Application of combination of resveratrol and tomato pectin in preparation of products for relieving obesity and regulating intestinal flora

Abstract

The invention discloses application of resveratrol and tomato pectin in preparation of a product for relieving obesity and regulating intestinal flora. The invention provides a composition. The composition is prepared from resveratrol and tomato pectin, the mass ratio of the resveratrol to the tomato pectin is (2-4): (3-5). The invention also discloses an application of the composition in preparation of products. Uses of the product: treatment of obesity; obesity is prevented; the intestinal flora is regulated. The invention has application and popularization values for solving the social problems of overweight and obesity.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology and relates to the application of resveratrol combined with tomato pectin in preparing products for alleviating obesity and regulating intestinal flora. Background Art

[0002] The global overweight and obesity rates are increasing rapidly and have become a serious problem threatening human health. The obesity epidemic has placed a heavy burden on the national economy due to its high medical costs. In developed countries such as Germany, the United States, and Italy, the government allocates 5% of the national economy to the treatment of obesity. Based on this, the country has put forward a number of plans and opinions to actively address the problem of obesity and overweight. The "Healthy China 2030" Outline clearly states that by 2030, the growth rate of the overweight and obese population needs to be significantly slowed. In 2024, the National Health Commission issued the "Dietary Guidelines for Adult Obesity (2024 Edition)" and the "Dietary Guidelines for Childhood and Adolescent Obesity (2024 Edition)" for the first time for obese people. The aim is to give full play to the advantages of modern nutrition and traditional Chinese and Western medicine in combining dietary nutrition, integrating food and medicine substances and new food ingredients into a reasonable diet, and emphasizing the importance of precise dietary intervention.

[0003] Current weight loss methods include: Exercise: Burning excess calories through exercise. This requires long-term training and can be difficult to maintain. Medication: Reduces appetite, increases satiety, or reduces fat absorption; these methods can have significant side effects, lead to dependency, and are costly. Surgery: Liposuction and gastrectomy are both expensive and carry potential complications. Therefore, dietary nutrition has become a first-line treatment for chronic diseases like obesity in countries like the United States and Canada.

[0004] Obesity is associated with changes in the composition of intestinal flora. Firmicutes and Bacteroidetes, which account for 98% of the human intestinal flora, are mainly involved in energy metabolism. The former helps the host absorb energy from the diet, and the latter participates in the metabolism of polysaccharides, steroids, etc. The ratio of Bacteroidetes to Firmicutes reflects how much the flora helps the body absorb calories, and is positively correlated with blood sugar concentration. A high ratio means that calories are easily absorbed and the body is prone to obesity. Studies have suggested that a long-term high-fiber diet can prevent obesity by promoting an increase in the proportion of intestinal flora associated with a lean body type, as well as increasing the richness and diversity of the intestinal flora. As research on the relationship between obesity and intestinal flora continues to deepen, regulating the structure of the intestinal flora through a nutritious diet has become a favorable means of improving obesity.

[0005] Tomatoes, a plant of the Solanaceae family originating in America, are the second most produced vegetable worldwide, with my country leading the world in tomato production. Approximately 80% of these tomatoes are processed into various tomato products, generating a significant amount of peel and pomace waste. Notably, tomato waste is rich in various functional components, such as tomato pectin, a natural polysaccharide. Summary of the Invention

[0006] The purpose of the present invention is to provide the use of resveratrol combined with tomato pectin in the preparation of products for alleviating obesity and regulating intestinal flora.

[0007] The invention provides a composition consisting of resveratrol and tomato pectin; the mass ratio of resveratrol to tomato pectin is 2-4:3-5.

[0008] Specifically, in the composition, the mass ratio of resveratrol to tomato pectin is 3:3-5.

[0009] Specifically, in the composition, the mass ratio of resveratrol to tomato pectin is 2-4:4.

[0010] Specifically, in the composition, the mass ratio of resveratrol to tomato pectin is 3:4.

[0011] The galacturonic acid content of any of the above tomato pectins is 50%-60%, and the esterification degree is 45%-50%.

[0012] The galacturonic acid content of any of the above tomato pectins is 52%-60%, and the degree of esterification is 46%-49%.

[0013] The galacturonic acid content of any of the above tomato pectins is 56.25%, and the degree of esterification is 47.40%.

