A nutritional preparation of a whole food matrix for improving intestinal flora and promoting intestinal mucosa repair and a preparation method thereof

Abstract

The present application provides a whole food matrix for improving intestinal flora and promoting intestinal mucosa repair and a preparation method thereof, and relates to the technical field of nutritional preparations.The nutritional preparation according to the present application comprises, by weight fraction, 20-50 parts of compound cereal powder, 1-20 parts of compound fruit and vegetable powder, 10-35 parts of plant fat powder, 5-25 parts of protein, 0.1-10 parts of prebiotics, 2-10 parts of compound minerals and 0.05-1.5 parts of compound vitamins, wherein the compound cereal powder, the compound fruit and vegetable powder and the prebiotics together serve as a source of total dietary fiber, the weight of soluble dietary fiber in the total dietary fiber accounts for more than 50% of the total weight of the total dietary fiber, the compound cereal powder comprises yam powder, and the yam powder accounts for 0.5-5% of the total mass of the compound cereal powder, the compound minerals comprise zinc, the content of zinc is 2-6 mg per 100 g of the nutritional preparation, and the compound fruit and vegetable powder is fruit and vegetable powder obtained by treating endogenous pectin gel particles.

Description

Technical Field

[0001] This invention relates to the technical field of nutritional preparations, specifically to a whole food matrix nutritional preparation and its preparation method that improves intestinal flora and promotes intestinal mucosal repair. Background Technology

[0002] In recent years, enteral nutrition (EN) has been increasingly widely used in clinical nutritional support, playing a vital role in maintaining nutritional status and supporting disease recovery. Meanwhile, a growing body of research indicates that gut microbiota structure and intestinal barrier function are crucial for overall health, and that gut microbiota imbalance and intestinal mucosal damage are closely related to various disease states. Therefore, balancing nutritional delivery with gut microbiota regulation and mucosal repair during nutritional support has gradually become a key focus in this field.

[0003] Currently, most enteral nutrition formulations widely used in clinical practice employ a "separation-purification-compounding" technical model. This involves separating and purifying macronutrients such as proteins, carbohydrates, and fats from natural foods, and then combining them with artificially synthesized vitamins and minerals. While this type of formulation has advantages in terms of clear composition, controllable dosage, and nutrient delivery, its highly purified design is gradually revealing limitations in regulating the intestinal microecology.

[0004] On the one hand, bioactive substances such as dietary fiber, plant polyphenols, and carotenoids, which are abundant in natural foods, are largely removed during the "separation-purification" process. Although these substances are not "essential nutrients" in the traditional sense, they are key metabolic substrates for maintaining gut microbiota homeostasis. Their absence makes it difficult for gut microbiota to obtain diverse nutritional sources, leading to a more homogeneous microbiota structure, a significant reduction in species diversity, and a decrease in the relative abundance of beneficial bacteria. This change directly weakens the function of the gut as an important immune organ, thereby easily inducing dysbiosis-related complications, including diarrhea, constipation, and secondary infections.

[0005] On the other hand, existing enteral nutrition formulations mainly focus on the supply of basic nutrients, with relatively insufficient support for functions related to intestinal mucosal repair. Processes such as intestinal epithelial cell renewal, mucus layer formation, and local immune regulation not only depend on the supply of basic nutrients but also involve the participation of various food-derived active ingredients. However, traditional enteral nutrition formulations have limited coverage in these aspects, making it difficult to achieve active regulation of intestinal barrier function.

[0006] In recent years, enteral nutrition based on the concept of "real food" has gradually gained attention, including homemade homogenized diets and commercial nutritional preparations containing ingredients derived from natural foods. Related studies have shown that nutritional preparations that retain the natural food matrix can provide dietary fiber and various plant-derived active ingredients, offering diverse metabolic substrates for gut microbiota, thereby regulating gut microbiota structure and influencing the gut environment to some extent. However, homemade homogenized diets suffer from unstable nutritional composition, a higher risk of microbial contamination, and complex preparation processes, making standardized application difficult.

[0007] In contrast, industrially produced nutritional supplements containing natural food-derived ingredients have certain advantages in terms of safety and stability. However, existing products still focus primarily on nutritional supplementation in their formulation design, with insufficient consideration given to the synergistic effects between multi-source carbohydrate structures, dietary fiber types, and plant-based active ingredients. This makes it difficult to simultaneously achieve effective regulation of the gut microbiota structure and support for intestinal mucosal repair. In particular, the fruit and vegetable powders added to existing products are mostly produced using ordinary spray-drying processes. The bioactive substances in these powders are significantly lost due to heat during this process. Furthermore, this type of dried powder causes the active substances to dissolve and release rapidly after entering the stomach, making them easily degraded and inactivated by gastric acid. It also easily triggers osmotic pressure shocks and rapid fermentation and gas production, resulting in poor intestinal tolerance and severely limiting their application in people with sensitive guts.

[0008] Therefore, under the premise of meeting basic nutritional needs, how to introduce multi-source food matrices and functional components through formulation design, while simultaneously addressing issues such as the destruction of active substances in ordinary fruit and vegetable powders, to regulate the gut microbiota and promote intestinal mucosal repair, has become a pressing technical problem. Based on this, it is necessary to develop a whole-food matrix nutritional formulation that can improve gut microbiota and promote intestinal mucosal repair, in order to overcome the aforementioned shortcomings of existing technologies. Summary of the Invention

[0009] This invention is made to solve the above-mentioned problems, and aims to provide a nutritional preparation and preparation method of a whole food matrix that improves intestinal flora and promotes intestinal mucosal repair. The nutritional preparation contains endogenous pectin gel microparticles of fruits and vegetables, which can not only meet the whole nutrition standards, but also actively repair the intestine and improve the flora by utilizing the integrity of natural foods. At the same time, it significantly improves intestinal tolerance through gel microparticle technology.

[0010] This invention provides a whole-food matrix nutritional preparation that improves gut microbiota and promotes intestinal mucosal repair. It is characterized by comprising, by weight: 20-50 parts of compound cereal powder, 1-20 parts of compound fruit and vegetable powder, 10-35 parts of plant fat powder, 5-25 parts of protein, 0.1-10 parts of prebiotics, 2-10 parts of compound minerals, and 0.05-1.5 parts of compound vitamins. The compound cereal powder, compound fruit and vegetable powder, and prebiotics collectively serve as sources of total dietary fiber, with soluble dietary fiber accounting for more than 50% of the total dietary fiber weight. The compound cereal powder includes yam powder, which accounts for 0.5%-5% of the total mass of the compound cereal powder. The compound minerals include zinc, with a zinc content of 2-6 mg / 100g of the nutritional preparation.

[0011] The nutritional preparation of whole food matrix that improves intestinal flora and promotes intestinal mucosal repair provided by the present invention may also have the following characteristics: the compound fruit and vegetable powder is a fruit and vegetable powder obtained by endogenous pectin gel microparticle technology.

[0012] The nutritional preparation of the whole food matrix that improves intestinal flora and promotes intestinal mucosal repair provided by the present invention may also have the following characteristics: The preparation process of the above-mentioned compound fruit and vegetable powder includes the following steps: Step S101, low-temperature enzyme inactivation: fresh fruits and vegetables are washed and cut, blanched at 85-90℃ for 10-20s, and pulped to form fruit and vegetable pulp; Step S102, ultra-high pressure micro-jet treatment: the fruit and vegetable pulp is circulated 3-5 times under a pressure of 120-160MPa to obtain a treated slurry, with the temperature ≤30℃ throughout the process; Step S103, deesterification treatment: the pH of the treated slurry is adjusted to 8.0-8.5, and stirred and reacted at 20-25℃ to obtain a deesterified slurry; Step S104, self-crosslinking treatment: the pH of the deesterified slurry is adjusted back to less than 7, calcium agent is added, stirred and allowed to stand to obtain a gel slurry; Step S105, freeze-drying treatment: the gel slurry is freeze-dried and pulverized to obtain compound fruit and vegetable powder.

[0013] The nutritional preparation of whole food matrix that improves intestinal flora and promotes intestinal mucosal repair provided by the present invention may further include one or more of the following: 0-5 parts meat meal, 0-1 part thickener and 0-2 parts acidity regulator.

[0014] The nutritional preparation of whole food matrix that improves intestinal flora and promotes intestinal mucosal repair provided by the present invention may also have the following characteristics: wherein the compound grain powder further includes one or more combinations of rice flour, enzymatically hydrolyzed rice flour, corn flour, millet flour and soybean flour.

[0015] The nutritional preparation of whole food matrix that improves intestinal flora and promotes intestinal mucosal repair provided by the present invention may also have the following characteristics: wherein the compound fruit and vegetable powder includes one or more of the following: pumpkin powder, apple powder, carrot powder, tomato powder, strawberry powder, broccoli powder, kale powder, pear powder, banana powder, blueberry powder, orange powder, pineapple powder, cucumber powder, spinach powder and purple sweet potato powder, preferably any one or more of the following: pumpkin powder, apple powder, carrot powder, strawberry powder and broccoli powder.

