Probiotic microcapsule with function of improving intestinal microecology and key technology

Abstract

The invention relates to the technical field of biological pharmacy, and discloses a probiotic microcapsule with a function of improving intestinal microecology, which comprises the following components in percentage by weight: lactobacillus rhamnosus: each gram of formula contains 1010 CFU of lactobacillus rhamnosus; lactobacillus acidophilus: each gram of the formula contains 109 CFU; clostridium butyricum: each gram of the formula contains 108 CFU; 5%-8% of a nano sodium alginate-chitosan compound; 3%-5% of peptide nanoparticles; 2%-4% of a natural polymer intelligent response film; 3%-6% of arabinoxylan oligosaccharide; 4%-7% of resistant dextran; 2%-4% of lentinan; and 0.1%-0.3% of resveratrol. Through a double-layer or three-layer embedding technology of the nano sodium alginate-chitosan compound and the intelligent response polymer film, the tolerance of the probiotics in gastric acid and cholate environments is remarkably enhanced, and targeted release of the probiotics in small intestines and colon is ensured.

Description

Technical Field

[0001] The present invention relates to the field of biopharmaceutical technology, in particular to probiotic microcapsules with the function of improving intestinal microecology and key technologies. Background Art

[0002] In recent years, probiotics have received extensive attention due to their significant effects in maintaining intestinal health, enhancing immune function and improving the digestive system. Probiotic products have been developed into various forms such as food, health products and medicines. However, there are some significant technical challenges in the clinical and practical application of existing probiotic preparations, mainly reflected in the problems of insufficient survival rate of probiotics in the gastrointestinal tract, poor stability, and uncontrollable release, which makes it difficult to fully exert their health benefits.

[0003] First, after entering the human body, probiotics must withstand the challenges of complex digestive fluid environments such as gastric acid and bile salts. Traditional probiotic preparations (such as powders, tablets or ordinary capsules) have extremely low survival rates in this acidic and bile salt environment. Most probiotics are killed in the stomach, resulting in their effective bacterial count being unable to reach the small and large intestines, thereby greatly weakening their effects on intestinal health. Although some studies have attempted to use simple single-layer coating technology to enhance the acid resistance of probiotics, such methods often lack targeted protection mechanisms and cannot provide long-lasting and effective protection in gastric acid and bile, resulting in a significant decrease in probiotic activity.

[0004] Secondly, probiotic products are easily inactivated by environmental factors (such as temperature, humidity and oxygen) during storage and transportation. The number of live bacteria in conventional probiotic preparations will decrease significantly after being stored for a period of time, especially in high temperature or humid environments, where the number of active probiotics is more difficult to guarantee, which directly affects the quality and efficacy of the product. Although some probiotic products improve stability by adding antioxidants or using moisture-proof packaging, these methods have limited effects and it is still difficult to maintain the high activity and functional integrity of probiotics during long-term storage.

[0005] In addition, existing probiotic preparations generally lack precise release control technology, resulting in disordered release of probiotics in the digestive tract and failure to achieve targeted colonization. Probiotics are often released quickly during the process of passing through the stomach and small intestine, lacking the ability to achieve optimal effects in target areas (such as the colon). Therefore, the effectiveness of existing probiotic products in specific intestinal segments is limited, and the potential of probiotics in promoting intestinal health, preventing diseases, and enhancing immune function cannot be fully realized.

[0006] To this end, those skilled in the art provide probiotic microcapsules and key technologies with the function of improving intestinal microecology to solve the problems raised in the above background technology. Summary of the invention

[0007] In view of the deficiencies of the prior art, the present invention provides probiotic microcapsules and key technologies with the function of improving intestinal microecology, which solve the problems of insufficient survival rate, poor stability and uncontrollable release in the gastrointestinal tract of the prior art.

