Plant prebiotics, composition and application

Abstract

The invention provides plant prebiotics, a composition and application, and belongs to the technical field of biological medicine. The plant prebiotics comprise plantain herb extract, lotus leaf extract, dandelion extract and mulberry extract, the weight ratio of the plantain herb extract to the lotus leaf extract to the dandelion extract to the mulberry extract is (2-4): (2-4): (1-3): (1-3), and the plantain herb extract and the lotus leaf extract are prepared by a synergistic reflux extraction method of alcohol extraction and water extraction in sequence. The dandelion extract and the mulberry extract are prepared by a water extraction method; the composition comprises plant prebiotics and probiotics, and traditional prebiotics can also be added. Each component of the plant prebiotics independently or jointly has the effect of inhibiting the growth of gastrointestinal pathogenic bacteria, and the plant prebiotics not only have the effects of inhibiting the gastrointestinal pathogenic bacteria, protecting the barrier function of intestinal mucosa and inhibiting the inflammatory response of a body, but also can enhance the effects of the probiotics, such as enhancing the effects of regulating the balance of intestinal flora and resisting inflammation of the probiotics.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a plant prebiotic, a composition and an application thereof. Background Art

[0002] The intestinal microenvironment includes a variety of dynamic microbial populations, such as bacteria, fungi, viruses, and protists, and the intestinal flora plays a vital role in human health. Studies have shown that disorders in the abundance and composition of intestinal flora caused by various reasons can cause changes in the intestinal microenvironment and damage the intestinal mucosal barrier, and induce intestinal and extraintestinal diseases, such as acute gastroenteritis, colon cancer, sepsis, or metabolic diseases.

[0003] Supplementing with microbial preparations is a direct way to improve intestinal flora disorders. Probiotics are one of the commonly used supplements and are widely recognized by food science and gastroenterology experts. The clinical efficacy of probiotics mainly includes adjusting the balance of intestinal flora, inhibiting the growth of harmful bacteria, increasing the concentration of beneficial metabolites and inhibiting the body's acute and chronic inflammatory response. The results of clinical studies and animal experiments have shown that probiotics have a certain inhibitory effect on the inflammatory response after gastrectomy for gastric cancer and the inflammation caused by chemotherapy after colorectal cancer surgery.

[0004] The existing application of probiotics for anti-inflammatory effects still has certain defects. For example, it has a certain improvement effect on colitis, but its anti-inflammatory effect is slightly worse than conventional drug treatment. Therefore, the anti-inflammatory effect of probiotics needs to be further improved. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a plant prebiotic, a composition and an application, aiming to solve at least one technical problem in the background technology.

[0006] The present invention is achieved in that: The first aspect of the present invention provides a plant prebiotic, which comprises a plantain extract, a lotus leaf extract, a dandelion extract and a mulberry extract; The plantain extract is prepared from the dried whole plant of plantain by a coordinated reflux extraction method of first extracting with alcohol and then extracting with water; The lotus leaf extract is prepared from dried lotus leaves by a coordinated reflux extraction method of first extracting with alcohol and then extracting with water; The dandelion extract is prepared by water extraction using the dried whole dandelion plant as raw material; The mulberry extract is prepared by water extraction using dry mulberry fruit as raw material; According to the weight ratio, the plant prebiotics include plantain extract: lotus leaf extract: dandelion extract: mulberry extract=2~4:2~4:1~3:1~3.

[0007] Furthermore, the preparation method of the plantain extract comprises the following steps: The dried whole plantago is crushed and sieved into dry powder, and then soaked in ethanol and heated under reflux for at least two times, and after solid-liquid separation, a plantago asiatica alcohol extract and a residue are obtained; After combining the plantago asiatica alcohol extracts, centrifugation, filtration, ethanol recovery under reduced pressure, resin adsorption and elution are sequentially performed to obtain a plantago asiatica alcohol extract eluate; The residue is mixed with water, heated under reflux and extracted at least once, and filtered to obtain a plantain water extract filtrate; The plantain alcohol extract eluate and the plantain water extract filtrate are respectively concentrated under reduced pressure, combined, concentrated and dried to obtain the plantain extract; The preparation method of the lotus leaf extract comprises the following steps: The dried lotus leaves are crushed and sieved into dry powder, and then soaked in ethanol and heated under reflux for at least two times, and after solid-liquid separation, a lotus leaf alcohol extract and a residue are obtained; After the lotus leaf alcohol extracts are combined, centrifugation, filtration, ethanol recovery under reduced pressure, resin adsorption and elution are performed in sequence to obtain a lotus leaf alcohol extract eluate; The residue is mixed with water, heated under reflux and extracted at least once, and filtered to obtain a lotus leaf water extract filtrate; The lotus leaf alcohol extract eluate and the lotus leaf water extract filtrate are concentrated under reduced pressure respectively, combined and then concentrated and dried to obtain the lotus leaf extract.