[0016] The nutritional preparation of whole food matrix for improving intestinal flora and promoting intestinal mucosal repair provided by the present invention may also have the following characteristics: wherein the plant fat powder includes: plant oils, including one or more combinations of medium-chain triglycerides, docosahexaenoic acid oils, eicosapentaenoic acid oils, arachidonic acid oils, 1,3-dioleoyl-2-palmitoyl triglycerides, rapeseed oil, low-erucic acid rapeseed oil, corn oil, soybean oil, sunflower seed oil, high-oleic sunflower seed oil, peanut oil, high-oleic peanut oil, palm oil, palm sap oil, palm kernel oil, olive oil, coconut oil, flaxseed oil, safflower seed oil, walnut oil, and peony seed oil; emulsifiers, including one or more combinations of sodium caseinate, casein, citrate fatty acid glycerides, mono- and diglycerides, and caprylic / capric glycerides; and wall materials, including one or more combinations of maltodextrin, glucose syrup, oligomaltose, and maltitol.

[0017] The whole food matrix nutritional preparation for improving intestinal flora and promoting intestinal mucosal repair provided by the present invention may also have the following characteristics: wherein the protein includes one or more combinations of whey protein, soy protein, casein, milk protein, rice protein, pea protein and yeast protein; the prebiotics include one or more combinations of fructooligosaccharides, galactooligosaccharides, inulin, pectin and β-glucan; the complex minerals include at least two of potassium, calcium, magnesium, iron, copper, manganese, iodine, selenium, sodium, phosphorus and chlorine; and the complex vitamins include at least two of vitamin A, vitamin D, vitamin E, vitamin K, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, folic acid, pantothenic acid, vitamin C, biotin, taurine and β-carotene.

[0018] The present invention also provides a method for preparing a nutritional preparation of a whole food matrix that improves gut microbiota and promotes intestinal mucosal repair. The method is characterized by the following steps: weighing each component according to the weight parts; dry mixing the weighed components; and purging the mixed components with nitrogen gas for protection to obtain the nutritional preparation.

[0019] The method for preparing a whole food matrix nutritional preparation that improves gut microbiota and promotes intestinal mucosal repair provided by the present invention may also have the following characteristics: dry mixing is carried out using an airflow mixer, and the mixing time is 5-15 minutes.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] This invention addresses a fundamental contradiction in existing nutritional formulations: while pursuing the purity of nutritional components, the overall synergistic effect of the food matrix is ​​sacrificed, leading to intestinal microecological imbalance and a lack of functional repair capabilities. Furthermore, ordinary fruit and vegetable powders suffer from poor intestinal tolerance and low bioavailability of active substances. This invention provides a nutritional formulation based on a whole food matrix that improves intestinal flora and promotes intestinal mucosal repair. It not only meets the standards for complete nutrition but also utilizes the integrity of natural foods to actively repair the intestines and improve the flora. At the same time, it significantly improves the bioavailability of active substances in fruit and vegetable powders and improves intestinal tolerance through endogenous pectin gel microparticle technology.

[0022] This invention constructs a synergistic system integrating "nutrient supply, gut microbiota regulation, and mucosal repair," utilizing a "dual-channel fermentation system" formed by compound grain powder, compound fruit and vegetable powder, and prebiotics. The compound fruit and vegetable powder employs endogenous pectin gel microparticle processing technology, combined with yam powder, zinc, and active ingredients from fruit and vegetable powders to form a "mucosal repair engine." This significantly optimizes the gut microbiota structure while inhibiting systemic and local intestinal inflammatory responses, thereby improving the gut microbiota and promoting intestinal mucosal repair in patients with gastrointestinal dysfunction while meeting basic nutritional needs.

[0023] Specifically, the present invention achieves the above effects through the following three aspects:

[0024] (1) Microbial colonization engine: dual-channel fermentation system

[0025] This invention constructs a composite carbon source system with different fermentation characteristics by regulating the ratio of insoluble dietary fiber (IDF) to soluble dietary fiber (SDF). The dietary fiber mainly originates from compound cereal powder, compound fruit and vegetable powder, and exogenous prebiotics. The dietary fiber system is composed of natural fiber components with different fermentation rates and different intestinal action sites, resulting in differentiated sequential fermentation effects: fructooligosaccharides act as a medium-speed fermentation prebiotic, primarily acting on the terminal small intestine and upper colon, rapidly colonizing beneficial bacteria; the compound fruit and vegetable powder contains natural pectin, hemicellulose, and water-soluble plant polysaccharides, serving as a medium-to-slow fermentation component, continuously and slowly releasing fermentation in the mid-colon, steadily supplying short-chain fatty acids; the slowly digestible starch, yam polysaccharides, and resistant starch components in the compound cereal powder resist upper digestion, reaching the distal colon for slow fermentation.

[0026] By combining IDF and SDF in a certain ratio, a substrate supply with different fermentation rates can be ensured in the intestine. Compared with the traditional concentrated burst fermentation of a single fiber, this invention forms a segmented, time-sequential, and gradient fermentation mechanism throughout the upper, middle, and lower colon, achieving homeostasis regulation of the gut microbiota and significantly improving microbial diversity and the proportion of beneficial bacteria. Insoluble dietary fiber, mainly derived from grains and some fruits and vegetables, is not easily degraded rapidly in the intestine and can act as a structural carrier to prolong the retention time of chyme in the intestine, providing a continuous interface for microbial fermentation. Simultaneously, its physical properties help maintain the distribution of intestinal contents, thus influencing the colonization environment of microorganisms to some extent. Soluble dietary fiber, mainly derived from pectin and other soluble polysaccharides and exogenous prebiotics in raw materials such as apple powder and pumpkin powder, has good swelling and fermentability, forming a gel-like structure in the intestine and serving as a preferred substrate for microorganisms, thereby initiating the fermentation process at an earlier stage. The differences in fermentation rate and mode of action between the two types of dietary fiber allow their combination to form a substrate release pattern similar to "two-stage fermentation" in the intestine, thereby improving overall fermentation efficiency and microbial metabolic activity.

[0027] Based on the aforementioned component characteristics, this composite system theoretically helps enhance the overall metabolic activity of the gut microbiota and promotes the growth dominance of certain probiotics. Specifically, oligosaccharides, pectin, and some resistant starch can be preferentially utilized by *Lactobacillus*, thereby increasing their relative abundance to some extent. Simultaneously, the short-chain fatty acids produced during the fermentation of dietary fiber and resistant starch can regulate the local pH environment of the gut and, together with polyphenols from fruits and vegetables and active ingredients from cruciferous vegetables, influence the growth and colonization of some unfavorable bacterial groups, such as *Helicobacter*, under certain conditions. Therefore, by controlling the ratio of insoluble to soluble dietary fiber and combining it with the synergistic design of multi-source grains and fruit and vegetable components, this invention theoretically constructs a composite substrate system with staged fermentation characteristics, thus providing a formulation basis for regulating the gut microbiota structure.

[0028] To achieve the above-mentioned effects, the present invention employs the following technical means: the weight of soluble dietary fiber accounts for more than 50% of the total weight of dietary fiber.

[0029] (2) Mucosal Repair Engine: Triple Barrier Reinforcement

[0030] Mechanism: Through multi-component synergy, the intestinal barrier is strengthened at three levels: physical, biochemical, and immune.

[0031] ① Physical barrier repair: Diosgenin and mucin in yam powder directly promote the proliferation of intestinal epithelial cells and the expression of tight junction proteins, accelerating the healing of mucosal damage.

[0032] ② Mucus layer strengthening and immune regulation: Pectin in fruit and vegetable powders stimulates goblet cells to secrete mucin, increasing the thickness of the mucus layer. The provided β-carotene is converted into vitamin A in the body, which helps repair the epithelium. Zinc ions directly participate in mucosal DNA synthesis and cell division, accelerating the healing of mucosal ulcers.

[0033] ③ Anti-inflammatory and antioxidant components: The glucosinolates in the fruit and vegetable powder are metabolized into sulforaphane, which activates the Nrf2 pathway and inhibits intestinal inflammatory factors (such as TNF-α and IL-6), controlling inflammation at its source. Lycopene can neutralize free radicals and reduce oxidative stress damage to the mucosa. Anthocyanins inhibit the NF-κB pathway and reduce the release of inflammatory mediators.

[0034] To achieve the above-mentioned effects, the present invention employs the following technical means: the compound cereal powder includes yam powder, and the yam powder accounts for 0.5%-5% of the total mass of the compound cereal powder; the compound minerals include zinc, and the zinc content is 2-6 mg / 100g of nutritional preparation.

[0035] (3) Endogenous pectin gel microparticle technology

[0036] This invention aims to break through the industry's reliance on exogenous carriers and provide a technology for preparing all-natural fruit and vegetable powders with zero exogenous carriers, zero encapsulating agents, and zero exogenous colloids. It regulates the molecular structure and aggregation state of the fruit and vegetable's own endogenous pectin through physical and mild physicochemical means, generating micron-sized gel particles with pH-intelligent response characteristics in situ. This achieves stable tolerance in the stomach and targeted sustained release in the intestines, while also regulating intestinal flora homeostasis and repairing intestinal mucosa. It meets the stringent raw material compliance and functional requirements of special medical foods.