[0014] The preparation method of any of the above-mentioned tomato pectin specifically comprises the following steps:

[0015] (1) Add pH 1.5 hydrochloric acid solution to tomato peel powder at a solid-liquid ratio of 1:20, mix well, and extract in an 85°C water bath for 1.5 h;

[0016] (2) After completing step (1), centrifuge at 10000 rpm for 10 min, collect the supernatant, and then filter with filter paper (the maximum particle size is 6 μm). 2 ) filter and collect the filtrate;

[0017] (3) taking the filtrate obtained in step (2), adding twice the volume of anhydrous ethanol, allowing to settle, filtering with a 400-mesh filter cloth and collecting the filter residue;

[0018] (4) The filter residue obtained in step (3) was washed with 70% ethanol solution, 90% ethanol solution and anhydrous ethanol in sequence, and then vacuum dried at 40° C. for 4 h to obtain a powdered product, which was further crushed and passed through a 40-mesh filter cloth to collect the powder, which was tomato pectin.

[0019] The material-liquid ratio of 1:20 means: add 20 ml of hydrochloric acid solution for every 1 g of tomato peel powder.

[0020] Preparation method of pH 1.5 hydrochloric acid solution: dilute commercially available 12 mol / L hydrochloric acid with water to 375 times the volume.

[0021] The preparation method of tomato peel powder is as follows: ripe tomato fruits are placed in boiling water for 1 minute, then taken out and the tomato skins are peeled, the tomato skins are freeze-dried, and then ground into powder to obtain the tomato peel powder.

[0022] The tomato fruit may specifically be the fruit of the tomato variety Tunhe 8501.

[0023] The uses of the composition are as follows (a) and / or (b) and / or (c):

[0024] (a) Treatment of obesity;

[0025] (b) prevention of obesity;

[0026] (c) Regulate intestinal flora.

[0027] The present invention also protects the use of any of the above compositions in preparing products;

[0028] The uses of the product are as follows (a) and / or (b) and / or (c):

[0029] (a) Treatment of obesity;

[0030] (b) prevention of obesity;

[0031] (c) Regulate intestinal flora.

[0032] The present invention also protects the use of resveratrol in preparing products;

[0033] The uses of the product are as follows (a) and / or (b) and / or (c):

[0034] (a) Treatment of obesity;

[0035] (b) prevention of obesity;

[0036] (c) Regulate intestinal flora.

[0037] The present invention also protects the use of tomato pectin in preparing products;

[0038] The uses of the product are as follows (a) and / or (b) and / or (c):

[0039] (a) Treatment of obesity;

[0040] (b) prevention of obesity;

[0041] (c) Regulate intestinal flora.

[0042] The tomato pectin is any of the above-mentioned tomato pectins.

[0043] The present invention also protects a product comprising any of the above compositions;

[0044] The uses of the product are as follows (a) and / or (b) and / or (c):

[0045] (a) Treatment of obesity;

[0046] (b) prevention of obesity;

[0047] (c) Regulate intestinal flora.

[0048] The present invention also protects a product comprising resveratrol;

[0049] The uses of the product are as follows (a) and / or (b) and / or (c):

[0050] (a) Treatment of obesity;

[0051] (b) prevention of obesity;

[0052] (c) Regulate intestinal flora.

[0053] The present invention also protects a product comprising tomato pectin;

[0054] The uses of the product are as follows (a) and / or (b) and / or (c):

[0055] (a) Treatment of obesity;

[0056] (b) prevention of obesity;

[0057] (c) Regulate intestinal flora.

[0058] The tomato pectin is any of the above-mentioned tomato pectins.

[0059] Any of the above treatments for obesity is embodied as follows (d1) and / or (d2) and / or (d3) and / or (d4) and / or (d5) and / or (d6):

[0060] (d1) inhibiting weight gain;

[0061] (d2) lowering blood lipids;

[0062] (d3) reduce fat content;

[0063] (d4) reduce fat cell volume;

[0064] (d5) inhibiting liver damage caused by obesity;

[0065] (d6) Inhibit intestinal flora imbalance caused by obesity.

[0066] Any of the above-mentioned blood lipid reduction is embodied as follows (e1) and / or (e2):

[0067] (e1) reduce serum total cholesterol levels;

[0068] (e2) Reduce serum total triglyceride levels.

