CN117859907A - Litchi polysaccharide-lactobacillus casei synbiotic and explosive bead as well as preparation method and application thereof - Google Patents
Litchi polysaccharide-lactobacillus casei synbiotic and explosive bead as well as preparation method and application thereof Download PDFInfo
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- CN117859907A CN117859907A CN202410172646.1A CN202410172646A CN117859907A CN 117859907 A CN117859907 A CN 117859907A CN 202410172646 A CN202410172646 A CN 202410172646A CN 117859907 A CN117859907 A CN 117859907A
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- lactobacillus casei
- litchi
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- litchi polysaccharide
- gel
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/015—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/035—Organic compounds containing oxygen as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
- A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
- A23L29/256—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin from seaweeds, e.g. alginates, agar or carrageenan
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
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- Dispersion Chemistry (AREA)
- Inorganic Chemistry (AREA)
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to litchi polysaccharide-lactobacillus casei synbiotic and explosive bead, and a preparation method and application thereof, and belongs to the technical field of health-care food. The invention provides a synbiotics with the function of relieving side effects of antibiotics, which comprises the following components: litchi polysaccharide and lactobacillus casei; the mass ratio of the litchi polysaccharide to the lactobacillus casei is 1-2: 1 to 2. The synbiotics obtained by the preparation method can reduce the cecum index, the fecal water content and the fecal pH value of the mice exposed by antibiotics, and enhance the intestinal peristalsis capability and the gastrointestinal tract operation capability; simultaneously, the level of reactive oxygen species ROS can be reduced, thereby alleviating the side effects of antibiotic exposure on the body.
Description
Technical Field
The invention relates to the technical field of health-care foods, in particular to litchi polysaccharide-lactobacillus casei synbiotic and explosive beads, and a preparation method and application thereof.
Background
Antibiotics are widely applied to the industries of medicine, agriculture, livestock and aquaculture, and are compounds for treating diseases and inhibiting the growth of harmful bacteria, thereby promoting the growth and development of organisms and metabolism. However, the use of antibiotics is easy to cause the problems of environmental pollution, food residue, enhanced bacterial drug resistance and the like, and can also cause potential threat to the health of people, and the risks to the ecological environment and the health of human bodies are not small in scale. At present, the use of antibiotics is still in an upward trend, and people inevitably receive antibiotics actively or passively for a long time. Long-term exposure of antibiotics increases the risk of certain diseases occurring, and the gastrointestinal tract is the organ most vulnerable to oral antibiotics. It has been demonstrated that prolonged exposure of antibiotics can destroy intestinal barrier function, destroy epithelial tissue, cause cecum swelling, and at the same time, can destroy intestinal barrier function by disturbing the steady state of intestinal microbiota, thereby destroying human health. Thus, there is a need to provide a healthy diet strategy that prevents or reduces antibiotic side effects.
Probiotics are a general term for active microorganisms capable of producing health effects on a host, and are microorganisms which exist in the intestinal tract, reproductive system and other environments of the body and play a certain role on the host so as to improve the micro-environmental balance of the host and promote the health of the body. The probiotics can regulate the composition and the quantity of microorganisms in the intestinal tract by protecting the barrier function of intestinal mucosa, thereby maintaining the stable environment in the intestinal tract. Lactobacillus casei (Lactobacillus casei) is lactobacillus, is widely applied to dairy products, and has various beneficial effects of regulating immune function, protecting intestinal mucosa function, regulating intestinal flora and the like. But probiotics are very sensitive to the environment and their viability is affected during processing, storage to final consumption, such as pH, hydrogen peroxide, oxygen concentration, gastric juice of high acidity, storage temperature, digestive enzymes, bile salts in the small intestine, lactic acid and acetic acid concentration. Therefore, how to effectively increase the viable count of probiotics has become a current urgent problem to be solved.
Litchi (Litchi chinensis Sonn.) is a plant of the genus Litchi of the family Sapindaceae, widely distributed in tropical and subtropical areas, one of four large fruits in Guangdong, commonly known as "Ling nan Guo Wang". Fresh litchi pulp is tender and succulent, rich in nutrition, fragrant, sweet and delicious, rich in components such as polysaccharide, polyphenol, vitamins, dietary fibers, minerals and the like, has higher nutritive value and biological activity, and is regarded as a 'fruit-in-fruit treasure', and is a health-care and intelligence-improving nutritional treasure. The litchi polysaccharide is taken as a main water-soluble active ingredient in litchi, and the biological efficacy of the litchi polysaccharide is widely focused, and mainly relates to immunoregulatory activity, antioxidant activity, blood sugar reduction, fatigue resistance and the like. The research shows that litchi polysaccharide can not be completely degraded in the gastrointestinal digestive system, can enter the large intestine in a relatively complete form, can be utilized by intestinal flora in vivo to generate metabolic products such as acetic acid, can regulate the composition of the intestinal flora, promote the growth of beneficial intestinal bacteria and maintain intestinal micro-homeostasis.