[0008] Furthermore, the preparation method of the dandelion extract is specifically performed as follows: the whole dried dandelion plant is crushed and sieved into dry powder, and then soaked in water and heated and refluxed for extraction at least three times; the refluxed extracts are combined, and then centrifuged to collect the supernatant, and concentrated and dried to obtain the active ingredient dry powder, which is the dandelion extract; The preparation method of the mulberry extract is specifically as follows: dry mulberry fruit is crushed and sieved into dry powder, soaked in water and then heated and refluxed for at least three times; the refluxed extracts are combined, centrifuged to collect the supernatant, concentrated and dried to obtain the active ingredient dry powder, which is the mulberry extract.

[0009] The second aspect of the present invention provides a composition, which comprises the above-mentioned plant prebiotics and probiotics.

[0010] Furthermore, the composition also includes traditional prebiotics, which include oligofructose and xylitol; in terms of weight ratio, oligofructose:xylitol=6~9:1~4.

[0011] Furthermore, according to the weight ratio, plant prebiotics: traditional prebiotics = 1:3~3:1.

[0012] Furthermore, the probiotics are selected from at least one of Lactobacillus acidophilus, Bifidobacterium infantis, Lactobacillus plantarum, Lactobacillus rhamnosus and Clostridium butyricum.

[0013] The third aspect of the present invention provides the use of the above-mentioned plant prebiotics in the preparation of a product having the effects of inhibiting gastrointestinal pathogens, regulating the balance of intestinal flora and / or anti-inflammatory effects, wherein the product is a dietary additive, a probiotic preparation, a health product or a medicine.

[0014] A fourth aspect of the present invention provides the use of the above-mentioned plant prebiotics in the preparation of a product having the effects of promoting probiotic colonization in the intestine and / or anti-inflammatory, wherein the product is a dietary additive, a probiotic preparation, a health product or a medicine.

[0015] The fifth aspect of the present invention provides the use of the above-mentioned composition, wherein the composition is used to prepare a product having the effect of regulating the balance of intestinal flora and / or anti-inflammatory effect, and the product is a probiotic preparation, a health product or a medicine.

[0016] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention provides a plant prebiotic that has the effect of inhibiting the growth of pathogenic bacteria alone or in combination. The plant prebiotic not only has the effect of protecting the intestinal mucosal barrier function and inhibiting the body's inflammatory response, but also can enhance the efficacy of probiotics.

[0017] 2. After the plant prebiotics of the present invention are combined with traditional prebiotics, they can further promote the colonization of probiotics such as Lactobacillus acidophilus, Bifidobacterium infantis, Lactobacillus plantarum, Lactobacillus rhamnosus and Clostridium butyricum in the intestine, and enhance the effect of probiotics in regulating the balance of intestinal flora and anti-inflammatory.

[0018] 3. The plant prebiotics of the present invention and the composition thereof combined with traditional prebiotics and probiotics can improve the balance of intestinal flora and restore the diversity of intestinal flora. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a graph showing changes in body weight of mice in each group during the experiment of Example 4 of the present invention; Figure 2 This is a graph showing changes in the disease activity index of each group of mice during the experiment of Example 4 of the present invention; Figure 3 This is a comparison of the spleen sizes of mice in each group at the end point of the experiment in Example 4 of the present invention; Figure 4 This is a comparison chart of the serum TNF-α levels of mice in each group at the end point of the experiment in Example 4 of the present invention; Figure 5 This is a comparison chart of the serum IL-1β levels of mice in each group at the end point of the experiment in Example 4 of the present invention; Figure 6This is a comparison chart of the serum IL-6 levels of mice in each group at the end point of the experiment in Example 4 of the present invention; Figure 7 This is a PCoA analysis diagram of the intestinal flora of each group of mice at the end point of the experiment in Example 4 of the present invention. DETAILED DESCRIPTION

[0020] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific implementation cases described herein are only used to explain the present invention and are not used to limit the present invention.

