression according to an embodiment of the present invention may be further added to a food, a health food, a dietary supplement or a drink. When the composition having the function of promoting GLUT4 gene expression according to an embodiment of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition may be further added to a carrier or other adjuvants well known in the art. The dosage form of the pharmaceutical composition can be, but is not limited to, a solution, a capsule, or a lozenge.

[0031] Although the present invention has been described with reference to the preferred embodiments, it will be apparent to those skilled in the art that a variety of modifications and changes in form and detail may be made without departing from the scope of the present invention defined by the appended claims.