Application of fermented ginseng zymolyte in preparation of product for improving inflammatory bowel disease

Abstract

The invention relates to application of fermented ginseng zymolyte in preparation of a product for improving inflammatory bowel disease, and belongs to the technical field of biological medicine. The invention provides application of fermented ginseng zymolyte in preparation of products for improving inflammatory bowel diseases, the fermented ginseng zymolyte contains characteristic peptide, and the amino acid sequence of the characteristic peptide is selected from at least one of SEQ ID NO.1-9. The effect of the fermented ginseng zymolyte for relieving DSS-induced ulcerative enteritis and TNBS-induced Crohn disease-like intestinal lesions is reported for the first time, so that the inflammatory bowel disease is improved.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to the application of fermented ginseng hydrolysate in the preparation of products for improving inflammatory bowel disease. Background Art

[0002] Ginseng (Panax ginseng CAMeyer) is a perennial herbaceous plant belonging to the genus Panax L. of the family Araliaceae. Modern medical research and clinical practice have shown that ginseng has significant clinical efficacy on diseases of the central nervous system, cardiovascular system, respiratory system, endocrine system and digestive system. In addition, ginseng is also commonly used to relieve fatigue, enhance memory and restore clinical symptoms such as physical weakness. Ginseng has a huge market space as a food, dietary supplement, functional food and medical product.

[0003] Inflammatory bowel disease (IBD) is an idiopathic intestinal inflammatory disease that affects the ileum, rectum, and colon. The etiology and pathogenesis have not yet been fully clarified. It is known that the inflammatory response caused by the abnormal response of the intestinal mucosal immune system plays an important role in the onset of IBD. It is believed that it is caused by the interaction of multiple factors, mainly including environmental, genetic, infectious and immune factors. The current treatment strategies are mainly drug therapy and surgical treatment. However, drug therapy can easily cause patients to become dependent, while surgical treatment is prone to complications. Therefore, it is very important to develop new safe and effective treatment strategies for patients with inflammatory bowel disease. Summary of the invention

[0004] The purpose of the present invention is to overcome the shortcomings of the prior art and provide an application of fermented ginseng hydrolysate in preparing a product for improving inflammatory bowel disease.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] The invention provides the use of fermented ginseng hydrolysate in preparing a product for improving inflammatory bowel disease.

[0007] The present invention reports for the first time that fermented ginseng hydrolysate has the efficacy of improving DSS-induced ulcerative enteritis and TNBS-induced Crohn's disease-like intestinal lesions, thereby improving inflammatory bowel disease. At the same time, the present invention finds through animal experiments that the fermented ginseng hydrolysate of the present invention has a better efficacy in improving inflammatory bowel disease than commercially available ginseng peptides.

[0008] As a preferred embodiment of the application of the present invention, the fermented ginseng hydrolysate contains characteristic peptides, and the amino acid sequence of the characteristic peptides is selected from at least one of SEQ ID NO.1-9. The present invention obtains fermented ginseng hydrolysate by enzymolysis of fermented ginseng, and through HPLC-MS / MS identification, it is found that the fermented ginseng hydrolysate contains characteristic peptides with amino acid sequences of SEQID NO.1-9, and animal experiments confirm that the 9 characteristic peptides have the effect of improving inflammatory bowel disease, and the characteristic peptides are the main active substances of the fermented ginseng hydrolysate.

[0009] As a preferred embodiment of the application of the present invention, the method for preparing the fermented ginseng hydrolysate comprises the following steps:

[0010] (1) Mixing fermented ginseng with water, adjusting the pH to 7.5-8.5, and heating in a water bath at 40-50° C. for 35-40 min to obtain an extract;

[0011] (2) adding protease to the extract obtained in step (1), performing enzymatic hydrolysis at 40-50° C. for 4-5 hours, and centrifuging to obtain an enzymatic hydrolyzate;

[0012] (3) The enzymatic hydrolysate obtained in step (2) is concentrated and dried to obtain a fermented ginseng enzymatic hydrolysate.

[0013] As a preferred embodiment of the application of the present invention, in step (1), the fermented ginseng is ginseng residue after microbial fermentation and filtration, and the ratio of the fermented ginseng to water is fermented ginseng: water = 1g: (25-35)mL.