[0021] Example 1 In this embodiment 1, a plant extract complex is used as a plant prebiotic, which includes plantain extract, lotus leaf extract, dandelion extract and mulberry extract. The weight ratio of each component is plantain extract: lotus leaf extract: dandelion extract: mulberry extract = 2~4: 2~4: 1~3: 1~3. In this embodiment, the ratio is set to 3:3:2:2, but it is not limited to the listed values, and other values ​​not listed in the numerical range are also applicable.

[0022] 1. The whole plantago was purchased from the medicinal material market in Zhangshu City, Jiangxi Province. The preparation steps of the plantago extract are as follows: (1) completely crushing the dried whole herb of Plantain and passing it through an 80-mesh sieve to obtain dry powder; (2) soaking each 100 g of dry powder in 1.5 L of 80% ethanol for 3 hours to completely soak the powder, then reflux extracting at 70 degrees Celsius for 2 hours, and filtering to obtain the first alcohol extract of plantain and the residue; (3) adding 1.5 L of 80% ethanol to the residue, extracting under reflux at 70 degrees Celsius for 2 hours, and filtering to obtain a second alcohol extract and a secondary residue of plantain; (4) combining the first alcohol extract and the second alcohol extract of plantain, and recovering ethanol under reduced pressure at a temperature of 60° C. and a pressure of 0.085 MPa to obtain a reduced pressure concentrated filtrate; (5) The filtrate was concentrated under reduced pressure and adsorbed with a macroporous resin, first eluted with double distilled water of 2 times the column volume, the water washing liquid was discarded, and then eluted with 60% ethanol by volume to obtain the plantain alcohol extraction eluate; (6) Add 1.0 L of distilled water to the secondary residue, heat to 100 °C and reflux for extraction for 3 hours, and filter through filter paper to obtain the plantain water extract filtrate; (7) Concentrating the eluate of the plantain alcohol extract and the filtrate of the plantain water extract under reduced pressure using a rotary evaporator at 55°C; (8) combining the plantain alcohol extract eluate and the plantain water extract filtrate after reduced pressure concentration; (9) Reduce pressure and concentrate to a clear paste at a temperature of 60°C and a pressure of 0.085 MPa. (10) Drying under reduced pressure at 70°C and 0.08 MPa to obtain a dry powder of the active ingredient, i.e., the plantain extract.

[0023] 2. Lotus leaves were purchased from Guangchang County, Jiangxi Province. The preparation steps of lotus leaf extract are as follows: (1) Completely crush the dried lotus leaves and pass them through an 80-mesh sieve to obtain dry powder; (2) soaking each 100 g of dry powder in 1.5 L of 80% ethanol for 3 hours to completely soak it, then reflux extracting it at 70 degrees Celsius for 2 hours, and filtering to obtain the first alcohol extract of lotus leaves; (3) Add 1.5 L of 80% ethanol to the residue, reflux extract at 70 degrees Celsius for 2 hours, and obtain the second alcohol extract and secondary residue of lotus leaves after filtration; (4) combining the first alcohol extract and the second alcohol extract of the lotus leaf, and recovering ethanol under reduced pressure at a temperature of 60° C. and a pressure of 0.085 MPa to obtain a reduced pressure concentrated filtrate; (5) The filtrate was concentrated under reduced pressure and adsorbed with a macroporous resin. It was first eluted with double distilled water of 2 times the column volume, the water washing liquid was discarded, and then eluted with 60% ethanol by volume to obtain the lotus leaf alcohol extract eluate; (6) Add 1.0 L of distilled water to the secondary residue, heat to 100 degrees Celsius and reflux for extraction for 3 hours, and filter through filter paper to obtain the lotus leaf water extract filtrate; (7) Using a rotary evaporator at 55°C, the lotus leaf alcohol extract eluate and the lotus leaf water extract filtrate were concentrated under reduced pressure; (8) combining the lotus leaf alcohol extract eluate and the lotus leaf water extract filtrate after reduced pressure concentration; (9) Concentrate under reduced pressure at 60°C and 0.085 MPa to a clear paste; (10) Drying under reduced pressure at 70°C and 0.08 MPa to obtain a dry powder of the active ingredient, namely the lotus leaf extract.

