Anti-depression composition containing difructose anhydride and application of anti-depression composition in functional preparation

Abstract

The invention discloses an anti-depression composition containing difructose anhydride and application of the anti-depression composition in a functional preparation, aiming at the current situation that related depressive symptoms of inflammatory bowel diseases lack targeted intervention, multi-target anti-depression intervention is realized through the difructose anhydride (DFA-I or DFA-III) composition for the first time: on the mechanism level, bifidobacterium / lactobacillus proliferation is selectively promoted, and the anti-depression effect is improved. The composition can be used for reducing TNF-alpha / IL-6, up-regulating hippocampus BDNF exon IX and inhibiting p75NTR, and synchronously reducing serum CORT and increasing the content of butyric acid in intestinal tracts. The invention provides an innovative solution with intestinal microecological regulation and control and neuroprotection functions for relieving IBD accompanied depression symptoms.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to an anti-depression composition containing difructose anhydride and application thereof in functional preparations. BACKGROUND

[0002] In recent years, inflammatory bowel disease (IBD) and its accompanying depressive symptoms have become an important challenge in the field of public health. The dextran sulfate sodium (DSS)-induced colitis model is a classic experimental system for studying the interaction mechanisms of intestinal inflammation and the gut-brain axis, and its pathological features include intestinal barrier damage, immune disorders, and central nervous system abnormalities, thereby inducing depressive behaviors. Current treatments for IBD focus on inflammation itself, while ignoring the accompanying emotional disorders, and there is an urgent need to develop intervention strategies with neuroregulatory functions.

[0003] Difructose anhydride I (DFA-I) and difructose anhydride III (DFA-III) are both metabolic derivatives of inulin. Studies have shown that DFA-III can improve intestinal health by promoting the proliferation of intestinal probiotics, enhancing mineral absorption, and regulating immune responses.

[0004] However, existing research on DFA-I and DFA-III is mostly limited to single intestinal health or metabolic regulation, and their role in the gut-brain axis has not been clearly defined, especially their regulatory effects and mechanisms on DSS-induced depressive behaviors. SUMMARY

[0005] The purpose of this section is to summarize some aspects of the embodiments of the present application and briefly introduce some preferred embodiments.

[0006] In view of the above and / or problems existing in the prior art, the present application is proposed.

[0007] Therefore, the purpose of the present application is to overcome the deficiencies in the prior art and provide an anti-depression composition containing difructose anhydride.

[0008] To solve the above technical problems, the present application provides the following technical solutions: an anti-depression composition containing difructose anhydride, wherein the difructose anhydride includes DFA-I or DFA-III.

[0009] As a preferred solution of the anti-depression composition of the present application, the composition exerts an anti-depression effect by regulating the gut microbiota-intestinal-brain axis pathway.

[0010] As a preferred scheme of the antidepressant composition, the composition increases the abundance of beneficial bacteria, reduces the abundance of conditional pathogenic bacteria, and significantly increases the level of intestinal butyric acid.

[0011] The beneficial bacteria include Bifidobacterium and Lactobacillus, and the conditional pathogenic bacteria include Clostridium.

[0012] As a preferred scheme of the antidepressant composition, the composition down-regulates the expression of hippocampal p75NTR mRNA, up-regulates the expression of BDNF exon IX, and reduces the level of serum corticosterone CORT.

[0013] As a preferred scheme of the antidepressant composition, the composition reduces the levels of serum pro-inflammatory factors TNF-α and IL-6, and increases the level of anti-inflammatory factor IL-10.

[0014] As a preferred scheme of the antidepressant composition, the composition comprises difructose anhydride, probiotics, and fructooligosaccharides, wherein the difructose anhydride accounts for 5-40% of the mass percentage of the composition.

[0015] As a preferred scheme of the antidepressant composition, the probiotics include at least one of Bifidobacterium or Lactobacillus.

[0016] Another object of the present application is to overcome the deficiencies in the prior art and provide the use of the antidepressant composition in the preparation of functional preparations, wherein the functional preparations are oral preparations or intestinal targeting preparations.

[0017] As a preferred scheme of the use, the dosage form of the functional preparation includes hard candies or granules.

