Coffee with anti-diarrhea function added with probiotics and application of coffee

Abstract

The invention provides coffee added with probiotics and having an anti-diarrhea function and application of the coffee, and belongs to the technical field of food. The coffee is prepared from Zhang (Lactobacillus casei Zhang) probiotic powder and Robursta coffee beans, and can be used for effectively relieving gastrointestinal discomfort and diarrhea symptoms caused by caffeine. The viable count of probiotics in the coffee is not less than 1 * 10 < 10 > CFU / g. By adjusting gastrointestinal motility and immune factors, the coffee can maintain the steady state of the intestinal tract and improve the problems of diarrhea, stomachache, frequent defecation and the like caused by caffeine intake. According to the anti-diarrhea coffee added with the probiotics and the application, the stimulation effect of caffeine on the stomach and intestine is remarkably improved, and the coffee has a remarkable application prospect.

Description

Technical Field

[0001] The invention relates to the technical field of food, and in particular to anti-diarrhea functional coffee with added probiotics and application thereof. Background Art

[0002] Diarrhea refers to bowel movements that are significantly more frequent than usual, with thin stools, increased water content, daily bowel movements exceeding 200g, or stools containing undigested food, pus, blood, or mucus. Diarrhea is often accompanied by symptoms such as an urgent need to defecate, anal discomfort, or incontinence. Studies have shown that the caffeine in coffee has a certain stimulating effect on the gastrointestinal tract. When a large amount of caffeine is consumed, it may cause faster intestinal peristalsis and faster food passage through the intestines, leading to diarrhea. In addition, some people are allergic to caffeine and may experience symptoms such as acid reflux, abdominal pain, diarrhea, nausea, or frequent bowel movements.

[0003] Probiotics are microorganisms that maintain the homeostasis of the intestinal microbial environment and can regulate constipation and diarrhea in the human body in both directions. Studies have shown that probiotics can restore normal intestinal peristalsis and defecation functions by regulating gastrointestinal regulatory peptides, thereby maintaining intestinal homeostasis. In addition, probiotics can also regulate immune factors and improve inflammation, thereby effectively relieving diarrhea symptoms and improving ulcerative colitis and irritable bowel syndrome.

[0004] The prior art also has the following deficiencies:

[0005] Although probiotics have been widely used to improve intestinal health, the prior art has not yet provided a solution that can effectively relieve the symptoms of gastrointestinal discomfort and diarrhea caused by caffeine. For people who are sensitive to caffeine or have weak gastrointestinal function, drinking ordinary coffee often causes significant gastrointestinal discomfort and diarrhea. In addition, although the existing low-caffeine or decaffeinated coffee reduces the intake of caffeine, it cannot fundamentally solve the gastrointestinal irritation caused by caffeine, and it also cannot meet consumers' dual needs for coffee flavor and function.

[0006] Therefore, the prior art lacks a functional coffee product that can effectively relieve the gastrointestinal discomfort and diarrhea symptoms caused by caffeine. Summary of the invention

[0007] The purpose of the present invention is to provide an anti-diarrhea functional coffee with added probiotics and its application. The anti-diarrhea functional coffee with added probiotics can regulate gastrointestinal regulatory peptides to enable normal intestinal peristalsis and normal defecation, restore intestinal peristalsis ability, maintain intestinal homeostasis, and relieve diarrhea symptoms caused by caffeine. At the same time, it can regulate immune factors, improve inflammation, effectively relieve diarrhea symptoms, and improve ulcerative colitis and irritable bowel syndrome.

[0008] To achieve the above object, the present invention provides an anti-diarrhea functional coffee with added probiotics, wherein the coffee comprises Lactobacillus casei Zhang probiotic powder and Robusta coffee beans.

[0009] Preferably, the viable count of Lactobacillus casei Zhang added to the coffee is not less than 1×10 10 CFU / g.

[0010] Preferably, the coffee is in the form of freeze-dried coffee, brewed coffee or solid beverage.

[0011] The present invention also provides a use of the anti-diarrhea functional coffee as described above in preparing food or beverage for regulating gastrointestinal function.