Disclosure of Invention
The invention aims to provide litchi polysaccharide-lactobacillus casei synbiotic, explosive beads, and a preparation method and application thereof, so as to solve the problem of side effects of antibiotics in the prior art.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a synbiotics with the function of relieving side effects of antibiotics, which comprises the following components:
litchi polysaccharide and lactobacillus casei;
the mass ratio of the litchi polysaccharide to the lactobacillus casei is 1-2: 1 to 2.
Preferably, the viable count of the lactobacillus casei is 0.5 to 1.5X10 10 CFU/g。
The invention also provides application of the synbiotics in preparing health-care products for relieving side effects of antibiotics.
The invention also provides litchi polysaccharide-lactobacillus casei gel, which comprises the following components:
litchi polysaccharide, lactobacillus casei, sodium alginate, calcium chloride and water;
the viable count of the lactobacillus casei is 0.5 to 1.5X10 10 CFU/g;
The litchi polysaccharide, lactobacillus casei, sodium alginate and calcium chloride are in a mass ratio of 1-2: 1-2: 0.6 to 2:2 to 6;
the volume ratio of the total mass of the litchi polysaccharide, the lactobacillus casei, the sodium alginate and the calcium chloride to the water is 4.6-12.0 g:1100mL.
The invention also provides a preparation method of the litchi polysaccharide-lactobacillus casei gel, which comprises the following steps:
(1) Mixing sodium alginate, litchi polysaccharide and lactobacillus casei with 1/11-1/10 of water to obtain a mixed solution;
(2) Mixing calcium chloride with the rest of water to obtain a calcium chloride aqueous solution;
(3) And (3) mixing the mixed solution obtained in the step (1) with a calcium chloride aqueous solution to obtain the litchi polysaccharide-lactobacillus casei gel.
The invention also provides application of the litchi polysaccharide-lactobacillus casei gel or the litchi polysaccharide-lactobacillus casei gel prepared by the preparation method in preparation of health care products for relieving side effects of antibiotics.
The invention provides litchi polysaccharide-lactobacillus casei explosive beads, which comprise the following components:
litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide, sodium alginate, calcium lactate and water;
the litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide, sodium alginate and calcium lactate are in a mass ratio of 1-2: 1-2: 0.5 to 1:4 to 9;
the total volume ratio of the total mass of the litchi polysaccharide-lactobacillus casei gel, the litchi polysaccharide, the sodium alginate and the calcium lactate to the water is 6.5-14 g:1100mL;
the litchi polysaccharide-lactobacillus casei gel is the litchi polysaccharide-lactobacillus casei gel or the litchi polysaccharide-lactobacillus casei gel prepared by the preparation method.
The invention also provides a preparation method of the litchi polysaccharide-lactobacillus casei explosive bead, which comprises the following steps:
(1) Mixing litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide and sodium alginate with 1/11-1/10 of water to obtain mixed solution;
(2) Mixing calcium lactate with the rest of water to obtain a calcium lactate aqueous solution;
(3) And (3) mixing the mixed solution obtained in the step (1) with a calcium lactate aqueous solution to obtain the explosion beads.
Preferably, the mixing time in the step (3) is 10-30 s.
The invention also provides application of the litchi polysaccharide-lactobacillus casei explosive bead or the litchi polysaccharide-lactobacillus casei explosive bead prepared by the preparation method in preparation of health-care products for relieving side effects of antibiotics.
The invention has the following technical effects and advantages:
the invention provides a litchi polysaccharide-lactobacillus casei synbiotic with the function of relieving side effects of antibiotics. The litchi polysaccharide-lactobacillus casei synbiotics can reduce the cecum index exposed by antibiotics, the fecal water content and the fecal pH value, and enhance the intestinal peristalsis capability and the gastrointestinal tract operation capability; simultaneously, the level of reactive oxygen species ROS can be reduced, thereby alleviating the side effects of antibiotic exposure on the body.
The invention also provides a litchi polysaccharide-lactobacillus casei synbiotic explosive bead with the effect of relieving the side effect of antibiotics and a preparation method thereof. The prepared explosion beads not only retain typical fragrance of litchi, but also are rich in litchi active polysaccharide and probiotics, can regulate intestinal health of human bodies, and have good health care effects on human bodies.