[0018] Another object of the present application is to overcome the deficiencies in the prior art and provide a preparation method of synbiotic hard candies containing DFA, comprising,

[0019] Preparation of microcapsule probiotics: mix the probiotic bacterial suspension and the sodium alginate solution at a volume ratio of 1:(5-20), add resistant starch equivalent to 3-10% of the mass of the sodium alginate solution, stir at 200-400 rpm for 10-15 min, then drop into an oil phase emulsion containing 0.4% (v / v) Tween-80 at a volume ratio of 1:4-7, cross-link and solidify with 1-3% calcium chloride solution for 30 min, centrifuge and wash, and then use low-temperature spray drying with an inlet air temperature of 40-50°C and an outlet air temperature of 25-30°C to obtain microcapsules with a water content of ≤5%;

[0020] Raw material solution: dissolve 5% to 40% DFA-I, 20% to 30% fructooligosaccharide, 25% to 35% maltodextrin in a buffer solution with pH 6.0 to 6.5 at a material to liquid ratio of 1:1.5, and stir at 60°C until completely transparent;

[0021] Vacuum boiling: dehydrate at 140 to 145°C and a vacuum degree of -0.08 to -0.1 MPa until the moisture content is less than or equal to 2%, and control the paste viscosity to be 500 to 600 mPa·s;

[0022] Probiotic embedding: when the paste is cooled to 80 to 85°C, add 5% to 10% of the total mass of microencapsulated probiotics, and stir at a high speed of 200 to 300 rpm to disperse, wherein the survival rate of live bacteria is greater than or equal to 90% after the microencapsulated probiotics are treated in the paste at 80 to 85°C for 5 minutes;

[0023] Molding and solidification: inject the paste into a mold at 40 to 45°C, degas under a vacuum of -0.05 MPa, and then gradually cool to 25°C to form a sugar body with a hardness of 50 to 70 N / cm 2 ;

[0024] Sterilization and packaging: after being cut into 5 to 8 g per piece, sterilize the surface by ultraviolet irradiation, package using a nitrogen-filled aluminum foil composite film, and store in the dark at 25°C or below.

[0025] Advantages of the present application:

[0026] (1) The present application discloses, for the first time, that DFA-I and DFA-III improve depressive behavior through a multi-target point synergistic mechanism of the gut-brain axis, and on this basis, develops a functional hard candy carrier that is resistant to high-temperature processing, thereby realizing the whole-chain intervention of microbiota regulation-metabolic intervention-neuroprotection.

[0027] (2) The present application discloses, for the first time, that a multi-target point antidepressant intervention is realized through a DFA-I or DFA-III composition: at the mechanism level, selectively promotes the proliferation of bifidobacteria / lactobacilli, reduces the pro-inflammatory factors TNF-α / IL-6, up-regulates hippocampal BDNF exon IX and inhibits p75NTR, and synchronously reduces serum CORT and increases the content of intestinal butyric acid.

[0028] (3) At the application level, the present application innovates a microcapsule process, combines DFA and fructooligosaccharide double prebiotics to increase the total amount of SCFAs, and realizes the synergistic effect of intestinal microbiota regulation-neuroinflammatory inhibition-behavior improvement through a hard candy carrier. BRIEF DESCRIPTION OF DRAWINGS

[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed in the embodiment description will be briefly introduced as follows. Among them:

[0030] Figure 1Figure 1 is a representative trajectory plot of the mice in the open field test in the example.

[0031] Figure 2 Figure 2 is a result plot of the elevated plus maze and tail suspension test in the example; wherein A is a representative heat map of the elevated plus maze test (the vertical direction in the figure is the open arm), B is the movement distance of the elevated plus maze test, C is the frequency of entering the open arm of the elevated plus maze test, D is the total time in the open arm of the elevated plus maze test, E is the total time of remaining still in the tail suspension test, the values are the average values ± standard deviations of 10 repetitions, the lowercase letters (a, b) indicate that the differences between groups are significant (p < 0.05), and the statistical significance analysis is performed by analysis of variance and Duncan's test.

