Potato extract as well as preparation method and application thereof

Abstract

The invention relates to the technical field of biological medicines, in particular to a potato extract as well as a preparation method and application thereof. The potato extract is extracted from potato tubers. Animal experiments prove that the potato extract can significantly promote growth and proliferation of AKK bacteria, improve the GLP-1 level in intestinal tracts and blood, effectively reduce fasting blood-glucose, improve sugar tolerance, relieve insulin resistance and improve obesity. The potato extract has the characteristics of easily available raw materials, simplicity and convenience in preparation, remarkable effect and the like, and has a good application prospect in preparation of healthy products for regulating intestinal flora and improving blood sugar and obesity.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to a potato extract, its preparation method, and its application. Background Technology

[0002] The gut microbiota is closely related to host health, and its structural dysregulation is significantly associated with various metabolic diseases, such as type 2 diabetes and obesity. Akkermansia (… Akkermansia muciniphila AKK bacteria (Acinetobacter akadina) are beneficial bacteria found in the intestinal mucosa of humans and animals, accounting for approximately 1-4% of the total gut microbiota. Recent studies have shown that decreased AKK abundance is closely associated with obesity, insulin resistance, type 2 diabetes, and inflammatory states. Conversely, increasing AKK abundance in the gut can improve intestinal barrier function and significantly improve host glucose metabolism and insulin sensitivity, as well as reduce body weight.

[0003] Currently, direct supplementation with live AKK bacteria or their inactivated formulations has become a potential treatment strategy, but it suffers from high production costs, stringent storage conditions, and poor bacterial stability. Therefore, selectively promoting the proliferation of innate AKK bacteria through dietary intervention or supplementation with specific prebiotics is a more feasible and promising strategy. However, reports on natural products with highly efficient and specific AKK bacteria proliferation-promoting functions are still limited.

[0004] Previous studies have shown that *Amycosis AKK* is associated with the secretion of enteropancreatic glucagon-like peptide-1 (GLP-1) (Yoon et al., 2021). GLP-1 is a hormone mainly produced by intestinal L cells. GLP-1 is a target for novel hypoglycemic and weight-loss drugs, which can activate GLP-1. 1 receptor enhances insulin secretion in a glucose concentration-dependent manner, inhibits glucagon secretion, and delays gastric emptying. It reduces food intake through central appetite suppression, thereby achieving effects such as lowering blood sugar and weight loss.

[0005] Potatoes are one of the world's most important food crops, rich in starch, protein, vitamins, and various bioactive substances. Currently, the deep processing and utilization of potatoes are mostly concentrated in the food industry, and the regulatory effects of specific bioactive components on gut microbiota, especially AKK bacteria, have not been reported.

[0006] Therefore, developing a potato extract that can specifically promote the proliferation of AKK bacteria and thus improve blood sugar levels is of great significance for developing novel adjunctive drugs or functional foods for diabetes. Summary of the Invention

[0007] In view of the above-mentioned shortcomings in the prior art, the purpose of the present invention is to provide a potato extract, its preparation method and application.

[0008] To achieve the above-mentioned objectives, the technical solution adopted by this invention is as follows: In a first aspect, a method for preparing potato extract is provided, comprising the following steps: S1. Soak the washed potato tubers in a sodium bisulfite solution, then crush and filter them; S2. After centrifuging the filtrate, adjust the pH to 2-6, let it stand, and then centrifuge again to collect the supernatant. S3. Dissolve the supernatant in NaCl and let it stand overnight; S4. Centrifuge the above mixture, discard the supernatant, and resuspend the precipitate in distilled water. S5. Concentrate and dry the resuspended liquid, and cut off the molecular weight to 5-10 kDa to obtain potato extract.

[0009] Furthermore, in step S1, the ratio of potato tubers to sodium bisulfite in liquid is 1.5-3.0:1.0.

[0010] Further, the method of step S2 is as follows: centrifuge the filtrate at 3000-7000 r / min for 5-15 min, obtain the supernatant, adjust the pH value to 2-6, let it stand for 10-30 min, centrifuge again and collect the supernatant.

[0011] Further, the method in step S3 is as follows: add 10-30% NaCl to the supernatant and stir continuously until all NaCl is dissolved, and let stand overnight at 4 ℃.

[0012] Secondly, a potato extract prepared by any of the above methods is provided.

[0013] Thirdly, the above-mentioned potato extract is provided for use in the preparation of a formulation for promoting the proliferation of Akkermansia.

[0014] Fourthly, the above-mentioned potato extract is provided for use in the preparation of formulations for increasing GLP-1 levels.

[0015] Fifthly, the invention provides the use of the aforementioned potato extract in the preparation of formulations for the prevention and adjunctive treatment of glucose metabolism disorders.

[0016] Furthermore, the formulation is a pharmaceutical composition or a functional food composition.

[0017] Furthermore, the pharmaceutical composition comprises a therapeutically effective amount of potato extract and a pharmaceutically acceptable carrier, and its dosage form includes tablets, capsules, granules, powders, and oral liquids; the functional food composition includes health products, nutritional supplements, and special medical foods.