[0008] To achieve the above objectives, the present invention is implemented by the following technical scheme: the probiotic microcapsules with the function of improving intestinal microecology include the following components in percentage by weight:

[0009] Lactobacillus rhamnosus: 10^10 CFU per gram of formula;

[0010] Lactobacillus acidophilus: 10^9 CFU per gram of formula;

[0011] Clostridium butyricum: 10^8 CFU per gram of formula;

[0012] Nano-alginate-chitosan complex: 5% to 8%;

[0013] Peptide nanoparticles: 3% to 5%;

[0014] Natural polymer intelligent response membrane: 2% to 4%;

[0015] Arabinoxylan oligosaccharides: 3% to 6%;

[0016] Resistant dextran: 4% to 7%;

[0017] Lentinan: 2% to 4%;

[0018] Resveratrol: 0.1%-0.3%;

[0019] Echinacea extract: 0.2%-0.4%;

[0020] Short-chain fatty acid precursor complex: 1% to 2%;

[0021] Menthol microcapsules: 0.5% to 1%;

[0022] Microcrystalline cellulose: 2% to 4%;

[0023] Lactitol: 1%~3%.

[0024] Preferably, the nano-alginate-chitosan complex is prepared by nano-encapsulation technology, and the complex has a dual protection structure, which can form a gel layer in the gastric acid environment to protect the probiotics, and gradually dissolve in the alkaline environment of the small intestine to release the probiotics.

[0025] Preferably, the peptide nanoparticles are pH-responsive nanoparticles, which can dissolve within a specific pH range (6.0-7.5) in the intestine and specifically bind to probiotic surface proteins.

[0026] Preferably, the natural polymer intelligent response membrane is composited with modified cellulose and polyglutamic acid, and the membrane can sense the pH value and temperature changes in the intestine, and is used to adjust the release rate of probiotics.

[0027] Preferably, the arabinoxylan oligosaccharides and resistant dextran are mixed as symbiotic nutritional regulating ingredients in a ratio of 1:1 to 1:2, which can specifically promote the proliferation of bifidobacteria and lactobacilli in the intestine and improve the intestinal flora structure by increasing the production of short-chain fatty acids.

[0028] The key technology of probiotic microcapsules with the function of improving intestinal microecology is used for probiotic microcapsules with the function of improving intestinal microecology, including the following steps:

[0029] S1: Lactobacillus rhamnosus, Lactobacillus acidophilus and Clostridium butyricum were cultured in a modified MRS medium to a high concentration of cells, collected by centrifugation, suspended in a cryoprotectant solution, pre-frozen to -80°C, and then dried in an ultra-low temperature freeze dryer at -60°C for 72 hours to obtain probiotic freeze-dried powder;

[0030] S2: Prepare a nano-alginate-chitosan composite solution, suspend the probiotic freeze-dried powder in it and mix it evenly, and use a double-nozzle electrostatic spray device to spray the suspension and the peptide nanoparticle solution into the calcium chloride solution at the same time to form nano-composite gel microspheres with a double protection structure;

[0031] S3: placing the nanocomposite gel microspheres in a smart responsive polymer solution prepared from modified cellulose and polyglutamic acid, and coating the polymer outer layer with a roller coating device for 30 minutes to ensure the formation of a stable outer protective film;

[0032] S4: A composite solution is prepared using arabinoxylan oligosaccharides, lentinan, resveratrol and echinacea extract, the wrapped microspheres are immersed in the composite solution, and a pulsed ultrasound method is used to promote the composite to penetrate into the microspheres to form a composite inclusion layer;

[0033] S5: Low-temperature spray drying is used to preliminarily dry the microcapsules, and then freeze drying is used to further remove moisture. At the same time, the temperature is gradually increased during the freeze drying process to achieve cross-linking and curing of the internal components and enhance the mechanical strength of the microcapsules;

[0034] S6: preparing a controlled-release coating solution of magnesium stearate and polyethylene glycol, spraying the controlled-release coating on the surface of the microcapsules using a fluidized bed spraying device, and coating at 45°C for 60 minutes to form a stable intestinal targeted controlled-release membrane layer;

[0035] S7: The coated probiotic microcapsules are sized, particles with uniform particle size are selected by vibration, and the particles are packed into moisture-proof sealed packaging bags, which are then filled with inert gas for sealed packaging.

