Application of green tea polyphenol in preparation of product for improving intestinal health

Abstract

The invention provides application of green tea polyphenol in preparation of a product for improving intestinal health, and belongs to the technical field of medicines. The invention provides application of green tea polyphenol in preparation of a product for improving intestinal health. The green tea polyphenols comprise epigallocatechin, epigallocatechin gallate, epicatechin and epicatechin gallate, and the green tea polyphenols are added into the green tea extract and the green tea extract. The green tea polyphenol can increase the length of the small intestine, increase the villus length of the small intestine, improve the intestinal barrier and increase the content of fructose transporter protein, so that the intestinal health can be remarkably improved. The green tea polyphenol provided by the invention can improve symptoms such as metabolic syndrome, intestinal injury diseases, endotoxemia and colitis while improving intestinal health, and can be applied to prevention and / or treatment of the metabolic syndrome, the intestinal injury diseases, the endotoxemia and the colitis.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and particularly relates to the application of green tea polyphenols in the preparation of products for improving intestinal health. Background Art

[0002] Obesity is a global epidemic and an important risk factor for metabolic disorders such as type 2 diabetes, insulin resistance, cardiovascular disease and various cancers. Excessive energy intake, sedentary lifestyle, and genetic and pathological factors are important factors leading to obesity-related metabolic disorders. Currently, drugs for the treatment of obesity include intestinal lipase inhibitors, such as orlistat, but the drug is prone to cause gastrointestinal adverse reactions such as flatulence and diarrhea, and its therapeutic effect is limited.

[0003] It is of great significance to provide a product that can improve gastrointestinal health and at the same time improve metabolic syndrome such as obesity. Summary of the invention

[0004] In view of the defects in the prior art, the purpose of the present invention is to provide a green tea polyphenol for use in the preparation of a product for improving intestinal health, wherein the green tea polyphenol can improve intestinal health and metabolic syndrome at the same time.

[0005] The objective of the present invention is achieved through the following technical solutions:

[0006] The invention provides application of green tea polyphenols in preparing products for improving intestinal health; the green tea polyphenols include epigallocatechin, epigallocatechin gallate, epicatechin and epicatechin gallate.

[0007] Preferably, a product is prepared to improve any one or two or more of metabolic syndrome, intestinal injury disease, endotoxemia and colitis.

[0008] Preferably, the metabolic syndrome includes any one or two or more of the following (1) to (5):

[0009] (1) Nonalcoholic fatty liver disease;

[0010] (2) Hyperlipidemia;

[0011] (3) Diabetes;

[0012] (4) Atherosclerosis;

[0013] (5) Obesity.

[0014] Preferably, the diabetes includes type 2 diabetes; the hyperlipidemia includes: hypercholesterolemia and / or hypertriglyceridemia.

[0015] Preferably, the improving intestinal health includes the following (I) to (IV):

[0016] (I) Increase the length of the small intestine;

[0017] (II) Increase the length of small intestinal villi;

[0018] (III) Improve intestinal barrier;

[0019] (IV) Increase the content of fructose transporter protein.

[0020] Preferably, the preparation method of green tea polyphenols comprises:

[0021] Mixing tea leaves with water and performing extraction twice to obtain tea extract;

[0022] After removing caffeine from the tea extract, ethyl acetate is used for extraction to obtain green tea polyphenols.

[0023] Preferably, the temperature of each leaching is 85-95° C. and the time of each leaching is 20-40 min.

[0024] Preferably, the method for removing caffeine from tea extract comprises: removing caffeine from tea extract using chloroform.

[0025] Preferably, the product includes any one or more of medicines, foods and health products.

[0026] Preferably, the product further comprises auxiliary materials.

[0027] Beneficial effects of the present invention

[0028] The present invention provides the use of green tea polyphenols in the preparation of products for improving intestinal health; the green tea polyphenols include epigallocatechin, epigallocatechin gallate, epicatechin and epicatechin gallate. The green tea polyphenols of the present invention can increase the length of the small intestine, increase the length of the small intestinal villi, improve the intestinal barrier and increase the content of fructose transporter, thereby significantly improving intestinal health. The present invention shows through the results of the examples that the green tea polyphenols can significantly reduce body weight while improving intestinal health; reduce fasting blood sugar, fasting triglycerides and total cholesterol content; reduce the weight of liver, epididymal fat and perirenal fat; reduce the content of inflammatory factor TNFα in the blood; reduce the protein expression of fatty acid synthase acetyl-CoA carboxylase, fatty acid synthase, and sterol regulatory element binding protein 1c in the liver, thereby reducing the synthesis of fatty acids, etc., thereby being beneficial to the prevention and / or treatment of metabolic syndrome, intestinal damage diseases, endotoxemia and colitis, etc. In summary, the green tea polyphenols provided by the present invention can improve intestinal health while also improving symptoms such as metabolic syndrome, intestinal damage diseases, endotoxemia and colitis, and can be used to prevent and / or treat metabolic syndrome, intestinal damage diseases, endotoxemia and colitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0030] Figure 1 This is a composition diagram of green tea polyphenols prepared in Example 1;