[0032] Figure 3 Figure 3 is the expression levels of TNF-α, IFN-γ, IL-6 and IL-10 in the serum of mice; wherein the lowercase letters a, b indicate that the differences between groups are significant (p < 0.05), and the statistical significance analysis is performed by analysis of variance and Duncan's test.

[0033] Figure 4 Figure 4 is a plot of the relative abundance differences of the intestinal microorganisms of mice; wherein A is Lactobacilus, B is Bifidobacterium, C is Clostridium, and D is Parabacteroides, the lowercase letters a, b indicate that the differences between groups are significant (p < 0.05), and the statistical significance analysis is performed by analysis of variance and Duncan's test.

[0034] Figure 5 Figure 5 is a plot of the short-chain fatty acid content in the intestine of mice, wherein A-F represent the levels of acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid and valeric acid in the intestine of mice, respectively.

[0035] Figure 6 Figure 6 is the mRNA expression levels of CORT in the serum of mice and p75NTR, proBDNF and BDNF exon IX in the hippocampus of mice. DETAILED DESCRIPTION

[0036] The specific embodiments of the present application are described below to facilitate the understanding of the present application for those skilled in the art, but it should be clear that the present application is not limited to the scope of the specific embodiments, and for those skilled in the art, as long as various changes are within the spirit and scope of the present application defined and determined by the appended claims, these changes are obvious, and all the inventions and creations using the concept of the present application are within the scope of protection.

[0037] Example 1

[0038] DFA-I and DFA-III improved the behavior of mice in the open field test:

[0039] 1.1 Animal treatment and grouping

[0040] Forty 6-week-old SPF male C57BL / 6J mice (Supplier: Beijing VitoLiu Hua Experimental Animal Technology Co., Ltd.) were selected, and the experimental protocol was approved by the Jiangnan University Experimental Animal Ethics Committee (JN. No 20210415-0831).

[0041] The experimental animals were acclimated in the barrier system (SYXK (Su) 2021-0056) for 7 days, and the feeding conditions were kept at a constant temperature (22 ± 1°C), constant humidity (55 ± 3%), and standard light cycle (12 h / 12 h), with free access to standard feed and drinking water.

[0042] Four groups of parallel controls (n = 10) were set up for the experiment: normal control group (CON, physiological saline), colitis model group (DSS, physiological saline), DFA-I intervention group (DFA-I, 30 g DFA-I / kg of food intake), and DFA-III intervention group (DFA-III, 30 g DFA-III / kg of food intake), all using gavage administration.

[0043] According to 3% of the daily food intake of mice (3.3 g), the daily intake of DFA is 0.099 g, so the daily gavage dose of DFA is 0.10 g (dissolved in 200 μL of sterile water).

[0044] The preparation of DFA-I and DFA-III was based on the method of Yu et al.[1] and used a multi-stage coupling process to prepare high-purity DFA from inulin raw materials.[1] Yu S, Li Q, Wang Z, et al. In vitro physiological properties of difructose anhydride I prepared from inulin by inulin fructotransferase [J]. Journal of Agricultural and Food Chemistry, 2025, 73(11): 6659-6667.

[0045] Experimental procedure:

[0046] Modeling period (days 8-14): Except for the CON group, the animals in the other groups continuously ingested 3% (w / v) dextran sulfate sodium (DSS) solution to establish a colitis model;

[0047] Intervention period (days 15-21): The corresponding substances were administered according to the experimental design;

[0048] Behavioral test period (days 22-25): behavioral tests were performed to assess depressive-like symptoms;

[0049] Sample collection (day 26): euthanasia was performed (cervical dislocation after 5% isoflurane anesthesia), and biological samples (serum / cecal content) were collected and transferred to -80°C freezer after quick freezing in liquid nitrogen.

[0050] 1.2 Open field test

[0051] Mice were placed individually in the corner of the open field box, and the time was started and their behavior was observed.

[0052] The observation time was set to 5 min. During this period, the movement trajectory of the animals was recorded using a camera system and video recording analysis software.

[0053] From Figure 1 It was observed that the total distance moved by the depression model group (DSS) showed a downward trend compared to the CON group, while the intervention groups (DFA-I and DFA-III) showed signs of recovery. The resting time of the DSS group increased significantly, reflecting their depressive behavior.