[0036] Preferably, in the step S2, a dual-nozzle electrostatic spray device is used to form microcapsules, thereby ensuring that a nanocomposite protective layer is formed on the surface of the probiotics, and the peptide nanoparticles are simultaneously wrapped during the spraying process to enhance the intestinal colonization effect of the microspheres.

[0037] Preferably, the outer layer of the smart response polymer film used in the step S3 is coated by a drum coating device, and the film can be adjusted according to the pH value and temperature changes in the intestine to ensure the release of probiotics.

[0038] Preferably, in step S4, pulsed ultrasound technology is used to carry out inclusion of prebiotics and antioxidant components to promote uniform penetration of the complex into the microspheres.

[0039] The application of probiotic microcapsules with the function of improving intestinal microecology is used to prevent and assist in the treatment of irritable bowel syndrome, inflammatory bowel disease, lactose intolerance, Helicobacter pylori infection and leaky gut related diseases, or to improve diarrhea, constipation and bloating symptoms caused by imbalance of intestinal flora.

[0040] The present invention provides probiotic microcapsules and key technologies that have the function of improving intestinal microecology. It has the following beneficial effects:

[0041] 1. The present invention uses a double-layer or triple-layer encapsulation technology of nano-sodium alginate-chitosan complex and an intelligent responsive polymer membrane to significantly enhance the tolerance of probiotics in gastric acid and bile salt environments, ensuring their targeted release in the small intestine and colon. The survival rate of the microcapsule probiotics in the gastric acid environment reaches 85%-90%, and the small intestine release rate reaches 70%-75%, which is significantly better than traditional preparations and effectively improves intestinal health.

[0042] 2. The present invention enhances the antioxidant capacity of probiotics and reduces the risk of inactivation during storage through the design of composite prebiotics and antioxidant inclusion. Experimental results show that the antioxidant inclusion technology increases the survival rate of probiotics under oxidative stress conditions to more than 85%, and extends the storage period to 240 days, ensuring the long-term activity and stability of probiotic products.

[0043] 3. The present invention introduces a controlled-release coating and an antibacterial immunomodulatory complex to achieve precise release of probiotics in the gastrointestinal tract, significantly improving bioavailability and health effects. The release rate of probiotic microcapsules in the small intestine reaches 82%, and has the effect of enhancing intestinal immune function, which is suitable for the prevention and adjuvant treatment of a variety of intestinal diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 It is a schematic diagram of the process of the present invention. DETAILED DESCRIPTION

[0045] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0046] Please refer to the attached Figure 1 :

[0047] Example 1: Preparation of double-layer embedded probiotic microcapsules

[0048] step:

[0049] Probiotic culture and freeze drying: Lactobacillus rhamnosus, Lactobacillus acidophilus and Clostridium butyricum were cultured in a modified MRS medium to a concentration of 10^10 CFU / mL, collected by centrifugation, suspended in a cryoprotectant solution, pre-frozen to -80°C, and then dried in an ultra-low temperature freeze dryer at -60°C for 72 hours to obtain probiotic freeze-dried powder.

[0050] Preparation of nanocomposite gel microspheres: Prepare a nano-alginate-chitosan composite solution, suspend the probiotic freeze-dried powder therein, and use a double-nozzle electrostatic spray device to spray the suspension into a 0.2M calcium chloride solution to form nanocomposite gel microspheres with a double protective structure.

[0051] Outer protective film coating: The nanocomposite gel microspheres are placed in a smart responsive polymer solution prepared from modified cellulose and polyglutamic acid, and the outer film is coated using a roller coating device for 30 minutes.

[0052] Low-temperature drying and molding: Low-temperature spray drying is used to preliminarily dry the microcapsules, and then freeze drying is used to further remove moisture to complete the preparation of probiotic microcapsules with a double-layer encapsulation structure.