[0018] The beneficial effects of this invention are as follows: This invention is the first to discover that, in a high-fat diet-induced mouse model, potato extract can significantly increase the relative abundance of AKK bacteria in the intestinal contents of diabetic mice and restore the balance of intestinal flora, which is closely related to its hypoglycemic effect.

[0019] The potato extract of this invention can significantly reduce fasting blood glucose, improve oral glucose tolerance, and alleviate insulin resistance. Mechanistic studies have shown that the potato extract preparation method provided by this invention is simple, uses widely available and inexpensive raw materials, and has high safety. It provides new raw material options and technical support for the development of hypoglycemic drugs or functional foods targeting the "microbiota-host" metabolic axis, and has broad application prospects. Attached Figure Description

[0020] Figure 1 The effects of potato extract on gut microbiota; Figure 2 The effect of potato extraction on the relative abundance of AKK bacteria; Figure 3 The effect of potato extract on the GLP-1 content in plasma; Figure 4 The effect of potato extract on the expression of the Gcg gene encoding GLP=1 in the colon; Figure 5 The changes in blood glucose levels in mice of different groups 2 hours after fasting glucose injection; Figure 6 Area under the blood glucose curve for different groups of mice 2 hours after fasting glucose injection; Figure 7 The effect of potato extract on fasting insulin in mice; Figure 8 The effect of potato extract on mouse body weight. Detailed Implementation

[0021] The specific embodiments of the present invention are described below to enable those skilled in the art to understand the present invention. However, it should be understood that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, various changes are obvious as long as they are within the spirit and scope of the present invention as defined and determined by the appended claims. All inventions utilizing the concept of the present invention are protected.

[0022] Example 1: Preparation of potato extract Weigh potato tubers, wash and shred them to increase the contact area and facilitate component dissolution. Soak them in sodium bisulfite solution at a material-to-liquid ratio of 2.5:1.0 to utilize its antioxidant properties to inhibit enzymatic browning and prevent microbial growth. Juice the tubers at room temperature, then centrifuge and filter to collect the filtrate, achieving solid-liquid separation and removing starch and fiber impurities. Adjust the pH to 3 and use the isoelectric point precipitation principle to remove some impurities. After standing for 20 minutes, centrifuge and collect the supernatant. Slowly add 25% NaCl to the supernatant while stirring continuously until all NaCl is dissolved. Place the tubers in a 4°C refrigerator overnight to precipitate the target protein through salting out. Centrifuge the precipitate and resuspend it in distilled water to remove salt. Finally, concentrate the precipitate using a hollow fiber ultrafiltration membrane to achieve a molecular weight cutoff of 10. The concentration of the potato extract was achieved by removing small molecule impurities and enriching the target components. The concentrated solution was then freeze-dried in a freeze dryer to maximize the preservation of product activity through low-temperature dehydration. The resulting potato extract sample was stored in a desiccator for later use, and its stability was ensured under a dry environment.

[0023] Example 2: Effects of potato extract on gut microbiota, GLP-1, blood glucose, and body weight in mice on a high-fat diet Animal experiments were conducted using the potato extract obtained in Example 1 to study its effects on gut microbiota, GLP-1, blood glucose, and body weight in vivo.

[0024] Male C57Bl / 6J mice (weighing 21-24 g) aged 6-8 weeks were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. They were housed according to SPF-grade animal husbandry standards. The animal experiments were reviewed and approved by the Ethics Committee of the Institute of Agricultural Product Processing, Chinese Academy of Agricultural Sciences. The ambient temperature was 25 ± 2°C, the humidity was 30-70%, and a continuous 12-hour light / 12-hour dark cycle was maintained. The ventilation rate in the animal room was 10-11 times / hour. After one week of acclimatization to a standard diet, the mice were randomly divided into three groups (n=6 per group): a standard diet group (Group C) – fed a standard diet and administered a daily dose of carrier distilled water via oral gavage; a high-fat, high-sugar diet group (Group M) – fed a high-fat, high-sugar diet and administered a daily dose of carrier distilled water via oral gavage; and a potato extract group (Group PES) – fed a high-fat, high-sugar diet and administered a daily dose of potato extract via oral gavage. The mice underwent the intervention for ten weeks, and their body weight was monitored weekly. Ten weeks after intervention, mouse feces were collected for 16sRNA detection. Subsequently, mice were fasted for 12 hours but allowed free access to water, and then administered glucose via gavage at a dose of 2 g / kg BW. Blood samples were collected from the tail tip of the mice at 0, 30, 60, 90, and 120 minutes after gavage, and blood glucose levels were measured using a glucometer. A curve was plotted, and the area under the curve (AUC) was calculated. After 10 weeks of intervention, the mice were sacrificed, and their serum and colon samples were collected for further assays.