[0054] The number of times the DSS group entered the central area decreased, and the time spent in the peripheral area increased, revealing the mouse's escape tendency. After DFA intervention, mice tended to explore the central area rather than staying in the peripheral area for a long time, indicating a decrease in their depression level.

[0055] Example 2

[0056] DFA-I and DFA-III improved depressive-like behavior in mice:

[0057] 2.1 Elevated plus maze test

[0058] The elevated plus maze was placed in a well-lit place with a camera above the arm joints, and the system was connected to the computer recording system.

[0059] Mice were placed gently in the central area of the maze facing away from the experimenter, facing the open arm, and the time was started. They were allowed to move freely in the maze, and the software automatically tracked the animal's trajectory, recording the number of entries, the time spent, the total exploration distance, and other indicators.

[0060] Based on Figure 2 The heat map of A showed that the CON group had exploration trajectories in both the open arm (vertical direction) and the closed arm, indicating normal risk exploration behavior. The DSS group's activity was concentrated in the closed arm (safe zone), and the open arm was only marginally explored, consistent with the characteristics of the depression model. After DFA intervention, the mice's continuous movement path in the open arm was extended, approaching the behavior pattern of the CON group, indicating an improvement in their depressive state.

[0061] From Figure 2 B-D mice total distance, open arm entry frequency and open arm stay time multiple quantitative index analysis found that the behavior pattern of DSS group is in line with the characteristics of animal behavior, DFA intervention group in the above indicators have improved, its behavior pattern tends to CON group level.

[0062] 2.2 tail suspension test

[0063] The mouse tail was fixed with a clip (about 1 cm from the tail tip), and it was inverted to a height of 15 cm from the ground;

[0064] The immobile time of the mouse was recorded, and the last 4 min of the 6 min duration test was recorded.

[0065] Combined Figure 2 E (tail suspension immobility time) behavioral data found that the tail suspension immobility time of the DSS model group was significantly prolonged, suggesting that it had severe despair behavior. The tail suspension immobility time of the DFA intervention group was lower than that of the model group, indicating that its depression state was restored.

[0066] Example 3

[0067] DFA-I and DFA-III improve the imbalance of immune homeostasis in mice:

[0068] The mouse blood was collected in a 1.5 mL centrifuge tube, centrifuged at 8000 x g for 15 min to collect serum, and the content of TNF-α, IFN-γ, IL-6 and IL-10 in mouse serum was determined according to the enzyme-linked immunosorbent kit instructions.

[0069] According to existing research, the expression levels of TNF-α, IFN-γ, IL-6 and IL-10 in mouse serum are closely related to the occurrence and development of depression, and its mechanism involves multiple aspects such as inflammatory response, neuroimmunomodulation and neural plasticity.

[0070] TNF-α and IL-6, as key pro-inflammatory factors, are often significantly increased in depression models. IL-10, as an anti-inflammatory factor, can inhibit the release of pro-inflammatory factors (such as TNF-α and IL-6), and reduce neuronal damage by regulating microglial cell activity. In the stress animal model, the serum IL-10 level is significantly reduced. IFN-γ is significantly increased in the early stage of stress, which exacerbates the inflammatory state by activating Th1-type immune response.

[0071] Figure 3 It is shown that the expression levels of TNF-α, IFN-γ and IL-6 in the serum of the DSS group are significantly increased, and the serum IL-10 level is significantly decreased, which is in line with its behavioral characteristics, and after DFA intervention, the serum indicators are improved, indicating that its depression pathology is relieved and recovered.

[0072] Example 4

[0073] DFA-I and DFA-III improve intestinal microbial imbalance in mice:

[0074] Four fecal samples were randomly selected from each group, and microbial genomic DNA was extracted using magnetic beads. The purity was verified by 1% agarose gel electrophoresis.

[0075] Universal primers 341F and 806R were used to amplify the 16S rDNA V3-V4 region by PCR. The purified product was subjected to double-end sequencing on the Illumina NovaSeq 6000 platform.

[0076] After removing low-quality sequences from the raw data, species annotation was performed using the MicrobiomeAnalyst platform, and STAMP software was used to compare the difference between groups (p<0.05 was significant).