[0031] Figure 2 This is a graph showing the changes in body weight of mice in each experimental group in Example 2;

[0032] Figure 3 This is a graph showing the fasting blood glucose levels of mice in each experimental group in Example 2;

[0033] Figure 4 This is a diagram of fasting triglyceride levels in mice in each experimental group in Example 2;

[0034] Figure 5 This is a graph showing the total cholesterol status of mice in each experimental group in Example 2;

[0035] Figure 6 This is a graph showing the organ weights of mice in each experimental group in Example 2;

[0036] Figure 7 This is a graph showing the TNFα status of mice in each experimental group in Example 2;

[0037] Figure 8 This is a diagram of the LPS status of mice in each experimental group in Example 2;

[0038] Fig. 9 This is a picture of the small intestine villi of mice in each experimental group in Example 2;

[0039] Fig.10 This is a diagram showing the length of the small intestine of mice in each experimental group in Example 2;

[0040] Fig.11 This is a diagram showing the length of the small intestinal villi in the duodenum of mice in each experimental group in Example 2;

[0041] Fig.12 This is a diagram showing the gene expression of target protein enolase (ENO1) in mice in each experimental group of Example 2;

[0042] Fig.13 This is a diagram showing the gene expression of fructose transporter GLUT5 in mice in each experimental group of Example 2;

[0043] Fig.14 This is a diagram showing the HIF-1α gene expression in mice in each experimental group in Example 2;

[0044] Fig.15 This is a diagram showing the Jam gene expression of mice in each experimental group in Example 2;

[0045] Fig.16 This is a diagram showing the occludin gene expression of mice in each experimental group in Example 2;

[0046] Fig.17 This is a diagram showing the expression of ZO-1 gene in mice in each experimental group in Example 2;

[0047] Fig.18 This is a graph showing the results of detecting the ACC protein expression in mice in each experimental group in Example 2;

[0048] Fig.19 This is a graph showing the results of detecting the expression of ACL protein in mice in each experimental group in Example 2;

[0049] Fig. 20 This is a graph showing the results of detecting the FAS protein expression in each experimental group of mice in Example 2;

[0050] Fig.21 This is a graph showing the results of detecting the Srebp1c protein expression level in mice in each experimental group in Example 2. DETAILED DESCRIPTION

[0051] The invention provides application of green tea polyphenols in preparing products for improving intestinal health; the green tea polyphenols include epigallocatechin, epigallocatechin gallate, epicatechin and epicatechin gallate.

[0052] In the present invention, the product includes any one or more of medicine, food and health care products; the product also includes auxiliary materials. The present invention has no special requirements for the auxiliary materials, and any conventional auxiliary materials in the art can be used.

[0053] As an optional embodiment of the present invention, the green tea polyphenols contain 15g / kg epigallocatechin (EGC), 613g / kg epigallocatechin gallate (EGCG), 40g / kg epicatechin (EC) and 202g / kg epicatechin gallate (ECG).

[0054] The present invention has no special limitation on the preparation method of the green tea polyphenols, and any conventional preparation method in the art can be used. As an optional embodiment of the present invention, the preparation method of the green tea polyphenols comprises the following steps: mixing tea leaves with water and performing two extractions to obtain a tea extract; after removing caffeine from the tea extract, extracting with ethyl acetate to obtain green tea polyphenols.

[0055] The present invention does not specifically limit the source of the tea leaves, and any conventional commercially available product in the art may be used. As an optional embodiment of the present invention, the tea leaves may be green tea; and the green tea may be West Lake Longjing. As an optional embodiment of the present invention, after obtaining the tea leaves, the present invention grinds the tea leaves into powder. The present invention does not specifically limit the grinding method, and any conventional grinding method in the art may be used. After obtaining the tea powder, the present invention mixes the tea leaves with water for two extractions. As an optional embodiment of the present invention, the tea leaves may be mixed with boiling water; the solid-liquid ratio of the tea leaves to water may be 1g:100mL; after obtaining the tea-water mixture, the present invention performs the first extraction. As an optional embodiment of the present invention, the temperature of the first leaching can be 85-95°C, or 85, 86, 87, 88, 89, 90, 91, 92, 93, 94 or 95°C; the time of the first leaching can be 20-40 minutes, or 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 minutes. After the first leaching is completed, the present invention performs solid-liquid separation on the leaching solution to obtain the first leaching solution and the leaching residue. After obtaining the leaching residue, the present invention can mix the leaching residue with water for a second leaching; or the leaching residue can be mixed with boiled water for a second leaching. The amount of water used in the second leaching of the present invention is preferably the same as the amount of water used for the first leaching. After obtaining the residue-water mixture, the present invention performs a second leaching; the temperature of the second leaching can be 90° C.; the time of the second leaching can be 30 minutes. After the second leaching is completed, the present invention obtains a second leaching solution. After obtaining the second leaching solution, the present invention preferably combines the first leaching solution and the second leaching solution to obtain a tea leaching solution.