[0037] Ordinary fruit and vegetable powders dissolve completely upon contact with water, with the carbon source concentrated in the proximal colon, resulting in explosive fermentation over 6-12 hours. This invention, however, uses endogenous pectin to form pH-responsive cross-linked gel microparticles. Under the strong acidity of the stomach, the cross-links tighten, maintaining the integrity of the solid particles. Dietary fiber is locked within the cross-linked network and does not dissolve all at once. Only a very small amount of small molecules slowly penetrate into the colon, where the cross-links break layer by layer from the outside in, releasing the substrate in a gradient. This achieves a uniform and stable generation of short-chain fatty acids over 24-48 hours, continuously providing energy to the intestinal mucosa and promoting repair.

[0038] To achieve the above-mentioned effects, the present invention employs the following technical means: the composite fruit and vegetable powder is a fruit and vegetable powder treated with endogenous pectin gel microparticle technology. Specifically, the present invention forms micron-sized intelligent gel microparticles through controlled deesterification of endogenous pectin in fruits and vegetables and in-situ self-crosslinking of endogenous ions. The main steps, functions, and effects of the preparation method are as follows:

[0039] 1. Low-temperature enzyme inactivation treatment: Wash and cut fresh fruits and vegetables, blanch at 85-90℃ for 10-20 seconds, pulp, inactivate polyphenol oxidase and pectin hydrolase, and retain native endogenous pectin.

[0040] 2. Physical decomposition achieved by ultra-high pressure micro-jet treatment: 120-160MPa pressure cycle for 3-5 times, with temperature ≤30℃ throughout, thereby breaking the long pectin chain, exposing carboxyl and hydroxyl active sites, and producing a particle size of 80-200nm;

[0041] 3. Precise pH gradient temperature control for endogenous deesterification: Short-time reaction at pH 8.0-8.5 and 20-25℃, with the deesterification rate precisely controlled at 35%-45%, converting high-ester pectin into medium- and low-ester pectin and obtaining pH-responsive characteristics;

[0042] 4. In-situ self-crosslinking of endogenous ions: Adding calcium ions, low-temperature static incubation + low-speed disturbance spontaneously forms 1-5μm gel microparticles with a particle size uniformity ≥90%;

[0043] 5. Directional low-temperature freeze-drying structure locking: Gradient directional freezing + vacuum freeze-drying locks in the microstructure of the gel, resulting in intelligent gel microparticle fruit and vegetable powder.

[0044] Through the above process, the intelligent gel microparticle composite fruit and vegetable powder obtained by the present invention has pH intelligent response characteristics: in the stomach environment of pH 1.2-2.0, the pectin carboxyl groups are ionized, the gel network shrinks and closes tightly, the structure is stable, and the active substances are not released; in the intestinal environment of pH 6.8-7.4, the carboxyl groups dissociate, the gel network slowly swells and dissociates, and the intestinal targeted sustained release is achieved.

[0045] In summary, the present invention can bring the following unique and beneficial effects:

[0046] 1. Whole grain and tuber food matrix, close to the composition of real food: This invention adopts a whole food source carbohydrate system of compound grain powder + endogenous pectin gel microparticle fruit and vegetable powder, which retains the integrity and synergistic effect of natural food matrix. It is different from the traditional "separation-purification-compounding" mode of nutritional preparations, and is closer to the nutritional composition of real food. It provides a variety of metabolic substrates for intestinal microorganisms and is conducive to maintaining the balance of intestinal microecology.

[0047] 2. Endogenous pectin gel microparticle technology to continuously provide energy to the intestinal mucosa: This invention applies endogenous pectin intelligent gel microparticle technology to whole food-based nutritional preparations. Through a pH-responsive gel network structure, it achieves stable tolerance in the stomach, targeted sustained release in the intestine, and continuous fermentation throughout the colon, resulting in good intestinal tolerance.

[0048] 3. Efficient delivery of active substances and enhanced mucosal repair: Intelligent gel microparticles protect active substances such as fruit and vegetable polyphenols and flavonoids from passing through the acidic environment of the stomach and are released in the colon to improve bioavailability; at the same time, slow-release fermentation increases butyric acid production, providing continuous energy to the whole colon. Combined with the mucosal repair effects of yam powder and zinc, a triple mucosal repair effect of "active substances + butyric acid + nutrients" is formed, which significantly promotes intestinal mucosal repair.

[0049] 4. Regulation of gut microbiota homeostasis and improvement of intestinal microecology: The insoluble fiber of compound grain powder, the slow-release fiber of gel microparticles, and the added prebiotics form a "dual-channel fermentation system" that provides carbon source for the gut microbiota from the proximal to the distal end of the colon. This avoids the gut microbiota fluctuations caused by the "explosive fermentation-rapid depletion" of ordinary prebiotics, thereby achieving gut microbiota homeostasis and significantly improving gut microbiota diversity and the proportion of beneficial bacteria.

[0050] 5. All-natural with zero additives, the special medical purpose compliant gel microparticles are formed entirely by the self-crosslinking of endogenous pectin from fruits and vegetables, with zero exogenous carriers, zero encapsulating agents, and zero exogenous colloids, fully complying with the stringent requirements for raw materials in the "General Rules for Foods for Special Medical Purposes".

[0051] 6. "Three-in-One" Synergistic Effect System: This invention constructs a three-in-one synergistic effect system of "nutritional supply-microbiota regulation-mucosal repair", which can not only meet basic nutritional needs, but also systematically improve intestinal health and promote intestinal mucosal repair. It is suitable for nutritional support for people with sensitive intestines and has broad clinical application prospects. Attached Figure Description

[0052] Figure 1 This is a graph showing the effects of different nutritional preparations on mouse body weight changes, colon length, and spleen index in Test Example 1 of the present invention. Figure 1 Figure A shows the weight changes of mice in each group; Figure 1 Figure B shows the comparison results of colon length in each group of mice; Figure 1 The graph in center C shows the comparison results of spleen indices in mice of different groups. Figure 1 The image in D shows typical photographs of the colon and spleen of mice in each group.

[0053] Figure 2 This is a graph showing the effects of different nutritional preparations on the degree of damage and inflammation in the colonic tissue of mice in Test Example 2 of the present invention. Figure 2 Image A shows a microscopic pathological image of colon tissue from each group of mice stained with HE (magnification ×100). Figure 2 Figure B shows the histopathological scoring results of the colon tissue of mice in each group.

[0054] Figure 3 The figure shows the effect of different nutritional preparations on the level of the inflammatory factor IL-6 in mice in Test Example 3 of the present invention.

[0055] Figure 4 This is a graph showing the effects of different nutritional preparations on the intestinal microecological diversity of mice in Test Example 4 of the present invention. Among them, Figure 4 Figure A shows the ACE analysis results of the gut microbiota of mice in each group; Figure 4 Figure B shows the results of Chao1 analysis of the gut microbiota of mice in each group; Figure 4 The middle section (C) shows the Shannon analysis results of the gut microbiota of each group of mice; Figure 4 Figure D shows the Simpson analysis results of the gut microbiota of mice in each group; Figure 4 Figure E shows the PCoA analysis results of the gut microbiota of mice in each group; Figure 4 The figure in the middle F is the NMDS analysis results of the intestinal flora of mice in each group. In the figure, Example 1 is Example 1, MBF is Comparative Example 1, and HC is the healthy control.

[0056] Figure 5 This is a graph showing the effects of different nutritional preparations on the genus-level composition of the intestinal flora in mice in Test Example 5 of the present invention. Among them, Figure 5 In Figure A, the relative abundance of gut microbiota at the genus level is shown for each group and individual mouse samples within each group. Figure 5 Plot B shows the average relative abundance of gut microbiota at the genus level for each group; Figure 5 The figure in middle C shows the comparison results of Lactobacillus levels among the groups; Figure 5 Figure D shows the comparison results of Helicobacter spp. levels among the groups. In the figure, Example 1 is Example 1, MBF is Comparative Example 1, and HC is the healthy control.

[0057] Figure 6 This is a graph showing the effects of different nutritional formulations on the small intestinal propulsion rate and fecal water content of mice in Test Example 6 of the present invention. Figure 6 Figure A shows the comparison results of small intestinal propulsion rate in mice of different groups; Figure 6 Figure B shows the comparison results of fecal water content in each group of mice; Figure 6 Figure C shows the effect of the absence of different components in the nutritional formulation of this application on the small intestinal propulsion rate in mice. Figure 6 Figure D shows the effect of the absence of different components in the nutritional formulation of this application on the water content of mouse feces.

[0058] Figure 7 This is a graph showing the effects of different proportions of soluble dietary fiber on the histopathological score of mouse colon tissue and on the levels of inflammatory factors in test example 7 of the present invention. Figure 7 Figure A shows the comparison of the effects of different proportions of soluble dietary fiber on the histopathological scores of mouse colon tissue; Figure 7 Figure B shows the comparison results of the effects of different proportions of soluble dietary fiber on the levels of inflammatory factors in mice. Detailed Implementation

[0059] To facilitate understanding of the technical means, creative features, objectives, and effects of this invention, the following embodiments, in conjunction with the accompanying drawings, specifically illustrate the nutritional formulation and preparation method of the whole food matrix for improving gut microbiota and promoting intestinal mucosal repair. In the following embodiments, reagents and materials not explicitly stated were obtained through general commercial channels, and experimental operations and conditions not described were performed in accordance with conventional practices and conditions in the art.

