JP7116413B2 - Sorting method - Google Patents

Description

The present invention relates to a method for selecting a subject to be administered Melinjo-derived resveratrol for promoting adiponectin multimerization.

Adiponectin is one of adipokines secreted into blood from adipocytes, and exists mainly as trimers, hexamers or multimers (HMW) in blood. Adiponectin has effects such as improving insulin resistance and anti-inflammatory effects, and is activated in vivo by forming multimers.

Disulfide bond-forming oxidoreductase A-like protein (DsbA-L) is a protein involved in adiponectin multimerization. So far, in vitro or mouse in vivo studies have reported that administration of trans-resveratrol extracted from grapes activated DsbA-L and promoted adiponectin multimerization. (Non-Patent Document 1).

Mohammadi-Sartang M. et al.,Pharmacological Research, 2017, Vol. 117, p394-405

However, human trials with trans-resveratrol have yielded inconsistent results regarding efficacy (Non-Patent Document 1). On the other hand, Melinjo-derived resveratrol has not been confirmed to have an effect on adiponectin. The present inventors found that Melinjo-derived resveratrol has a high multimerization promoting effect in a specific subject group.

An object of the present invention is to provide a method for selecting a subject having a more effective adiponectin multimerization-promoting effect by ingestion of Melinjo-derived resveratrol.

The present invention is a method for selecting a subject to be administered Melinjo-derived resveratrol for promoting adiponectin multimerization, wherein the subject is (1) genotype G in DsbA-L rs1917760 (-1308G / T) / T type or T / T type, and / or (2) when the DsbA-L mRNA expression level in peripheral blood mononuclear cells is less than or equal to a predetermined value, administer Melinjo-derived resveratrol to the subject A method is provided comprising the step of screening for a subject. By the above method, subjects having a higher adiponectin multimerization-promoting effect by ingestion of Melinjo-derived resveratrol are selected as subjects for administration.

The present invention also provides a method for predicting the adiponectin multimerization-promoting effect of ingestion of Melinjo-derived resveratrol, wherein the subject (1) has a DsbA-L rs1917760 (-1308G/T) genotype of G/T type or T / T type, and / or (2) when the DsbA-L mRNA expression level in peripheral blood mononuclear cells is a predetermined value or less, the subject is high due to ingestion of Melinjo-derived resveratrol Provided is a method comprising the step of predicting that it has an adiponectin multimerization-promoting effect. By the above method, it is possible to predict whether or not ingestion of Melinjo-derived resveratrol has a high adiponectin multimerization-promoting effect without requiring actual administration.

The present invention further provides a method for determining the properties of a subject, wherein the subject having an adiponectin multimerization-promoting effect due to ingestion of resveratrol derived from Melinjo is (1) a gene in DsbA-L rs1917760 (-1308G/T) A person whose type is G/T type or T/T type, and (2) a person whose DsbA-L mRNA expression level in peripheral blood mononuclear cells is a predetermined value or less. provide a way. By the above method, it is possible to determine the physiological properties of a subject having a high adiponectin multimerization-enhancing effect by ingestion of Melinjo-derived resveratrol.

The above-described determination method comprises the step of measuring the blood total adiponectin concentration and high-molecular-weight adiponectin concentration after ingestion of Melinjo-derived resveratrol, and based on the blood-total adiponectin concentration and high-molecular-weight adiponectin concentration, the above A step of evaluating the adiponectin multimerization-promoting effect of ingestion of Melinjo-derived resveratrol in the subject.

The present invention also contains as an active ingredient at least one selected from the group consisting of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D, and (1) the genotype in DsbA-L rs1917760 (-1308G/T) is G/ Provided is an adiponectin multimerization-promoting agent for a subject who is T-type or T/T-type and/or (2) has a DsbA-L mRNA expression level in peripheral blood mononuclear cells of a predetermined value or less. The adiponectin multimerization-promoting agent according to the present invention has an excellent adiponectin multimerization-promoting effect on the above object.

INDUSTRIAL APPLICABILITY The present invention provides a method for selecting subjects having more effective adiponectin multimerization-enhancing action by ingestion of Melinjo-derived resveratrol.

DETAILED DESCRIPTION OF THE INVENTION Embodiments for carrying out the present invention will be described in detail below. However, the present invention is not limited to the following embodiments.

