ANTIDIABETIC COMPOSITION COMPRISING GINGER EXTRACT OBTAINED FROM MICROWAVE-PROCESSED GINGER AND METHOD OF PREVENTING OR TREATING DIABETIS MELLITUS
20220133835 · 2022-05-05
Assignee
Inventors
- Jungyeob Ham (Gangneung-si, KR)
- Taejung Kim (Gangneung-si, KR)
- Young Tae Park (Gangneung-si, KR)
- Pilju Choi (Gangneung-si, KR)
- Ji-yool KIM (Gangneung-si, KR)
- Cheol Hee YOON (Gangneung-si, KR)
- Haneul JU (Gangneung-si, KR)
Cpc classification
A23L33/105
HUMAN NECESSITIES
A61K2236/10
HUMAN NECESSITIES
International classification
Abstract
Provided are a pharmaceutical composition for preventing or treating diabetes and a method of preventing or treating diabetes in a subject by using the pharmaceutical composition, the pharmaceutical composition including a ginger extract, as an active ingredient, obtained by microwave irradiation, under pressure, of ginger, an extract thereof, or a combination thereof.
Claims
1. A pharmaceutical composition for preventing or treating diabetes, the pharmaceutical composition comprising: a ginger extract, as an active ingredient, obtained by microwave irradiation, under pressure, of ginger, an extract thereof, or a combination thereof, wherein in the ginger extract, a content of [6]-shogaol, [8]-shogaol, and [10]-shogaol is 20 wt % or more greater than a content of [6]-gingerol, [8]-gingerol, and [10]-gingerol, wherein the microwave irradiation is performed under a pressure in a range of about 2 atm to about 100 atm.
2. The pharmaceutical composition of claim 1, wherein the extract used in the microwave irradiation is extracted by using a solvent after the microwave irradiation.
3. The pharmaceutical composition of claim 1, wherein the microwave irradiation is performed at a temperature of about 120° C. to about 200° C. for about 30 minutes to about 90 minutes.
4. The pharmaceutical composition of claim 1, wherein the extract used in the microwave irradiation is a crude extract of ginger from water, a C.sub.1-C.sub.4 alcohol, or a mixture thereof; a solvent fraction of n-hexanet, methylene chloride, ethyl acetate, butanol, acetone, or a mixture thereof of the crude extrac; or a purified product of the solvent fraction.
5. The pharmaceutical composition of claim 1, wherein, in the ginger extract, a content of [6]-shogaol, [8]-shogaol, and [10]-shogaol is 80 wt % or greater of a total content of [6]-shogaol, [8]-shogaol, [10]-shogaol, [6]-gingerol, [8]-gingerol, and [10]-gingerol.
6. The pharmaceutical composition of claim 1, wherein, in the ginger extract, a content of [6]-shogaol is 80 wt % or greater of a total content of [6]-shogaol, [8]-shogaol, [10]-shogaol, [6]-gingerol, [8]-gingerol, and [10]-gingerol.
7. The pharmaceutical composition of claim 1, wherein, in the ginger extract, a content of [6]-shogaol, [8]-shogaol, and [10]-shogaol is 80 wt % or greater of a total content of [6]-shogaol, [8]-shogaol, [10]-shogaol, [6]-gingerol, [8]-gingerol, and [10]-gingerol.
8. The pharmaceutical composition of claim 1, wherein, in the ginger extract, a content of [6]-shogaol, [6]-gingerol, and [8]-gingerol is in a range of about 7.5 wt % to about 12.5 wt %; about 2.5 wt % to about 7.5 wt %; or about 80 wt % to about 90 wt %, based on a total content of [6]-shogaol, [8]-shogaol, [10]-shogaol, [6]-gingerol, [8]-gingerol, and [10]-gingerol.
9. The pharmaceutical composition of claim 1, wherein the ginger extract is a fraction obtained by fractionating a crude extract with ethanol.
10. The pharmaceutical composition of claim 9, wherein the ethanol is an about 70% to about 90% aqueous solution.
11. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition promotes insulin secretion ability of cells.