[0014] As a preferred embodiment of the application of the present invention, in step (1), the pH is 8.0, and the mixture is heated in a water bath at 45°C for 30 minutes.

[0015] As a preferred embodiment of the application of the present invention, in step (2), the protease is trypsin, and the added amount of the protease is 0.70-0.75 mg of protease per 1 g of fermented ginseng, and the concentration of the protease is 1250-1300 U / mg.

[0016] As a preferred embodiment of the application of the present invention, in step (2), the enzymatic hydrolysis condition is enzymatic hydrolysis at 50° C. for 5 hours.

[0017] As a preferred embodiment of the application of the present invention, in step (2), the enzymatic hydrolysate is bathed in 85-90° C. water for 30 min before centrifugation.

[0018] As a preferred embodiment of the application of the present invention, in step (3), the concentration is reduced pressure concentration, and the drying is freeze drying.

[0019] As a preferred embodiment of the application of the present invention, in step (3), the conditions for reduced pressure concentration are 65-70°C and -(0.09-0.10)MPa.

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

[0021] The present invention obtains fermented ginseng hydrolysate by enzymolysis of fermented ginseng, and identifies the fermented ginseng hydrolysate as containing characteristic peptides with amino acid sequences of LY, MM, LR, AP, LAR, RLDFR, SALAFR, LLLLGH and LLLLLHG by HPLC-MS / MS. The present invention has been confirmed by animal experiments to have good anti-inflammatory effects, can treat ulcerative enteritis and Crohn's disease, improve inflammatory bowel disease, and has the potential to treat inflammatory bowel disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the liquid quality BPC diagram of the fermented ginseng hydrolysate in Example 1 of the present invention;

[0023] Figure 2 is the secondary mass spectrum of the LY peptide segment in Example 1 of the present invention;

[0024] Figure 3 is the secondary mass spectrum of the MM peptide segment in Example 1 of the present invention;

[0025] Figure 4 is the secondary mass spectrum of the LR peptide segment in Example 1 of the present invention;

[0026] Figure 5 is the secondary mass spectrum of the LAR peptide segment in Example 1 of the present invention;

[0027] Figure 6 is the secondary mass spectrum of the SALAFR peptide segment in Example 1 of the present invention;

[0028] Figure 7 is the secondary mass spectrum of the RLDFR peptide segment in Example 1 of the present invention;

[0029] Figure 8 is the secondary mass spectrum of the LLLLGH peptide segment in Example 1 of the present invention;

[0030] Fig. 9 is the secondary mass spectrum of the LLLLLHG peptide segment in Example 1 of the present invention;

[0031] Fig.10 This is the secondary mass spectrum of the AP peptide segment in Example 1 of the present invention. DETAILED DESCRIPTION

[0032] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0033] Unless otherwise specified, other materials, reagents, etc. used in the examples can be obtained from commercial sources.

[0034] The ginseng species used in the fermented ginseng hydrolysate and fermented ginseng of the present invention are not limited. The fermented ginseng used in the following embodiments and comparative examples is obtained by subjecting ginseng to microbial fermentation according to the method of Example 1 in the technology disclosed in CN 116334147 A, and centrifugation is performed after the fermentation is completed, and the filter residue obtained is the fermented ginseng.

[0035] SPF male C57BL / 6 mice were purchased from Guangdong Medical Experimental Animal Center (SCXK (Yue) 2022-0002).

[0036] The techniques not described in detail in the following embodiments, comparative examples and effect examples are all commonly used techniques in the art. Please refer to "Molecular Biology Experiment Manual" (Ma Wenli, People's Military Medical Publishing House), "Molecular Biology Experiment (Second Edition)" (Zhejiang University Press), and "Cell Biology Experiment" (Yang Hongbing, Hou Lixia, Zhang Yuxi, Higher Education Press).