[0069] Any of the above-mentioned inhibition of liver damage caused by obesity is manifested as the following (f1) and / or (f2) and / or (f3) and / or (f4) and / or (f5):

[0070] (f1) reduce serum alanine aminotransferase levels;

[0071] (f2) reduce serum aspartate aminotransferase levels;

[0072] (f3) reduce the total cholesterol content in the liver;

[0073] (f4) reduce the total triglyceride content in the liver;

[0074] (f5) Inhibits lipid droplet accumulation in the liver.

[0075] Exemplarily, any of the above-mentioned reducing fat content refers to reducing white fat content.

[0076] Exemplarily, any of the above-mentioned reductions in fat cell volume refers to reductions in fat cell volume in white fat.

[0077] Exemplarily, any of the above white fat is epididymal fat and / or inguinal fat and / or perirenal fat.

[0078] Regulating intestinal flora means: the abundance of Verrucomicrobiota is upregulated.

[0079] Regulating the intestinal flora refers to the downregulation of Lachnospiraceac abundance and Colidextribacter abundance.

[0080] Regulating the intestinal flora means: the abundance of unclassified_f_Lachnospiraceac is downregulated and the abundance of Colidextribacter is downregulated.

[0081] Regulating the intestinal flora means: downregulation of Lachnospiraceac abundance and downregulation of Lachnospiraceac abundance, downregulation of Colidextribacte abundance and downregulation of Alistipes abundance, and upregulation of Akkermansia abundance.

[0082] Regulating intestinal flora means: the abundance of norank_f_Lachnospiraceac is downregulated, the abundance of unclassified_f_Lachnospiraceac is downregulated, the abundance of Colidextribacte is downregulated, the abundance of Alistipes is downregulated, and the abundance of Akkermansia is upregulated.

[0083] Any of the above products is suitable for obese animals.

[0084] Any of the above-mentioned regulating intestinal flora refers to regulating the intestinal flora of obese animals.

[0085] Any of the above obesity is caused by a high-fat diet.

[0086] The animal is a mammal.

[0087] The mammal is a human or a mouse.

[0088] Any of the above products may be food, medicine or health care products.

[0089] The results of this study show that resveratrol combined with pectin effectively reduces obesity and hepatic lipid accumulation induced by a high-fat diet, including lowering body weight, blood lipids, and blood glucose levels, and improving liver damage. Furthermore, the combined intervention significantly improved the intestinal flora structure disorder caused by a high-fat diet, leading to a decrease in the abundance of norank_f_Lachnospiraceac, unclassified_f_Lachnospiraceac, Colidextribacte, and Alistipes, and an increase in the abundance of Akkermansia.

[0090] The present invention has application and promotion value in solving the social problems of overweight and obesity. BRIEF DESCRIPTION OF THE DRAWINGS

[0091] Figure 1 These are the results related to weight changes during the feeding process in step 3 of Example 2.

[0092] Figure 2 This is the relevant result of the fat tissue detection in step 2 of step 4 of Example 2.

[0093] Figure 3 The results of weighing the liver, taking the liver triglyceride content and the liver total cholesterol content in step 3 of step 4 of Example 2 are shown.

[0094] Figure 4 These are the results of HE staining and Oil Red O staining of the liver in step 3 of step 4 of Example 2.

[0095] Figure 5This is the result of the intestinal flora α diversity and β diversity analysis in step 4 of Example 2.

[0096] Figure 6 This is the result of the relative abundance of the top 10 communities at the intestinal microbial phylum level.

[0097] Figure 7 This is the result of the relative abundance of the top 10 communities at the intestinal microbial phylum level.

[0098] Figure 8 The results are the relative abundance of intestinal flora at the genus level.

[0099] Figure 9 The results are the relative abundance of intestinal flora at the genus level.

[0100] Figure 10 The results are the relative abundance of intestinal flora at the genus level.

[0101] Figure 11 The results are the relative abundance of intestinal flora at the genus level. DETAILED DESCRIPTION

[0102] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0103] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in literature in the field or in accordance with product specifications. Materials and reagents used in the following examples are commercially available unless otherwise specified. Quantitative experiments in the following examples were performed in triplicate, and the results were averaged. Statistical analysis in the examples was performed using SPSS 18.0, using ANOVA and Dunnett's test for multiple comparisons. Data are presented as mean ± standard error; a P < 0.05 indicates statistical significance. Images were processed using Graphpad 9.0. SPF: Specific pathogens free. Concentrated sulfuric acid refers to commercially available 95%-98% sulfuric acid. Resveratrol (CAS No. 501-36-0): Sigma-Aldrich. The tomato fruit used in the examples is from the tomato variety Tunhe 8501, registered under GPD Tomato (2018) 650307. Tomato skin refers to the exocarp of the tomato.