[0053] Summary: The double-layer encapsulation technology of nano-alginate-chitosan complex and intelligent response polymer membrane was adopted. Through the core protection of nano-composite gel microspheres, probiotics can form a gel layer in the gastric acid environment to resist the erosion of gastric acid and bile salts, thereby greatly improving the survival rate of probiotics. At the same time, the outer layer of intelligent response membrane can control the release of probiotics according to the pH value and temperature changes in the intestine, ensuring that the probiotics are gradually released in the small intestine and colon and give full play to their biological effects. The experimental results show that the design of this double-layer encapsulation structure significantly improves the survival rate (85%) and small intestine release rate (70%) of probiotics in a simulated gastrointestinal environment, and at the same time prolongs the storage period of probiotics. Compared with the traditional single-layer encapsulation technology, this double-layer encapsulation technology not only enhances the acid and bile resistance of probiotics, but also optimizes the release rate and stability of probiotics, significantly improving their colonization ability in the intestine, and providing a more stable and long-term guarantee for the health benefits of probiotics. 。

[0054] Example 2: Preparation of three-layer embedded probiotic microcapsules

[0055] step:

[0056] Probiotics culture and freeze drying: Lactobacillus rhamnosus, Lactobacillus acidophilus and Clostridium butyricum were cultured and freeze dried according to the steps of Example 1.

[0057] Preparation of nanocomposite gel microspheres: Prepare a nano-alginate-chitosan composite solution, add 2% pectin solution, and use a double-nozzle electrostatic spray device to spray the suspension into a 0.2M calcium chloride solution to form nanocomposite gel microspheres.

[0058] Intermediate chitosan-gelatin composite layer coating: the nanocomposite gel microspheres are placed in a composite solution of chitosan and gelatin, and a roller coating device is used to coat the intermediate layer membrane.

[0059] Coating of the outer layer of smart response film: According to the steps of Example 1, a smart response film is coated on the outer surface of the middle layer film to form a three-layer protection structure.

[0060] Multi-step drying and molding: Multi-step drying is carried out by combining spray drying and freeze drying to enhance the structural stability of the microcapsules.

[0061] Summary: The protective effect and function of probiotic microcapsules are further improved through three-layer encapsulation technology. In addition to the basic design of nano-alginate-chitosan complex and intelligent responsive polymer membrane, a chitosan-gelatin composite layer is added to provide additional mechanical strength and acid and bile salt resistance protection. The design of this intermediate layer can provide a stronger physical barrier in the gastric acid environment, prevent the loss and inactivation of probiotics, and gradually dissolve in specific parts of the intestine to release probiotics. Experimental results show that the three-layer encapsulation structure makes the survival rate of probiotics in the gastric acid environment reach 90%, and the small intestine release rate is increased to 75%. Compared with double-layer encapsulation, three-layer encapsulation significantly improves the stability and tolerance of microcapsules and extends the storage period of probiotics to 200 days. This design not only provides additional protection, but also improves the colonization and lasting effect of probiotics in the intestine, which is very suitable for the development of probiotic preparations that are more sensitive to the gastrointestinal environment.

[0062] Example 3: Probiotic microcapsules containing composite prebiotics and antioxidants

[0063] step:

[0064] Probiotics culture and freeze drying: Lactobacillus rhamnosus, Lactobacillus acidophilus and Clostridium butyricum were cultured and freeze dried according to the steps of Example 1.

[0065] Preparation of nanocomposite gel microspheres: Nanocomposite gel microspheres were prepared according to the steps of Example 1.

[0066] Outer protective film coating: Coat the outer layer of the smart responsive polymer film according to the steps of Example 1.

[0067] Composite inclusion of prebiotics and antioxidant ingredients: a composite solution of arabinoxylan oligosaccharides, lentinan, resveratrol and echinacea extract was prepared, the wrapped microspheres were immersed in it, and pulsed ultrasound (40kHz, 100W, 15 minutes) was used to promote the penetration of the complex into the microspheres to form a composite inclusion layer.

[0068] Drying and forming: Use low temperature spray drying and freeze drying methods for drying and solidification.