12. A food composition for preventing or ameliorating diabetes, the pharmaceutical composition comprising: a ginger extract obtained by microwave irradiation, under pressure, of ginger, an extract thereof, or a combination thereof, wherein in the ginger extract, a content of [6]-shogaol, [8]-shogaol, and [10]-shogaol is 20 wt % or more greater than a content of [6]-gingerol, [8]-gingerol, and [10]-gingerol, wherein the microwave irradiation is performed under a pressure in a range of about 2 atm to about 100 atm.
13. A method of preventing or treating diabetes in a subject, the method comprising administering the pharmaceutical composition to the subject.
14. The method of claim 13, wherein the extract used in the microwave irradiation is extracted by using a solvent after the microwave irradiation.
15. The method of claim 13, wherein the microwave irradiation is performed at a temperature of about 120° C. to about 200° C. for about 30 minutes to about 90 minutes.
16. The method of claim 13, wherein the extract used in the microwave irradiation is a crude extract of ginger from water, a C1-C4 alcohol, or a mixture thereof; a solvent fraction of n-hexanet, methylene chloride, ethyl acetate, butanol, acetone, or a mixture thereof of the crude extract or a purified product of the solvent fraction
17. The method of claim 13, wherein, in the ginger extract, a content of [6]-shogaol, [8]-shogaol, and [10]-shogaol is 80 wt % or greater of a total content of [6]-shogaol, [8]-shogaol, [10]-shogaol, [6]-gingerol, [8]-gingerol, and [10]-gingerol.
18. The method of claim 13, wherein, in the ginger extract, a content of [6]-shogaol is 80 wt % or greater of a total content of [6]-shogaol, [8]-shogaol, [10]-shogaol, [6]-gingerol, [8]-gingerol, and [10]-gingerol.
19. The method of claim 13, wherein, in the ginger extract, a content of [6]-shogaol, [8]-shogaol, and [10]-shogaol is 80 wt % or greater of a total content of [6]-shogaol, [8]-shogaol, [10]-shogaol, [6]-gingerol, [8]-gingerol, and [10]-gingerol.
20. The method of claim 13, wherein, in the ginger extract, a content of [6]-shogaol, [6]-gingerol, and [8]-gingerol is in a range of about 7.5 wt % to about 12.5 wt %; about 2.5 wt % to about 7.5 wt %; or about 80 wt % to about 90 wt %, based on a total content of [6]-shogaol, [8]-shogaol, [10]-shogaol, [6]-gingerol, [8]-gingerol, and [10]-gingerol.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0035] The above and other aspects, features, and advantages of certain embodiments of the disclosure will be more apparent from the following description taken in conjunction with the accompanying drawings, in which:
[0036]
[0037]
[0038]
[0039]
DETAILED DESCRIPTION
[0040] Reference will now be made in detail to embodiments, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to like elements throughout. In this regard, the present embodiments may have different forms and should not be construed as being limited to the descriptions set forth herein. Accordingly, the embodiments are merely described below, by referring to the figures, to explain aspects of the present description. As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items. Expressions such as “at least one of,” when preceding a list of elements, modify the entire list of elements and do not modify the individual elements of the list.
[0041] The present disclosure will be described in more detail through the following Examples. However, these Examples are for illustrative purposes only, and the present disclosure is not intended to be limited by these Examples.
Example 1: Preparation of Processed Ginger Product by Simple Heat-Drying
[0042] A processed ginger product was prepared from ginger through simple processing as follows. Specifically, 100 g of undried ginger was cut and steamed at a temperature of 100° C. for 6 hours and 12 hours, and then dried to obtain steamed ginger. 800 mL of 80% by volume ethanol aqueous solution was added to the simple-processed ginger as described above, and the mixture was refluxed and extracted at a temperature of 70° C. for 3 hours. Then, the solvent was evaporated under reduced pressure, followed by drying of the residue to thereby obtain a simple-processed ginger extract in powder form.
Example 2: Preparation of Microwave-Processed Ginger Product Under Reduced Pressure
[0043] The processing in Example 2 was performed in the same manner as the ginger processing method for increasing a shogaol content by microwave irradiation under pressure of a simple ginger extract according to KR 10-1514793.