[0024] 3. The whole dandelion was purchased from the medicinal material market in Zhangshu City, Jiangxi Province. The specific extraction steps of dandelion extract are as follows: (1) completely crushing the whole dried dandelion plant and passing it through an 80-mesh sieve to obtain dry powder; (2) Soak every 100 g of dry powder in 1.0 L of distilled water for 3 hours to completely soak it, heat it to 85 degrees Celsius and reflux it for 2 hours, and filter it to obtain the first extract; (3) Add 1.0 L of distilled water to the residue again, heat to 85 degrees Celsius and reflux for 2 hours, filter, and repeat the reflux extraction of the residue once more to obtain the second extract and the third extract; (4) The three extracts were combined, centrifuged at 800 g for 5 minutes, the supernatant was collected, filtered and concentrated to 1 / 5 to 1 / 2 of the volume using a low-pressure rotary evaporator; (5) Freeze-drying with a vacuum freeze dryer to obtain a dry powder of the active ingredient, which is the dandelion extract.

[0025] 4. Dried mulberry fruit was purchased from Beijing Tong Ren Tang. The specific extraction steps of mulberry extract are as follows: (1) Completely crush the dried mulberry fruits and pass them through an 80-mesh sieve to obtain dry powder; (2) Soak every 100 g of dry powder in 1.0 L of distilled water for 3 hours to completely soak it, heat it to 60 degrees Celsius and reflux it for 3 hours, and filter it to obtain the first extract and the residue; (3) Add 1.0 L of distilled water to the residue again, heat to 60 degrees Celsius and reflux for extraction for 3 hours, filter, and repeat the reflux extraction of the residue once more to obtain the second extract and the third extract; (4) The three extracts were combined, centrifuged at 800 g for 5 minutes, the supernatant was collected, filtered and concentrated to 1 / 5 to 1 / 2 of the volume using a low-pressure rotary evaporator; (5) Freeze-drying using a vacuum freeze dryer to obtain a dry powder of the active ingredient, namely, the mulberry extract.

[0026] The above active ingredient dry powders can be stored at 4°C or -20°C for long-term storage.

[0027] The present invention found that four plant extracts have antibacterial function, and the antibacterial ability of the four plant extracts and the composite was detected by the Oxford cup method, as follows: Four common gastrointestinal pathogens (Escherichia coli E. coli O157, Staphylococcus aureus S. aureus, Shigella S. castellani, Salmonella S. enteritidis) were activated with LB plate medium and then expanded with LB liquid medium. The cell concentration of the four pathogenic bacteria was adjusted to 10 8 CFU / mL, take 200μL of bacterial solution and spread it on LB plate; after the bacterial solution is blown dry, carefully put a sterile Oxford cup in the plate, add 200μL of four plant extracts and their complexes to the Oxford cup, and dissolve them in sterile aqueous solution at a concentration of 2mg / ml, and culture at 37℃ for 24h; measure the diameter of the inhibition zone in the plate. In the plant extract complex, the weight ratio of plantain extract: lotus leaf extract: dandelion extract: mulberry extract = 3:3:2:2. Gentamycin (2mg / ml) was used as a positive control. The experiment was repeated 3 times for each group. The diameter of the inhibition zone is shown in Table 1 below, in mm.

[0028] Table 1

[0029] As can be seen from Table 1, the four plant extracts and their complexes of the present invention have inhibitory effects on four common gastrointestinal pathogens, and compared with conventional antibiotics (such as gentamicin), their effects can reach more than 37% of the antibacterial effect of the drugs, and the complex of the four plant extracts can achieve more than 43% of the antibacterial effect of the drugs.

[0030] Example 2 This embodiment 2 is a composition of plant prebiotics and probiotics.

[0031] The present invention also finds that plant prebiotics can be used as synergists and are mixed with probiotics to prepare a composition. The probiotics are selected from at least one of Lactobacillus acidophilus, Bifidobacterium infantis, Lactobacillus plantarum, Lactobacillus rhamnosus and Clostridium butyricum. Specifically, the corresponding probiotics commonly used in the art can be used, and the above-mentioned probiotics are made into freeze-dried powder after high-density fermentation.