[0060] Example 1

[0061] The whole food matrix nutritional preparation for improving gut microbiota and promoting intestinal mucosal repair in this embodiment comprises, by weight: 20-50 parts of compound grain powder, 1-20 parts of compound fruit and vegetable powder, 10-35 parts of vegetable fat powder, 5-25 parts of protein, 0.1-10 parts of prebiotics, 2-10 parts of compound minerals, 0.05-1.5 parts of compound vitamins, 0-5 parts of meat meal, 0-1 part of thickener, and 0-2 parts of acidity regulator, or one or more of these components in combination.

[0062] The core technology of this invention lies in the fact that compound grain powder, compound fruit and vegetable powder and prebiotics are used together as sources of total dietary fiber, and the weight of soluble dietary fiber accounts for more than 50% of the total dietary fiber.

[0063] The compound cereal powder must include yam powder, and the yam powder accounts for 0.5%-5% of the total mass of the compound cereal powder; the compound mineral powder must include zinc, and the zinc content is 2-6mg / 100g of nutritional preparation.

[0064] Specifically, the specific components of the whole food matrix nutritional preparation for improving gut microbiota and promoting intestinal mucosal repair in this embodiment are as follows:

[0065] 38.5 parts of compound grain powder (37 parts rice powder, 1.5 parts yam powder), 11 parts of compound fruit and vegetable powder (3 parts apple powder, 2 parts strawberry powder, 4 parts pumpkin powder, 1 part broccoli powder, 1 part tomato powder), 30 parts of vegetable fat powder, 10 parts of soy protein isolate, 3 parts of fructooligosaccharides, 7 parts of compound minerals (including 0.005 parts zinc), and 0.5 parts of compound vitamins.

[0066] The composite mineral is a combination of potassium, calcium, magnesium, iron, zinc, copper, manganese, iodine, phosphorus, selenium and sodium, with the mass ratio of each component being 0.5:0.45:0.1:0.006:0.005:0.0005:0.0008:0.0001:0.3:0.0001:0.3.

[0067] The multivitamin contains vitamin A, vitamin D, vitamin E, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, folic acid, pantothenic acid, vitamin C, and biotin. The mass ratio of each component is 0.0002:0.000007:0.006:0.0005:0.0005:0.0005:0.000008:0.046:0.003:0.0001:0.002:0.00001.

[0068] In this embodiment, the compound fruit and vegetable powder is obtained through endogenous pectin gel microparticle processing technology, and the process includes the following steps:

[0069] Step S101, Low-temperature enzyme inactivation treatment: Wash and cut fresh fruits and vegetables, blanch at 90℃ for 15 seconds, and then pulp to form fruit and vegetable pulp.

[0070] Step S102, Ultra-high pressure micro-jet treatment: The fruit and vegetable pulp obtained in step S101 is treated with an ultra-high pressure micro-jet device to obtain a treated slurry. The treatment conditions are 140MPa pressure for 3 cycles, temperature ≤28℃ throughout the process, and output particle size D50 of about 150nm.

[0071] Step S103, deesterification treatment: The pH of the treated slurry was finely adjusted to 8.2 with 0.5 mol / L sodium carbonate solution, and the reaction was stirred at a constant temperature of 22℃ for 40 min. The reaction was terminated with dilute hydrochloric acid to obtain a deesterified slurry. The pectin deesterification rate was tested to be 38%.

[0072] Step S104, self-crosslinking treatment: The pH of the deesterified slurry was adjusted to 5.5, and 1.1 g of calcium chloride was added per liter of slurry. The mixture was stirred at low speed (80 rpm) for 2 hours, and then allowed to stand at 4°C for 12 hours. During this process, the endogenous pectin in the deesterified slurry spontaneously crosslinked with calcium ions to form gel microparticles, resulting in a gel slurry. Testing showed that the average particle size of the microparticles in the gel slurry was 2.8 μm, with a particle size uniformity of 92%.

[0073] Step S105, freeze-drying: The gel slurry is pre-frozen in a gradient manner using a freeze dryer (-10℃ / 2h → -40℃ / 12h) at a vacuum of 40Pa for 36h. The freeze-dried solid is then pulverized using a low-temperature airflow to obtain gel microparticle fruit and vegetable powder, which is used as the composite fruit and vegetable powder in this embodiment.

[0074] To compare the gel microparticle fruit and vegetable powder obtained by the above process with conventional fruit and vegetable powder, this embodiment also prepared spray-dried fruit and vegetable powder by conventional spray drying process. The main process of this spray drying process is as follows:

[0075] Step 1, Low-temperature enzyme inactivation treatment: Wash and cut fresh fruits and vegetables, blanch them at 90℃ for 15 seconds, and then blend them into a fruit and vegetable pulp.

[0076] Step 2, sterilization: Boil the slurry at 80℃ for 15 minutes.

[0077] Step 3, Homogenization: Homogenization at 10 MPa makes the material particles more dispersed.

[0078] Step 4, spray drying: inlet temperature 160℃, outlet temperature 90℃.

[0079] The performance of gel microparticle fruit and vegetable powder and spray-dried fruit and vegetable powder were tested separately. The main testing methods are as follows:

[0080] 1. Gastric swelling rate

[0081] 1.1 Principle

[0082] By simulating the gastric fluid environment, the volume expansion factor of fruit and vegetable powder gel microparticles over a certain period of time was measured to evaluate their gastric tolerance. The lower the swelling rate, the more stable the microparticle structure in the stomach, and the better the gastric tolerance.

[0083] 1.2 Reagents and Instruments

[0084] Simulated gastric juice (SGF): pH 1.2, prepared by adding 2.0g of sodium chloride and 7ml of hydrochloric acid, then diluting with water to 1000ml.

[0085] 10ml graduated centrifuge tubes with stoppers

[0086] Constant temperature water bath shaker (37±1℃)

[0087] Analytical balance (accuracy 0.1 mg)

[0088] low-speed centrifuge

[0089] 1.3 Measurement Procedure

[0090] a. Accurately weigh 0.200g (W0) of the fruit and vegetable powder sample and place it in a 10ml stoppered graduated centrifuge tube;

[0091] b. Add 5.0 ml of simulated gastric fluid preheated to 37°C, and gently shake to disperse the sample evenly;

[0092] c. Place the centrifuge tubes in a 37°C constant temperature water bath shaker and incubate them by shaking at 100 rpm.

[0093] d. After incubating for 2 hours, remove the product and centrifuge at 3000 rpm for 5 minutes;

[0094] e. Read the volume V (ml) of the precipitate layer at the bottom of the centrifuge tube, which is the volume after swelling.

[0095] 1.4 Calculation Formula

[0096] Swelling rate (%) = (V - V0) / V0 × 100%

[0097] Where: V is the volume of precipitate after swelling (ml), and V0 is the initial volume of the sample (ml), which is calculated by dividing the sample mass by the bulk density.

[0098] 2. Intestinal release rate

[0099] 2.1 Principle

[0100] The dynamic dialysis method was used to simulate the gastric and intestinal fluid environments sequentially, and the cumulative release of active substances (with total polyphenols as the indicator) at different time points was measured to evaluate the intestinal targeted sustained-release performance of gel microparticles.

[0101] 2.2 Reagents and Instruments

[0102] Simulated gastric juice (SGF): pH 1.2, same as 10.1.2

[0103] Simulated intestinal fluid (SIF): pH 6.8. Dissolve 6.8g of potassium dihydrogen phosphate in 500ml of water, adjust the pH to 6.8 with 0.1mol / L sodium hydroxide solution, and dilute with water to 1000ml.

[0104] Dialysis bags (molecular weight cutoff 8000–14000 Da)

[0105] Intelligent dissolution tester (paddle method)

[0106] UV-Vis spectrophotometer

[0107] Constant temperature water bath (37±0.5℃)

[0108] 2.3 Measurement Procedure

[0109] a. Accurately weigh 0.500g of fruit and vegetable powder sample, place it in a pretreated dialysis bag, add 2ml of simulated gastric fluid, and clamp both ends tightly;

[0110] b. Place the dialysis bag into the dissolution cup, add 500ml of simulated gastric fluid preheated to 37℃ as the release medium, paddle speed 50rpm, temperature 37℃;

[0111] c. Incubate in gastric juice for 2 hours, and take 2 ml samples at 0.5 h, 1 h and 2 h respectively (add an equal amount of isothermal fresh medium at the same time).

[0112] d. After the gastric juice stage, transfer the dialysis bag to a dissolution vessel containing 500 ml of preheated simulated intestinal fluid and continue incubation for 8 hours;

[0113] e. During the intestinal fluid stage, 2 ml samples were taken at 1 h, 2 h, 4 h, 6 h, and 8 h (with simultaneous fluid replacement);

[0114] f. The total polyphenol content in the samples at each time point was determined by the Folin-phenol method.