[0056] As an optional embodiment of the present invention, after obtaining the tea extract, the present invention preferably rotary evaporates the tea extract to obtain a concentrated tea extract. The present invention does not specifically limit the method of rotary evaporation, and any conventional rotary evaporation method in the art can be used. As an optional embodiment of the present invention, the temperature of the rotary evaporation can be 60°C, the speed of the rotary evaporation can be 150rpm, the vacuum degree of the rotary evaporation can be 10mbar, and the time of the rotary evaporation can be 2h. The present invention removes most of the water in the tea extract by the rotary evaporation. After the rotary evaporation is completed, the present invention obtains a concentrated tea extract. As an optional embodiment of the present invention, the volume of the obtained concentrated tea extract can be 500mL.

[0057] After obtaining the concentrated tea extract, the present invention preferably removes caffeine from the concentrated tea extract and then uses ethyl acetate for extraction to obtain green tea polyphenols. The present invention does not specifically limit the method for removing caffeine, and any conventional removal method in the art can be used. As an optional embodiment of the present invention, the method for removing caffeine includes using chloroform to remove caffeine from the tea extract. The present invention uses chloroform to remove caffeine from the tea extract, including: mixing the concentrated tea extract with an equal volume of chloroform and then letting it stand. The standing of the present invention can be carried out at room temperature; the standing temperature can be 4 to 20°C, or 4, 8, 10, 12, 14, 16, 18 or 20°C; the standing time can be 30 minutes. After the standing is completed, the present invention obtains the discarded chloroform part to obtain a tea polyphenol water extract.

[0058] After obtaining the tea polyphenol water extract, the present invention extracts the tea polyphenol water extract with ethyl acetate to obtain green tea polyphenols. The present invention preferably extracts the tea polyphenol water extract after mixing it with an equal volume of ethyl acetate. The present invention has no special restrictions on the extraction method, and any conventional extraction method in the art can be used. As an optional embodiment of the present invention, the extraction can be carried out under normal temperature and pressure conditions; the extraction temperature can be 4 to 20°C, or 4, 8, 10, 12, 14, 16, 18 or 20°C; the extraction time can be 10 minutes. After the extraction is completed, the present invention obtains an extract. As an optional embodiment of the present invention, after obtaining the extract, the present invention preferably further includes rotary evaporating the extract to obtain green tea polyphenols. The present invention has no special restrictions on the rotary evaporation method, and any conventional evaporation method in the art can be used. As an optional embodiment of the present invention, the rotary evaporation temperature can be 40°C, the rotary evaporation speed can be 150rpm; the rotary evaporation vacuum can be 10mbar. After the rotary evaporation is completed, the present invention obtains green tea polyphenols.

[0059] The green tea polyphenols prepared by the preparation method of the technical scheme of the present invention contain 15g / kg epigallocatechin (EGC), 613g / kg epigallocatechin gallate (EGCG), 40g / kg epicatechin (EC), and 202g / kg epicatechin gallate (ECG).

[0060] In the present invention, the green tea polyphenols can improve intestinal health. In the present invention, the improvement of intestinal health includes any one or two or more of the following (I) to (IV):

[0061] (I) Increase the length of the small intestine;

[0062] (II) Increase the length of small intestinal villi;

[0063] (III) Improve intestinal barrier;

[0064] (IV) Increase the content of fructose transporter protein.

[0065] In the present invention, the green tea polyphenols can increase the length of the small intestine. The present invention shows through the results of the examples that the green tea polyphenols provided by the present invention can significantly increase the length of the small intestine compared with the group without intragastric administration of green tea polyphenols.

[0066] In the present invention, the green tea polyphenols can increase the length of the small intestinal villi. In the present invention, the green tea polyphenols can significantly upregulate the expression of the fructose transporter GLUT5, the target protein ENO1 of the hypoxia-inducible factor, and the HIF-1α gene, thereby increasing the length of the small intestinal villi. The present invention shows through the results of the examples that the green tea polyphenols provided by the present invention can significantly increase the length of the small intestinal villi relative to the group not gavaged with green tea polyphenols.

[0067] In the present invention, the green tea polyphenols can improve the intestinal barrier. In the present invention, the green tea polyphenols can significantly increase the expression of tight junction proteins, thereby protecting the intestinal mucosal barrier and achieving the effect of improving the intestinal barrier. The present invention shows through the results of the examples that the green tea polyphenols can significantly upregulate the expression of tight junction proteins Jam, ZO-1 and occludin.