[0012] Therefore, the present invention adopts the above-mentioned anti-diarrhea functional coffee and application with added probiotics, and has beneficial technical effects: probiotic coffee regulates gastrointestinal regulatory peptides, so that the intestines can move normally, defecate normally, restore intestinal peristalsis ability, maintain intestinal homeostasis, and relieve diarrhea symptoms caused by caffeine. At the same time, it regulates immune factors, improves inflammation, effectively relieves diarrhea symptoms, and improves ulcerative colitis and irritable bowel syndrome. It can be seen that this anti-diarrhea functional coffee with added probiotics can significantly maintain the homeostasis of intestinal flora, improve gastrointestinal discomfort symptoms caused by improper diet / ingestion of irritating food, and relieve diarrhea symptoms, and has great application prospects in the preparation of products with diarrhea / gastrointestinal discomfort symptoms. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The changes in the weight and moisture content of mouse feces on the 7th and 14th days; Figure 1 (a) is the feces weight of mice on day 7; Figure 1 (b) is the feces weight of mice on day 14; Figure 1 (c) shows the change of water content in mouse feces on the 7th day; Figure 1 (d) shows the change of water content in mouse feces on day 14;

[0014] Figure 2 is the level of changes in gastrointestinal regulatory peptides in mice; among them, Figure 2 (a) shows the change of motilin level in mice; Figure 2 (b) in the figure shows the change level of gastrin; Figure 2 (c) in the figure is the change level of vasoactive intestinal peptide;

[0015] Figure 3 is a diarrhea severity scale; Figure 3 (a) shows the change level of mouse feces consistency score; Figure 3 (b) in the table represents the level of change in severity of diarrhea;

[0016] Figure 4 for bowel movement frequency;

[0017] Figure 5 is the colon turnover rate. DETAILED DESCRIPTION

[0018] The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

[0019] Unless otherwise defined, technical or scientific terms used in the present invention shall have the common meanings understood by one having ordinary skills in the field to which the present invention belongs.

[0020] If no specific experimental steps or conditions are specified in the following examples, the conventional experimental steps or conditions described in the literature in this field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially. The RCM solid culture medium and RCM liquid culture medium involved in the following examples were purchased from Beijing Luqiao Technology Co., Ltd.

[0021] The method for detecting the number of viable bacteria involved in the following embodiments is: using the national standard "GB 4789.35-2016 National Food Safety Standard Food Microbiology Detection Lactic Acid Bacteria Detection".

[0022] A single colony of Lactobacillus casei Zhang was selected and inoculated into RCM liquid culture medium, and anaerobically cultured at 37°C for 24 hours to activate the activated solution; the activated solution was inoculated into RCM liquid culture medium at an inoculum amount of 2% (v / v), and anaerobically cultured at 37°C for 20 hours to obtain a bacterial solution; the bacterial solution was centrifuged at 12000g for 10 minutes to obtain a bacterial mud; the bacterial mud was washed with physiological saline, and then centrifuged at 12000g for 10 minutes to obtain bacterial cells; the bacterial cells and the freeze-dried protective agent glycerol were mixed at a mass ratio of freeze-dried protective agent: bacterial cells = 1:1 to obtain a mixed solution; the mixed solution was freeze-dried to obtain bacterial powder (the number of viable bacteria in the bacterial powder was 1×10 10 CFU / g); mixing the bacterial powder with Robusta coffee powder to obtain a mixture; pouring the mixture into a filling device, filling and plastic sealing, and obtaining freeze-dried black coffee containing live bacteria of Lactobacillus casei Zhang (the live bacteria content of Lactobacillus casei Zhang is 15 billion / 1.5g).

[0023] Embodiment 1

[0024] Improvement effect of probiotic coffee on diarrhea in mice.