[0104] Example 1. Preparation of tomato pectin

[0105] 1. Preparation of tomato pectin

[0106] 1. Put the ripe tomato fruit into boiling water for 1 minute, then take out the tomato skin, peel it, freeze-dry it, and then grind it into powder to make tomato skin powder.

[0107] 2. Add pH 1.5 hydrochloric acid solution to the tomato peel powder with a solid-liquid ratio of 1:20. Mix well and extract in an 85°C water bath for 1.5 hours.

[0108] The material-liquid ratio of 1:20 means: add 20 ml of hydrochloric acid solution for every 1 g of tomato peel powder.

[0109] Preparation method of pH 1.5 hydrochloric acid solution: dilute commercially available 12 mol / L hydrochloric acid with water to 375 times the volume.

[0110] 3. After completing step 2, centrifuge at 10000 rpm for 10 min, collect the supernatant, and then filter with Whatman No. 3 filter paper (the maximum particle size is 6 μm). 2 ) was filtered and the filtrate was collected.

[0111] 4. Take the filtrate obtained in step 3, add twice the volume of anhydrous ethanol, let it settle, filter with a 400-mesh filter cloth and collect the residue.

[0112] 5. Take the filter residue obtained in step 4, wash it with 70% ethanol aqueous solution, 90% ethanol aqueous solution and anhydrous ethanol in sequence, then vacuum dry it at 40°C for 4 hours to obtain a powdered product, further grind it and pass it through a 40-mesh filter cloth to collect the powder, which is tomato pectin.

[0113] 2. Identification of tomato pectin

[0114] 1. Galacturonic acid content in tomato pectin

[0115] 0.15% carbazole reagent: Weigh 0.15 g of carbazole, dissolve it in chromatographic grade ethanol and dilute to 100 mL.

[0116] To prepare a standard curve: Take eight 25mm x 200mm test tubes, add 6mL of concentrated sulfuric acid to each, cool on ice, then add 1mL of galacturonic acid standard solution. Mix thoroughly, cool on ice, and then place in a boiling water bath for 10 minutes. Rapidly cool to room temperature, then add 1mL of 0.15% carbazole reagent. Shake well, let stand at room temperature for 30 minutes, and then measure the absorbance at 530nm. Galacturonic acid standard solution: Prepare a commercially available galacturonic acid standard solution with deionized water to a concentration of 1g / L. Then dilute with deionized water to obtain solutions with concentrations of 10mg / L, 20mg / L, 30mg / L, 40mg / L, 50mg / L, 60mg / L, and 70mg / L, respectively. Deionized water is used as the zero concentration standard solution. Draw a standard curve with galacturonic acid concentration and absorbance as variables.

[0117] Pectin suspension: Take the tomato pectin prepared in step 1 and add deionized water to obtain a pectin suspension with a concentration of 40 mg / L.

[0118] Galacturonic acid determination method (sulfate carbazole method): Add 3 mL of concentrated sulfuric acid to 0.5 mL of pectin suspension, mix thoroughly, and place in a boiling water bath for 10 minutes. Then, quickly cool to 25°C in an ice-water bath. Add 0.5 mL of 0.15% carbazole reagent, shake well, and let stand at 25°C for 30 minutes. Measure the absorbance at 530 nm. Substitute the absorbance value into the standard curve to obtain the galacturonic acid concentration in the pectin suspension (in mg / L).

[0119] Galacturonic acid content in tomato pectin (%) = (galacturonic acid concentration / 40) × 100.

[0120] 2. Degree of esterification in tomato pectin

[0121] Pectin suspension: Take the tomato pectin prepared in step 1 and add deionized water to obtain a pectin suspension with a concentration of 5 g / L.

[0122] Determination of esterification degree (titration method): Take 20 mL of pectin suspension and add 80 mL of decarbonated water, then add 3 drops of phenolphthalein, and then titrate with 0.05 mol / L sodium hydroxide aqueous solution until it turns pink (the volume of the sodium hydroxide aqueous solution used is V1 mL); then, add 20 mL of 0.5 mol / L sodium hydroxide aqueous solution to the solution, shake vigorously for 15 minutes, then add 0.5 mol / L hydrochloric acid aqueous solution to completely neutralize the 20 mL of sodium hydroxide solution, and shake until the pink color disappears; then titrate again with 0.05 mol / L sodium hydroxide aqueous solution until it turns pink (the volume of the sodium hydroxide aqueous solution used is V2 mL).