[0068] Furthermore, the green tea polyphenols of the present invention also have the effect of inhibiting lipid synthesis. The present invention shows through the results of the examples that the green tea polyphenols of the present invention can reduce the protein expression of fatty acid synthase acetyl-CoA carboxylase, fatty acid synthase, and sterol regulatory element binding protein 1c in the liver, thereby reducing the synthesis of fatty acids.

[0069] In the present invention, the green tea polyphenols can be used to prepare products for improving any one or two or more of metabolic syndrome, intestinal damage diseases, endotoxemia and colitis.

[0070] In the present invention, the green tea polyphenols can be used to prepare products for improving metabolic syndrome. In the present invention, the metabolic syndrome includes any one or more of the following (1) to (5):

[0071] (1) Nonalcoholic fatty liver disease;

[0072] (2) Hyperlipidemia;

[0073] (3) Diabetes;

[0074] (4) Atherosclerosis;

[0075] (5) Obesity.

[0076] In the present invention, the green tea polyphenols can be used to prepare products for improving non-alcoholic fatty liver disease. In the present invention, the green tea polyphenols can be used to prepare products for preventing and / or treating non-alcoholic fatty liver disease. The present invention shows through the results of the examples that the green tea polyphenols can significantly reduce the content of the inflammatory factor TNF-α, increase the expression of the fructose transporter GLUT5, and reduce the protein expression of the fatty acid synthase acetyl-CoA carboxylase, fatty acid synthase, and sterol regulatory element binding protein 1c, thereby reducing fatty acids, which is beneficial to improve and / or alleviate non-alcoholic fatty liver disease.

[0077] In the present invention, the green tea polyphenols can be used to prepare products for improving hyperlipidemia. In the present invention, the green tea polyphenols can be used to prepare products for preventing and / or treating hyperlipidemia. In the present invention, the hyperlipidemia includes hypertriglyceridemia and / or hypercholesterolemia. The present invention shows through the results of examples that the green tea polyphenols can significantly reduce the increase in total cholesterol caused by a high-fat diet, and maintain its content at a normal level. The present invention shows through the results of examples that the green tea polyphenols can significantly reduce the increase in fasting triglycerides caused by a high-fat diet, and maintain its content at a normal level.

[0078] In the present invention, the green tea polyphenols can be used to prepare products for improving diabetes. In the present invention, the green tea polyphenols can be used to prepare products for preventing and / or treating diabetes. In the present invention, the diabetes includes type 2 diabetes. The present invention shows through the results of the examples that the green tea polyphenols can significantly reduce the increase in fasting blood sugar caused by a high-fat diet and maintain it at a normal level.

[0079] In the present invention, the green tea polyphenols can be used to prepare products for improving atherosclerosis. In the present invention, the green tea polyphenols can be used to prepare products for preventing and / or treating atherosclerosis. The present invention shows through the results of the examples that the green tea polyphenols can regulate lipid metabolism, inhibit lipid synthesis, reduce the content of triglycerides and cholesterol in plasma, and thus help improve atherosclerosis.

[0080] In the present invention, the green tea polyphenols can be used to prepare products for improving obesity. The present invention shows through the results of examples that the green tea polyphenols can regulate lipid metabolism, inhibit lipid synthesis, and thus be beneficial for anti-obesity. The present invention shows through the results of examples that the green tea polyphenols can significantly reduce the increase in body weight caused by a high-fat diet and significantly reduce the body weight of mice.

[0081] In the present invention, the green tea polyphenols can be used to prepare products for improving endotoxemia. The present invention shows through the results of the examples that the green tea polyphenols can be used to reduce LPS, thereby helping to improve endotoxemia.

[0082] In the present invention, the green tea polyphenols can be used to prepare products for improving colitis. The present invention shows through the results of the examples that the green tea polyphenols can be used to improve intestinal health, increase the length of the small intestine, increase the length of the small intestinal villi, improve the intestinal barrier, reduce the content of the inflammatory factor TNF-α, increase the target protein enolase (ENO1) gene expression and increase the HIF-1α gene expression, thereby helping to improve colitis.

[0083] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0084] Example 1

[0085] Extract green tea polyphenols, the specific method is as follows:

[0086] Tea leaves (West Lake Longjing tea leaves purchased from Ying Kee Tea House in Hong Kong) were ground into powder, and boiled distilled water was added at a material-liquid ratio of 1 g:100 mL to perform two extractions to obtain tea extract.

[0087] Specifically, 20 g of tea powder is added to 2 L of boiling distilled water, maintained at 90° C., and soaked for 30 minutes to complete the first extraction, and solid-liquid separation is performed to obtain the first extract and the remaining residue of the extraction; the remaining residue of the extraction is added to 2 L of boiling distilled water, maintained at 90° C., and soaked for 30 minutes to complete the second extraction, and solid-liquid separation is performed to obtain the second extract; the first extract and the second extract are combined to obtain the tea extract.