The present invention provides a method of selecting a subject to be administered Melinjo-derived resveratrol for promoting adiponectin multimerization. Promoting adiponectin multimerization specifically means increasing the proportion of high molecular weight (HMW) adiponectin concentration in the total adiponectin concentration in blood. High-molecular-weight adiponectin refers to adiponectin that forms multimers larger than hexamers.

Melinjo is a plant belonging to the Gnetumaceae native to Indonesia, and its scientific name is Gnetum Gnemon L. is. Melinjo-derived resveratrol is a general term for resveratrols contained in Melinjo, and specifically refers to gnetin C, gnetin L, gnemonoside A, gnemonoside C and gnemonoside D. Melinjo-derived resveratrol preferably contains at least one selected from the group consisting of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D. Melinjo-derived resveratrol is characterized by a longer blood residence time than trans-resveratrol, and is preferable because it exhibits more stable efficacy.

Gnemonoside A, gnemonoside C and gnemonoside D are gnetin C glycosides. The structural formulas of the above compounds are shown below.

In gnetin C, R ₁ , R ₃ and R ₄ are hydrogen atoms, and R ₂ is a hydroxyl group in the above general formula (1). In gnetin L, R ₁ is a group represented by —OCH ₃ , R ₂ and R ₄ are hydrogen atoms, and R ₃ is a hydroxyl group in the above general formula (1). In gnemonoside A, R ₁ and R ₃ are hydrogen atoms, R ₂ is a group represented by —OGlc, and R ₄ is Glc in the above general formula (1). In gnemonoside C, R ₁ , R ₃ and R ₄ are hydrogen atoms, and R ₂ is a group represented by —OGlc in the above general formula (1). Gnemonoside D is a group represented by the above general formula (1), in which R ₁ and R ₃ are hydrogen atoms, R ₂ is a hydroxyl group, and R ₄ is —Glc. Glc indicates a glucose residue (D-glucopyranosyl).

As a raw material containing Melinjo-derived resveratrol, for example, a Melinjo extract can be used. A Melinjo extract can be obtained, for example, by extracting Melinjo seeds with a solvent such as water, ethanol, methanol, or acetone. The Melinjo extract may be one that has undergone further treatments such as fractionation and purification.

In the selection method according to this embodiment, the subject is (1) genotype G/T or T/T in DsbA-L rs1917760 (-1308G/T), and/or (2) peripheral blood single If the expression level of DsbA-L mRNA in nuclear cells is equal to or less than a predetermined value, the subject is selected as a subject to be administered Melinjo-derived resveratrol. A subject corresponding to at least one of the above (1) and (2) is highly likely to have a high adiponectin multimerization promoting effect by administration of Melinjo-derived resveratrol.

Targets selected in the selection method according to the present embodiment will be described. DsbA-L rs1917760 (-1308G/T) has genotypes of G/G type, G/T type and T/T type. In the selection method according to this embodiment, if the subject has a genotype of G/T or T/T in DsbA-L rs1917760 (-1308G/T), the subject is administered Melinjo-derived resveratrol. selected as targets for Melinjo-derived resveratrol is likely to have a high adiponectin multimerization-promoting effect in subjects with G/T or T/T genotypes.

Genotyping for DsbA-L rs1917760 (-1308G/T) can be performed by known methods, and can be analyzed using, for example, commercially available kits. Samples such as blood and saliva can be used as test samples for genotyping. The selection method according to this embodiment further includes a step of extracting DNA from a test sample collected from a subject, analyzing the DNA sequence in the DNA to determine the genotype of DsbA-L rs1917760 (-1308G/T) of the subject. A step of determining may be included.

Melinjo-derived resveratrol is likely to have a high adiponectin multimerization-promoting effect in subjects with low DsbA-L mRNA expression levels in peripheral blood mononuclear cells. In the selection method according to the present embodiment, when the DsbA-L mRNA expression level in the peripheral blood mononuclear cells of the subject is a predetermined value or less, the subject is selected as a subject to be administered Melinjo-derived resveratrol. .

The DsbA-L mRNA expression level in peripheral blood mononuclear cells can be measured by a known method. The selection method according to this embodiment may further comprise the step of measuring the expression level of DsbA-L mRNA in peripheral blood mononuclear cells of the subject. The step of measuring the expression level of DsbA-L mRNA in peripheral blood mononuclear cells of a subject is, for example, the step of extracting RNA in peripheral blood mononuclear cells from a blood sample collected from the subject, cDNA transcribed from the RNA and measuring the DsbA-L mRNA expression level based on the obtained cDNA level.