[0044] Specifically, 10 g of raw ginger dry extract was added to 50 mL of a 50 vol % aqueous solution of ethanol in an 80 mL container of a pressurized microwave irradiator (CEM, Model No. 909150, USA). The container was sealed, and the processing temperature was set to a temperature of 140° C. Microwaves were irradiated for 30 minutes at 100 W (frequency of 2,455 MHz) in a fixed state. After completion of processing, lyophilization was performed to obtain microwave-processed ginger product in powder form. The pressure during microwave irradiation was 6 bar.
Example 3: Preparation of Fraction of Ginger Extract
[0045] The raw ginger fraction used in Example 2 was obtained by a reversed-phase silica gel flash chromatography method.
[0046] Specifically, octadecyl-silica (C18 ODS) was filled into a glass column having a diameter of 3 cm, to a height of 20 cm, and 2 g of the raw ginger extract, i.e., the ginger extract obtained in Example 1, was added to a specific thickness. After sequentially flowing 200 mL of each of 20%, 40%, 60%, and 80% ethanol aqueous solutions, 100% ethanol, and 100% acetone thereto, the eluent was distilled under reduced pressure to obtain fractions of 840 mg, 233 mg, 225 mg, 115 mg, 131 mg, and 49 mg, respectively.
[0047]
Example 4: Preparation of Fraction of Processed Ginger Product
[0048] The microwave-processed ginger fraction used in Example 4 was obtained by using the processed ginger product prepared in Example 2 through a reversed-phase silica gel flash chromatography method.
[0049] Specifically, octadecyl-silica (C18 ODS) was filled into a glass column having a diameter of 3 cm, to a height of 20 cm, and 2 g of the microwave-processed ginger product was added to a specific thickness. After sequentially flowing 200 mL of each of 20%, 40%, 60%, and 80% ethanol aqueous solutions, 100% ethanol, and 100% acetone thereto, the eluent was distilled under reduced pressure to obtain fractions of 428 mg, 323 mg, 369 mg, 145 mg, 167 mg, and 67 mg, respectively.
[0050]
Example 5: Preparation of Single Components of [6]-Gingerol, [8]-Gingerol, [10]-Gingerol, [6]-Shogaol, [8]-Shogaol, and [10]-Shogaol
[0051] From the ginger extracts and the microwave-processed ginger products of Examples 1 and 2, [6]-gingerol, [8]-gingerol, [10]-gingerol, [6]-shogaol, [8]-shogaol, and [10]-shogaol compounds were separated.
[0052] Through Prep-HPLC (stationary phase: Phenomenex Luna C18(2) column, particle size of 10 μm, a column size of 250×21.20 mm), the eluent was developed from 30% to 100% of initial acetonitrile for 60 to 90 minutes at a flow rate of 8 mL/min to thereby obtain 6 peaks appearing at UV of 282 nm. For Peaks 1 to 6, through reversed-phase semi-preparative high-performance liquid chromatography (stationary phase: Phenomenex Gemini 6 Phenyl column, particle size of 5 μm, column size of 250×10 mm), the eluent was developed from 50% to 100% of initial methanol for 60 to 90 minutes at a flow rate of 4 mL/min to thereby obtain [6]-gingerol, [8]-gingerol, [10]-gingerol, [6]-shogaol, [8]-shogaol, and [10]-shogaol compounds.
[0053] The compounds were identified through nuclear magnetic resonance spectrometer (NMR), mass spectroscopy, or the like, and [6]-gingerol, [8]-gingerol, [10]-gingerol, [6]-shogaol, [8]-shogaol, and [10]-shogaol were confirmed by referring to existing literature information. Chemical formulae of the separated compounds are the same as Formula 1 and Formula 2 above.
Experimental Example 1: In Vitro Insulin Secretion Evaluation of Ginger Extract and Fraction Thereof, Steamed Ginger Extract, Microwave-Processed Ginger Product and Fraction Thereof, and Single Ingredients
[0054] 1. Experiment Method
[0055] Insulin secretion ability was evaluated using a mouse insulinoma cell line INS-1 (Biohermes, Shanghai, China) as follows.