[0088] After obtaining the tea extract, it was subjected to rotary evaporation to remove part of the aqueous solution. The conditions used for the rotary evaporation were: vacuum degree 10 mbar, 60° C., rotation speed 150 rpm, and about 2 h until most of the water was removed to obtain about 500 mL of green tea aqueous extract.

[0089] The green tea water extract was fully mixed with an equal volume of chloroform and allowed to stand at room temperature for 30 minutes to remove most of the caffeine, and the chloroform portion was discarded to obtain the tea polyphenols water extract.

[0090] An equal volume of ethyl acetate is added to the tea polyphenol water extract for extraction. The extraction is carried out at room temperature and pressure for 10 minutes. After the extraction is completed, rotary evaporation is performed. The rotary evaporation conditions are: vacuum degree 10 mbar, 40°C, and rotation speed 150 rpm until the ethyl acetate is evaporated to dryness. After the rotary evaporation is completed, green tea polyphenols are obtained.

[0091] Unless otherwise stated, all chemicals used were purchased from Sigma-Aldrich (Hong Kong, China). High performance liquid chromatography (HPLC) system (Agilent 1100 series, Agilent Technologies, Inc.) was used to qualitatively and quantitatively analyze the composition of green tea polyphenols.

[0092] The analysis results of green tea polyphenol composition are as follows Figure 1 shown. Figure 1 Among them, 1 is epigallocatechin (EGC); 2 is epigallocatechin gallate (EGCG); 3 is epicatechin (EC); 4 is epicatechin gallate (ECG); and 5 is caffeine.

[0093] The quantitative results showed that green tea polyphenols contained 15g / kg epigallocatechin (EGC), 613g / kg epigallocatechin gallate (EGCG), 40g / kg epicatechin (EC), and 202g / kg epicatechin gallate (ECG).

[0094] Example 2

[0095] Example 1: Effect verification of green tea polyphenols prepared

[0096] 1. Experimental Grouping and Process

[0097] Experimental mice: 6-week-old C57BL / 6 healthy male mice weighing about 21g; the mice had free access to food and water, and after 1 week of adaptive feeding, they were randomly divided into 5 groups, with 9 mice in each group, namely: LFD group (NC group), HFD group, HFHF group, HFD+GP group and HFHF+GP group.

[0098] Among them, the LFD group was fed with conventional feed;

[0099] The HFD group was fed a high-fat diet;

[0100] The HFHF group was fed a high-fat diet containing fructose;

[0101] The HFD+GP group was fed with a high-fat diet and gavaged once a day with 400 mg / kg of the green tea polyphenol solution prepared in Example 1, wherein the mass concentration of the green tea polyphenol solution was 40 mg / mL and the solvent was normal saline;

[0102] The HFHF+GP group was fed a high-fat diet containing fructose and gavaged once a day with 400 mg / kg of the green tea polyphenol solution prepared in Example 1.

[0103] Each of the above experimental groups, LFD group, HFD group and HFHF group, was given an equal amount of normal saline according to body weight when the HFD+GP group and HFHF+GP group were administered.

[0104] The composition of the feeds fed to each experimental group is shown in Table 1.

[0105] Table 1 Feed composition and energy table corresponding to each experimental group

[0106]

[0107] Sources of feed products: casein (Foodchem); L-cystine, dextrose, fructose, choline bitartrate, all from Sigma; corn starch, lard, corn oil, all from the market; cellulose powder, AIN 93 vitamin mixture, AIN 76 mineral mixture, all from MPBiomedicals

[0108] The mice were fed with the above method for 12 weeks. After 12 weeks of feeding, they were deprived of food and water overnight and then killed.

[0109] 2. Body weight changes of mice in each experimental group

[0110] 1) The first week of adaptive feeding was taken as the first week. From the first week to the 13th week, the weight of mice in each experimental group was measured once a week at the same time and recorded. The results are shown in Tables 2 to 3 and Figure 2 shown.

[0111] Table 2 Body weight changes of mice in each experimental group (unit: g)

[0112]

[0113]

[0114]

[0115] Table 3 Body weight changes of mice in each experimental group (mean)

[0116] LFD HFD HFD+GP HFHF HFHF+GP 1 week 20.86 21.50 21.50 21.49 21.50 2 weeks 21.84 22.34 21.81 22.83 22.17 3 weeks 22.64 23.52 22.99 23.79 22.95 4 weeks 22.72 24.21 23.63 24.64 23.43 5 weeks 23.53 25.27 24.28 25.87 23.73 6 weeks 23.82 26.59 24.59 26.84 24.21 7 weeks 24.65 27.33 24.77 28.19 24.48 8 weeks 25.08 28.37 24.60 28.41 24.48 9 weeks 26.14 29.75 24.39 29.56 24.76 10 weeks 26.58 31.45 24.63 30.52 24.69 11 weeks 27.89 31.99 25.24 31.27 24.88 12 weeks 27.79 31.54 25.21 31.23 24.53 13 weeks 27.76 31.00 25.06 31.32 25.34