In the selection method according to this embodiment, the predetermined value of the DsbA-L mRNA expression level in peripheral blood mononuclear cells may be, for example, the median value or average value of the mRNA expression levels in a plurality of subjects.

By the selection method according to the present embodiment, it is possible to select subjects having an adiponectin multimerization-promoting effect due to ingestion of Melinjo-derived resveratrol. Subjects who do not fall under any of the above (1) and (2) have a high possibility that even if Melinjo-derived resveratrol is ingested, the adiponectin multimerization promoting effect is not exhibited or is very low, and Melinjo-derived resveratrol Can be excluded from administration of veratrol. By using the selection method according to the present embodiment, it is possible to avoid unnecessary ingestion of Melinjo-derived resveratrol by a subject with poor adiponectin multimerization-promoting effect by administration of Melinjo-derived resveratrol.

In the selection method according to this embodiment, the subject may be a mammal such as a human.

(Prediction method)
(1) DsbA-L rs1917760 (-1308G/T) genotype is G/T type or T/T type, and/or (2) DsbA-L mRNA expression level in peripheral blood mononuclear cells is predetermined If it is below the value, there is a high possibility that ingestion of Melinjo-derived resveratrol will exhibit a high adiponectin multimerization promoting effect. Therefore, the present invention also provides a method for predicting the adiponectin multimerization-promoting effect of ingestion of Melinjo-derived resveratrol, wherein the subject (1) has a DsbA-L rs1917760 (-1308G/T) genotype of G / T type or T / T type, and / or (2) when the DsbA-L mRNA expression level in peripheral blood mononuclear cells is a predetermined value or less, the subject ingests Melinjo-derived resveratrol to have a high adiponectin multimerization-promoting effect. The same aspect as the selection method described above can be applied to the aspect of determining whether or not the object corresponds to the above (1) and (2).

(Object property judgment method)
The present invention also provides a method for determining the properties of a subject, wherein the subject having an adiponectin multimerization-promoting effect by ingestion of Melinjo-derived resveratrol is (1) the gene in DsbA-L rs1917760 (-1308G/T) A person whose type is G/T type or T/T type, and (2) a person whose DsbA-L mRNA expression level in peripheral blood mononuclear cells is a predetermined value or less. provide a way. By the above method, the subject's genotype or DsbA-L mRNA expression level can be determined without directly measuring the subject's genotype or DsbA-L mRNA expression level.

The determination method according to the present embodiment includes, for example, a step of measuring the total adiponectin concentration and/or high-molecular-weight adiponectin concentration in the blood after ingestion of Melinjo-derived resveratrol, and the ratio of the high-molecular-weight adiponectin concentration to the total adiponectin concentration. A step such as a step of measuring may be included.

The determination method according to the present embodiment further includes the step of measuring the ratio of the high-molecular-weight adiponectin concentration in the total adiponectin concentration before ingestion of Melinjo-derived resveratrol, and the high-molecular-weight adiponectin concentration in the total adiponectin concentration before and after ingestion. and comparing the proportions.

The determination method according to the present embodiment includes the step of evaluating the adiponectin multimerization-promoting effect of ingestion of Melinjo-derived resveratrol in the subject, based on the measured blood total adiponectin concentration and high-molecular-weight adiponectin concentration. It may further contain: In the evaluation step, for example, after ingestion of Melinjo-derived resveratrol, when the ratio of high-molecular-weight adiponectin concentration in the total adiponectin concentration in the blood increases by a certain amount or more, for example, when it increases by 6% or more, the subject can be evaluated as having a high adiponectin multimerization-promoting effect.

In the determination method according to the present embodiment, when the subject has an adiponectin multimerization promoting effect due to ingestion of Melinjo-derived resveratrol, the subject is 1) the gene at DsbA-L rs1917760 (-1308G/T) It can be determined that at least one of a person whose type is G/T type or T/T type, and (2) a person whose DsbA-L mRNA expression level in peripheral blood mononuclear cells is a predetermined value or less. .

The blood total adiponectin concentration and high-molecular-weight adiponectin concentration can be measured by known methods. Measurement of blood total adiponectin concentration and high-molecular-weight adiponectin concentration after ingestion of Melinjo-derived resveratrol was performed after ingestion of Melinjo-derived resveratrol was continued for 3 days or more, 1 week or more, 2 weeks or more, or 1 month or more. may be broken.