[0069] Summary: The functionality and stability of probiotic microcapsules are further enhanced through the inclusion technology of composite prebiotics and antioxidants. Arabinoxylan oligosaccharides, lentinan, resveratrol and echinacea extracts are introduced into the microcapsule inclusion system, and pulsed ultrasound technology is used to promote the uniform penetration of these active ingredients to form a composite inclusion layer. The addition of prebiotics provides a good symbiotic environment and nutritional support for probiotics, and promotes the proliferation of probiotics in the intestine. The antioxidant ingredients effectively resist the challenges brought by oxidative stress, ensuring that probiotics remain highly active during long-term storage and use. Experimental data show that the survival rate of probiotics under oxidative stress conditions in this composite inclusion design reached 85%, which is significantly better than the group without added antioxidants. At the same time, the small intestine release rate reached 78%, and the total probiotic survival rate was 83%, showing excellent functionality and stability, which is suitable for the development of health products with high requirements for antioxidants and immune regulation.

[0070] Example 4: Preparation of multifunctional controlled-release probiotic microcapsules

[0071] step:

[0072] Probiotics culture and freeze drying: Probiotics were cultured and freeze dried according to the steps of Example 1.

[0073] Preparation of nanocomposite gel microspheres: Nanocomposite gel microspheres were prepared according to the steps of Example 1.

[0074] Smart response film coating: The smart response film was coated according to the steps of Example 1.

[0075] Formation of controlled-release coating: A controlled-release coating solution of magnesium stearate and polyethylene glycol (PEG) was prepared and sprayed on the surface of the microcapsules using a fluidized bed spraying device at 45° C. for 60 minutes to form a stable controlled-release film layer.

[0076] Drying and molding: The drying step of Example 1 is followed to obtain multifunctional controlled-release probiotic microcapsules.

[0077] Summary: The introduction of multifunctional controlled-release technology enables the precise targeted release of probiotic microcapsules in different intestinal areas. By adding a controlled-release coating (such as magnesium stearate and polyethylene glycol) on the basis of the outer intelligent response membrane, the release behavior of probiotics in the gastrointestinal tract is significantly optimized. The controlled-release membrane can reduce the premature release of probiotics in the gastric acid environment, and gradually release them in the small intestine and colon, ensuring that the probiotics can exert their maximum effectiveness in the most suitable location. The experimental results show that the probiotic release rate of the multifunctional controlled-release microcapsules in the gastric acid environment is only 10%, while the release rate in the small intestine reaches 82%, which greatly improves the targeted release effect and overall utilization of probiotics. At the same time, the overall probiotic survival rate is as high as 81%, showing the significant advantages of this controlled-release design in different environments, and is particularly suitable for probiotic preparations that require precise targeting, such as products for the treatment and prevention of specific intestinal diseases.

[0078] Example 5: Antibacterial and immunomodulatory composite probiotic microcapsules

[0079] step:

[0080] Probiotics culture and freeze drying: Probiotics were cultured and freeze dried according to the steps of Example 1.

[0081] Preparation of nanocomposite gel microspheres: Nanocomposite gel microspheres were prepared according to the steps of Example 1.

[0082] Smart response film coating: The smart response film was coated according to the steps of Example 1.

[0083] Antibacterial and immunomodulatory composite layer coating: prepare a menthol microcapsule and short-chain fatty acid precursor complex solution, immerse the coated microspheres therein, and use a roller coating device to coat for 30 minutes to form an antibacterial and immunomodulatory composite layer.

[0084] Drying and forming: Use low temperature spray drying and freeze drying methods for drying and solidification.

[0085] Summary: By adding antibacterial and immunomodulatory complexes (such as menthol microcapsules and short-chain fatty acid precursors) to the probiotic microcapsule system, additional intestinal health functions and immune enhancement effects are provided. This composite layer can not only be gradually released in the intestine, provide a mild antibacterial effect, and inhibit the proliferation of harmful bacteria such as Escherichia coli and Helicobacter pylori, but also promote the optimization of intestinal immune response through the action of short-chain fatty acid precursors. Experimental data show that this composite design enables the survival rate of probiotics in the gastric acid environment to reach 87%, the small intestine release rate is 80%, and the antibacterial effect is obvious. The overall survival rate is 82%, and it shows excellent stability under storage conditions (storage period can reach 220 days). This design is particularly suitable for the development of functional foods and drugs that hope to enhance immunity while maintaining the balance of intestinal microecology, demonstrating the innovative value of probiotic preparations in multifunctional composite applications.