[0056] (1) INS-1 Cell Culture
[0057] Cell line INS-1 (Biohermes, Shanghai, China) is a mouse insulinoma species and has insulin secretion ability. INS-1 cells were cultured in RPMI1640 (Cellgro, Manassas, Va., USA) medium supplemented with 10% fetal bovine serum (FBS), 100 units/mL penicillin G, 100 μg/mL streptomycin, 10 mM HEPES, 2 mM L-glutamine, 1 mM sodium pyruvate and 0.05 mM 2-mercapto ethanol under a condition of a temperature of 37° C., 90% air, and 10% CO.sub.2. Among the above-mentioned components, reagents not mentioned by the manufacturer were purchased from Gibco BRL, Life Technologies.
[0058] (2) Measurement of INS-1 Cell Toxicity
[0059] The cultured INS-1 cells were inoculated in each well of a 96-well plate in the number of 1×10.sup.4 in a medium of 100 μL and cultured for 24 hours to stabilize the cells. Next, the ginger extract, fraction thereof, and single compounds were added to a specific concentration, followed by culturing for 24 hours. Next, 10 μL of a CCK-8 reagent (Dojindo Laboratories, Japan) was added to each well, followed by culturing at a temperature of 37° C. After 1 hour, cell viability was measured by measuring absorbance at a wavelength of 450 nm on a BIO-TEK (Winooski, Vt., USA) microplate reader. The specific concentrations of the ginger extract and a fraction thereof were set to 1 μg/m L, 2.5 μg/mL, and 5 μg/mL, and the specific concentrations of the single compound were set to 1 μM, 2.5 μM, and 5 μM.
[0060] Table 1 shows toxicity test results of the ginger extract, an extract of ginger steamed for 6 hours, and an extract of ginger steamed for 12 hours, the fraction of the ginger extract, the microwave-processed ginger product, and the fraction thereof for INS-1 cells, which are mouse insulinoma cells.
TABLE-US-00001 TABLE 1 Classification Concentration (μg/mL) Cell viability (%) Ginger extract 1 99.5 ± 2.0 2.5 97.8 ± 0.2 5 98.4 ± 3.7 6 hours-steamed ginger 1 101.5 ± 1.3 2.5 98.8 ± 1.7 5 95.3 ± 1.2 12 hours-steamed ginger 1 97.4 ± 2.7 2.5 98.4 ± 3.3 5 98.6 ± 3.2 Microwave-processed 1 97.0 ± 4.8 ginger product 2.5 101.4 ± 3.5 5 102.8 ± 3.3 Ginger extract F1 1 104.3 ± 4.8 2.5 101.0 ± 3.3 5 101.7 ± 2.9 Ginger extract F2 1 102.7 ± 2.0 2.5 102.7 ± 4.8 5 101.9 ± 3.3 Ginger extract F3 1 91.6 ± 1.8 2.5 93.8 ± 3.1 5 102.5 ± 0.3 Ginger extract F4 1 105.4 ± 2.7 2.5 100.3 ± 2.6 5 103.5 ± 1.0 Ginger extract F5 1 99.9 ± 2.7 2.5 100.3 ± 3.0 5 101.3 ± 2.0 Ginger extract F6 1 99.4 ± 2.1 2.5 100.0 ± 2.5 5 100.0 ± 2.1 Microwaved-processed 1 102.3 ± 1.0 ginger product F1 2.5 103.5 ± 3.5 5 105.6 ± 3.6 Microwaved-processed 1 98.6 ± 3.6 ginger product F2 2.5 97.0 ± 0.4 5 99.9 ± 3.3 Microwaved-processed 1 101.3 ± 4.1 ginger product F3 2.5 99.1 ± 3.2 5 98.5 ± 4.3 Microwaved-processed 1 107.3 ± 4.5 ginger product F4 2.5 100.2 ± 3.8 5 102.4 ± 4.5 Microwaved-processed 1 105.6 ± 3.8 ginger product F5 2.5 100.5 ± 4.2 5 98.8 ± 2.7 Microwaved-processed 1 96.5 ± 3.7 ginger product F6 2.5 97.3 ± 3.2 5 102.3 ± 3.8
[0061] Table 2 shows toxicity test results of [6]-gingerol, [8]-gingerol, [10]-gingerol, [6]-shogaol, [8]-shogaol, and [10]-shogaol, which are single compounds contained in ginger, and gliclazide, for an INS-1 cell, which is a mouse insulinoma cell.