(Adiponectin multimerization promoter)
The present invention also provides an adiponectin multimerization promoter containing at least one selected from the group consisting of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D as an active ingredient. The adiponectin multimerization-promoting agent according to the present embodiment has (1) genotype in DsbA-L rs1917760 (-1308G/T) is G/T type or T/T type, and/or (2) peripheral blood For subjects whose DsbA-L mRNA expression level in mononuclear cells is below a predetermined value.

Since the adiponectin multimerization accelerator according to the present embodiment contains at least one selected from the group consisting of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D as an active ingredient, especially the above (1) and (2) Adiponectin multimerization can be promoted by ingesting the adiponectin multimerization-enhancing agent in at least one subject.

Gnetin C, gnemonoside A, gnemonoside C and gnemonoside D may be commercially available products or may be naturally derived. Gnetin C, gnemonoside A, gnemonoside C and gnemonoside D may be derived, for example, from Melinjo. As the raw material containing at least one selected from the group consisting of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D, for example, Melinjo extract can be used. A Melinjo extract can be obtained, for example, by extracting Melinjo seeds with a solvent such as water, ethanol, methanol, or acetone. The Melinjo extract may be one that has undergone further treatments such as fractionation and purification.

Since the adiponectin multimerization-promoting agent according to the present embodiment contains the above-mentioned active ingredient, in particular the DsbA-L mRNA expression level in peripheral blood mononuclear cells is below a predetermined value, the DsbA-L mRNA expression level It has an increasing effect. Therefore, the adiponectin multimerization-promoting agent according to the present embodiment can also be said to be a DsbA-L mRNA expression-promoting agent targeting those whose DsbA-L mRNA expression level in peripheral blood mononuclear cells is a predetermined value or less. . The predetermined value may be, for example, the median value or average value of the mRNA expression levels in multiple subjects.

The adiponectin multimerization accelerator according to the present embodiment contains the above active ingredient, for example, 0.2% by mass or more, 0.4% by mass or more, 0.5% by mass or more, based on the total amount of the adiponectin multimerization accelerator. It may be contained in an amount of 1% by mass or more or 3% by mass or more, preferably 5% by mass or more, and more preferably 10% by mass or more. The adiponectin multimerization accelerator according to the present embodiment contains, for example, 100% by mass or less, 95% by mass or less, 90% by mass or less, 80% by mass or less, 70% by mass of the above-mentioned active ingredient with respect to the total amount of the adiponectin multimerization accelerator. % or less, 50.0 mass % or less, 40.0 mass % or less, 30.0 mass % or less, 20.0 mass % or less, or 10.0 mass % or less.

In the adiponectin multimerization accelerator according to the present embodiment, the active ingredient is preferably a mixture of two or more selected from the group consisting of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D, and a mixture of three or more. and more preferably all of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D are included.

The content of gnetin C is preferably 10 to 20% by mass, more preferably 12 to 16% by mass, based on the total amount of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D. The content of gnemonoside A is preferably 45 to 60% by mass, more preferably 50 to 55% by mass, based on the above total amount. The content of gnemonoside C is preferably 5 to 15% by mass, more preferably 7 to 12% by mass, based on the above total amount. The content of gnemonoside D is preferably 20 to 30% by mass, more preferably 22 to 27% by mass, based on the above total amount.

The adiponectin multimerization promoter according to the present embodiment can be used, for example, in a dose of 1 mg to 1 g per day for an adult with a body weight of 60 kg, preferably in a dose of 10 to 500 mg, in terms of the amount of active ingredient. The dose can be appropriately set within the above range depending on factors such as the health condition of the person taking the drug, administration method, and combination with other agents.

The adiponectin multimerization-enhancing agent according to this embodiment may be orally administered (ingested) or parenterally administered. The adiponectin multimerization promoter of the present invention may be administered once a day, or multiple times such as twice a day, three times a day, etc., as long as the amount of the active ingredient per day is within the range described above. may be administered in separate doses.

The adiponectin multimerization-enhancing agent according to this embodiment can be suitably used for long-term administration. Long-term administration of the adiponectin multimerization-enhancing agent makes the adiponectin multimerization-enhancing effect even more pronounced. The adiponectin multimerization-enhancing agent according to the present embodiment is preferably administered over a period of 1 week or more, more preferably over a period of 2 weeks or more, and is administered over a period of 4 weeks or more. is more preferred, and administration over a period of 3 months or longer is even more preferred.