[0032] The plant prebiotics prepared in Example 1 were mixed with the probiotic freeze-dried powder, and every 2 g of plant prebiotics was mixed with no less than 1×10 9 If there are multiple probiotic complexes, the content of each probiotic complex shall not be less than 1×10 9 CFU.

[0033] Example 3 This embodiment 3 is a composition of plant prebiotics, traditional prebiotics and probiotics.

[0034] Traditional prebiotics include oligofructose and xylitol, and the weight ratio of oligofructose: xylitol is 6-9:1-4. In the specific embodiment of the present invention, the ratio of the components is 8:2, but it is not limited to the listed values. Other values ​​not listed in the numerical range are also applicable. There are three purposes for adding traditional prebiotics to plant prebiotics: (1) as a flavor enhancer, to adjust the taste and further expand the application of plant prebiotics in the food field; (2) to avoid the potential risk of allergies caused by common prebiotic inulin, while exerting its own prebiotic function and avoiding the addition of other additives to affect the efficacy of plant prebiotics; (3) as a synergist of plant prebiotics, it synergistically promotes the colonization of probiotics in the intestine and anti-inflammatory effects in combination with plant prebiotics.

[0035] The weight ratio of plant prebiotics to traditional prebiotics is 1:3 to 3:1. In a specific embodiment of the present invention, the weight ratio of plant prebiotics to traditional prebiotics is 1:1, but is not limited to the listed values, and other values ​​not listed within the numerical range are also applicable.

[0036] The prepared plant prebiotics, traditional prebiotics and probiotic freeze-dried powder were mixed, and every 2g of total prebiotics was mixed with no less than 1×10 9 If there are multiple probiotic complexes, the content of each probiotic complex shall not be less than 1×10 9 CFU.

[0037] Example 4 This Example 4 uses animal experiments to verify the efficacy of the products of Examples 1 to 3, specifically using a mouse colitis model for verification.

[0038] Source of mice and feeding environment: C57BL / 6J female mice, 6 weeks old, were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd. and kept in the Animal Center of Nanchang University. The animal use license number is SYXK (Gan) 2021-0004. The feeding environment is an SPF-level animal room with standard environmental conditions, that is, a specific pathogen-free animal room. The feeding conditions are strictly controlled as follows: L / D = 12h / 12h, temperature = 23℃±1℃, humidity = 40%~60%. Animals have free access to standard diets and drinking water; this experiment was approved by the Nanchang University Experimental Animal Welfare Ethics Committee.

[0039] Animal experimental grouping and corresponding treatment: divided into 7 groups, each with 6 mice.

[0040] ① Blank control group (Group C): The mice were allowed to eat freely and were gavaged with sterile water for 10 days (once a day, 200 μL / time).

[0041] ② Model group (M group): 2% DSS solution (dextran sulfate sodium) was used for induction for 7 days. Sterile saline was used for gavage during the induction period. After the induction, the mice were gavaged with sterile water for 3 days (1 time / day, 200 μL / time).

[0042] ③ Probiotic group (PRO group): 2% DSS solution (dextran sulfate sodium) was used for induction for 7 days. During the induction period, the mice were gavaged with the probiotic compound preparation solution (once a day, 200 μL / time). After the induction, the mice were gavaged with the probiotic compound preparation solution for 3 days (once a day, 200 μL / time). The probiotic compound preparation solution was a solution formed by mixing Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium infantis and Clostridium butyricum in a weight ratio of 1:1:1:1:1 and then dissolved in water. The concentration of each probiotic was not less than 10 9 CFU / ml.

[0043] ④ Plant prebiotic group (PE group): 2% DSS solution (dextran sulfate sodium) was induced for 7 days. During the induction period, the mice were gavaged with plant prebiotic solution (1 time / day, 200μL / time). After the induction, the mice were gavaged with plant extract solution for 3 days (1 time / day, 200μL / time). The plant prebiotic solution is a solution formed by dissolving plantain extract, lotus leaf extract, dandelion extract, and mulberry extract in sterile water at a weight ratio of 3:3:2:2. The dosage for mice is 0.4g plant prebiotic mass / kg mouse body weight. Taking a mouse weighing 20g as an example, the concentration of the plant prebiotic solution is 40mg / mL.