TABLE-US-00002 TABLE 2 Classification Concentration (μM) Cell viability (%) [6]-gingerol 1 98.1 ± 2.7 2.5 98.7 ± 2.3 5 98.2 ± 3.5 [8]-gingerol 1 97.4 ± 1.0 2.5 96.1 ± 2.2 5 96.4 ± 1.4 [10]-gingerol 1 99.3 ± 1.6 2.5 100.6 ± 3.3 5 100.7 ± 3.5 [6]-shogaol 1 101.1 ± 0.4 2.5 101.8 ± 0.7 5 101.2 ± 0.2 [8]-shogaol 1 94.0 ± 2.6 2.5 96.3 ± 1.9 5 92.4 ± 1.7 [10]-shogaol 1 101.9 ± 2.2 2.5 99.8 ± 3.9 5 99.6 ± 3.3 Glyclazide 1 99.7 ± 0.2 2.5 96.3 ± 0.9 5 91.1 ± 1.4
[0062] Cell viability in Tables 1 and 2 was calculated as follows.
[0063] Cell viability (%)=A/B×100
[0064] A: absorbance of sample to which a reagent is not added
[0065] B: absorbance of sample to which a reagent is added
[0066] As shown in Tables 1 and 2, the ginger extract, the extract ginger steamed for 6 hours, the extract ginger steamed for 12 hours, the fraction of the ginger extract, the microwave-processed ginger product and a fraction thereof, single compounds contained in ginger, such as [6]-gingerol, [8]-gingerol, [10]-gingerol, [6]-shogaol, [8]-shogaol, and [10]-shogaol, were found not to exhibit toxicity against INS-1 cells.
[0067] (3) Measurement of Ability of Glucose Stimulated Insulin Secretion (GSIS) of INS-1 Cell
[0068] 5×10.sup.5 cultured INS-1 cells were inoculated in each well of the 12-well plate in the same medium of 2 mL and cultured for 24 hours under the same condition as described in Section (1) INS-1 cell culture to stabilize the cells. Then, 114 mM sodium chloride (NaCl), 4.4 mM potassium chloride (KCl), 1 mM magnesium sulfate (MgSO.sub.4), 1.28 mM calcium chloride (CaCl.sub.2)), 29.5 mM sodium hydrogen carbonate (NaHCO.sub.3), 10 mM HEPES; Gibco BRL Life Technologies), and 0.1% bovine serum albumin (BSA) were mixed, and the cells were washed twice with 2 mL of Krebs-Ringer buffer with a hydrogen ion concentration of pH 7.4. Thereafter, 2 mL of Krebs-Ringer buffer was added to the washed cells and incubated for 1 hour. Then, the ginger extract, each fraction, and each compound were incubated for 30 minutes in 1.8 mL of Krebs-Ringer buffer at a specific concentration. The specific concentrations of the ginger extract and a fraction thereof were set to 1 μg/m L, 2.5 μg/mL, and 5 μg/mL, and the specific concentrations of the single compound were set to 1 μM, 2.5 μM, and 5 μM.
[0069] 0.2 mL of Krebs-Ringer buffers containing 33 mM and 167 mM glucose were each added to the wells and incubated for 1 hour. Thereafter, after centrifugation at 12,000 revolutions per minute (rpm) for 10 minutes at a temperature of 4° C., the supernatant was taken, and the amount of insulin was measured with a Rat insulin RIA kit (Gentaur Molecular Products, Belgium). The stimulation index (SI) is a value obtained by dividing a value measured at a high glucose concentration, that is, the amount of insulin secreted by stimulation of a 16.7 mM glucose concentration by a value measured at a low glucose concentration, that is, the amount of insulin secreted by stimulation of a 3.3 mM glucose concentration. The measured SIs are shown in Tables 3 and 4. As a positive control, gliclazide was used at concentrations of 2.5 μM, 5.0 μM, 10 μM, and 20 μM.
[0070] Table 3 shows insulin secretion ability test results of the ginger extract, the extract of ginger steamed for 6 hours, and the extract of ginger steamed for 12 hours, the fraction of the ginger extract, the microwave-processed ginger product, and the fraction thereof for INS-1 cells, which are mouse insulinoma cells.