[0115] 2.4 Calculation Formula

[0116] Cumulative release rate (%) = M t / M0 × 100%

[0117] Where: M t M0 represents the cumulative mass of polyphenols released at time t (mg), and M0 represents the initial mass of total polyphenols in the sample (mg).

[0118] 3. Polyphenol retention rate

[0119] 3.1 Principle

[0120] Using the total polyphenol content in fresh fruit and vegetable raw materials as a benchmark, the residual content of total polyphenols in the finished fruit and vegetable powder was determined, the retention rate was calculated, and the protective effect of the process on active substances was evaluated. The Folin-Ciocalteu colorimetric method was used to determine the total polyphenol content.

[0121] 3.2 Reagents and Instruments

[0122] Folin-phenol reagent

[0123] 7.5% sodium carbonate solution

[0124] Gallic acid standard

[0125] 60% ethanol solution (by volume)

[0126] Ultraviolet-Vis spectrophotometer (765nm wavelength)

[0127] ultrasonic cleaner

[0128] low-speed centrifuge

[0129] 3.3 Preparation of Standard Curve

[0130] Accurately weigh 10.0 mg of gallic acid standard dried to constant weight, dissolve it in distilled water and dilute to 100 ml to obtain a 100 μg / ml standard stock solution;

[0131] Pipette 0, 0.2, 0.4, 0.6, 0.8, and 1.0 ml of the standard stock solution into 10 ml volumetric flasks, and add distilled water to each flask to a final volume of 5 ml.

[0132] Add 0.5 ml of Folin-phenol reagent, shake well, and let stand for 3 minutes;

[0133] Add 1.5 ml of 7.5% sodium carbonate solution, dilute to the mark with distilled water, and shake well;

[0134] After being placed at 25℃ in the dark for 60 minutes, the absorbance was measured at a wavelength of 765nm.

[0135] A standard curve was plotted with gallic acid concentration on the x-axis and absorbance on the y-axis.

[0136] 3.4 Sample determination procedure

[0137] Fresh sample determination: Take fresh fruit and vegetable raw materials, homogenize them, accurately weigh 2.0g, add 20ml of 60% ethanol, extract by sonication for 30min, centrifuge at 4000rpm for 10min, and collect the supernatant. Extract twice in the same way, combine the supernatants, and make up to 50ml. Measure the absorbance according to the standard curve method and calculate the total polyphenol content C0 (mg / g fresh weight).

[0138] Finished product testing: Accurately weigh 0.200g of fruit and vegetable powder sample, add 20ml of 60% ethanol, extract ultrasonically for 30min, centrifuge and collect the supernatant, extract twice using the same method, combine and dilute to 50ml. Measure the absorbance and calculate the total polyphenol content C1 (mg / g dry powder).

[0139] Conversion factor: The starch yield of fresh fruits and vegetables (the number of grams of dry powder obtained from 100g of fresh fruits and vegetables), denoted as K (g / 100g).

[0140] 3.5 Calculation Formula

[0141] Polyphenol retention rate (%) = (C1 × K / 100) / C0 × 100%

[0142] Simplified to: Polyphenol Retention Rate (%) = Total Polyphenol Content of Finished Product / Equivalent Total Polyphenol Content of Fresh Sample × 100%

[0143] In this embodiment, the gel microparticle fruit and vegetable powder and the spray-dried fruit and vegetable powder were tested using the above-described performance testing process, and the results are shown in Table 1 below:

[0144] Table 1 Performance Test Results

[0145]

[0146] This embodiment also provides a method for preparing a whole food matrix nutritional preparation that improves gut microbiota and promotes intestinal mucosal repair, including the following steps:

[0147] S201: Weigh each component according to the aforementioned weight proportions.

[0148] S202: The weighed components are dry-mixed using an airflow mixer for 10 minutes.

[0149] S203: Nitrogen gas is introduced into the mixed components for protection to obtain a nutrient preparation.

[0150] Comparative Example 1

[0151] This comparative example provides a nutritional preparation with low soluble dietary fiber. The difference between the comparative example and Example 1 is that maltodextrin (14 parts) is used in equal amounts to replace the compound fruit and vegetable powder and prebiotics in Example 1. All other components and preparation methods are the same as in Example 1.

[0152] In this comparative example, since the total dietary fiber comes only from the natural dietary fiber in the compound grain powder, the proportion of soluble dietary fiber is significantly lower than 50%. At the same time, since compound fruit and vegetable powder was not used, it also lacks the natural active substances in fruits and vegetables (such as phenols, flavonoids, terpenoids, etc.).

[0153] Comparative Example 2

[0154] This comparative example provides a nutritional preparation that does not contain yam or zinc. The difference from Example 1 is that millet flour (1.5 parts) is used to replace the yam powder in Example 1 in equal amounts, and the compound minerals do not include zinc. All other components and preparation methods are the same as in Example 1.

[0155] Comparative Example 3

[0156] This comparative example provides a nutritional preparation with 30% soluble dietary fiber. The difference from Example 1 is that the amount of compound fruit and vegetable powder added is 6.5 parts (containing apple powder, strawberry powder, pumpkin powder, broccoli powder, and tomato powder in the same proportion as in Example 1), the amount of fructooligosaccharides added is 2 parts, and the amount of compound cereal powder added is 44 parts. Other components and preparation methods are the same as in Example 1.

[0157] Comparative Example 4

[0158] This comparative example provides a nutritional preparation with 20% soluble dietary fiber. The difference from Example 1 is that the amount of compound fruit and vegetable powder added is 4.5 parts (containing apple powder, strawberry powder, pumpkin powder, broccoli powder, and tomato powder in the same proportion as in Example 1), the amount of fructooligosaccharides added is 2 parts, and the amount of compound cereal powder added is 44 parts. Other components and preparation methods are the same as in Example 1.

[0159] Test Example 1

[0160] This test case investigated the effects of different nutritional formulations on changes in body weight, colon length, and spleen index in C57BL / 6 mice.

[0161] The experimental method for this test case is as follows:

[0162] 1. LPS-induced systemic inflammation mouse model

[0163] Principle: LPS (lipopolysaccharide) is a major component of the cell wall of Gram-negative bacteria. It can be recognized by the host immune system as a pathogen-associated molecular pattern (PAMP), thereby triggering a strong innate immune response.

[0164] step:

[0165] a. Five-week-old C57BL / 6 mice (hereinafter referred to as mice) were randomly divided into four groups: a healthy control group, Example 1 group, Comparative Example 1 group, and Comparative Example 2 group. After one week of acclimatization feeding, except for the healthy control group, the mice in the other groups were injected intraperitoneally with 3 mg / kg LPS once. The healthy control group mice were injected intraperitoneally with an equal volume of physiological saline.

[0166] b. Observe and record symptoms such as weight loss, diarrhea, and mental state.

[0167] 2. Preparation and feeding of Example 1, Comparative Example 1, and Comparative Example 2

[0168] a. Weigh 8g of Example 1 and add it to 30ml of sterile double-distilled water and mix well;

[0169] b. Weigh out 18g of the comparative example and add it to 30ml of sterile double-distilled water and mix well;

[0170] c. Weigh out 28g of the comparative example and add it to 30ml of sterile double-distilled water and mix well.

[0171] The prepared nutrient solutions were administered to each group of mice via gavage daily at a volume of 0.2 mL / 10g body weight, continuing until the end of the experiment. Mice had free access to water.

[0172] Healthy control mice were not given any nutritional supplements by gavage, but ate and drank normally.

[0173] 3. Mouse fecal collection

[0174] Before the end of the experiment, use the natural defecation method (suitable for short-term experiments): Place the mice individually in a clean cage (lined with filter paper) and observe for 30-60 minutes, collecting 2-3 fresh feces (avoiding urine contamination). If the amount of feces is insufficient, stimulate and gently press the mouse's lower abdomen with a warm, moist cotton ball to assist defecation. Immediately after collecting the feces, place them in liquid nitrogen for flash freezing and store at -80°C for later use.

[0175] 4. Colon sampling

[0176] a. The mouse was euthanized by cervical dislocation, and the intact colon and rectum from the ileocecal junction to the anus were quickly removed by laparotomy. The colon and rectum were placed in ice-cold PBS solution and gently rinsed to remove the intestinal contents.

[0177] b. Measurement data

[0178] Gently straighten the intestines, avoiding stretching and deformation, and measure the total length of the colon and rectum.

[0179] c. Remove about 1 cm of the rectum at the end, then cut off 1 cm of the colon and immerse it in 4% paraformaldehyde.

[0180] 5. Spleen collection and weighing in mice

[0181] a. After colon sampling, the mouse spleen was removed by laparotomy, placed in ice-cold PBS solution, and then blotted dry with filter paper.

[0182] b. Quickly weigh the spleen using an electronic balance and take a picture.

[0183] Figure 1 This is a graph showing the effects of different nutritional preparations on mouse body weight changes, colon length, and spleen index in Test Example 1 of the present invention. Figure 1 Figure A shows the weight changes of mice in different groups; Figure 1 Figure B shows the comparison of colon length in different groups of mice; Figure 1 The graph in center C shows the comparison results of spleen indices in different groups of mice. Figure 1 In the middle D, typical photographs of the colon and spleen of mice from different groups are shown. (*** indicates p<0.001; ** indicates p<0.01; * indicates p<0.05; no indication means no difference between groups, p>0.05, and subsequent test cases are consistent.)