[0117] From Tables 2 to 3 and Figure 2As shown, the body weight of the HFD and HFHF groups increased significantly relative to the LFD group, indicating that high-fat diet and high-fat diet containing fructose can significantly increase the body weight of mice. The body weight of the HFD+GP group decreased significantly relative to the HFD group, and even decreased below the LFD group; the body weight of the HFHF+GP group decreased significantly relative to the HFHF group, and even decreased below the LFD group. This indicates that the green tea polyphenols can significantly reduce the weight gain caused by a high-fat diet or a high-fat diet containing fructose. This indicates that green tea polyphenols have an anti-obesity effect and can be used for weight loss, prevention and / or treatment of obesity, etc.

[0118] 2) After 12 weeks of feeding, the fasting blood glucose of mice in each experimental group was tested. Blood was collected from the tail tip of the mice. The blood glucose meter was Bayer Contour Plus. The test method was in accordance with the product manual. The results are shown in Table 4 and Figure 3 shown.

[0119] Table 4 Fasting blood glucose of mice in each experimental group

[0120] Group Mean SD (Standard Deviation) LFD 7.02 1.10 HFD 7.93 0.79 HFD+GP 6.80 0.83 HFHF 8.66 0.85 HFHF+GP 7.60 0.93

[0121] From Table 4 and Figure 3 It can be seen that the fasting blood glucose of the HFD and HFHF groups was significantly higher than that of the LFD group, indicating that high-fat diet and high-fat diet containing fructose can significantly increase the fasting blood glucose of mice. The blood glucose of the HFD+GP group was significantly lower than that of the HFD group; the blood glucose of the HFHF+GP group was significantly lower than that of the HFHF group, and the HFD+GP group and the HFHF+GP group were able to reduce blood glucose to normal levels. This shows that green tea polyphenols have the effect of lowering blood glucose and can be used to prevent and / or treat diseases such as hyperglycemia and diabetes, such as type 2 diabetes.

[0122] 3) After 12 weeks of feeding, the fasting triglyceride levels of mice in each experimental group were tested. When the mice were killed, blood was collected from the heart, heparin was added to the blood, and the serum was collected by centrifugation at 1000g for 10 minutes. The triglyceride determination kit of Nanjing Jiancheng Bioengineering Institute was used, and the detection method was carried out according to the instructions. The fasting triglyceride results of mice in each experimental group are shown in Table 5 and Figure 4 shown.

[0123] Table 5 Fasting triglycerides of mice in each experimental group

[0124] Group Mean SD (Standard Deviation) LFD 1.41 0.12 HFD 1.35 0.26 HFD+GP 1.14 0.18 HFHF 1.36 0.21 HFHF+GP 1.06 0.13

[0125] From Table 5 and Figure 4It can be seen that there is no significant difference in fasting triglyceride content between the HFD and HFHF groups and the LFD group. However, the fasting triglyceride content of the HFD+GP group was significantly lower than that of the HFD group, and the fasting triglyceride content of the HFHF+GP group was significantly lower than that of the HFHF group. This shows that green tea polyphenols have the effect of lowering fasting triglycerides and can significantly reduce hypertriglyceridemia caused by a high-fat diet.

[0126] 4) After 12 weeks of feeding, the total cholesterol of mice in each experimental group was tested. When the mice were killed, blood was collected from the heart, heparin was added to the blood, and the serum was collected by centrifugation at 1000g for 10 minutes. The cholesterol determination kit of Nanjing Jiancheng Bioengineering Institute was used, and the detection method was carried out according to the instructions. The total cholesterol results of mice in each experimental group are shown in Table 6 and Figure 5 shown.

[0127] Table 6 Total cholesterol of mice in each experimental group

[0128] Group Mean SD (Standard Deviation) LFD 318.67 105.61 HFD 461.42 143.34 HFD+GP 321.55 88.96 HFHF 435.21 209.83 HFHF+GP 288.87 68.00

[0129] From Table 6 and Figure 5 It can be seen that the total cholesterol in the HFD and HFHF groups was significantly higher than that in the LFD group, indicating that high-fat diet and fructose high-fat diet can significantly increase the total cholesterol content of mice. The total cholesterol content of the HFD+GP group was significantly reduced compared with the HFD group and returned to normal levels; the total cholesterol content of the HFHF+GP group was significantly reduced compared with the HFHF group and returned to normal levels. This shows that green tea polyphenols can significantly reduce total cholesterol, can significantly reduce hypercholesterolemia in mice caused by a high-fat diet, and can be used to treat diseases such as atherosclerosis.

[0130] 5) After 12 weeks of feeding, the organ weights of mice in each experimental group were weighed. The results are shown in Table 7 and Figure 6 shown.