[0077] Lactobacillus is known as a "psychobiotic". Lactobacillus maintains interferon-gamma (IFN-γ) homeostasis, inhibits microglial cell overactivation, reduces neural inflammation, and thus enhances the adaptability to chronic stress. In addition, Lactobacillus can promote the production of short-chain fatty acids (SCFAs) such as butyric acid, repair the intestinal barrier, and reduce the level of systemic inflammation.

[0078] Bifidobacterium produces 5-hydroxytryptamine precursors (such as indole derivatives) by metabolizing tryptophan, which directly or indirectly affects the balance of central neurotransmitters. Bifidobacterium inhibits the NF-κB pathway by increasing SCFAs (such as butyric acid), reducing intestinal and central inflammation. Clostridium contains pathogenic bacteria (such as Clostridium perfringens). Pathogenic bacteria activate NLRP3 inflammasomes by releasing toxins (such as lipopolysaccharides), exacerbating intestinal leakage and neural inflammation. Studies have shown that Parabacteroides may proliferate abnormally in patients with depression, and its metabolic products (such as succinic acid) may activate Th17 immune responses, increase intestinal permeability, and promote the release of pro-inflammatory factors (such as IL-17, TNF-α). In addition, Parabacteroides indirectly affects mood regulation by degrading hormones such as testosterone.

[0079] Based on Figure 4 16S rRNA gene sequencing analysis found that the beneficial bacteria (Lactobacillus, Bifidobacterium) in the intestinal flora of the model group mice were significantly reduced, and the conditional pathogenic bacteria (Clostridium, Parabacteroides) were significantly increased, which was consistent with the intestinal flora composition of the depression mouse model. After DFA intervention, the flora disorder phenomenon was significantly reversed, especially DFA-I specifically promoted the proliferation of Bifidobacterium, restored the intestinal flora imbalance, indicating that DFA had a specific intervention effect on the intestinal flora.

[0080] Example 5

[0081] DFA-I and DFA-III improved the intestinal microbial metabolic disorder in mice:

[0082] The mouse feces collected one day before dissection were placed in a freeze dryer, 50 mg of the freeze-dried fecal sample was placed in a 2 mL centrifuge tube, 500 μL of saturated sodium chloride solution was added, and it was placed at room temperature for 30 min. A tissue grinding homogenizer was used for sufficient homogenization, 40 μL of 50% sulfuric acid solution (v / v) was added for acidification, and it was shaken for 30 s on a vortex oscillator. 800 μL of diethyl ether was added in a fume hood to extract short-chain fatty acids, and it was shaken for 30 s on a vortex oscillator. After shaking, it was centrifuged at 12,000 g at 4°C for 15 min.

[0083] The supernatant was taken, the supernatant was added to a 2 mL centrifuge tube containing 0.25 g of anhydrous sodium sulfate, and it was placed for 10 min. After centrifugation at 12,000 g at 4°C for 15 min, the supernatant was taken to a gas phase vial, and it was analyzed on the machine. The short-chain fatty acid analysis was performed using a GC-2010 Plus gas chromatography system equipped with an Rtx-Wax capillary column (30 m x 0.32 mm x 0.25 μm) and an FID detector. The standard samples included acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, and valeric acid, and they were quantitatively detected using an external standard method.

[0084] Short-chain fatty acids (SCFAs) are metabolic products produced by intestinal flora through fermentation of dietary fiber, mainly including acetic acid, propionic acid, and butyric acid.

[0085] Recent studies have shown that SCFAs play a key role in the occurrence and development of depression by regulating the microbe-gut-brain axis, and its mechanisms cover multiple aspects such as neurotransmitter regulation, immune regulation, and metabolic regulation.

[0086] Combined with the composition of intestinal flora in Example 4 and Figure 5 The data of medium-chain fatty acid determination showed that DSS intervention significantly inhibited the generation of all SCFAs by affecting intestinal flora metabolism, which was closely related to the occurrence of depressive behavior, and the content of intestinal SCFAs was effectively restored after DFA intervention.

[0087] Among them, the significant increase in acetic acid and butyric acid also echoed the proliferation of beneficial bacteria in Example 4.