[0025] The experimental process is as follows:

[0026] 1.1 Experimental design

[0027] 40 6-week-old male C57 mice were kept under strict temperature control of 22±2℃ and humidity of 45±10%. During the experiment, the mice were fed with ordinary feed and were allowed to eat and drink freely. After the adaptation period, the follow-up experiment was started. The 40 mice were randomly divided into 4 groups (weight range 20±2g): control group (Control), model group (Model), model + probiotic coffee group (Pro), model + ordinary coffee group (Pla), 10 mice in each group. The specific intervention plan is as follows: During the modeling period, the control group was gavaged with 0.1mL of normal saline every day, and the other groups were gavaged with senna solution according to 0.2mL / 10g of mouse weight, twice a day for 03 consecutive days. During the intervention period, the control group was gavaged with 0.1 mL of saline every day for 1 hour and then continued to gavage with 0.1 mL of saline. The model group was gavaged with 0.1 mL of saline 1 hour after gavage with senna solution. The other groups were gavaged with probiotic coffee or ordinary coffee (0.1 mL / 10 g) 1 hour after gavage with senna solution. The gavage was continued for 14 days, once a day. On the 18th day, all mice were bled from the eyeballs and killed by cervical dislocation. Soft or watery stools; poor mental state, dull hair, and obvious contamination around the anus indicate that the model was successful.

[0028] 1.2. Experimental observation and sample collection.

[0029] Daily monitoring: record the mouse's weight, food intake, water intake, stool morphology (whether diarrhea occurs), etc.

[0030] Fecal moisture content: Fresh feces discharged on the day were collected, and the feces were weighed after each collection to obtain the wet weight, and then placed in an oven (80°C for 8 hours) to measure the dry weight. The number of feces particles and dry and wet weights of each group of mice were summarized.

[0031] Feces moisture content (%) = (feces wet weight - feces dry weight) / feces wet weight × 100%.

[0032] Time of first black stool discharge: On the last day of modeling (day 3) and the day of sacrifice (day 18), mice were gavaged with 0.2 mL of activated carbon solution. The timing was started from the gavage of activated carbon solution, and the time of first black stool discharge was recorded.

[0033] Small intestinal propulsion rate: On the day of killing mice, 1% activated carbon solution (10 mL / kg) was gavage, and the mice were killed 20 minutes later. The whole intestinal tract (from stomach to cecum) was taken, and the percentage of carbon powder propulsion length to the total small intestinal length was measured: small intestinal propulsion rate (%) = black juice propulsion length (cm) / total small intestinal length (cm) × 100%.

[0034] 1.3. Sample collection time.

[0035] Feces: Feces of each mouse was collected on days 0, 7, and 14 of the experiment and stored in a -80°C refrigerator.

[0036] Blood: Blood was collected from the eyeball on the 15th day, and serum was separated and stored at -80°C for the detection of inflammatory indicators.

[0037] Intestinal tissue: After sacrifice, the whole intestine was removed and segmented (duodenum, ileum, cecum, colon) and fixed in 4% paraformaldehyde solution. Some of them were stored in liquid nitrogen for molecular biology detection.

[0038] 1.4. Measurement of serum gastrointestinal regulatory peptides and immune factors.

[0039] The levels of gastrointestinal regulatory peptides (motilin, gastrin, and vasoactive intestinal peptide) in mouse serum were measured using an enzyme-linked immunosorbent assay (ELISA) kit. Similarly, the levels of immune factors such as interleukin-6 (IL-6), interleukin-10 (IL-10), and interleukin-1β (IL-1β) in mouse serum were measured using ELISA kits.

[0040] 1.5. Detection of gene expression in colon tissue.

[0041] Total RNA was extracted from the proximal colon tissue of mice, and the expression of Claudin-1, tight junction protein ZO-1, aquaporin 3 (AQP3), and PYY genes was detected by reverse transcription real-time quantitative polymerase chain reaction.

[0042] 1.6. Experimental results

[0043] Depend on Figure 1-Figure 2 It can be seen that motilin and gastrin are excitatory peptides, while vasoactive intestinal peptide has an inhibitory effect. The levels of gastrointestinal regulatory peptides (motilin, gastrin and vasoactive intestinal peptide) in mouse serum were determined using an enzyme-linked immunosorbent assay (ELISA) kit. The results showed that the detection levels of motilin and gastrin in the probiotic coffee group mice were significantly reduced, and the feces weight and moisture content were significantly lower than those in the model and ordinary coffee groups.

[0044] As shown in Table 1, the levels of pro-inflammatory factors IL-6 and IL-1β in mouse serum were significantly reduced, while the level of anti-inflammatory factor IL-10 was significantly increased.