[0037] In the following embodiments and effect examples, "V" refers to valine (Val), "P" refers to proline (Pro), "G" refers to glycine (Gly), "H" refers to histidine (His), "R" refers to arginine (Arg), "K" refers to lysine (Lys), "I" refers to isoleucine (Ile), "F" refers to phenylalanine (Phe), "L" refers to leucine (Leu), "W" refers to tryptophan (Trp), "A" refers to alanine (Ala), "M" refers to methionine (Met), "C" refers to cysteine ​​(Cys), "N" refers to asparagine (Asn), "S" refers to serine (Ser), "Q" refers to glutamine (Gln), "Y" refers to tyrosine (Tyr), "D" refers to aspartic acid (Asp), "E" refers to glutamic acid (Glu), and "T" refers to threonine (Thr).

[0038] In the following examples, comparative examples and effect examples, the LY characteristic peptide refers to the characteristic peptide whose amino acid sequence is shown in SEQ ID NO.1, MM refers to the characteristic peptide shown in SEQ ID NO.2, LR characteristic peptide refers to the characteristic peptide shown in SEQ ID NO.3, LAR characteristic peptide refers to the characteristic peptide shown in SEQ ID NO.4, SALAFR characteristic peptide refers to the characteristic peptide shown in SEQ ID NO.5, RLDFR characteristic peptide refers to the characteristic peptide shown in SEQ ID NO.6, LLLLGH characteristic peptide refers to the characteristic peptide shown in SEQ ID NO.7, LLLLLHG characteristic peptide refers to the characteristic peptide shown in SEQ ID NO.8, and AP characteristic peptide refers to the characteristic peptide shown in SEQ ID NO.9.

[0039] Example 1

[0040] This embodiment provides a fermented ginseng hydrolyzate and a preparation method thereof, the preparation method comprising the following steps:

[0041] S1. Mix the fermented ginseng with water at a solid-liquid ratio of 1 g:30 mL, adjust the pH to 8.0 with 2M NaOH solution, and heat in a water bath at 45°C for 40 min to obtain an extract;

[0042] S2. Add 0.70 mg of trypsin (concentration of 1300 U / mg) per 1 g of fermented ginseng residue, perform enzymolysis at 45°C for 4 h, place in a water bath at 85°C for 30 min, cool to room temperature, centrifuge at 10000 rpm for 10 min, collect the supernatant, and obtain an enzymolysis solution;

[0043] S2. The enzymatic hydrolysate obtained in step S1 is concentrated under reduced pressure at 65° C. and −0.09 MPa, and freeze-dried to obtain the fermented ginseng hydrolysate.

[0044] Example 2

[0045] The fermented ginseng hydrolysate obtained in Example 1 was characterized, and the specific scheme is as follows:

[0046] 1. The peptide sequence of the fermented ginseng hydrolysate obtained in Example 1 was identified.

[0047] The amino acid sequence of the fermented ginseng hydrolysate prepared in Example 1 was identified using the HPLC-MS / MS method. The specific detection method is as follows:

[0048] The samples were separated and detected using an X500 LC-ESI-Q-TOF high-resolution liquid chromatography-mass spectrometer (AB SCIEX, USA). The liquid chromatography and mass spectrometry control software used was SCIEX OS2.0 integrated version (AB SCIEX, USA); the chromatographic column used was 1×100mm HSS T3 (1.8μm, Waters, USA).

[0049] The mobile phase consisted of 0.1% (v / v) formic acid aqueous solution (A) and acetonitrile (B), and the elution method was: 0-4.00min5.0% B, 4.00-6.00min5.0-10.0% B, 6.00-30.00min10.0-40.0% B, 30.00-34.00min40.0-90.0% B, 34.00-40.00min90% B, 40.00-42.00min90.0-5.0% B, 42.00-52.00min5.0% B, flow rate 0.05mL / min, injection volume 1μL, column temperature 40°C, the percentages of the above mobile phases mean volume percentages, volume of mobile phase B / total volume of mobile phase A and mobile phase B.

[0050] Mass spectrometry detection method: scanning cycle 0.642s, ESI ion source temperature 500℃, positive ion mode, spray voltage 5500V, TOF primary scanning range 100-1200Da, secondary scanning range 50-1200Da, working mode IDA, maximum candidate ion number 4, dynamic exclusion turned on, and the rest of the parameters used the default values ​​of the proteomics method. Note that the dead volume should be minimized before using the instrument. The identification results are shown in Figure 1-10 and Table 1.