[0086] Summary: Double-layer and triple-layer encapsulation technologies provide basic probiotic protection and controlled release capabilities, which are suitable for routine intestinal health regulation; the combination of compound prebiotics and antioxidants further improves the stability, proliferation ability and antioxidant properties of probiotics; and the multifunctional controlled release system and antimicrobial immunomodulatory composite design significantly optimize the targeted release and long-term effects of probiotics for different health needs and treatment goals.

[0087] Test experiments and specific test data

[0088] Experimental design

[0089] Survival test: Testing the survival rate of probiotics in a simulated gastrointestinal environment.

[0090] Controlled release test: Testing the release behavior of probiotics under different pH conditions.

[0091] Antioxidant performance testing: Testing the antioxidant protection of probiotics under storage conditions.

[0092] Test experimental data

[0093]

[0094] Comparative Experimental Design and Data

[0095] Comparative experiment 1: Effects of different numbers of encapsulation layers on the activity of probiotics

[0096] Embedding layer number Gastric acid survival rate (%) Small intestine release rate (%) Storage period (days) Single layer 60％ 50％ 100 Double Layer 85％ 70％ 180 Three-layer 90％ 75％ 200

[0097] Comparative experiment 2: Effect of antioxidant addition on the stability of probiotics

[0098]

[0099] Comparative experiment 3: Effect of controlled-release agent on the release behavior of probiotics

[0100] Controlled release type Gastric acid release rate (%) Small intestine release rate (%) Overall survival rate (%) No controlled release agent 20％ 60％ 55％ Magnesium Stearate 10％ 82％ 81％

[0101] Summarize

[0102] Through testing and comparative experiments, the results show that the probiotic microcapsule preparation of the present invention has significant advantages in gastric acid tolerance, intestinal colonization effect, antioxidant protection and controlled release performance. Experimental data proves that by optimizing the number of embedding layers, adding antioxidants and designing a controlled release system, the stability and bioavailability of probiotics in different environments can be effectively improved, highlighting the innovation and application value of the present invention.

[0103] Animal experiment design: evaluating the effect of probiotic microcapsules in regulating intestinal microecology and enhancing immune function

[0104] Experimental animals

[0105] Experimental subjects: Male C57BL / 6 mice

[0106] Quantity: 60 (average weight 20±2g)

[0107] Grouping: Randomly divided into three groups, 20 in each group

[0108] Control group (Group A): given normal saline

[0109] Traditional formula group (Group B): given traditional Lactobacillus rhamnosus probiotic preparation (no embedding design, containing 10^10 CFU per gram)

[0110] The microcapsule group (Group C) of the present invention was administered with the multi-layer embedded probiotic microcapsules (containing 10^10 CFU per gram) of the present invention.

[0111] Experimental methods

[0112] Dosage:

[0113] Each group of mice was intragastrically administered with a dose of 0.2 mL / 10 g body weight daily for 4 weeks.

[0114] Monitoring indicators:

[0115] Body weight changes: The mice were weighed once a week to assess weight gain.

[0116] Intestinal flora analysis: On days 0, 14, and 28 of the experiment, stool samples were collected for 16S rRNA gene sequencing analysis to evaluate changes in beneficial bacteria (such as Lactobacillus and Bifidobacterium) and harmful bacteria (such as Escherichia coli).

[0117] Detection of inflammatory factor levels: After the experiment, the concentrations of TNF-α, IL-6 (inflammatory factor), and IL-10 (anti-inflammatory factor) in serum were measured by enzyme-linked immunosorbent assay (ELISA).

[0118] Intestinal barrier function testing: evaluate the expression of tight junction proteins (such as ZO-1, Occludin) in small intestinal tissue.

[0119] Immune organ index: After the experiment, the spleen and thymus of the mice were removed and the immune organ index (organ weight / body weight) was calculated.

[0120] Experimental period: 4 weeks.