[0044] ⑤Compound prebiotic group (PRE group): 2% DSS solution (dextran sulfate sodium) was induced for 7 days. During the induction period, mice were gavaged with compound prebiotic solution (1 time / day, 200μL / time). After the induction, mice were gavaged with plant prebiotic combination solution for 3 days (1 time / day, 200μL / time). The compound prebiotic solution is a solution formed by mixing plant prebiotics and traditional prebiotics in a weight ratio of 1:1 and dissolving in sterile water. Among them, the plant prebiotics are the same as those in the PE group, and the traditional prebiotics are oligofructose and xylitol mixed in a weight ratio of 4:1. The dosage of mice is 0.4g plant prebiotic mass / kg mouse body weight. Taking a mouse weighing 20g as an example, the amount of plant prebiotics in this group of solutions is 8mg, and its concentration is 40mg / mL.

[0045] ⑥ Probiotics + plant prebiotics group (PRO + PE group): 2% DSS solution (dextran sulfate sodium) was used for induction for 7 days. During the induction period, mice were gavaged with probiotics + plant prebiotics solution (1 time / day, 200μL / time). After the induction, mice were gavaged with probiotics + plant extract solution for 3 days (1 time / day, 200μL / time). Among them, the composition of probiotics was the same as that of the PRO group, and the composition of plant prebiotics was the same as that of the PE group. The probiotics + plant prebiotics solution was a solution formed by dissolving plant prebiotics and probiotic dry powder in sterile water, and the concentration of each probiotic was not less than 10 9 CFU / ml, the dosage for mice is 0.4g plant extract mass / kg mouse body weight. Taking the mouse weight of 20g as an example, the dosage of plant prebiotics in this group of solutions is 8mg, and its concentration is 40mg / mL.

[0046] ⑦ Probiotics + compound prebiotics group (PRO+PRE group): 2% DSS solution (dextran sulfate sodium) was used for induction for 7 days, during which mice were gavaged with probiotics + compound prebiotics solution (1 time / day, 200μL / time), and then mice were gavaged with probiotics + plant prebiotics combination solution for 3 days (1 time / day, 200μL / time). The composition of probiotics was the same as that of the PRO group, and the composition of compound prebiotics was the same as that of the PRE group. The probiotics + compound prebiotics solution was a solution formed by dissolving compound prebiotics and probiotic dry powder in sterile water, and the concentration of each probiotic was not less than 10 9 CFU / ml, the dosage for mice is 0.4g plant prebiotics / kg mouse weight. Taking the weight of mice as 20g, the dosage of plant prebiotics in this group of solutions is 8mg, and its concentration is 40mg / mL.

[0047] All mice in each group were euthanized on the 10th day of the experiment.

[0048] 1. Changes in symptoms and signs of mice during the experiment During the 10-day experiment, the body weight of each group of mice was measured every day, and the ratio of the body weight to the initial body weight before the experiment was used as the body weight change. Figure 1 As shown by Figure 1 It can be seen that: the body weight of mice in group C basically maintained a steadily increasing trend; the body weight of mice in group M decreased rapidly after induction administration, and increased slowly after stopping induction administration; the body weight of mice in group PRO and group PE decreased rapidly after induction administration, but the degree of decrease was lower than that of mice in group M, and their body weight increased slowly about 1 day after stopping induction administration, and the degree of increase was higher than that of mice in group M, and the final body weight of group PE>group PRO>group M; while the body weight of mice in group PRO+PRE, group PRO+PE, and group PRE decreased after administration, but the degree of decrease was relatively gentle, and their body weight increased rapidly after stopping induction administration. At the end of the experiment, the body weights of mice in the PRO+PRE group (102.08%±0.04%), PRO+PE group (97.87%±0.99%), PRE group (98.90%±1.07%), PE group (97.10%±0.76%), and PRO group (96.31%±1.32%) were significantly higher than those in the M group (93.51%±1.83%), which was statistically significant (P<0.001). Figure 1 The average value of the ratio of the measured body weight to the initial body weight of each group of mice was plotted.