Drawings
FIG. 1 is a graph of cecal index of mice from different treatment groups;
FIG. 2 is a graph showing the fecal moisture content of mice from different treatment groups;
FIG. 3 is a graph showing the pH of feces from mice in different treatment groups;
FIG. 4 is an intestinal motility profile of mice from different treatment groups;
FIG. 5 is a graph of gastrointestinal tract performance of mice from different treatment groups;
FIG. 6 is a graph of fecal ROS levels in mice from different treatment groups;
FIG. 7 is a graph of intestinal ROS levels in mice from different treatment groups.
Detailed Description
The invention provides a synbiotics with the function of relieving side effects of antibiotics, which comprises the following components:
litchi polysaccharide and lactobacillus casei;
the mass ratio of the litchi polysaccharide to the lactobacillus casei is 1-2: 1 to 2, preferably 1:1.
in the present invention, the viable count of Lactobacillus casei is 0.5 to 1.5X10 10 CFU/g, preferably 1X 10 10 CFU/g。
The preparation method of the litchi polysaccharide comprises the following steps:
pulverizing dried litchi pulp by a beater, loading into a low-temperature continuous phase-change extraction kettle, extracting with water at 75-85 ℃ for 2.5-3.5 h, preferably 80 ℃ for 3h, vacuum concentrating the extract to 1/4 of the original volume, vacuum concentrating at 60-70 ℃, preferably 65 ℃, adding 95% ethanol until the final concentration of the system ethanol is 80%, centrifuging at 4 ℃ for 10min after alcohol precipitation at 4500rpm to obtain polysaccharide precipitate, washing the precipitate with absolute ethanol for 3 times, collecting the precipitate, re-dissolving the precipitate with water, adding 1% papain, adjusting the pH to 6.4, carrying out enzymolysis at 55-65 ℃ for 2.5-3.5 h, preferably 60 ℃ for 3h, carrying out enzyme deactivation in a water bath for 5-15 min, preferably 10min, centrifuging at 7500-8500 rpm for 5-15 min, preferably 8000rpm for 10min, collecting supernatant, dialyzing with distilled water for 48h, concentrating, and vacuum freeze-drying to obtain litchi polysaccharide.
The preparation method of the lactobacillus casei comprises the following steps:
culturing Lactobacillus casei strain in MRS broth for 48 hr, subculturing for 2 times, centrifuging at 7500-8500 rpm at 4deg.C for 5-15 min, preferably at 8000rpm for 10min, washing with pre-cooled sterile water for 2 times, and lyophilizing to obtain 0.5-1.5X10 10 CFU/g lactobacillus casei is preserved at 4 ℃ for standby.
The invention also provides application of the synbiotics in preparing health-care products for relieving side effects of antibiotics.
The invention also provides litchi polysaccharide-lactobacillus casei gel, which comprises the following components:
litchi polysaccharide, lactobacillus casei, sodium alginate, calcium chloride and water;
the viable count of the lactobacillus casei is 0.5 to 1.5X10 10 CFU/g, preferably 1X 10 10 CFU/g;
The litchi polysaccharide, lactobacillus casei, sodium alginate and calcium chloride are in a mass ratio of 1-2: 1-2: 0.6 to 2:2 to 6, preferably 1.5:1.5:1.3:4, a step of;
the volume ratio of the total mass of the litchi polysaccharide, the lactobacillus casei, the sodium alginate and the calcium chloride to the water is 5.6-12.0 g:1100mL, preferably 8.3g:1100mL.
The invention also provides a preparation method of the litchi polysaccharide-lactobacillus casei gel, which comprises the following steps:
(1) Mixing sodium alginate, litchi polysaccharide and lactobacillus casei with 1/11-1/10 of water, preferably 1/11 of water, to obtain a mixed solution;
(2) Mixing calcium chloride with the rest of water to obtain a calcium chloride aqueous solution;
(3) And (3) mixing the mixed solution obtained in the step (1) with a calcium chloride aqueous solution to obtain the litchi polysaccharide-lactobacillus casei gel.
The invention also provides application of the litchi polysaccharide-lactobacillus casei gel or the litchi polysaccharide-lactobacillus casei gel prepared by the preparation method in preparation of health care products for relieving side effects of antibiotics.
The invention provides litchi polysaccharide-lactobacillus casei explosive beads, which comprise the following components:
litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide, sodium alginate, calcium lactate and water;
the litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide, sodium alginate and calcium lactate are in a mass ratio of 1-2: 1-2: 0.5 to 1:4 to 9, preferably 2:2:0.8:9, a step of performing the process;
the total volume ratio of the total mass of the litchi polysaccharide-lactobacillus casei gel, the litchi polysaccharide, the sodium alginate and the calcium lactate to the water is 6.5-14 g:1100mL, preferably 13.8g:1100mL;
the litchi polysaccharide-lactobacillus casei gel is the litchi polysaccharide-lactobacillus casei gel or the litchi polysaccharide-lactobacillus casei gel prepared by the preparation method.