[0121] Experimental Results

[0122] 1. Weight change (mean ± standard deviation, g):

[0123] Time (day) Control group (A) Traditional formula group (B) Microcapsule group (C) of the present invention 0 20.1±1.5 20.2±1.4 20.3±1.3 14 22.5±1.7 23.1±1.6 23.9±1.5 28 23.0±1.8 24.0±1.7 25.2±1.6

[0124] 2. Changes in intestinal flora (increase rate of Lactobacillus and decrease rate of Escherichia coli):

[0125]

[0126]

[0127] 3. Level of inflammatory factors (pg / mL):

[0128] Group TNF-α IL-6 IL-10 Control group (A) 45.2±5.3 38.1±4.8 20.5±2.7 Traditional formula group (B) 32.8±4.1 27.5±3.6 28.4±3.1 Microcapsule group (C) of the present invention 22.1±3.7 18.9±2.9 35.7±3.4

[0129] 4. Intestinal barrier function detection (tight junction protein expression level, unit: relative expression):

[0130] Group ZO-1 Occludin Control group (A) 1.0±0.1 1.0±0.1 Traditional formula group (B) 1.5±0.2 1.4±0.2 Microcapsule group (C) of the present invention 2.2±0.3 2.0±0.3

[0131] 5. Immune organ index (organ weight / body weight, mg / g):

[0132] Group Spleen index Thymus index Control group (A) 3.5±0.3 1.2±0.2 Traditional formula group (B) 4.2±0.4 1.6±0.3 Microcapsule group (C) of the present invention 5.0±0.5 1.9±0.3

[0133] Experimental Summary

[0134] The experimental results show that the probiotic microcapsules (GroupC) of the present invention are superior to the traditional probiotic formula (GroupB) and the control group (GroupA) in many aspects. Specifically, the microcapsules of the present invention show significant advantages in the weight gain of mice, the proliferation of beneficial intestinal bacteria (Lactobacillus) and the inhibition of harmful bacteria (Escherichia coli), proving its excellent efficacy in regulating the balance of intestinal flora; in terms of inflammatory factor levels, the microcapsule group of the present invention shows lower TNF-α and IL-6 concentrations and higher anti-inflammatory factor IL-10 concentrations, indicating its superiority in reducing inflammatory response and enhancing immune function; in addition, the intestinal barrier function test shows that the expression level of tight junction proteins in the microcapsule group of the present invention is significantly improved, reflecting a better intestinal barrier repair ability; the immune organ index (spleen index and thymus index) is also significantly higher than that of other groups, further verifying its significant effect in enhancing the function of the immune system. In general, the high-activity probiotic microcapsules of the present invention are significantly superior to traditional probiotic preparations in regulating intestinal health, enhancing immune function and improving digestive function, showing its innovation and application potential in the field of probiotics.

[0135] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A probiotic microcapsule having the function of improving intestinal microecology, characterized in that: The composition includes the following weight percentages: Lactobacillus rhamnosus: 10^10 CFU per gram of formula; Lactobacillus acidophilus: 10^9 CFU per gram of formula; Clostridium butyricum: 10^8 CFU per gram of formula; Nano-alginate-chitosan complex: 5% to 8%; Peptide nanoparticles: 3% to 5%; Natural polymer intelligent response membrane: 2% to 4%; Arabinoxylan oligosaccharides: 3% to 6%; Resistant dextran: 4% to 7%; Lentinan: 2% to 4%; Resveratrol: 0.1%-0.3%; Echinacea extract: 0.2%-0.4%; Short-chain fatty acid precursor complex: 1% to 2%; Menthol microcapsules: 0.5% to 1%; Microcrystalline cellulose: 2% to 4%; Lactitol: 1%~3%.

2. The probiotic microcapsule having the function of improving intestinal microecology according to claim 1, characterized in that: The nano-alginate-chitosan complex is prepared by nano-encapsulation technology. The complex has a dual protection structure, can form a gel layer to protect probiotics in a gastric acid environment, and gradually dissolves in an alkaline environment of the small intestine to release the probiotics.

3. The probiotic microcapsule having the function of improving intestinal microecology according to claim 1, characterized in that: The peptide nanoparticles are pH-responsive nanoparticles, which can dissolve within a specific pH range (6.0-7.5) of the intestinal tract and specifically bind to the surface proteins of probiotics.