[0088] Example 6

[0089] DFA-I and DFA-III improved the integrity of the blood-brain barrier in mice:

[0090] The mouse blood was collected in a 1.5 mL centrifuge tube, centrifuged at 8000 x g for 15 min to collect the serum, and the content of corticosterone CORT in the mouse serum was determined according to the enzyme-linked immunoassay kit instructions. The total RNA in the hippocampus was extracted by Trizol reagent, and the separated RNA was quantitatively measured by ultramicro UV spectrophotometer. According to the determination results, only high-quality RNA samples (OD260 / 280 = 1.8-2.2, OD260 / 230≥2.0) were used for subsequent experiments.

[0091] The RNA sample was taken, and cDNA was synthesized using a reverse transcription kit. The qPCR amplification reaction was performed using a full-automatic fluorescence quantitative PCR instrument. GAPDH was used as an internal reference, and the mRNA relative expression level of the target protein in the hippocampus was determined by 2 -ΔΔCt The detailed information of the qRT-PCR primer sequence is shown in Table 1.

[0092] Table 1

[0093]

[0094] Corticosterone (CORT) is the main glucocorticoid in mice, and the increase of its serum level is a characteristic feature of hypothalamic-pituitary-adrenal axis (HPA axis) dysfunction. In various depression models, the serum CORT level of mice is significantly increased, and is positively correlated with depressive-like behaviors (such as prolongation of static time in the tail suspension test, etc.). p75NTR is a low-affinity neurotrophic factor receptor, which is widely involved in the regulation of neural plasticity, neuronal survival and apoptosis. proBDNF (brain-derived neurotrophic factor precursor) tends to promote cell apoptosis through p75NTR receptor. In mouse models of depression, the expressions of p75NTR and proBDNF are usually increased. The expression of BDNF is controlled by multiple exons, among which exonIX is crucial for the overall BDNF expression.

[0095] It is found that in animal models of depression, the expression level of BDNF exonIX mRNA is usually significantly decreased, which is related to weakened neural plasticity, cognitive impairment and depressive behavior.

[0096] Figure 6The results showed that CORT levels in the DSS group were significantly elevated, simulating the over-activation of the HPA axis commonly seen in depressed mice. High CORT can inhibit hippocampal neurogenesis and exacerbate neuronal apoptosis. After DFA intervention, the CORT levels in the DFA group fell close to those in the CON group, suggesting that it may inhibit the over-reaction of the HPA axis by regulating the gut-brain axis (such as vagus nerve signals mediated by short-chain fatty acids). In addition, the DFA group significantly reduced p75NTR expression, which may be related to the anti-inflammatory effects of short-chain fatty acids (such as propionic acid) that block the apoptosis signal triggered by neurotrophin imbalance through the NF-κB pathway. DFA intervention can also promote the restoration of hippocampal proBDNF / mBDNF balance. The above studies suggest that DFA can inhibit the over-activation of the HPA axis by regulating the production of short-chain fatty acids by intestinal flora, and reshape the neurotrophin balance, ultimately improving depressive-like behavior.

[0097] Example 7

[0098] Process for making DFA-containing hard candy (synbiotic) products:

[0099] Example 7 provides an innovative preparation process for DFA-containing synbiotic hard candy, with a formula that uses 5% to 40% of difructose anhydrides (DFA-I or DFA-III) as the core functional ingredient, 5% to 10% of microencapsulated bifidobacteria or lactobacilli (prepared by combining the multi-layer embedding technology of sodium alginate and resistant starch with low-temperature spray drying process), 20% to 30% of oligofructose, and 25% to 35% of maltodextrin as auxiliary materials.

[0100] The specific method is as follows:

[0101] Step (1): Preparation of microencapsulated probiotics

[0102] 1.1 Mix the bifidobacteria or lactobacilli suspension with the sodium alginate solution to form a primary embedding solution.

[0103] The volume ratio of the bacterial suspension to the sodium alginate solution is 1: (5-20), i.e., the ratio of the volume of the bacterial suspension to the volume of the sodium alginate solution ranges from 1:5 to 1:20.