The adiponectin multimerization promoter according to the present embodiment is, in addition to one or more selected from the group consisting of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D, gnetin L, resveratrol such as trans-resveratrol It may contain types. The adiponectin multimerization promoter according to the present embodiment may contain, for example, Melinjo seed extract and separately added trans-resveratrol. The content of trans-resveratrol in the adiponectin multimerization accelerator may be, for example, 0.1 to 20% by mass, or 1 to 10% by mass. The content of trans-resveratrol in the adiponectin multimerization accelerator according to the present embodiment may be 0 to 1% by mass, or may be 0 to 0.5% by mass.

The adiponectin multimerization promoting agent according to the present embodiment may contain only the above-mentioned active ingredients, or may contain only the melinjo extract, unless the effects of the present invention are hindered. It may further contain other components. Other ingredients include, for example, pharmaceutically acceptable ingredients (e.g., excipients, binders, lubricants, disintegrants, emulsifiers, surfactants, bases, solubilizers, suspending agents). , Food acceptable ingredients (e.g., minerals, vitamins, flavonoids, quinones, polyphenols, amino acids, nucleic acids, essential fatty acids, cooling agents, binders, sweeteners, disintegrants, lubricants, coloring agents , fragrances, stabilizers, preservatives, sustained release modifiers, surfactants, solubilizers, wetting agents).

The adiponectin multimerization accelerator according to the present embodiment may be in any form such as solid, liquid, paste, etc. Tablets (uncoated tablets, sugar-coated tablets, effervescent tablets, film-coated tablets, chewable tablets, lozenges, etc.) ), capsules, pills, powders (powders), fine granules, granules, liquids, suspensions, emulsions, syrups, pastes, injections (when used, distilled water or amino acid or electrolyte infusions (including the case where it is prepared as a liquid agent by blending it with an infusion solution such as ). These various formulations can be prepared, for example, by mixing the above-mentioned active ingredients and, if necessary, other ingredients and molding them into the above dosage forms. The adiponectin multimerization promoter according to this embodiment is preferably stored in a light-shielding container.

The adiponectin multimerization promoter according to the present embodiment can be used as a pharmaceutical, quasi-drug, or food composition itself, or added to a pharmaceutical, quasi-drug, or food composition. The food composition is preferably a food in which the tertiary function of food (physical conditioning function) is emphasized. Examples of foods in which the tertiary functions of foods are emphasized include health foods, foods with function claims, dietary supplements, foods with nutrient function claims, supplements, and foods for specified health uses.

A pharmaceutical, quasi-drug, or food composition comprising the adiponectin multimerization enhancer according to the present embodiment, or a pharmaceutical, quasi-drug, or food composition comprising the adiponectin multimerization enhancer according to the present embodiment, It may be for promoting adiponectin multimerization, for example, improvement of insulin resistance, suppression of TNF-α production, and type 2 diabetes, arteriosclerosis, coronary artery disease, renal failure, myocardial infarction, hypertension, liver fibrosis, etc. An indication such as that there is a preventive or ameliorative effect may be attached.

A food composition containing an adiponectin multimerization promoter according to the present embodiment may contain, for example, 0.2 to 10.0% by mass of the active ingredient. The food composition includes the following, and the above-mentioned adiponectin multimerization promoter is mixed with the intermediate product during the manufacturing process or the final product to obtain the food composition used for the above purpose. Can be: soft drinks such as coffee, juice and tea drinks, milk drinks, lactic acid drinks, yogurt drinks, carbonated drinks, and alcoholic beverages such as sake, western liquor, fruit wine and honey wine; spreads such as custard cream Paste such as fruit paste; Western confectionery such as chocolate, donut, pie, cream puff, gum, jelly, candy, cookie, cake and pudding; and frozen desserts such as sherbet; cooked foods such as curry, beef bowl, porridge, miso soup, soup, meat sauce, pasta, pickles and jam;

EXAMPLES The present invention will now be described more specifically based on examples. However, the present invention is not limited to the following examples.