[0184] Mouse weight changes: such as Figure 1 As shown in Figure A, there was no significant difference in body weight change between the mice in Example 1 group and Comparative Example 1 and Comparative Example 2 groups.

[0185] Colon length: as Figure 1 China B and Figure 1 As shown in Figure D, there were no significant changes in colon length among the groups, and no significant difference compared with the healthy controls (p>0.05).

[0186] Spleen index: such as Figure 1 As shown in Figure C, the spleen index (spleen weight / mouse body weight × 100%) showed no significant difference among the various nutritional intervention groups (Example 1, Comparative Example 1, Comparative Example 2), but was significantly higher than that of the healthy control group (p < 0.001). Figure 1 As shown in Figure D, there was no significant difference in spleen length among the groups, but the spleen index was significantly increased in the nutritional preparation intervention group, indicating that the LPS-induced inflammation model was successfully established.

[0187] Test Example 2

[0188] This test case examines the effects of different nutritional preparations on colonic tissue damage and intestinal mucosal repair in mice.

[0189] The experimental method for this test case is as follows:

[0190] 1. HE staining of colon tissue

[0191] a. Material collection and fixation: The distal colon segment of the mouse was cut off with scissors and fixed in formalin.

[0192] b. Dehydration and Clearing: Gradient alcohols are used as dehydrating agents to remove water from the colon tissue. The tissue is then placed in xylene, a clearing agent, to replace the alcohol in the tissue.

[0193] c. Paraffin embedding: The transparent tissue block is placed in melted paraffin wax and embedded after the paraffin wax has completely penetrated the tissue.

[0194] d. Sectioning and mounting: Fix the embedded tissue block in a microtome and cut it into 5-8 μm thick sections. Place the cut sections in hot water to flatten them and mount them onto glass slides, then dry them in a 45°C oven.

[0195] e. Dewaxing: Immerse the sections in the following solutions in sequence: Xylene I for 10 minutes → Xylene II for 10 minutes → Anhydrous ethanol for 15 minutes → Anhydrous ethanol II for 5 minutes → 95% ethanol for 5 minutes → 90% ethanol for 5 minutes → 80% ethanol for 5 minutes → 70% ethanol for 5 minutes → Wash with distilled water.

[0196] f. Staining: Colon sections were stained in hematoxylin solution for 5 minutes, rinsed with tap water, and then differentiated and blued using 1% hydrochloric acid alcohol and 0.6% ammonia solution, respectively, followed by rinsing with tap water. Finally, the sections were stained in eosin solution for 1-3 minutes.

[0197] g. Dehydration and sealing: Dehydration and transparency are achieved using gradient alcohols and xylene, followed by sealing with neutral resin.

[0198] h. Observation: Observe and take pictures using a microscope.

[0199] HE staining assesses the degree of colonic tissue damage and inflammation. The Histological score includes inflammation (0-5 points), crypt damage (0-4 points), ulcers (0-3 points), and edema (0-1 points). The total score is the sum of the scores of each item. The higher the score, the stronger the inflammation and the greater the degree of mucosal damage.

[0200] Figure 2 This is a graph showing the effects of different nutritional preparations on the degree of damage and inflammation in the colonic tissue of mice in Test Example 2 of the present invention. Figure 2 Image A shows a microscopic pathological image of colon tissue from each group of mice stained with HE (magnification ×100). Figure 2 Figure B shows the histopathological scoring results of the colon tissue of mice in each group.

[0201] like Figure 2As shown in Figure A, the colonic mucosa of healthy control mice was intact, with glands arranged neatly and no obvious inflammatory cell infiltration. After LPS induction, all intervention groups showed varying degrees of colonic damage, but the mucosal edema, villous structure disorder, inflammatory infiltration, and crypt deformation in Example 1 group were significantly less than those in Comparative Example 1 and Comparative Example 2, indicating that Example 1 can effectively repair intestinal mucosal damage. In addition, the intestinal mucosal repair function was significantly reduced in the group lacking soluble dietary fiber (Comparative Example 1) and the group lacking yam and zinc (Comparative Example 2), which also indicates that soluble dietary fiber and yam + zinc have a synergistic effect, and both are indispensable in promoting intestinal mucosal repair.

[0202] like Figure 2 As shown in Figure B, the quantitative scoring results indicate that the total score for colitis in Example 1 group was approximately 5 points; the total score for Comparative Example 1 group was between 5 and 6 points; and the total score for Comparative Example 2 group was close to 6 points. Although there was no statistically significant difference between the scores of Example 1 group and the two comparative example groups (p>0.05), its score was lower than that of the two comparative example groups, suggesting that Example 1 may have a certain effect in alleviating intestinal mucosal inflammation.

[0203] It is worth noting that Comparative Example 1, containing yam powder and zinc (mucosal repair components) but lacking soluble dietary fiber, had a higher colitis score than Example 1. Comparative Example 2, while containing soluble dietary fiber (prebiotics + compound fruit and vegetable powder), lacked yam powder and zinc, and thus had the highest score. Example 1, containing both mucosal repair components (yam powder, zinc) and soluble dietary fiber components (prebiotics, compound fruit and vegetable powder), had the lowest score. This result is consistent with the design concept of the synergistic effect of the "microbial colonization engine + mucosal repair engine" of this invention: providing only mucosal repair components (such as Comparative Example 1) or only soluble dietary fiber (such as Comparative Example 2) is insufficient to achieve the best repair effect; only the synergistic effect of both can alleviate intestinal mucosal inflammation and promote intestinal repair.

[0204] Test Example 3

[0205] This test case investigated the effects of different nutritional formulations on the level of the inflammatory cytokine IL-6 in mice. IL-6 is an important pro-inflammatory cytokine released after damage to the intestinal mucosa; higher IL-6 levels indicate stronger active inflammation of the mucosa.

[0206] The experimental method for this test case is as follows:

[0207] After the experiment, mice were anesthetized, blood was collected from their eyes, and after resting for 2 hours, the blood was centrifuged at 3000 rpm for 10 minutes. The serum was collected in clean centrifuge tubes and stored at -20°C. The level of IL-6 in the blood was detected using an ELISA kit.

[0208] Figure 3The figure shows the effect of different nutritional preparations on the level of inflammatory factors in mice in Test Example 3 of the present invention (ns indicates no statistical difference between groups, p>0.05).

[0209] like Figure 3 As shown, the serum IL-6 level in the healthy control group mice was low, while the IL-6 level in the comparative group 1 was significantly higher than that in the healthy control group (p < 0.05), indicating that the inflammatory response in this group of mice was more pronounced. In contrast, the IL-6 level in the mice in the Example 1 group was not significantly different from that in the healthy control group (p > 0.05), indicating that the inflammation level was effectively controlled and the active mucosal inflammation was reduced.

[0210] The above results indicate that Example 1 has a stronger inhibitory effect on inflammation levels than Comparative Example 1, and can maintain the IL-6 level in LPS-induced inflammatory mice at a level close to that of the healthy control group. This is consistent with the design concept of the anti-inflammatory and antioxidant components in the "mucosal repair engine" of this invention (if the glucosinolates in the fruit and vegetable powder are metabolized into sulforaphane to activate the Nrf2 pathway and anthocyanins to inhibit the NF-κB pathway). The fruit and vegetable powder of Example 1 is rich in a variety of anti-inflammatory active ingredients, which synergistically inhibit the release of inflammatory factors such as IL-6; while Comparative Example 1, due to the lack of fruit and vegetable powder, has insufficient anti-inflammatory components, weakened anti-inflammatory effect, and significantly increased IL-6 level.

[0211] Test Example 4

[0212] This test case investigated the effects of different nutritional formulations on the diversity of the gut microbiota in mice. In the gut, gut microbiota homeostasis is fundamental to mucosal repair.

[0213] The experimental method for this test case is as follows:

[0214] 1. Gut microbiota testing

[0215] a. Collect intestinal contents from mouse models

[0216] b. DNA Extraction: Genomic DNA was extracted from the samples using a DNA extraction kit, and the DNA concentration was then detected using agarose gel electrophoresis and NanoDrop2000. Using the genomic DNA as a template, PCR was performed using barcode-specific primers and Takara's TksGflex DNA Polymerase, depending on the selected sequencing region, to ensure amplification efficiency and accuracy.

[0217] c. PCR amplification of the 16S rRNA gene

[0218] (1) Primer selection (amplification of V3-V4 region):

[0219] Forward primer: 343F 5′-TACGGRAGGCAGCAG-3′ Reverse primer: 798R 5′AGGGTATCTAATCCT-3′

[0220] (2) Prepare the reaction system (25 μL):

[0221]

[0222] (3) Set up the amplification program:

[0223] 94℃ for 5 min

[0224] 94℃ 30 sec, 56℃ 30 sec, 72℃ 20 sec (26 cycles)

[0225] 72℃ for 5 min

[0226] (4) PCR products were obtained after the PCR program was completed.

[0227] c. Library construction and sequencing

[0228] (1) Purification of PCR products: PCR products were purified using AMPure XP magnetic beads to remove primers, unreacted nucleotides and other impurities from the PCR reaction.