[0025] β-diversity analysis reflects the diversity and variability of the microbial community. The gut microbiota of mice was measured to investigate the impact of this invention's application on the gut microbiota of mice on a high-fat diet. Figure 1 and 2 As shown, compared to group C, the abundance of AKK bacteria in group M mice was significantly reduced, while potato extract intervention (PES group) significantly increased the abundance of AKK bacteria in high-fat diet mice from 0.05% to 19.06%. This indicates that potato extract has prebiotic properties, significantly increases the level of probiotic AKK, and is beneficial in improving symptoms related to glucose metabolism and obesity.

[0026] The levels of GLP-1 in serum and colon were measured to investigate the effect of this invention's application examples on GLP-1 content in mice on a high-fat diet. Figure 3As shown, compared to group C, the serum GLP-1 level in the high-fat diet model group (group M) was significantly reduced, indicating that the effect of high-fat diet on insulin resistance was reduced in mice. Administration of potato extract (PES group) significantly increased the serum GLP-1 level in high-fat diet mice, increasing it by 80.5%. Given that the colon is a major source of GLP-1, probiotics such as AKK bacteria have been reported to increase GLP-1 levels, and the potato extract in this invention increases AKK bacteria levels; therefore, the gene Gcg encoding GLP-1 in the colon was detected. Figure 4 As shown, the potato extract of the present invention can significantly enhance the expression of GLP-1 mRNA. This indicates that the potato extract increases serum GLP-1 levels by stimulating colonic GLP-1 expression.

[0027] An oral glucose tolerance test was conducted and fasting insulin levels were measured to investigate the effect of the obtained potato extract on blood glucose. Figure 5-6 As shown, compared to the blank control group (Group C), the high-fat diet group (Group M) led to impaired insulin tolerance, while potato extract intervention (PES group) increased the rate of blood glucose decline in high-fat diet mice, increased the magnitude of blood glucose reduction, and significantly reduced the area under the blood glucose line, suggesting that potato extract can improve high-fat diet-induced glucose intolerance. Figure 7 As shown, potato extract can lower insulin levels and improve insulin resistance caused by a high-fat diet.

[0028] During the intervention period, changes in mouse body weight were monitored to explore the effect of this invention on improving obesity induced by a high-fat diet. Figure 8 As shown, compared to the blank control group (group C), mice in the high-fat diet group (group M) gained weight. Intervention with potato extract (PES group) significantly reduced obesity induced by the high-fat diet and decreased mouse weight.

[0029] In summary, animal experiments have demonstrated that the prepared potato extract significantly promotes the growth and proliferation of AKK bacteria, increases GLP-1 levels in the gut and blood, effectively reduces fasting blood glucose, improves glucose tolerance, alleviates insulin resistance, and improves obesity. This potato extract is characterized by readily available raw materials, simple preparation, and significant effects, and shows promising application prospects in the preparation of health products that regulate gut microbiota and improve blood sugar and obesity.

[0030] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, it is intended that all variations falling within the meaning and scope of equivalents of the claims be included within the present invention.

[0031] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A method for preparing a potato extract, characterized in that, Includes the following steps: S1. Soak the washed potato tubers in a sodium bisulfite solution, then crush and filter them; S2. After centrifuging the filtrate, adjust the pH to 2-6, let it stand, and then centrifuge again to collect the supernatant. S3. Dissolve the supernatant in NaCl and let it stand overnight; S4. Centrifuge the above mixture, discard the supernatant, and resuspend the precipitate in distilled water. S5. Concentrate and dry the resuspended liquid, and cut off the molecular weight to 5-10 kDa to obtain potato extract.

2. The method for preparing potato extract according to claim 1, characterized in that, In step S1, the ratio of potato tubers to sodium bisulfite in liquid is 1.5-3.0:1.

0.

3. The method for preparing potato extract according to claim 1, characterized in that, The specific method for step S2 is as follows: centrifuge the filtrate at 3000-7000 r / min for 5-15 min, obtain the supernatant, adjust the pH value to 2-6, let it stand for 10-30 min, centrifuge again, and collect the supernatant.

4. The method for preparing potato extract according to claim 1, characterized in that, The specific method for step S3 is as follows: add 10-30% NaCl to the supernatant and stir continuously until all NaCl is dissolved, then let it stand overnight at 4 ℃.

5. The potato extract prepared by the method according to any one of claims 1-4.

6. The use of the potato extract of claim 5 in the preparation of a formulation for promoting the proliferation of Akkermansia.

7. The use of the potato extract of claim 5 in the preparation of formulations for increasing GLP-1 levels.

8. The use of the potato extract according to claim 5 in the preparation of formulations for the prevention and adjunctive treatment of glucose metabolism disorders.

9. The application according to any one of claims 6-8, characterized in that, The preparation is a pharmaceutical composition or a functional food composition.

10. The application according to claim 9, characterized in that, The pharmaceutical composition comprises a therapeutically effective amount of the potato extract and a pharmaceutically acceptable carrier, and its dosage forms include tablets, capsules, granules, powders, and oral liquids; the functional food composition includes health products, nutritional supplements, and special medical foods.

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