<Test method>
Twenty-eight male healthy adults who met the following eligibility criteria for constitution and lifestyle were included in this study.
1) No diseases affecting drug metabolism and excretion, such as gastrointestinal, heart, lung and liver diseases, and congenital metabolic disorders.
2) Never smoked.
3) Not taking functional foods related to fat or sugar metabolism on a daily basis.
4) You do not drink red wine on a daily basis.

Melinjo-derived resveratrol test food containing 150 mg of Melinjo seed extract powder per tablet, of which 30 mg as resveratrols, was prepared. Table 1 shows the amounts of resveratrols contained in 1.0 g of the Melinjo seed extract powder. As a placebo test meal, one containing no Melinjo seed extract powder was prepared. The Melinjo-derived resveratrol test food and placebo test food used contain dextrin, cellulose, sucrose fatty acid ester and gelatin as additives.

Subjects were randomly assigned to two groups of 14 subjects each, a test group and a placebo group, respectively. The test group was given two tablets of the Melinjo-derived resveratrol test meal once a day, and the placebo group was given two tablets of the placebo test meal once a day. The subjects were instructed not to take alcohol, grapefruit, and resveratrol-containing food from two days before the test meal to the end of the test. Subjects continued to take each test meal (before breakfast) for 14 days. Blood was collected from the subjects immediately before the first oral administration of the test meal and on the 14th day of oral administration.

(Measurement of adiponectin concentration)
Using serum extracted from the collected blood samples, total adiponectin (μg/ml) and HMW adiponectin (μg/ml) protein levels of each subject were measured. Also, the proportion (%) of HMW adiponectin in the total adiponectin of each subject was calculated. An ELISA kit was used for the measurement (Human Total Adiponectin/Acrp30 and Human HMW Adiponectin/Acrp30 Quantikine ELISA Kits, R&D Systems).

(Gene expression level)
RNA in peripheral blood mononuclear cells (PBMC) was extracted from the collected blood samples using RNA iso Blood (Takara Bio Inc.). Thereafter, the yield of total RNA was calculated from the absorbance of UV (260 nm), and it was confirmed that the ratio of OD260/OD280 was 1.7 or more, indicating that highly pure RNA was obtained. Furthermore, using Prime Script (registered trademark) RT reagent Kit with gDNA Eraser Perfect Real Time (Takara Bio Inc.), genome removal reaction (2 minutes at 42°C), reverse transcription reaction (15 minutes at 37°C, 5 minutes at 85°C) seconds, ∞ at 4° C.; 1 cycle) to obtain complementary DNA (cDNA).

Using the obtained cDNA, the gene expression level of DsbA-L was measured. For the measurement, Step One Plus real-time PCR system version 2.1 (Applied Biosystems) was used, and quantitative TaqMan polymerase chain reaction (PCR) method was used. Relative mRNA amounts of target genes DsbA-L were calculated by the comparative Ct method (ΔΔCt method). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal standard gene.

(Genotyping)
Flexi Gene DNA kit (Qiagen) was used for DNA extraction from peripheral blood. Using the extracted DNA, DsbA-L rs1917760 (-1308G/T) and ADIPOQ rs3774261, rs182052 genetic polymorphisms were detected using commercially available probes and primers (TaqMantm Genotyping Master Mix (Applied Biosystems); TaqMantm SNP Genotyping Assay (ID C 27479710 10; C 2412785 10) (Applied Biosystems) using the TaqMan PCR method.

(Statistical analysis)
Numeric variables were compared using one-way ANOVA, Student's t-test, or Mann-Whitney U-test. Fisher's exact test was used to compare categorical variables. Multiple regression analysis was used for multivariate analysis, and the degree of influence was indicated by the partial regression coefficient (standard error). SPSS software package for Windows (version 23.0, IBM) was used for the above statistical analysis, and P<0.05 was considered significant.

Table 2 shows the clinical background of the subjects at the start of observation.

Table 3 shows the subject's DsbA-L genotype and ADIPOQ genotype. The genotype frequency of each gene was consistent with previous reports in Japanese. In addition, each genotype frequency satisfied the Hardy-weinberg equilibrium (P>0.05 for each).

<Results>
(Laboratory test values before and after oral administration of the test meal)
BMI, total protein, albumin, AST, ALT, γ-GT, total bilirubin, creatinine, urea nitrogen, uric acid, HDL-C, LDL-C, triglycerides, and glycoalbumin in both placebo and test groups , no significant change was observed before and after oral administration of the test meal.