[0051] Table 1 Identification results of characteristic peptide sequences in fermented ginseng hydrolysate

[0052]

[0053]

[0054] like Figure 1-10 As shown in Table 1, the present invention uses HPLC-MS / MS technology to identify 9 characteristic amino acid sequences from the fermented ginseng hydrolysate, including 4 dipeptides (LY, MM, LR, AP), 1 tripeptide (LAR), 1 pentapeptide (RLDFR), 2 hexapeptides (SALAFR, LLLLGH), and 1 septapeptide (LLLLLHG), which can be used as quality control standards for the fermented ginseng hydrolysate.

[0055] 2. Determination of the ginsenoside content of the fermented ginseng hydrolysate obtained in Example 1

[0056] The ginsenoside contents in ginseng, fermented ginseng and fermented ginseng hydrolysate were determined by HPLC.

[0057] HPLC conditions: Waters UPLC liquid chromatograph, chromatographic column: Waters XBridge BEH Shield C18 2.1 mm × 100 mm × 1.7 μm, column temperature: 40 ° C, detection wavelength: 203 nm, flow rate: 0.6 mL / min, injection volume: 2 μ L, mobile phase consists of water (A) and acetonitrile (B), elution method: 0-0.5 min 15% B, 40.5-14.5 min 15-30% B, 14.5-15.5 min 30-32% B, 15.5-18.5 min 32-38% B, 18.5-24.0 min 38-43% B, 24.0-27.0 min 43.0-55.0% B, 27.0-31.0 min, 55% B, 31.0-35.0 min, 55-70% B, 35.0-38.0min, 70-90%B. The percentages of the above mobile phases are all volume percentages.

[0058] Preparation of reference solution: Weigh 10.35 mg of ginsenoside Rg1, 10.25 mg of ginsenoside Re, 10.08 mg of ginsenoside Rb1, 10.07 mg of ginsenoside Rd, 10.70 mg of ginsenoside Rc and 10.02 mg of ginsenoside Rb2 respectively, add appropriate amount of methanol to dissolve and make the volume to 5 mL, then shake well; add appropriate amount of methanol to dissolve 3.16 mg of ginsenoside Rf to make the volume to 1.5 mL, then shake well. Accurately pipette 1 mL of the above solution into a 20 mL volumetric flask, dilute to the mark with methanol and shake well to obtain reference solutions with ginsenoside Rg1 concentrations of 103.5 μg / mL, ginsenoside Re concentrations of 102.5 μg / mL, ginsenoside Rb1 concentrations of 100.8 μg / mL, ginsenoside Rd concentrations of 100.7 μg / mL, ginsenoside Rf concentrations of 105.35 μg / mL, ginsenoside Rc concentrations of 107.0 μg / mL, and ginsenoside Rb2 concentrations of 100.2 μg / mL.

[0059] Preparation of sample solution: Take about 1g each of ginseng powder (passed through No. 4 sieve), fermented ginseng, and fermented ginseng hydrolysate, accurately weigh, place in a 20mL volumetric flask, add appropriate amount of methanol, treat with ultrasound (500W, 37kHz) for 30 minutes, take out, leave overnight, ultrasound (500W, 37kHz) for 30min, cool and add methanol to the scale, shake well, filter with a 0.22μm organic filter membrane, and take the filtrate to obtain.

[0060] The results of ginsenoside detection are shown in Table 2.

[0061] Table 2 Ginsenoside detection results of different samples

[0062] project Ginseng(%) Fermented ginseng residue (%) Fermented ginseng hydrolysate (%) <![CDATA[Ginsenoside Rb1]]> 0.316 0.0697 0.0858 <![CDATA[Ginsenoside Rb2]]> 0.215 0.0724 0.0761 Ginsenoside Rc 0.258 0.0646 0.0874 Ginsenoside Rd 0.107 0.1251 0.1246 Ginsenoside Re 0.395 Not detected Not detected Ginsenoside Rf 0.119 0.0455 0.0587 <![CDATA[Ginsenoside Rg1]]> 0.264 Not detected Not detected Total saponins 1.674 0.3773 0.4326

[0063] As shown in Table 2, the content of ginsenoside in fermented ginseng and fermented ginseng hydrolysate is extremely low, indicating that the main active ingredients of fermented ginseng hydrolysate are characteristic peptides with amino acid sequences as shown in SEQ ID NO.1-9.