[0049] The disease activity index scores (DAI scores) of the seven groups of mice during the experiment were further compared. The disease activity index consists of three parts: blood in stool score, diarrhea score, and weight loss score. The results are as follows: Figure 2 As shown. Figure 2 It can be seen that the PRO+PRE group, PRO+PE group, PRE group, PRO group, and PE group can all improve the disease activity index of mice, among which the PRO+PRE group and PRO+PE group have better effects, and the PRO+PRE group has the best effect. Figure 2 The mean DAI score of each group of mice was plotted.

[0050] 2. Comparison of mouse colon length and spleen mass at the end of the experiment At the end of the experiment, the colon length and spleen weight of each group of mice were measured. The data are shown in Table 2. Figure 3 shown.

[0051] Table 2

[0052] As shown in Table 2, at the end of the experiment, the colon of mice in group M was significantly shorter than that of mice in group C, while the PRO+PRE group, PRO+PE group, PRE group, PRO group, and PE group could significantly improve the colon length of mice, among which the PRO+PRE group and PRO+PE group had better effects, and the PRO+PRE group had the best effect. The P value of group M was calculated by comparing group M with group C, and the P values ​​of group PRO, group PE, group PRE, group PRO+PE, and group PRO+PRE were all obtained by comparing with group M.

[0053] Figure 3 From left to right in the figure are the spleens of mice after treatment in group C, group M, group PRO, group PE, group PRE, group PRO+PE, and group PRO+PRE. At the end of the experiment, the spleens of mice in group M were significantly larger than those in group C, indicating that the level of inflammation in mice was significantly increased. The spleens of mice in groups PRO+PRE, PRO+PE, PRE, PRO, and PE were all reduced after treatment, among which the effects of groups PRO+PRE and PRO+PE were better, and the effect of group PRO+PRE was the best.

[0054] 3. Differences in serum inflammatory indicators TNF-α, IL-6, and IL-1β in mice at the end of the experiment The differences in serum inflammatory indicators TNF-α, IL-1β and IL-6 of mice in each group at the end of the experiment were as follows Figure 4 , Figure 5 and Figure 6 shown.

[0055] Depend on Figures 4 to 6It can be seen that at the end of the experiment, the inflammatory indicators TNF-α, IL-6, and IL-1β of mice in group M were significantly higher than those of mice in group C. After intervention, the inflammation levels of mice in groups PRO+PRE, PRO+PE, PRE, PE, and PRO were significantly reduced. Among them, the effects of reducing inflammation levels in groups PRO+PRE and PRO+PE were better, and the effect of PRO+PRE was the best. Figures 4 to 6 In the table, *** indicates P < 0.001, ### indicates P < 0.001 (all compared with group M).

[0056] 4. Effect of intestinal flora diversity in mice At the end of the experiment, mouse feces were collected and 16S rRNA high-throughput sequencing was performed to observe the effects of plant prebiotics and probiotics on the diversity of mouse intestinal flora.

[0057] Among them, the α-diversity indices such as the Shannon index, Simpson index, and Chao1 index are shown in Table 3. The Shannon index and Chao1 index increase with the increase of species richness, while the Simpson index decreases with the increase of species richness.

[0058] Table 3

[0059] From the data in Table 3, we can see that the Shannon index and Chao1 index of mice in group M were significantly lower than those in group C, and the Simpson index was significantly increased. After intervention, the Shannon index and Chao1 index of mice in group PRO+PRE, group PRO+PE, group PRE, group PE, and group PRO were significantly increased and the Simpson index was significantly decreased compared with group M, indicating that the intervention can help restore the richness of intestinal flora to a certain extent. The P value of group M was calculated by comparing group M with group C, and the P values ​​of group PRO, group PE, group PRE, group PRO+PE, and group PRO+PRE were all obtained by comparing with group M.

[0060] The β diversity index of the intestinal flora of each group of mice was further analyzed, and PCoA analysis of the intestinal flora was performed at the OTU level, that is, principal coordinates analysis (PCoA), such as Figure 7 As shown. Figure 7 It can be seen that the community structure of intestinal flora in M ​​group mice showed significant changes compared with that in C group mice, and intervention in PRO+PRE group, PRO+PE group, PRE group, PE group, and PRO group can help restore the normal community structure of intestinal flora.