TABLE-US-00003 TABLE 3 Classification Concentration (μg/mL) Stimulation index (SI) Ginger extract 1 5.1 ± 0.1 2.5 7.7 ± 0.2 5 6.4 ± 0.4 6 hours-steamed ginger 1 6.1 ± .02 2.5 3.7 ± 0.0 5 2.5 ± 0.1 12 hours-steamed ginger 1 2.2 ± 0.5 2.5 2.7 ± 0.0 5 4.2 ± 0.0 Microwave-processed 1 6.5 ± 0.6 ginger product 2.5 6.0 ± 0.2 5 12.4 ± 0.4 Ginger extract F1 1 1.2 ± 0.0 2.5 1.4 ± 0.1 5 4.3 ± 0.3 Ginger extract F2 1 2.3 ± 0.0 2.5 2.9 ± 0.1 5 6.2 ± 0.1 Ginger extract F3 1 3.2 ± 0.3 2.5 2.3 ± 0.0 5 2.7 ± 0.1 Ginger extract F4 1 1.3 ± 0.0 2.5 2.8 ± 0.1 5 4.1 ± 0.0 Ginger extract F5 1 5.7 ± 0.1 2.5 6.7 ± 0.3 5 10.8 ± 0.6 Ginger extract F6 1 2.4 ± 0.0 2.5 3.1 ± 0.0 5 3.9 ± 0.2 Microwaved-processed 1 3.1 ± 0.1 ginger product F1 2.5 4.5 ± 0.0 5 5.4 ± 0.0 Microwaved-processed 1 4.9 ± 0.0 ginger product F2 2.5 6.2 ± 0.2 5 7.4 ± 0.0 Microwaved-processed 1 1.2 ± 0.0 ginger product F3 2.5 1.5 ± 0.0 5 6.4 ± 0.1 Microwaved-processed 1 2.1 ± 0.0 ginger product F4 2.5 8.5 ± 1.1 5 18.1 ± 0.5 Microwaved-processed 1 2.3 ± 0.0 ginger product F5 2.5 3.4 ± 0.2 5 3.0 ± 0.0 Microwaved-processed 1 1.0 ± 0.0 ginger product F6 2.5 5.0 ± 0.0 5 5.3 ± 0.1
[0071] Table 4 shows insulin secretion ability test results of [6]-gingerol, [8]-gingerol, [10]-gingerol, [6]-shogaol, [8]-shogaol, and [10]-shogaol, which are single compounds contained in ginger, and gliclazide for INS-1 cells, which are mouse insulinoma cells.
TABLE-US-00004 TABLE 4 Classification Concentration (μM) Stimulation index (SI) [6]-gingerol 1 2.4 ± 0.0 2.5 2.2 ± 0.1 5 2.3 ± 0.1 [8]-gingerol 1 1.1 ± 0.0 2.5 1.5 ± 0.0 5 2.1 ± 0.0 [10]-gingerol 1 1.7 ± 0.2 2.5 1.7 ± 0.1 5 1.5 ± 0.1 [6]-shogaol 1 3.0 ± .0 2.5 3.5 ± 0.1 5 4.8 ± 0.0 [8]-shogaol 1 1.3 ± 0.1 2.5 1.5 ± 0.1 5 1.8 ± 0.1 [10]-shogaol 1 1.9 ± 0.0 2.5 1.4 ± 0.0 5 1.4 ± 0.1 Glyclazide 1 1.3 ± 0.0 2.5 1.9 ± 0.1 5 3.0 ± 0.1
[0072] Table 5 shows results of measuring the content of a single component using ultra performance liquid chromatography (UPLC) for processed ginger products F1 to F6. Each single component was prepared at 10 ppm, 25 ppm, 50 ppm, 100 ppm, and 250 ppm, and a verification line was prepared after UPLC analysis. After that, by analyzing the fraction of the processed ginger product according to the same method, the content of each of [6]-gingerol, [8]-gingerol, [10]-gingerol, [6]-shogaol, [8]-shogaol, and [10]-shogaol was analyzed The results thereof are shown in Table 5 as mg per 1 g of the fraction.