[0064] Example 3

[0065] Dextran sulfate sodium (DSS) enters the gastrointestinal tract through drinking water, directly damages colonic epithelial cells, destroys the intestinal mucosal barrier, and thus induces intestinal inflammation. This inflammatory response is similar to the pathological characteristics of human ulcerative colitis. Therefore, this example uses DSS to induce ulcerative colitis in mice, and based on this model, evaluates the improvement effect of the fermented ginseng hydrolysate in Example 1 on inflammatory bowel disease. The specific scheme is as follows:

[0066] The experiment was conducted after 6-week-old SPF C57BL / 6 male mice were adaptively raised for one week at an ambient temperature of 20-25°C, a humidity of 40% to 60%, and a daily light intensity of 12 h (7:00-19:00).

[0067] The mice were randomly divided into a normal control group, a DSS model group, a low-dose (250 mg / kg) group of fermented ginseng hydrolysate, a medium-dose (500 mg / kg) group of fermented ginseng hydrolysate, a high-dose (1000 mg / kg) group of fermented ginseng hydrolysate, and a mesalazine positive drug group (200 mg / kg), with 5 mice in each group. Except for the normal control group, the mice in the remaining groups were modeled in the following manner: drinking water containing 3% DSS was given to drink freely, while the mice in the normal group drank distilled water freely. From the date of modeling, the fermented ginseng hydrolysate and mesalazine prepared in Example 1 were gavaged every day, while the normal control group and the model group were given an equal volume of physiological saline. On the 7th day after modeling, the mice were euthanized and the colon tissue was isolated.

[0068] 1. Evaluate the enteritis symptoms of mice according to the disease activity index (DAI) scoring criteria, including the following three items:

[0069] Weight change scores: 0 (no change), 1 (1-5% loss), 2 (5-10% loss), 3 (>10% loss);

[0070] Stool shape score: 0 (normal), 1 (soft stool), 2 (loose stool), 3 (diarrhea);

[0071] Bleeding score: 0 (no blood), 1 (mild), 2 (moderate), 3 (severe).

[0072] It was calculated according to the following formula: DAI = (weight change score + stool shape score + bleeding score) / 3.

[0073] The DAI scoring results are shown in Table 3.

[0074] Table 3 Colon DAI scores and colon lengths of DSS-induced model mice in different treatment groups

[0075]

[0076] As shown in Table 3, the colon DAI scores of mice in the DSS group were significantly increased compared with those in the normal control group, indicating that ulcerative colitis was successfully induced. In contrast, different doses of fermented ginseng hydrolysate can significantly reduce the colon DAI scores of colitis mice, among which under high dose conditions (1000 mg / kg), its protective effect on the colon is better than that of the positive drug mesalazine. It can be seen that the fermented ginseng hydrolysate of the present invention can enhance resistance to ulcerative colitis and relieve intestinal inflammatory lesions.

[0077] 2. DSS can cause severe local inflammatory reactions in the colon, leading to massive infiltration of immune cells and release of inflammatory mediators, resulting in edema, ulcers and extensive tissue damage in the colon tissue. Moreover, persistent inflammation can lead to fibrosis of the colon wall, and the formation of fibrotic tissue makes the colon stiff and contracted. In addition, repeated damage and regeneration of epithelial cells can disrupt the normal structure of the colon. The combined effect of the above-mentioned lesions can shorten the colon, so the length of the colon is an important indicator for evaluating colon inflammatory lesions. The colon lengths of mice in different treatment groups were measured and statistically analyzed, and the results are shown in Table 3.

[0078] As shown in Table 4, as the dosage of fermented ginseng hydrolysate increases, the colon length of DSS-induced model mice also increases, and the colon length of the high-dose group is similar to that of the positive drug group, indicating that the fermented ginseng hydrolysate of the present invention has the effect of improving colon shortening caused by inflammatory bowel disease.