[0104] 1.2 Add resistant starch for secondary embedding

[0105] The amount of resistant starch added is usually 3% to 10% of the total mass of the sodium alginate solution (i.e. 3 to 10 g of resistant starch is added per 100 mL of sodium alginate solution). The resistant starch is uniformly mixed with the primary embedding solution (sodium alginate-bacterial body mixture) at a stirring speed of 200 to 400 rpm for 10 to 15 min to ensure uniform dispersion of the starch. This embedding layer can enhance the digestion resistance and slow-release effect of the microcapsules. The mixed solution is dropped into soybean oil (containing 0.4% (v / v) Tween-80 as an emulsifying agent) at a volume ratio of the droplet to the oil phase of 1:4 to 1:7 to form uncrosslinked sodium alginate droplets by emulsification.

[0106] 1.3 Solidification of the microcapsule structure by ion crosslinking technology

[0107] The emulsified uncrosslinked droplets are transferred to a calcium chloride solution to form a gel network through an ion exchange reaction between calcium ions and sodium alginate, and the solidification of the microcapsules is finally completed. The specific parameters are as follows:

[0108] Calcium chloride solution concentration: 1% to 3% (w / v) for inducing sodium alginate gelation;

[0109] Crosslinking time: 30 min for static solidification to ensure sufficient crosslinking;

[0110] Centrifugal separation conditions: centrifugal speed 1000 rpm for 5 min, and the wet capsules are collected and washed with physiological saline for 3 times to remove residual reagents.

[0111] 1.4 Dry microcapsules are prepared by low-temperature spray drying (inlet air temperature 40 to 50°C and outlet air temperature 25 to 30°C) to control the moisture content to be ≤5% and ensure the survival rate of the bacterial bodies to be ≥90%.

[0112] Step (2): Pretreatment of raw materials and sugarization

[0113] 2.1 Weigh the ingredients according to the formula: double fructose anhydride (DFA-I / III) 5% to 40%, fructooligosaccharide 20% to 30%, and maltodextrin 25% to 35%.

[0114] 2.2 Dissolve the raw materials in deionized water (material to liquid ratio 1:1.5), adjust the pH to 6.0 to 6.5 (phosphate buffer), and stir to dissolve at 60°C until completely transparent.

[0115] Step (3): Vacuum thin-film sugar boiling

[0116] 3.1 Inject the sugar solution into a vacuum thin-film sugar boiling machine, set the temperature to 140 to 145°C, the vacuum degree to -0.08 to -0.1 MPa, and quickly dehydrate to a moisture content of ≤2%.

[0117] 3.2 Real-time monitoring of sugar paste viscosity, end-point viscosity control at 500-600 mPa-s (Brookfield viscometer, 60 rpm, 25°C).

[0118] Step (4): Probiotic embedding and blending

[0119] 4.1 When the sugar paste is cooled to 80-85°C, add the microencapsulated probiotics (5-10%), and mix evenly at high speed (200-300 rpm). After the double-embedded microcapsules are treated at 80-85°C for 5 min, the survival rate of live bacteria is ≥90%, which is significantly higher than that of unembedded bacteria (<10%).

[0120] Step (5): Low-temperature casting

[0121] 5.1 Preheat the mold to 40-45°C, vacuum degas the sugar paste after injection (-0.05 MPa, 10 s) to eliminate bubbles and ensure a light transmittance of ≥80%.

[0122] 5.2 Cooling tunnel gradient cooling (45°C→25°C, humidity 30%), sugar body hardness 50-70 N / cm 2 (Texture analyzer measurement).

[0123] Step (6): Cutting and packaging

[0124] 6.1 Hard candy is cut into 5-8 g / piece, and the surface is sterilized by ultraviolet irradiation (254 nm, 10 s).

[0125] 6.2 Packaged with aluminum foil composite film filled with nitrogen, stored in the dark at 25°C or below, to ensure that the number of live probiotic bacteria is ≥1×10 8 CFU / g.

[0126] The present application combines the traditional sugar refining-boiling process of hard candy with DFA functional modification: the DFA thermal stability is maintained in a weakly acidic environment with a pH of 6.0-6.5 during the sugar refining stage, and finally a stable sugar body with a hardness of 50-70 N / cm 2 and a light transmittance of ≥80% is formed by mold casting at 40-45°C. The finished product has the dual health benefits of regulating intestinal flora (synergistic effect of double prebiotics) and low glycemic index.