[0229] (2) Add adapter and index: Use the Illumina Nextera XT Index Kit to add adapter and index sequence to the purified PCR product.

[0230] (3) Library quantification: Qubit + Agilent Bioanalyzer (detecting fragment size).

[0231] Specifically, to ensure library quality and sequencing accuracy, quantification and quality assessment were performed after library construction. The concentration of the DNA library was determined using a Qubit real-time quantitation system to ensure sufficient DNA for subsequent sequencing. Simultaneously, fragment size analysis of the library was performed using an Agilent Bioanalyzer to assess the size distribution of DNA fragments and ensure that the fragment sizes met the requirements of the sequencing platform.

[0232] (4) Sequencing platform: Illumina MiSeq / NovaSeq (paired-end 250 / 300 bp).

[0233] Specifically, the constructed libraries are sequenced using Illumina MiSeq or NovaSeq sequencing platforms. These platforms employ paired-end sequencing technology, reading sequence information from both ends of the DNA fragment, typically with a sequencing length of 250 or 300 base pairs (bp).

[0234] 2. Diversity Analysis

[0235] a. Alpha diversity (within a single sample)

[0236] The intestinal flora of mice in each nutritional preparation group and the healthy control group were analyzed for ACE, abundance (Chao1), diversity (Shannon), and dominance (Simpson).

[0237] b. β-diversity (differences between samples)

[0238] β-diversity analysis was performed on the gut microbiota of mice in each nutritional preparation group and the healthy control group, including principal coordinate analysis (PCoA) based on Bray-Curtis distance and UniFrac distance and non-metric multidimensional scaling (NMDS).

[0239] Figure 4 This is a graph showing the effects of different nutritional preparations on the intestinal microecological diversity of mice in Test Example 4 of the present invention. Among them, Figure 4 Figure A shows the ACE analysis results of the gut microbiota of mice in each group; Figure 4 Figure B shows the results of Chao1 analysis of the gut microbiota of mice in each group; Figure 4 The middle section (C) shows the Shannon analysis results of the gut microbiota of each group of mice; Figure 4 Figure D shows the Simpson analysis results of the gut microbiota of mice in each group; Figure 4 Figure E shows the PCoA analysis results of the gut microbiota of mice in each group; Figure 4 The figure in the middle (F) shows the NMDS analysis results of the gut microbiota of mice in each group. Figure 4 In this context, FBF represents Example 1, MBF represents Comparative Example 1, and HC represents a healthy control.

[0240] 16S rRNA was sequenced from mouse feces, such as... Figure 4 China A and Figure 4 As shown in Figure B, ACE and Chao1 index analyses both revealed the highest microbial richness in the healthy control group. Comparative Example 1 showed the lowest richness, with significant differences compared to the other groups. Example 1 exhibited moderate richness and the lowest intra-group dispersion, indicating a uniform intervention effect.

[0241] like Figure 4As shown in Figure C, Shannon index analysis revealed that Comparative Example 1 had the lowest diversity, significantly lower than the healthy control group. Example 1 group showed no significant difference in diversity compared to the healthy control group, but was significantly higher than Comparative Example 1.

[0242] like Figure 4 As shown in Figure D, Simpson index analysis revealed that Example 1 group had the highest uniformity, followed by the healthy control group, and Comparative Example 1 group had the lowest. Comparative Example 1 group had significantly lower uniformity than the other groups and exhibited the largest intra-group dispersion.

[0243] Integrating four alpha diversity indices (ACE, Chao1, Shannon, and Simpson) revealed that the gut microbiota richness and evenness were highest in the healthy control group, followed by Example 1, which in turn was highest in Comparative Example 1. This indicates that the intervention in Example 1 improved the richness and diversity of gut microbiota, reconstructed the gut microbiota, and nourished the intestinal epithelium with secreted short-chain fatty acids and other nutrients, promoting continuous mucosal healing. The intervention effect of Example 1 was significantly better than that of Comparative Example 1.

[0244] like Figure 4 As shown in Figures E and F, both NMDS and PCoA analyses based on Bray-Curtis distance consistently revealed that the gut microbiota structure of Example 1 group was significantly separated from that of Comparative Example 1 group and the healthy control group. Comparative Example 1 group also showed significant intragroup variation in PCoA. These results indicate that the intervention in Example 1 group had a significant remodeling effect on the gut microbiota.

[0245] Integrating α and β diversity reveals that, compared to the healthy control group, the intervention in Example 1 reduced richness while increasing evenness, forming a "simplified but balanced" microbial community state with high consistency within the group. Example 1's intervention effect on intestinal microecological diversity is significantly better than that of Comparative Example 1. The addition of gel microparticles, fruit and vegetable powder, and more than 50% soluble dietary fiber significantly improves the diversity of intestinal microecology.

[0246] Test Example 5

[0247] This test case examines the effects of different nutritional formulations on the phylum and genus levels of the intestinal flora in mice.

[0248] To investigate the specific changes in the gut microbiota, this test case analyzed the main components of the intestinal microbiota of each group of mice at the phylum level. All groups were dominated by Firmicutes and Bacteroidetes at the phylum level, and there was no statistically significant difference in the F / B ratio (Firmicutes / Bacteroidetes) among the groups.

[0249] Figure 5 This is a graph showing the effects of different nutritional preparations on the genus-level composition of the intestinal flora in mice in Test Example 5 of the present invention. Among them, Figure 5In Figure A, the relative abundance of gut microbiota at the genus level is shown for each group and individual mouse samples within each group. Figure 5 Plot B shows the average relative abundance of gut microbiota at the genus level for each group; Figure 5 The figure in middle C shows the comparison results of Lactobacillus levels among the groups; Figure 5 Figure D shows the comparison results of Helicobacter genus levels among the groups. Figure 5 In this context, FBF represents Example 1, MBF represents Comparative Example 1, and HC represents a healthy control.

[0250] Plot a clustering histogram at the genus level, such as Figure 5 As shown in Figures A and B. Wherein, Figure 5 Figure A shows the relative abundance and clustering relationships of gut microbiota at the genus level in individual mouse samples within each group, reflecting the differences between individuals within each group. Figure 5 Figure B shows the average relative abundance of each group after merging, which facilitates comparison of the bacterial community composition characteristics between groups.

[0251] based on Figure 5 The clustering results of groups A and B were used to further compare the relative abundance of dominant bacterial genera among the groups.

[0252] Lactobacillus: such as Figure 5 As shown in Figure C, the relative abundance of Lactobacillus in the intestines of mice in Example 1 group was significantly higher than that in Comparative Example 1 group, indicating that the intervention in Example 1 can effectively promote the colonization of this beneficial bacterium, Lactobacillus.

[0253] Helicobacter: such as Figure 5 As shown in Figure D, the relative abundance of Helicobacter spp. in the intestines of mice in Example 1 group was significantly lower than that in healthy controls and Comparative Example 1 group. Some species of Helicobacter spp. are associated with intestinal inflammation, and their reduced abundance is beneficial in alleviating intestinal inflammatory responses.

[0254] The above results indicate that by regulating the ratio of soluble to insoluble dietary fiber, and combining the slow-release dietary fiber of the gel microparticle composite fruit and vegetable powder with the precise delivery of active substances in the fruit and vegetable powder, this invention can selectively promote the growth of beneficial bacteria such as Lactobacillus at the genus level, and inhibit the colonization of potentially harmful bacteria such as Helicobacter, thereby optimizing the intestinal flora structure and repairing the flora barrier. This can continuously protect the newly formed intestinal mucosa from further invasion and damage, which is consistent with the design concept of the aforementioned flora colonization engine.

[0255] Test Example 6

[0256] Intestinal motility and fecal physicochemical properties are external phenotypes of intestinal barrier function recovery. This test case investigated the effects of different nutritional formulations on small intestinal propulsion rate and fecal water content in mice to evaluate the impact of interventions on the recovery of intestinal barrier function.

[0257] The experimental procedure for this test case is as follows:

[0258] 1. Measurement of small intestinal propulsion rate

[0259] a. Administration of marker via gavage

[0260] After the mice were fixed, they were administered activated charcoal suspension (0.2 mL / 10 g) by gavage according to their body weight.

[0261] Record time: Start timing from the completion of gavage.

[0262] b. Execution and Sampling

[0263] Timing: 30 minutes after gavage.

[0264] Mice were euthanized by cervical dislocation, and the small intestine from the pylorus to the ileocecal junction was quickly removed by laparotomy and placed in ice-cold saline.

[0265] c. Measurement data

[0266] Gently straighten the small intestine to avoid stretching and deformation.

[0267] Measure the total length of the small intestine (Ltotal) and the distance from the front edge of the charred powder to the pylorus (Lcharred).

[0268] Calculate the small intestinal propulsion rate:

[0269] Propulsion rate (%) = (L carbon / L total) × 100%

[0270] 2. Determination of water content in mouse feces

[0271] a. Fecal sample collection

[0272] Natural defecation method (suitable for short-term experiments):

[0273] Mice were placed alone in clean cages (lined with filter paper) and observed for 30-60 minutes. Fresh fecal pellets were then collected.