[0123]

[0124] After detection in step 1 and step 2, the galacturonic acid (GalA) content of the tomato pectin prepared in step 1 was 56.25±3.84%, and the degree of esterification was 47.40±0.79%.

[0125] Example 2: Animal Experiment

[0126] 1. Preparation of experimental animals and animal feed

[0127] Experimental animals: SPF-grade 5-week-old healthy C57BL / 6J male mice, purchased from Weitonglihua Biotechnology Co., Ltd.

[0128] Animal feed was commercially available from Beijing Huafukang Biotechnology Co., Ltd. The composition and energy percentage of low-fat and high-fat feeds are shown in Table 1. The material composition and energy content of low-fat and high-fat feeds are shown in Table 2. The low-fat feed was H10010 feed, a purified feed with a 10% energy content. The high-fat feed was H10060 feed, a high-fat purified feed with a 60% functional content.

[0129] Table 1 Composition and energy ratio of low-fat feed and high-fat feed

[0130]

[0131] Table 2 Material composition and energy of low-fat feed and high-fat feed

[0132]

[0133] 2. Group feeding of experimental animals

[0134] Forty SPF-grade 5-week-old healthy C57BL / 6J male mice were adaptively fed for one week and then randomly divided into 5 groups, with 8 mice in each group (4 mice per cage).

[0135] The group feeding method is as follows:

[0136] Blank control group (NCD): free access to low-fat diet and oral administration of normal saline, single oral administration volume of 10 μL / g body weight;

[0137] High-fat diet group (HFD) was fed with high-fat diet ad libitum and gavaged with normal saline, with a single gavage volume of 10 μL / g body weight;

[0138] The resveratrol-high-fat diet group (HFDR) was fed a high-fat diet ad libitum and gavaged with a resveratrol solution (resveratrol solution: resveratrol dissolved in saline to a concentration of 30 mg / mL, prepared freshly). A single gavage volume of 10 μL / g body weight was used. This group was also referred to as the resveratrol intervention group.

[0139] The tomato pectin-high-fat diet group (HFDT) received a high-fat diet ad libitum and was gavaged with a tomato pectin suspension (tomato pectin suspension prepared in step 1 of Example 1, added to physiological saline to a concentration of 40 mg / mL, prepared immediately before use). The single gavage volume was 10 μL / g body weight. This HFDT group is also referred to as the tomato pectin intervention group.

[0140] The resveratrol-tomato pectin-high-fat diet group (HFDRT) received a high-fat diet ad libitum and was gavaged with a resveratrol-tomato pectin mixture (resveratrol-tomato pectin mixture: resveratrol and tomato pectin prepared in step 1 of Example 1, added to physiological saline, and mixed to a resveratrol concentration of 30 mg / mL and tomato pectin content of 40 mg / mL, prepared immediately before use). The single gavage volume was 10 μL / g body weight. The resveratrol-tomato pectin-high-fat diet group is also referred to as the combined intervention group.

[0141] The total feeding time of the groups was 15 weeks, with oral gavage once a day (8:30-9:30 am every day). During this period, the mice had free access to feed and water.

[0142] 3. Weight changes during feeding

[0143] In step 2, body weight was measured at group 0 time, the 7th day of group feeding (i.e., the 1st week), the 14th day of group feeding (i.e., the 2nd week), the 21st day of group feeding (i.e., the 3rd week), the 28th day of group feeding (i.e., the 4th week), the 35th day of group feeding (i.e., the 5th week), the 42nd day of group feeding (i.e., the 6th week), the 49th day of group feeding (i.e., the 7th week), the 56th day of group feeding (i.e., the 8th week), the 63rd day of group feeding (i.e., the 9th week), the 70th day of group feeding (i.e., the 10th week), the 77th day of group feeding (i.e., the 11th week), the 84th day of group feeding (i.e., the 12th week), the 91st day of group feeding (i.e., the 13th week), the 98th day of group feeding (i.e., the 14th week), and the 105th day of group feeding (i.e., the 15th week). The average value of each group was taken, the feed quality was measured, and the feed consumption (g) of that week was calculated.