[0079] 3. Detection of the inflammatory level of mouse colon tissue. Add appropriate amount of RIPA protein lysis buffer to mouse colon tissue, homogenize with an automatic homogenizer for 2 minutes, place on ice to continue lysis for 30 minutes, centrifuge at 12000 rpm for 10 minutes at 4°C, collect the supernatant, and use ELISA kits to detect the content of TNF-α, IL-6 and IL-1β. The results are shown in Table 4.

[0080] Table 4 Effects of different treatment groups on the levels of inflammatory factors in colon tissue of DSS-induced model mice

[0081]

[0082] As shown in Table 4, compared with the normal control group mice, a large amount of inflammatory factors accumulated in the colon tissue of the DSS group mice, and the fermented ginseng hydrolysate of the present invention can significantly reduce the content of TNF-α, IL-6 and IL-1β in the colon tissue, promote the elimination of inflammatory mediators, and the therapeutic effect of 1000 mg / kg of fermented ginseng hydrolysate is better than that of the positive drug mesalazine.

[0083] Example 4

[0084] 2,4,6-trinitrobenzenesulfonic acid (TNBS) is administered rectally together with ethanol. TNBS binds to proteins in the colon to form haptens, stimulating the immune system to produce a strong T-cell-mediated inflammatory response. Ethanol increases the contact and binding of TNBS with colon tissue by destroying the intestinal epithelial barrier. This model can simulate chronic inflammation and immune response, which is similar to the pathological mechanism of human Crohn's disease. This example uses TNBS to induce Crohn's disease-like intestinal lesions in mice, and based on this model, evaluates the improvement effect of the enzymatic fermentation of ginseng peptides described in the present invention on inflammatory bowel disease. The specific scheme is as follows:

[0085] The experiment was conducted after 6-week-old SPF C57BL / 6 male mice were adaptively raised for one week at an ambient temperature of 20-25°C, a humidity of 40% to 60%, and a daily light intensity of 12 h (7:00-19:00).

[0086] The mice were randomly divided into a normal control group, a TNBS model group, a low-dose (250 mg / kg) group of fermented ginseng hydrolysate, a medium-dose (500 mg / kg) group of fermented ginseng hydrolysate, a high-dose (1000 mg / kg) group of fermented ginseng hydrolysate, and a mesalazine positive drug group (200 mg / kg), with 5 mice in each group. Except for the normal control group, the mice in the remaining groups were modeled in the following manner: the mice were anesthetized with sodium pentobarbital, 100 μL of TNBS ethanol solution (containing 2 mg of TNBS) was slowly injected through the anus, and the mice were inverted for 15 minutes after enema. From the date of modeling, the fermented ginseng hydrolysate and mesalazine prepared in Example 1 were gavaged every day, while the normal control group and the model group were given an equal volume of normal saline. On the 7th day after modeling, the mice were euthanized and the colon tissue was separated. The colon DAI scores of each treated mouse and the levels of inflammatory factors TNF-α, IL-6 and IL-1β were evaluated according to the method of Example 3. The results are shown in Table 5.

[0087] Table 5 Effects of different treatment groups on colon DAI scores and inflammatory factor levels in TNBS model mice

[0088]

[0089]

[0090] As shown in Table 5, the colon DAI scores of mice in the TNBS group increased significantly compared with those in the normal control group, indicating that Crohn's disease-like intestinal lesions were successfully induced. Compared with the model group, different doses of fermented ginseng hydrolysate can significantly reduce the colon DAI scores of mice with colitis, and 1000 mg / kg of fermented ginseng hydrolysate produces better enteritis resistance than the positive drug mesalazine, which is consistent with the results under the DSS model, indicating that the fermented ginseng hydrolysate provided by the present invention is a new type of potent intestinal inflammatory lesion inhibitor. At the same time, the fermented ginseng hydrolysate of the present invention can dose-dependently reduce the content of TNF-α, IL-6 and IL-1β in the colon tissue of model mice, and has a potent anti-inflammatory effect.