[0127] It should be noted that the above examples are only used to illustrate the technical solutions of the present application and are not limiting. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced equivalently without departing from the spirit and scope of the present application, and they should be included in the scope of the present application.

Claims

1. An antidepressant composition containing difructose anhydride, characterized in that: The difructose anhydride includes type I difructose anhydride DFA-I or type III difructose anhydride DFA-III.

2. The antidepressant composition according to claim 1, wherein: The composition exerts an antidepressant effect by regulating the intestinal flora-gut-brain axis pathway.

3. The antidepressant composition according to claim 2, wherein: The composition increases the abundance of beneficial bacteria, reduces the abundance of opportunistic pathogens, and significantly increases intestinal butyrate levels; The beneficial bacteria include Bifidobacterium and Lactobacillus; and the opportunistic pathogens include Clostridium and Parabacteroides.

4. The antidepressant composition according to claim 3, wherein: The composition downregulates the expression of p75NTRmRNA in the hippocampus, upregulates the expression of BDNF exon IX, and reduces the level of serum corticosterone CORT.

5. The antidepressant composition according to claim 4, wherein: The composition reduces serum pro-inflammatory factors TNF-α and IL-6 and increases anti-inflammatory factor IL-10.

6. The antidepressant composition according to any one of claims 1 to 5, characterized in that: The composition comprises disulphose anhydride, probiotics and oligofructose, wherein the mass percentage of disulphose anhydride in the composition is 5% to 40%.

7. The antidepressant composition according to claim 6, wherein: The probiotics include at least one of Bifidobacterium or Lactobacillus, etc., wherein the probiotics are added in the form of microcapsules, accounting for 5% to 10% of the total mass.

8. Use of the antidepressant composition according to any one of claims 1 to 7 in the preparation of a functional preparation, characterized in that: The functional preparation is an oral preparation or an intestinal targeted preparation.

9. The use according to claim 8, characterized in that: The dosage form of the functional preparation includes hard candy or granules.

10. A method for preparing synbiotic hard candy containing DFA, characterized by: include, Preparation of microcapsule probiotics: a probiotic suspension is mixed with a sodium alginate solution at a volume ratio of 1:(5-20), resistant starch is added at a mass ratio of 3% to 10% of the mass of the sodium alginate solution, and after stirring at 200-400 rpm for 10-15 minutes, an oil phase containing 0.4% (v / v) Tween-80 is dropwise added at a volume ratio of 1:4-7 for emulsification, and cross-linked and cured with a 1% to 3% calcium chloride solution for 30 minutes. After centrifugal washing, the mixture is spray-dried at a temperature of 40-50°C for inlet air and 25-30°C for outlet air to produce microcapsules with a moisture content of ≤5%. Raw material dissolution: dissolve 5% to 40% by mass of disfructose anhydride, 20% to 30% of oligofructose, and 25% to 35% of maltodextrin in a pH 6.0 to 6.5 buffer solution at a material-liquid ratio of 1:1.5, and stir at 60°C until completely transparent; Vacuum sugar boiling: dehydrate at 140-145°C and -0.08-0.1 MPa vacuum to a moisture content of ≤2%, and control the viscosity of the sugar paste to 500-600 mPa·s; Embedding probiotics: When the sugar mass is cooled to 80-85°C, add 5%-10% of the total mass of microcapsule probiotics, stir and disperse at a high speed of 200-300 rpm, and after the microcapsule probiotics are treated in the sugar mass at 80-85°C for 5 minutes, the survival rate of the viable bacteria is ≥90%; Molding and curing: inject the sugar paste into the mold at 40-45℃, degas in a vacuum at -0.05MPa and then gradually cool it down to 25℃ to form a hardness of 50-70N / cm 2 of glycosomes; Sterilization packaging: Cut into 5-8g / pill, sterilize the surface by ultraviolet radiation, package with nitrogen-filled aluminum foil composite film and store below 25℃ away from light.

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