[0144] Weight statistics are shown in Figure 1 A. At time 0, there was no significant difference in body weight between the groups. Starting from week 4, the body weight of the HFD group was significantly higher than that of the NCD group. By week 6, the body weight of the HFD group was 1.2 times that of the NCD group, reaching the obesity standard, indicating the successful establishment of a high-fat obesity model.

[0145] At week 15, the weight gain of mice in each group relative to time 0 was calculated. Weight gain = (weight at week 15 - weight at time 0) ÷ weight at time 0 × 100%. Figure 1 B. The HFD group had a 108% increase in weight gain, 1.45 times that of the NCD group. Tomato pectin alone did not significantly reduce the weight of obese mice, but both resveratrol alone (HFDR) and the combined resveratrol and tomato pectin (HFDRT) significantly reduced the weight of obese mice. The HFDR group had a 79.03% increase in weight gain, while the combined intervention group had a 65.82% increase.

[0146] At week 15, the daily energy intake of each mouse in each group was calculated. Energy intake = (the amount of feed consumed by the mice in the group in that week × the energy of the feed in grams) ÷ (7 × 8). Figure 1 There were no significant differences among the HFD, HFDR, HFDT, and HFDRT groups, indicating that the weight loss effects of resveratrol intervention alone and resveratrol-tomato pectin combined intervention were not caused by affecting the appetite and food intake of mice.

[0147] 4. Testing of various indicators

[0148] After completing step 2, collect mouse feces. After completing step 2, fast for 12 hours, remove the eyeballs to collect blood, separate the serum, and then dissect the mice. After dissection, collect epididymal fat, perirenal fat, inguinal fat, scapular fat, liver, and ileum. Some tissues were immersed in 10% formalin and stained with H&E. Some tissues were frozen at -80°C for biochemical analysis.

[0149] 1. Blood biochemistry test

[0150] Long-term high-fat diets can lead to elevated blood glucose and blood lipids. A fully automated blood biochemistry analyzer (Hitachi, Japan) was used to measure serum levels of total cholesterol (TC), total triglycerides (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). The results are shown in Table 3. Compared with the NCD group, the serum TC and TG levels of mice in the HFD group were significantly increased. Compared with the HFD group, the serum TC and TG levels of mice in the HFDR, HFDT, and HFDRT groups were significantly decreased, with the HFDRT group showing the greatest decrease. Compared with the NCD group, the serum ALT and AST levels of mice in the HFD group were significantly increased. Compared with the HFD group, the serum ALT and AST levels of mice in the HFDR, HFDT, and HFDRT groups were significantly decreased, with the HFDRT group showing the greatest decrease.

[0151] Table 3

[0152]

[0153] 2. Fat tissue detection

[0154] Obesity causes changes in the weight, morphology, and function of adipose tissue. Epididymal fat, perirenal fat, and inguinal fat are all white fat. Scapular fat is brown fat.

[0155] Epididymal fat (Epi-WAT), inguinal fat (Ing-SAT) and perirenal fat (Per-WAT) were weighed separately. Figure 2 A. Compared with the NCD group, the white fat weight of mice in the HFD group was significantly increased. Compared with the HFD group, the white fat weight of mice in the HFDRT group was significantly decreased.

[0156] Scapular fat and epididymal fat were collected, paraffin sections were made and HE staining was performed. Figure 2 B. The diameter of adipocytes in epididymal fat was counted using Image J software. The results are shown in Figure 2 Adipocytes in the NCD group were smaller and more structurally intact. Compared with the NCD group, adipocytes in the HFD group were significantly larger and irregularly arranged. Resveratrol alone (HFDR), tomato pectin alone (HFDT), and a combination of resveratrol and tomato pectin (HFDRT) all significantly reduced adipocyte size, with the combined treatment having the greatest effect.

[0157] 3. Detect the degree of liver damage

[0158] A high-fat diet can cause lipid accumulation in the liver, further leading to liver damage. Alanine aminotransferase (ALT) and lactate dehydrogenase (AST) are primarily found in liver cells. When liver tissue is damaged and liver cells die, they are released into the blood. Therefore, they are two important enzymes that reflect liver damage.