[0274] b. Weigh the wet weight (W0).

[0275] Quickly weigh the fresh feces using an electronic balance.

[0276] Record the initial wet weight (W0) of each fecal pellet (unit: mg).

[0277] c. Drying feces

[0278] Oven drying method:

[0279] Place the feces in aluminum foil or a weighing dish.

[0280] Dry in an oven at 65℃ for 6 hours (until constant weight, i.e., the difference between two weighings is < 0.1 mg).

[0281] d. Weigh the dry weight (W1)

[0282] Remove the dried feces, cool it to room temperature, and weigh it (W1).

[0283] e. Calculate the water content

[0284] Moisture content (%) = (W0 − W1) / W0 × 100%

[0285] Figure 6 This is a graph showing the effects of different nutritional formulations on the small intestinal propulsion rate and fecal water content of mice in Test Example 6 of the present invention. Figure 6 Figures A and B show the effects of the absence of different components in the nutritional formulation of this application on the small intestinal propulsion rate and the water content of mouse feces, respectively.

[0286] like Figure 6 As shown in Figures A and B, compared to the formulations of Comparative Examples 1 and 2, the dietary treatment of the formulation in Example 1 significantly improved the small intestinal propulsion rate and fecal water content in mice, indicating that the formulation in Example 1 has a stronger effect on promoting intestinal motility.

[0287] Both the lack of dietary fiber (Comparative Example 1) and the lack of yam + zinc significantly affected the small intestinal propulsion rate and fecal water content in mice, indicating that the combination of "dietary fiber + yam + zinc" in Example 1 has a synergistic promoting effect on intestinal motility.

[0288] Test Example 7

[0289] This test case investigated the effects of different proportions of soluble dietary fiber on intestinal mucosal repair and the level of the inflammatory factor IL-6. The experimental method used was the same as that used in test cases 2 and 3.

[0290] Figure 7 This is a graph showing the effects of different proportions of soluble dietary fiber on the histopathological score of mouse colon tissue and on the levels of inflammatory factors in test example 7 of the present invention. Figure 7 Figure A shows the comparison of the effects of different proportions of soluble dietary fiber on the histopathological scores of mouse colon tissue; Figure 7 Figure B shows the comparison results of the effects of different proportions of soluble dietary fiber on the levels of inflammatory factors in mice.

[0291] like Figure 7 As shown in Figures A and B, the nutritional preparation with 50% soluble dietary fiber in Example 1 significantly enhanced the intestinal mucosal repair capacity and inhibited the inflammatory response in mice compared with the control preparations with 20% (Comparative Example 4) or 30% (Comparative Example 3) soluble dietary fiber, indicating that 50% soluble dietary fiber is the preferred range for achieving the effects of promoting repair and inhibiting inflammation.

[0292] Based on the experimental results of Test Examples 1-7 above, it can be concluded that the nutritional preparation prepared in this invention (Example 1) constructs a synergistic system integrating "nutritional supply - flora regulation - mucosal repair," utilizing a "dual-channel fermentation system" formed by compound grain powder, gel microparticle compound fruit and vegetable powder, and prebiotics, combined with yam powder, zinc, and active ingredients from fruit and vegetable powder to form a "mucosal repair engine." This significantly optimizes the intestinal flora structure (increasing the abundance of Lactobacillus and decreasing the abundance of Helicobacter, forming a "simplified and balanced" flora state), while inhibiting systemic and local intestinal inflammatory responses (reducing IL6 levels and alleviating colonic pathological damage). Thus, under the premise of meeting basic nutritional needs, it improves the intestinal flora of patients in disease states such as intestinal inflammation and promotes intestinal mucosal repair.

[0293] Those skilled in the art should understand that this invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to this invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. A nutritional preparation based on a whole food matrix that improves gut microbiota and promotes intestinal mucosal repair, characterized in that, By weight, it includes: 20-50 parts compound cereal powder, 1-20 parts compound fruit and vegetable powder, 10-35 parts vegetable fat powder, 5-25 parts protein, 0.1-10 parts prebiotics, 2-10 parts compound minerals, and 0.05-1.5 parts compound vitamins. The compound cereal powder, the compound fruit and vegetable powder, and the prebiotics together serve as sources of total dietary fiber, with soluble dietary fiber accounting for more than 50% of the total dietary fiber by weight. The compound grain powder includes yam powder, and the yam powder accounts for 0.5%-5% of the total mass of the compound grain powder. The compound minerals include zinc, and the zinc content is 2-6 mg / 100g of nutritional preparation.

2. The nutritional preparation of a whole food matrix for improving intestinal flora and promoting intestinal mucosal repair according to claim 1, characterized in that: in, The composite fruit and vegetable powder is obtained by processing fruit and vegetable powder through endogenous pectin gel microparticle technology.

3. The nutritional preparation of a whole food matrix for improving intestinal flora and promoting intestinal mucosal repair according to claim 2, characterized in that: in, The preparation process of the compound fruit and vegetable powder includes the following steps: Step S101, Low-temperature enzyme inactivation: Wash and cut fresh fruits and vegetables, blanch at 85-90℃ for 10-20 seconds, and then pulp to form fruit and vegetable pulp; Step S102, Ultra-high pressure micro-jet treatment: The fruit and vegetable pulp is circulated 3-5 times under a pressure of 120-160MPa to obtain the treated pulp, with the temperature ≤30℃ throughout the process; Step S103, Deesterification treatment: Adjust the pH of the treated slurry to 8.0-8.5, and stir the reaction at 20-25℃ to obtain a deesterified slurry; Step S104, self-crosslinking treatment: adjust the pH of the deesterified slurry to less than 7, add calcium agent, stir and let stand to obtain gel slurry; Step S105, freeze-drying: freeze-dry and pulverize the gel slurry to obtain the composite fruit and vegetable powder.

4. The nutritional preparation of a whole food matrix for improving intestinal flora and promoting intestinal mucosal repair according to claim 1, characterized in that, Also includes: A combination of one or more of the following: 0-5 parts meat meal, 0-1 parts thickener, and 0-2 parts acidity regulator.

5. The nutritional preparation of a whole food matrix for improving intestinal flora and promoting intestinal mucosal repair according to claim 1, characterized in that: in, The compound grain powder also includes one or more combinations of rice flour, enzymatically hydrolyzed rice flour, corn flour, millet flour, and soybean flour.

6. The nutritional preparation of a whole food matrix for improving intestinal flora and promoting intestinal mucosal repair according to claim 1, characterized in that: in, The compound fruit and vegetable powder includes one or more of the following: pumpkin powder, apple powder, carrot powder, tomato powder, strawberry powder, broccoli powder, kale powder, pear powder, banana powder, blueberry powder, orange powder, pineapple powder, cucumber powder, spinach powder, and purple sweet potato powder.

7. The nutritional preparation of a whole food matrix for improving intestinal flora and promoting intestinal mucosal repair according to claim 1, characterized in that: in, The plant fat powder includes: Vegetable oils, including one or more of the following: medium-chain triglycerides, docosahexaenoic acid (DHA) oils, eicosapentaenoic acid (EPA) oils, arachidonic acid (RAA) oils, 1,3-dioleoyl-2-palmitoyl triglycerides, rapeseed oil, low-erucic acid rapeseed oil, corn oil, soybean oil, sunflower seed oil, high-oleic sunflower seed oil, peanut oil, high-oleic peanut oil, palm oil, palm sap oil, palm kernel oil, olive oil, coconut oil, flaxseed oil, safflower seed oil, walnut oil, and peony seed oil; Emulsifiers include one or more combinations of sodium caseinate, casein, glyceryl citrate, mono- and diglycerides of fatty acids, and caprylic and capric glycerides. Wall materials include one or more of maltodextrin, glucose syrup, oligomaltose, and maltitol.

8. The nutritional preparation of a whole food matrix for improving intestinal flora and promoting intestinal mucosal repair according to claim 1, characterized in that: in, The proteins include one or more combinations of whey protein, soy protein, casein, milk protein, rice protein, pea protein, and yeast protein. The prebiotics include one or more combinations of fructooligosaccharides, galactooligosaccharides, inulin, pectin, and β-glucan. The complex minerals also include at least two of the following: potassium, calcium, magnesium, iron, copper, manganese, iodine, selenium, sodium, phosphorus, and chlorine. The multivitamin includes at least two of the following: vitamin A, vitamin D, vitamin E, vitamin K, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, folic acid, pantothenic acid, vitamin C, biotin, taurine, and beta-carotene.

9. A method for preparing a nutritional preparation of a whole food matrix that improves gut microbiota and promotes intestinal mucosal repair, used to prepare the nutritional preparation of a whole food matrix that improves gut microbiota and promotes intestinal mucosal repair as described in any one of claims 1-8, characterized in that, Includes the following steps: Weigh each component according to the weight proportions specified in claim 1; The weighed components are then dry-mixed. The mixed components are then protected by nitrogen gas to obtain a nutrient preparation.

10. The method for preparing a nutritional preparation of a whole food matrix that improves intestinal flora and promotes intestinal mucosal repair according to claim 9, characterized in that: in, The dry mixing is carried out using an airflow mixer, and the mixing time is 5-15 minutes.

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