(Adiponectin levels before and after oral administration of the test meal)
There was no significant effect on the total adiponectin amount, the HMW adiponectin amount and the HMW adiponectin ratio in any group before and after oral administration of the test meal. The Mann-Whitney U test was used to calculate the significance probability.

(Inter-group comparison of changes in adiponectin due to oral administration of test meals, univariate analysis)
Changes in total adiponectin amount (μg/ml), HMW adiponectin amount (μg/ml) and ratio (%) before and after melinjo-derived resveratrol test oral administration were compared with those in the placebo group. The Mann-Whitney U test was used to calculate the significance probability. In the test group, there was no significant difference in the amount of change in total adiponectin amount (μg/ml), HMW adiponectin amount (μg/ml), and HMW adiponectin ratio (%) compared to the placebo group before and after oral administration of the test meal. rice field.

(Inter-group comparison of changes in adiponectin due to oral administration of test meals, multivariate analysis)
A multivariate analysis was performed on changes in the total adiponectin amount (μg/ml), the HMW adiponectin amount (μg/ml) and the ratio (%) due to oral administration of the test diet. Tables 4, 5 and 6 show the results. Multiple regression analysis was used for analysis. Before and after taking the test meal, HMW adiponectin (%) was significantly increased in the test group compared to the placebo group. On the other hand, no significant effect was observed on the total adiponectin amount (μg/ml) and the HMW adiponectin amount (μg/ml).

(Influence of genotype, multivariate analysis)
The effect of genotype on the amount of change in the HMW/total adiponectin ratio due to oral administration of the test diet was examined by multivariate analysis (multiple regression analysis). Table 7 shows the results. In DsbA-L rs1917760 (-1308G/T) G/T or T/T type carriers, HMW adiponectin (%) was significantly increased by oral administration of the test diet, and DsbA-L rs1917760 (-1308G/T) G No significant effect was observed in /G type carriers. On the other hand, when stratified by ADIPOQ genotype, no difference was observed in the effect of oral administration of resveratrols.

Each group was stratified by the median amount of DsbA-L mRNA in PBMCs, and HMW adiponectin (%) before and after administration of the test food was examined. Multiple regression analysis was used for analysis. Table 8 shows the results. In the group with DsbA-L mRNA levels lower than the median, oral administration of the test diet significantly increased HMW adiponectin (%). On the other hand, no effect was observed in the group in which the DsbA-L mRNA level was higher than the median value.

The amount of change in DsbA-L mRNA before and after oral administration of the test meal was analyzed using multiple regression analysis. Table 9 shows the results. In all subjects, no effect on DsbA-L mRNA by resveratrol oral administration was observed, but in the group with a low amount of DsbA-L mRNA, compared with the placebo group in the test group, significantly DsbA-L mRNA quantity increased.

Claims (3)

A method for selecting a subject to be administered Melinjo-derived resveratrol for promoting adiponectin multimerization,
The subject is (1) genotype G/T or T/T at DsbA-L rs1917760 (-1308G/T) and/or (2) DsbA-L mRNA expression in peripheral blood mononuclear cells A method comprising selecting the subject as a subject to be administered Melinjo-derived resveratrol when the amount is equal to or less than a predetermined value, wherein the predetermined value is the median value of the mRNA expression levels in a plurality of subjects. . A method for predicting the adiponectin multimerization-promoting effect of ingestion of Melinjo-derived resveratrol,
if the subject is (1) genotyped at DsbA-L rs1917760 (−1308G/T) as G/T or T/T , compared to a subject whose genotype is G/G; and / or (2) when the DsbA-L mRNA expression level in peripheral blood mononuclear cells is a predetermined value or less, compared with a subject whose mRNA expression level exceeds the predetermined value, the subject is Melinjo-derived less A method, comprising the step of predicting that intake of veratrol has a high adiponectin multimerization-promoting effect , wherein the predetermined value is an intermediate value of the mRNA expression levels in a plurality of subjects . containing at least one selected from the group consisting of gnetin C, gnemonoside A, gnemonoside C and gnemonoside D as an active ingredient,
(1) DsbA-L rs1917760 (-1308G/T) genotype is G/T type or T/T type, and/or (2) DsbA-L mRNA expression level in peripheral blood mononuclear cells is multiple an adiponectin multimerization-enhancing agent for a subject whose mRNA expression level is below the median value in the subject of