4. The probiotic microcapsule having the function of improving intestinal microecology according to claim 1, characterized in that: The natural polymer intelligent response membrane is composed of modified cellulose and polyglutamic acid. The membrane can sense the pH value and temperature changes in the intestine and is used to adjust the release rate of probiotics.

5. The probiotic microcapsule having the function of improving intestinal microecology according to claim 1, characterized in that: The arabinoxylan oligosaccharides and resistant dextran are mixed as symbiotic nutritional regulating components in a ratio of 1:1 to 1:2, which can specifically promote the proliferation of bifidobacteria and lactobacilli in the intestine and improve the intestinal flora structure by increasing the production of short-chain fatty acids.

6. Key technology of probiotic microcapsules with intestinal microecological function improvement, used for the probiotic microcapsules with intestinal microecological function improvement according to any one of claims 1 to 5, characterized in that: The following steps are involved: S1: Lactobacillus rhamnosus, Lactobacillus acidophilus and Clostridium butyricum were cultured in a modified MRS medium to a high concentration of cells, collected by centrifugation, suspended in a cryoprotectant solution, pre-frozen to -80°C, and then dried in an ultra-low temperature freeze dryer at -60°C for 72 hours to obtain probiotic freeze-dried powder; S2: Prepare a nano-alginate-chitosan composite solution, suspend the probiotic freeze-dried powder in it and mix it evenly, and use a double-nozzle electrostatic spray device to spray the suspension and the peptide nanoparticle solution into the calcium chloride solution at the same time to form nano-composite gel microspheres with a double protection structure; S3: placing the nanocomposite gel microspheres in a smart responsive polymer solution prepared from modified cellulose and polyglutamic acid, and coating the polymer outer layer with a roller coating device for 30 minutes to ensure the formation of a stable outer protective film; S4: A composite solution is prepared using arabinoxylan oligosaccharides, lentinan, resveratrol and echinacea extract, the wrapped microspheres are immersed in the composite solution, and a pulsed ultrasound method is used to promote the composite to penetrate into the microspheres to form a composite inclusion layer; S5: Low-temperature spray drying is used to preliminarily dry the microcapsules, and then freeze drying is used to further remove moisture. At the same time, the temperature is gradually increased during the freeze drying process to achieve cross-linking and curing of the internal components and enhance the mechanical strength of the microcapsules; S6: preparing a controlled-release coating solution of magnesium stearate and polyethylene glycol, spraying the controlled-release coating on the surface of the microcapsules using a fluidized bed spraying device, and coating at 45°C for 60 minutes to form a stable intestinal targeted controlled-release membrane layer; S7: The coated probiotic microcapsules are sized, particles with uniform particle size are selected by vibration, and the particles are packed into moisture-proof sealed packaging bags, which are then filled with inert gas for sealed packaging.

7. The key technology of probiotic microcapsules with the function of improving intestinal microecology according to claim 6 is characterized in that: In the S2 step, a double-nozzle electrostatic spray device is used to form microcapsules, thereby ensuring that a nanocomposite protective layer is formed on the surface of the probiotics, and the peptide nanoparticles are simultaneously wrapped during the spraying process to enhance the intestinal colonization effect of the microspheres.

8. The key technology of probiotic microcapsules with the function of improving intestinal microecology according to claim 6 is characterized in that: The smart response polymer outer layer film used in the S3 step is coated by a drum coating device. The film can be adjusted according to the pH value and temperature changes in the intestine to ensure the release of probiotics.

9. The key technology of probiotic microcapsules with the function of improving intestinal microecology according to claim 6 is characterized in that: In the step S4, pulsed ultrasound technology is used to carry out inclusion of prebiotics and antioxidant components to promote uniform penetration of the complex into the microspheres.

10. Application of probiotic microcapsules with the function of improving intestinal microecology, according to any one of claims 1 to 5, characterized in that: Used for the prevention and adjunctive treatment of irritable bowel syndrome, inflammatory bowel disease, lactose intolerance, Helicobacter pylori infection and leaky gut-related diseases, or for improving symptoms of diarrhea, constipation and bloating caused by imbalance of intestinal flora.

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