[0131] Table 7 Organ weight of mice in each experimental group (g)

[0132]

[0133] From Table 7 and Figure 6 It can be seen that the epididymal fat and perirenal fat in the HFD group and HFHF group were significantly increased compared with the LFD group, indicating that a high-fat diet can significantly increase epididymal fat and perirenal fat. The epididymal fat and perirenal fat in the HFD+GP group were significantly reduced compared with the HFD group. The epididymal fat and perirenal fat in the HFHF+GP group were significantly reduced compared with the HFHF group. This shows that green tea polyphenols can reduce the weight of epididymal fat and perirenal fat, and can be used to treat metabolic syndrome, obesity and other diseases.

[0134] 6) After 12 weeks of feeding, the TNFα of mice in each experimental group was detected. The TNFα detection kit of Suzhou Yikesai Biotechnology Co., Ltd. was used to measure the TNFα in the serum. The results are shown in Table 8 and Figure 7 shown.

[0135] Table 8 TNFα of mice in each experimental group

[0136] Group Mean SD (Standard Deviation) LFD 83.22 14.13 HFD 135.40 40.85 HFD+GP 104.60 10.08 HFHF 94.45 16.83 HFHF+GP 71.46 15.49

[0137] From Table 8 and Figure 7 It can be seen that compared with the LFD group, TNFα in the HFD group was significantly increased, indicating that a high-fat diet can significantly increase TNFα. Compared with the HFD group, the HFHF group did not significantly increase TNFα, indicating that fructose can alleviate the increase in TNFα caused by a high-fat diet. The TNFα in the HFD+GP group was significantly reduced compared with the HFD group. Although TNFα was not significantly increased in the HFHF group compared with the LFD group, the HFHF group could still significantly reduce TNFα. This shows that green tea polyphenols can reduce the content of inflammatory factor TNFα in the blood and have an anti-inflammatory effect.

[0138] 7) After 12 weeks of feeding, the LPS of mice in each experimental group was detected by using the LPS detection kit of Suzhou Yikesai Biotechnology Co., Ltd. to measure the LPS in the serum. The results are shown in Table 9 and Figure 8 shown.

[0139] Table 9 LPS of mice in each experimental group

[0140] Group Mean SD (Standard Deviation) LFD 1.36 0.43 HFD 1.83 0.18 HFD+GP 1.5 0.07 HFHF 1.40 0.16 HFHF+GP 1.12 0.16

[0141] From Table 9 and Figure 8 It can be seen that compared with the LFD group, the LPS in the HFD group was significantly increased, indicating that a high-fat diet can significantly increase LPS. Compared with the HFD group, the LPS in the HFHF group was not significantly increased, indicating that fructose can alleviate the increase in LPS caused by a high-fat diet. The LPS in the HFD+GP group was significantly reduced compared with the HFD group. Although the LPS in the HFHF group was not significantly increased compared with the LFD group, the HFHF group could still significantly reduce the LPS. This shows that green tea polyphenols can reduce LPS and can be used to fight endotoxemia.

[0142] 8) Measurement of small intestine and small intestinal villi length

[0143] The entire small intestine of the mouse was taken and measured in length. The duodenum was taken, rolled into a "Swiss roll", fixed in 4% paraformaldehyde, embedded, fixed, sectioned, and H&E stained. After being photographed under a 400x microscope, morphological differences were analyzed and artificial villi were analyzed. Each intestinal segment was further divided into 4 quadrants, and 10 complete villi were measured from the distal edge of the crypt to the top of the villi in each quadrant. The average villus length of the entire intestinal segment was then calculated using ImageJ.

[0144] The small intestinal villi of mice in each experimental group are shown in Fig. 9 The length of small intestine and villus of mice in each experimental group are shown in Table 10 and Figures 10-11 shown. Fig.10 is the length of small intestine of mice in each experimental group; Fig.11 is the length of the small intestinal villi in the duodenum of mice in each experimental group.

[0145] Table 10 Small intestine length and small intestinal villus length of mice in each experimental group

[0146]

[0147] From Table 10 and Figures 9-11 It was found that after taking green tea polyphenols, the length of the small intestine of obese mice increased significantly. In addition, compared with the HFD group, the length of the small intestinal villi of HFD+GP increased significantly. Similarly, the length of the small intestinal villi of HFHF+GP increased significantly compared with that of HFHF. This shows that green tea polyphenols are beneficial to improve intestinal health.

[0148] 9) qRT-PCR verification of gene expression of fructose transporter, villus growth-related genes and tight junction proteins in mouse duodenum

[0149] RNA was extracted from the duodenum of mice using the Trizol method and reverse transcribed into cDNA, and then qRT-PCR was performed to verify the relevant genes, including the fructose transporter GLUT5, the villus growth-related gene hypoxia-inducible factor-1α (HIF-1α) and its target protein enolase (ENO1), and the tight junction proteins Jam, ZO-1, and occludin.