TABLE-US-00005 TABLE 5 [6]- [8]- [10]- [6]- [8]- [10]- Sample gingerol gingerol gingerol shogaol shogaol shogaol Total Microwave- — — — — — — — processed ginger Processed product F1 MW ginger — — — — — — — Processed product F2 MW ginger — — — — — — — Processed product F3 MW ginger 17.0 7.2 — 118.9 — — 143.1 Processed product F4 Microwave- — — 3.9 41.9 24.3 36.6 106.7 processed ginger Processed product F5 Microwave- — — — 0.5 — — 0.5 processed ginger Processed product F6
[0073] As shown in Tables 3 and 4, INS-1 cells secreted insulin concentration-dependently in the presence of the ginger extract, the extract of ginger steamed for 6 hours, the extract of ginger steamed for 12 hours, the fraction of the ginger extract, the microwave-processed ginger product and a fraction thereof, and single compounds contained in ginger, such as [6]-gingerol, [8]-gingerol, [10]-gingerol, [6]-shogaol, [8]-shogaol, and [10]-shogaol. In particular, when comparing the ginger extract, the steamed ginger extracts, and the microwave-processed ginger product, the insulin secretion ability of the microwave-processed ginger product improved significantly. When comparing the fractions, the fraction of the processed ginger product in 80% ethanol (processed ginger product F4) was found to have the best insulin secretion ability. As a result of component analysis through UPLC, the highest activity was achieved when the content of [6]-shogaol, [8]-shogaol, and [10]-shogaol was 83% of the total gingerol and shogaol content in the fraction. In particular, when the content of [6]-shogaol was 80% or higher, high activity was achieved. Based on this result, from the results of Table 4 showing the insulin secretion ability of gingerols and shogaols, which are single components contained in ginger, [6]-shogaol was found to promote insulin secretion in INS-1 cells, as compared with other components.
[0074] (2) Results of Oral Glucose Tolerance Test (OGTT)
[0075] ICR mice were used for OGTT experiments. ICR mice were fasted for 12 hours before the experiment, blood was collected from the caudal vein of mice that had been fasted for 12 hours, and the fasting blood glucose level was measured as the initial blood glucose level. Thereafter, experiments were conducted with a group administered with glucose (2 g/Kg) (control), a group administered with glucose and a raw ginger extract (100 mg/kg, not processed), and a group administered with a microwave-processed ginger product (100 mg/kg, processed). After oral administration for each concentration, glucose was orally administered 1 hour later, and measurement was performed by using a blood glucose meter from the caudal vein at 0, 15, 30, 60, and 120 minutes.
[0076]
[0077] Next, to test the efficacy of a single component of ginger, blood was collected from the caudal vein of mice that had been fasted for 12 hours, and the fasting blood glucose level was measured as the initial blood glucose level. Thereafter, experiments were conducted with a group administered with glucose (2 g/Kg) (control), a group administered with glucose and 6-gingerol (100 mg/kg), and a group administered with glucose and 6-shogaol (100 mg/kg). After oral administration for each concentration of 6-gingerol and 6-shogaol, glucose was orally administered 1 hour later, and measurement was performed by using a blood glucose meter from the caudal vein at 0, 15, 30, 60, and 120 minutes.
[0078]
[0079] From the above results, when a ginger extract was irradiated with microwaves, [6]-gingerol, [8]-gingerol, and [10]-gingerol were dehydrated to [6]-shogaol, [8]-shogaol, and [10]-shogaol, respectively, to thereby obtain a processed ginger product with an increased content of shogaol. In addition, it was found that the microwave-processed ginger products, in particular, the fraction (processed ginger product F4) with a high [6]-shogaol content and the [6]-shogaol single compound, stimulate the pancreatic cells to promote insulin secretion.
[0080] As apparent from the foregoing description, the antidiabetic composition according to an aspect may be used in preventing, improving, or treating diabetes in a subject.
[0081] The method of preventing or treating diabetes in a subject according to one aspect of the present disclosure may effectively prevent or treat diabetes.
[0082] It should be understood that embodiments described herein should be considered in a descriptive sense only and not for purposes of limitation. Descriptions of features or aspects within each embodiment should typically be considered as available for other similar features or aspects in other embodiments. While one or more embodiments have been described with reference to the figures, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the disclosure as defined by the following claims.