[0091] In summary, the fermented ginseng hydrolysate of the present invention has excellent efficacy in improving inflammatory bowel disease.

[0092] Example 5

[0093] In order to clarify the therapeutic effect of the characteristic peptides in the fermented ginseng hydrolysate of Example 1 on inflammatory bowel disease, characteristic peptides LY, MM, LR, AP, LAR, RLDFR, SALAFR, LLLLGH and LLLLLHG with amino acid sequences as shown in SEQ ID NO.1-9 were synthesized by solid phase synthesis, and the colon DAI scores of DSS-induced ulcerative colitis model mice were evaluated according to the method of Example 3. The characteristic peptides were gavaged to mice at a dose of 100 mg / kg. The DAI score results are shown in Table 6.

[0094] Table 6 Effects of different characteristic peptides on colon DAI scores in DSS-induced model mice

[0095]

[0096]

[0097] As shown in Table 6, all nine characteristic peptides have the effect of improving DSS-induced ulcerative colitis, indicating that the nine characteristic peptides play a key role in improving the effect of inflammatory bowel disease and are the main active ingredients of fermented ginseng hydrolysate.

[0098] Example 6

[0099] The inflammatory bowel disease efficacy of three commercially available ginseng peptides and the fermented ginseng hydrolysate of the present invention were compared. The colon DAI scores of the three ginseng peptides in DSS-induced ulcerative colitis model mice were evaluated according to the method of Example 3. The dosage of the ginseng peptides and the fermented ginseng hydrolysate of the present invention were both 1000 mg / kg. The results are shown in Table 7.

[0100] Table 7 Effects of different ginseng peptides or fermented ginseng hydrolysates on colon DAI scores in DSS-induced model mice

[0101]

[0102] As shown in Table 9, although the three commercially available ginseng peptides have a certain effect on improving enteritis and can reduce the DAI score of the mouse colon, the effect is significantly weaker than that of the fermented ginseng hydrolysate of the present invention (which can reduce the DAI score to 0.800±0.380), which indicates that the fermented ginseng hydrolysate provided by the present invention has a strong effect on improving enteritis and is superior to other similar products on the market.

[0103] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. Application of fermented ginseng hydrolysate in the preparation of products for improving inflammatory bowel disease.

2. The use according to claim 1, characterized in that The fermented ginseng hydrolysate contains a characteristic peptide, and the amino acid sequence of the characteristic peptide is selected from at least one of SEQ ID NO.1-9.

3. The use according to claim 1, characterized in that: The method for preparing the fermented ginseng hydrolysate comprises the following steps: (1) Mixing fermented ginseng with water, adjusting the pH to 7.5-8.5, and heating in a water bath at 40-50° C. for 35-40 min to obtain an extract; (2) adding protease to the extract obtained in step (1), performing enzymatic hydrolysis at 40-50° C. for 4-5 hours, and centrifuging to obtain an enzymatic hydrolyzate; (3) The enzymatic hydrolysate obtained in step (2) is concentrated and dried to obtain a fermented ginseng enzymatic hydrolysate.

4. The use according to claim 3, characterized in that: In step (1), the fermented ginseng is ginseng residue after microbial fermentation and filtration, and the ratio of the fermented ginseng to water is fermented ginseng: water = 1g: (25-35)mL.

5. The use according to claim 3, characterized in that: In step (1), the pH is 8.0 and the mixture is heated in a water bath at 45°C for 30 min.

6. The use according to claim 3, characterized in that: In step (2), the protease is trypsin, and the amount of the protease added is 0.70-0.75 mg of protease per 1 g of fermented ginseng, and the concentration of the protease is 1250-1300 U / mg.

7. The use according to claim 3, characterized in that: In step (2), the enzymatic hydrolysis condition is at 50° C. for 5 h.

8. The use according to claim 3, characterized in that: In step (2), the enzymatic hydrolysate is bathed in 85-90° C. water for 30 min before centrifugation.

9. The use according to claim 3, characterized in that: In step (3), the concentration is reduced pressure concentration, and the drying is freeze drying.

10. The use according to claim 9, characterized in that: In step (3), the conditions for reduced pressure concentration are 65-70°C and -(0.09-0.10)MPa.

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