[0159] The liver was weighed and the results were shown in Figure 3 A. Take the liver and test the triglyceride content and total cholesterol content. The results are shown in Figure 3 B and Figure 3C. Triglyceride (TG) assay kit: Nanjing Jiancheng Bioengineering Institute, catalog number A110-1-1, operated according to the instructions. Total cholesterol (TCH / T-CHO) assay kit: Nanjing Jiancheng Bioengineering Institute, catalog number A111-1-1, operated according to the instructions. Compared with the NCD group, the liver weight of mice in the HFD group was significantly increased. Compared with the HFD group, resveratrol intervention alone (HFDR), tomato pectin intervention alone (HFDT), and resveratrol and tomato pectin combined intervention (HFDRT) all significantly reduced liver weight. Compared with the NCD group, the TC and TG levels in the liver of mice in the HFD group were significantly increased. Compared with the HFD group, resveratrol intervention alone (HFDR), tomato pectin intervention alone (HFDT), and resveratrol and tomato pectin combined intervention (HFDRT) all significantly reduced TC and TG levels in the liver, with the combined intervention group showing the best effect. The results showed that resveratrol intervention alone (HFDR), tomato pectin intervention alone (HFDT) and resveratrol and tomato pectin combined intervention (HFDRT) all had protective effects on the liver, and the combined intervention had the best improvement effect.

[0160] Liver tissue was taken and paraffin sections were made, and HE staining and Oil Red O staining were performed respectively. Figure 4 . HE staining results showed that compared with the NCD group, the HFD group mice had significant and widely distributed lipid droplet accumulation in the liver; compared with the HFD group, resveratrol intervention alone (HFDR), tomato pectin intervention alone (HFDT) and resveratrol and tomato pectin combined intervention (HFDRT) could significantly reduce the lipid droplet accumulation in the liver. Oil red O staining results showed that compared with the NCD group, the HFD group mice had obvious red lipid droplet staining; compared with the HFD group, resveratrol intervention alone (HFDR), tomato pectin intervention alone (HFDT) and resveratrol and tomato pectin combined intervention (HFDRT) could significantly reduce the lipid droplet accumulation in the liver. The results showed that the combined intervention had the best effect, with almost no lipid droplet accumulation, which was close to the section results of the NCD group.

[0161] 4. Detect intestinal flora

[0162] The test sample is mouse feces.

[0163] (1) Analysis of α-diversity and β-diversity of intestinal flora

[0164] The intestinal flora is the main factor affecting host metabolism, so 16S rRNA sequencing was further performed to analyze the intestinal flora structure. By performing α-diversity typing on the sample microorganisms, the richness and diversity of the microbial community can be understood. The Shannon index and Simpson index reflect the diversity of the microbial community. The larger the Shannon value, the higher the community diversity, while the larger the Simpson index, the lower the community diversity. The results of the Shannon index are shown in Figure 5 The results of Simpson index are shown in Figure 5 B. Compared with the NCD group, the Shannon index in the HFD group was significantly decreased, and the Simpson index was significantly increased. Compared with the HFD group, all three interventions significantly increased the Shannon index and decreased the Simpson index, improving the richness of the intestinal microbial community in mice, with the combined intervention having the best effect.

[0165] Further PCA analysis of the intestinal flora structure was performed, and the results are shown in Figure 5 The five groups of samples were almost completely separated, indicating that the microbial structures of the samples in each group were significantly different. Among them, the samples of the pectin intervention group (HFDT) were closer to those of the HFD group, indicating that the intestinal flora structure was more similar, while the HFDR and HFDRT groups were farther away from the HFD group, indicating that resveratrol and combined intervention can significantly improve the intestinal flora structure of high-fat obese mice.

[0166] (2) Relative abundance of mouse intestinal flora at the phylum level

[0167] The relative abundance of the top 10 communities at the intestinal microbial phylum level is as follows Figure 6 and Figure 7 As shown, the dominant phyla were Firmicutes, Bacteroidetes, and Verrucomicrobiota, accounting for over 90% of the bacterial community. Significant changes occurred in the bacterial community structure across the different treatment groups. Compared with the NCD group, the relative abundance of Firmicutes in the HFD group was significantly increased, while the relative abundance of Verrucomicrobiota was significantly decreased. The relative abundance of Firmicutes in the pectin-only intervention group (HFDT) was significantly decreased, while the relative abundance of Bacteroidetes was significantly higher than in the other groups. Compared with the HFD group, the relative abundance of Verrucomicrobiota was significantly increased in the combined intervention group (HFDRT).