[0045] Statistical results show that coffee with added probiotics can significantly reduce coffee-induced diarrhea symptoms in mice.

[0046] Table 1 Changes in the levels of immune factors in mouse serum

[0047]

[0048] Embodiment 2

[0049] The improving effect of probiotic coffee on diarrhea in humans.

[0050] The experimental process is as follows:

[0051] 1.1 Experimental design

[0052] This example is a randomized double-blind placebo-controlled experiment, recruiting 80 patients with chronic diarrhea (18-60 years old, regardless of gender). All patients had diarrhea symptoms for at least 6 months before enrollment, and at least 25% of the time in the past three months was loose stool or watery stool (Bristol stool typing 5, 6, 7). Patients who met the inclusion criteria were randomly divided into a probiotic coffee group (Pro) and a regular coffee group (Pla), 40 people in each group, and continued to intervene for 28 days, 1 (1.5g) per day. There is no difference between probiotic coffee and regular coffee in appearance, taste, etc. During the trial, all patients stopped taking drugs and irritating foods that affect gastrointestinal motility.

[0053] This study has been approved by the Ethics Committee, and all participants participated voluntarily with full knowledge and signed the informed consent form. The relevant regulations of the Ethics Committee were strictly followed during the study to ensure that the rights and safety of the participants were fully protected.

[0054] 1.2. Experimental observation and sample collection.

[0055] Information such as stool consistency (Bristol score), daily bowel movement frequency, bowel movement urgency, and diarrhea severity were recorded on a bowel movement diary card. On day 28, a colonic transit study was performed. Twenty capsules of radiopaque markers were swallowed at 8:00 a.m. on the day of the examination, and an abdominal X-ray, including the diaphragm and pubic bone, was taken 8 hours later. The amount of marker excretion was calculated.

[0056] Colon emptying rate = (20-number of markers remaining in the intestine) / 20*100.

[0057] The diarrhea severity score, defecation frequency, stool consistency, and average score of colon emptying rate were calculated on days 0, 14, and 28. Blood samples were collected from patients on days 0 and 28 to test intestinal permeability, including lipopolysaccharide, D-lactic acid, and dimethylamine; serum IL-6, IL-4, and TNF-α immune indicators were detected using ELISA kits.

[0058] 1.3 Experimental results

[0059] Depend on Figure 3-Figure 5 It can be seen that the frequency of defecation in the experimental population decreased and the severity of diarrhea was reduced. Statistical results show that coffee with added probiotics can significantly inhibit intestinal peristalsis caused by coffee and relieve diarrhea.

[0060] As shown in Table 2, TNF-α is a pro-inflammatory factor, and the detection level in the probiotic coffee group was significantly lower than that in the control group and the ordinary coffee group.

[0061] Statistical results show that coffee with added probiotics can significantly reduce coffee-induced diarrhea symptoms in the human body.

[0062] Table 2 Changes in blood indexes of experimental population

[0063]

[0064] It is worth noting that the contents not elaborated in detail in the present invention are all prior art and are well known to those skilled in the art.

[0065] Therefore, the present invention adopts the above-mentioned anti-diarrhea functional coffee with added probiotics and its application. The anti-diarrhea functional coffee with added probiotics can regulate gastrointestinal regulatory peptides to achieve normal intestinal peristalsis and normal defecation, restore intestinal peristalsis ability, maintain intestinal homeostasis, and relieve diarrhea symptoms caused by caffeine. At the same time, it can regulate immune factors, improve inflammation, effectively relieve diarrhea symptoms, and improve ulcerative colitis and irritable bowel syndrome.

[0066] 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 it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims

1. An anti-diarrhea functional coffee with added probiotics, characterized in that: The coffee contains Lactobacillus casei Zhang probiotic powder and Robusta coffee beans.

2. The anti-diarrhea functional coffee according to claim 1, characterized in that: The viable count of Lactobacillus casei Zhang added to the coffee is not less than 1×10 10 CFU / g.

3. The anti-diarrhea functional coffee according to claim 2, characterized in that: The coffee may be in the form of freeze-dried coffee, brewed coffee or solid beverage.

4. Use of the anti-diarrhea functional coffee according to any one of claims 1 to 3 in preparing food or beverages for regulating gastrointestinal function.

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