[0061] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A plant prebiotic, characterized in that: The plant prebiotics include plantain extract, lotus leaf extract, dandelion extract and mulberry extract; The plantain extract is prepared from the dried whole plant of plantain by a coordinated reflux extraction method of first extracting with alcohol and then extracting with water; The lotus leaf extract is prepared from dried lotus leaves by a coordinated reflux extraction method of first extracting with alcohol and then extracting with water; The dandelion extract is prepared by water extraction using the dried whole dandelion plant as raw material; The mulberry extract is prepared by water extraction using dry mulberry fruit as raw material; According to the weight ratio, the plant prebiotics include plantain extract: lotus leaf extract: dandelion extract: mulberry extract=2~4:2~4:1~3:1~3.

2. A plant prebiotic according to claim 1, characterized in that: The preparation method of the plantain extract comprises the following steps: The dried whole plantago is crushed and sieved into dry powder, and then soaked in ethanol and heated under reflux for at least two times, and after solid-liquid separation, a plantago asiatica alcohol extract and a residue are obtained; After combining the plantago asiatica alcohol extracts, centrifugation, filtration, ethanol recovery under reduced pressure, resin adsorption and elution are sequentially performed to obtain a plantago asiatica alcohol extract eluate; The residue is mixed with water, heated under reflux and extracted at least once, and filtered to obtain a plantain water extract filtrate; The plantain alcohol extract eluate and the plantain water extract filtrate are respectively concentrated under reduced pressure, combined, concentrated and dried to obtain the plantain extract; The preparation method of the lotus leaf extract comprises the following steps: The dried lotus leaves are crushed and sieved into dry powder, and then soaked in ethanol and heated under reflux for at least two times, and after solid-liquid separation, a lotus leaf alcohol extract and a residue are obtained; After the lotus leaf alcohol extracts are combined, centrifugation, filtration, ethanol recovery under reduced pressure, resin adsorption and elution are performed in sequence to obtain a lotus leaf alcohol extract eluate; The residue is mixed with water, heated under reflux and extracted at least once, and filtered to obtain a lotus leaf water extract filtrate; The lotus leaf alcohol extract eluate and the lotus leaf water extract filtrate are concentrated under reduced pressure respectively, combined and then concentrated and dried to obtain the lotus leaf extract.

3. A plant prebiotic according to claim 1, characterized in that: The preparation method of the dandelion extract is specifically as follows: the whole dried dandelion plant is crushed and sieved into dry powder, and then soaked in water and heated and refluxed for at least three times; the refluxed extracts are combined, and then the supernatant is collected by centrifugation, and concentrated and dried to obtain the active ingredient dry powder, which is the dandelion extract; The preparation method of the mulberry extract is specifically as follows: dry mulberry fruit is crushed and sieved into dry powder, soaked in water and then heated and refluxed for at least three times; the refluxed extracts are combined, centrifuged to collect the supernatant, concentrated and dried to obtain the active ingredient dry powder, which is the mulberry extract.

4. A composition, characterized in that The composition comprises the plant prebiotics and probiotics according to any one of claims 1 to 3.

5. The composition according to claim 4, characterized in that: The composition also includes traditional prebiotics, which include oligofructose and xylitol; in terms of weight ratio, oligofructose:xylitol=6-9:1-4.

6. The composition according to claim 5, characterized in that In terms of weight ratio, plant prebiotics: traditional prebiotics = 1:3~3:

1.

7. The composition according to any one of claims 4 to 6, characterized in that The probiotics are selected from at least one of Lactobacillus acidophilus, Bifidobacterium infantis, Lactobacillus plantarum, Lactobacillus rhamnosus and Clostridium butyricum.

8. The use of a plant prebiotic according to any one of claims 1 to 3, characterized in that: The plant prebiotics are used to prepare products with the effects of inhibiting gastrointestinal pathogens, regulating the balance of intestinal flora and / or anti-inflammatory effects.

9. The use of a plant prebiotic according to any one of claims 1 to 3, characterized in that: The plant prebiotics are used to prepare products with the effects of promoting colonization of probiotics in the intestine and / or anti-inflammatory effects.

10. Use of the composition according to any one of claims 4 to 7, characterized in that: The composition is used for preparing products with the functions of regulating the balance of intestinal flora and / or anti-inflammatory effects.

Images