[0150] The gene expressions of fructose transporter, villus growth-related genes and tight junction proteins in the duodenum of mice in each experimental group are shown in Tables 11 and Figures 12 to 17 shown. Fig.12 This is a diagram of the gene expression of target protein enolase (ENO1) in mice of each experimental group; Fig.13 This is a diagram showing the gene expression of fructose transporter GLUT5 in mice in each experimental group; Fig.14 Figure 2 shows the expression of HIF-1α gene in mice of each experimental group; Fig.15 This is a graph showing the Jam gene expression in mice in each experimental group; Fig.16This is a graph showing the occludin gene expression in mice in each experimental group; Fig.17 Figure 2 is the expression of ZO-1 gene in mice of each experimental group.

[0151] Table 11 Gene expression of fructose transporter, villus growth-related genes and tight junction proteins in the duodenum of mice in each experimental group

[0152]

[0153] From Table 11 and Figures 12 to 17 It was found that the green tea polyphenols treatment group upregulated the expression of fructose transporter GLUT5 and hypoxia inducible factor target protein ENO1, thereby increasing the villi of the small intestine. In addition, the expression of tight junction proteins increased after green tea polyphenols treatment, thereby protecting the intestinal mucosal barrier and maintaining intestinal health. This shows that green tea polyphenols have a good effect on maintaining the integrity of the colon barrier structure and can be used to treat intestinal damage diseases such as colitis.

[0154] 10) Western blotting to verify the expression of fatty acid metabolism-related proteins in mouse liver

[0155] Liver tissue samples were homogenized using RIPA buffer, and after centrifugation, the total protein concentration of the supernatant was measured, and equal amounts of protein (80 μg) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel, and then blotted onto polyvinylidene fluoride (PVDF) membranes. After sealing with 50 mL / L skim milk solution, the membranes were incubated with primary and secondary antibodies, and the protein expression was quantified using Image Lab (BioRad) after taking pictures.

[0156] The results of the detection of fatty acid metabolism-related proteins in mice in each experimental group are shown in Table 12 and Figures 18 to 21 As shown. Fig.18 The results of the detection of ACC protein expression in mice in each experimental group are shown in Figure 2. Fig.19 This is the result of detecting the expression of ACL protein in mice in each experimental group; Fig. 20 The results of FAS protein expression detection in mice of each experimental group are shown in the figure; Fig.21 This is the result of detecting the Srebp1c protein expression in mice in each experimental group.

[0157] Table 12 Detection results of fatty acid metabolism related proteins in mice in each experimental group

[0158]

[0159] The results showed that green tea polyphenols can reduce the protein expression of fatty acid synthase acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and sterol regulatory element-binding protein 1c (SREBP1c) in the liver, thereby reducing the synthesis of fatty acids. It can be seen that green tea polyphenols can inhibit lipid synthesis and show anti-obesity activity. They can be used to lower blood lipids and relieve non-alcoholic fatty liver inflammation.

[0160] In summary, the green tea polyphenols provided by the present invention are used to treat diseases such as obesity, metabolic syndrome, non-alcoholic steatohepatitis, diabetes, atherosclerosis, endotoxemia and colitis.

[0161] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of green tea polyphenols in the preparation of products for improving intestinal health; the green tea polyphenols include epigallocatechin, epigallocatechin gallate, epicatechin and epicatechin gallate.

2. The application according to claim 1, characterized in that: Prepare products for improving any one or two or more of metabolic syndrome, intestinal injury disease, endotoxemia and colitis.

3. The application according to claim 2, characterized in that: The metabolic syndrome includes any one or two or more of the following (1) to (5): (1) Nonalcoholic fatty liver disease; (2) Hyperlipidemia; (3) Diabetes; (4) atherosclerosis; (5) Obesity.

4. The use according to claim 3, characterized in that: The diabetes mellitus includes type 2 diabetes mellitus; the hyperlipidemia includes hypercholesterolemia and / or hypertriglyceridemia.

5. The use according to claim 1, characterized in that: The improvement of intestinal health includes the following (I) to (IV): (I) Increase the length of the small intestine; (II) Increase the length of small intestinal villi; (III) Improve intestinal barrier; (IV) Increase the content of fructose transporter.

6. The use according to claim 1, characterized in that: The preparation method of green tea polyphenols comprises: Mixing tea leaves with water and performing extraction twice to obtain tea extract; After removing caffeine from the tea extract, ethyl acetate is used for extraction to obtain green tea polyphenols.

7. The use according to claim 6, characterized in that: The temperature of each extraction is 85-95°C; the time of each extraction is 20-40 minutes.

8. The use according to claim 6, characterized in that: The method for removing caffeine from tea extract comprises: removing caffeine from tea extract by using chloroform.

9. The use according to claim 1, characterized in that: The products include any one or more of medicines, foods and health products.

10. The use according to claim 9, characterized in that: The product also includes auxiliary materials.

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