[0168] (3) Relative abundance of genus levels in mouse intestinal flora

[0169] The relative abundance of the top 10 communities at the genus level of intestinal flora is as follows Figure 8 、 Figure 9 、 Figure 10 and Figure 11 As shown in the figure. The dominant genera were Blautia, Akkermansia, Bacteroides, norank_f_Muribaculaceae, unclassified_f_Lachnospiraceac, and Lactobacillus. Significant changes occurred in the microbial community structure across the different treatment groups. Compared with the NCD group, the relative abundance of unclassified_f_Lachnospiraceac, Lactobacillus, and Colidextribacte was significantly increased in the HFD group, while the relative abundance of probiotics (such as Akkermansia) was significantly decreased. Compared with the HFD group, resveratrol alone (HFDR) significantly decreased the abundance of norank_f_Muribaculaceae, unclassified_f_Lachnospiraceac, Colidextribacte, and Alistipes, and increased the abundance of Akkermansia. Compared with the HFD group, pectin alone (HFDT) significantly decreased the relative abundance of unclassified_f_Lachnospiraceac and Colidextribacter. Compared with the HFD group, the combined intervention (HFDRT) significantly downregulated the relative abundance of norank_f_Lachnospiraceac, unclassified_f_Lachnospiraceac, Colidextribacte and Alistipes and upregulated the relative abundance of Akkermansia. In addition, the combined intervention had better effects on inhibiting harmful bacteria and promoting beneficial bacteria than either single intervention, indicating that the combined intervention can significantly regulate the disorder of intestinal flora structure caused by obesity.

[0170] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be practiced over a wide range of parameters, concentrations, and conditions without departing from the spirit and scope of the present invention and without unnecessary experimentation. Although specific embodiments have been given herein, it should be understood that further modifications may be made to the present invention. In summary, this application is intended to encompass any variations, uses, or improvements to the present invention, including those made by conventional techniques known in the art that depart from the scope of the present invention. Applications of the essential features may be made within the scope of the following claims.

Claims

1. A composition comprising resveratrol and tomato pectin; the mass ratio of resveratrol to tomato pectin is 2-4:3-5.

2. Use of the composition according to claim 1 in preparing a product; The uses of the product are as follows (a) and / or (b) and / or (c): (a) Treatment of obesity; (b) prevention of obesity; (c) Regulate intestinal flora.

3. Application of resveratrol in product preparation; The uses of the product are as follows (a) and / or (b) and / or (c): (a) Treatment of obesity; (b) prevention of obesity; (c) Regulate intestinal flora.

4. Application of tomato pectin in product preparation; The uses of the product are as follows (a) and / or (b) and / or (c): (a) Treatment of obesity; (b) prevention of obesity; (c) Regulate intestinal flora.

5. A product comprising the composition of claim 1; The uses of the product are as follows (a) and / or (b) and / or (c): (a) Treatment of obesity; (b) prevention of obesity; (c) Regulate intestinal flora.

6. A product comprising resveratrol; The uses of the product are as follows (a) and / or (b) and / or (c): (a) Treatment of obesity; (b) prevention of obesity; (c) Regulate intestinal flora.

7. Products comprising tomato pectin; The uses of the product are as follows (a) and / or (b) and / or (c): (a) Treatment of obesity; (b) prevention of obesity; (c) Regulate intestinal flora.

8. The use according to any one of claims 2 to 4 or the product according to any one of claims 5 to 7, characterized in that: The treatment of obesity is embodied as follows (d1) and / or (d2) and / or (d3) and / or (d4) and / or (d5) and / or (d6): (d1) inhibiting weight gain; (d2) lowering blood lipids; (d3) reduce fat content; (d4) reduce fat cell volume; (d5) inhibiting liver damage caused by obesity; (d6) Inhibit intestinal flora imbalance caused by obesity.

9. The use or product according to claim 8, characterized in that: The blood lipid reduction is embodied as follows (e1) and / or (e2): (e1) reduce serum total cholesterol levels; (e2) Reduce serum total triglyceride levels.

10. The use or product according to claim 8, wherein: The inhibition of liver damage caused by obesity is embodied as follows (f1) and / or (f2) and / or (f3) and / or (f4) and / or (f5): (f1) reduce serum alanine aminotransferase levels; (f2) reduce serum aspartate aminotransferase levels; (f3) reduce the total cholesterol content in the liver; (f4) reduce the total triglyceride content in the liver; (f5) Inhibits lipid droplet accumulation in the liver.

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