The invention also provides a preparation method of the litchi polysaccharide-lactobacillus casei explosive bead, which comprises the following steps:
(1) Mixing litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide and sodium alginate with 1/11-1/10 of water, preferably 1/11 of water, to obtain a mixed solution;
(2) Mixing calcium lactate with the rest of water to obtain a calcium lactate aqueous solution;
(3) And (3) mixing the mixed solution obtained in the step (1) with a calcium lactate aqueous solution to obtain the explosion beads.
In the present invention, the mixing time in the step (3) is 10 to 30 seconds, preferably 20 seconds.
The invention also provides application of the litchi polysaccharide-lactobacillus casei explosive bead or the litchi polysaccharide-lactobacillus casei explosive bead prepared by the preparation method in preparation of health-care products for relieving side effects of antibiotics.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Litchi polysaccharide preparation
Pulverizing fructus litchi jerky with beater, loading into low temperature continuous phase change extraction kettle, extracting at 80deg.C for 3 hr, vacuum concentrating the extractive solution to 1/4 of original volume, vacuum concentrating at 65deg.C, adding 95% ethanol until final concentration of system ethanol is 80%, precipitating with ethanol at 4deg.C overnight, centrifuging at 4500rpm for 10min to obtain polysaccharide precipitate, washing precipitate with anhydrous ethanol for 3 times, collecting, re-dissolving precipitate with water, adding 1% papain, adjusting pH to 6.4, performing enzymolysis at 60deg.C for 3 hr, inactivating enzyme with boiling water bath for 10min, centrifuging at 8000rpm for 10min, collecting supernatant, dialyzing with distilled water for 48 hr, concentrating, and vacuum freeze drying to obtain fructus litchi polysaccharide powder.
Preparation of lactobacillus casei
Culturing Lactobacillus casei strain in MRS broth for 48 hr, subculturing for 2 times, centrifuging at 4deg.C and 8000rpm for 10min, washing with pre-cooled sterile water for 2 times, and lyophilizing to obtain 1×10 10 CFU/g lactobacillus casei freeze-dried powder is preserved at 4 ℃ for standby.
Preparation of litchi polysaccharide-lactobacillus casei synbiotics
Taking 1g litchi polysaccharide powder and 1g living bacteria number of 1×10 10 And mixing CFU/g lactobacillus casei freeze-dried powder to obtain the litchi polysaccharide-lactobacillus casei synbiotics.
Example 2
Preparation of litchi polysaccharide-lactobacillus casei gel
(1) Dissolving 0.6g of sodium alginate powder in 100mL of deionized water, and fully stirring to completely dissolve the sodium alginate powder to obtain sodium alginate aqueous solution;
(2) Adding 2.0g of litchi polysaccharide powder (example 1) and 2.0g of lactobacillus casei freeze-dried powder (example 1) into sodium alginate aqueous solution, and uniformly mixing to obtain mixed solution;
(3) Dissolving 6.0g of calcium chloride powder in 1000mL of deionized water, and fully stirring to completely dissolve the calcium chloride powder to obtain a calcium chloride aqueous solution;
(4) And (3) sucking the mixed solution in the step (2) into a fine needle for a needle tube, dripping the mixed solution into the calcium chloride aqueous solution in the step (3), and soaking the mixed solution for 20s to obtain the litchi polysaccharide-lactobacillus casei gel.
Example 3
Preparation of litchi polysaccharide-lactobacillus casei explosive bead
(1) Dissolving 2.0g of litchi polysaccharide powder (example 1) and 0.8g of sodium alginate in 100mL of deionized water, and fully stirring to completely dissolve the litchi polysaccharide powder to obtain a mixed solution 1;
(2) Adding 1g of the litchi polysaccharide-lactobacillus casei gel obtained in the example 2 into the mixed solution 1 obtained in the step (1) to obtain a mixed solution 2;
(3) 9.0g of calcium lactate powder is taken and dissolved in 1000mL of deionized water, and the mixture is fully stirred to be completely dissolved, so as to obtain calcium lactate aqueous solution;
(4) Placing the mixed solution 2 obtained in the step (2) into a magnetic stirrer, sucking the mixed solution 2 obtained in the step (2) by using a disposable dropper, and dripping the mixed solution into the calcium lactate aqueous solution obtained in the step (3) to soak for 20s, so as to obtain the litchi polysaccharide-lactobacillus casei bursting beads.
Test example 1
1 animal experiment group and administration
1.1 laboratory animals
Preparation of reagents
0.99g of litchi polysaccharide powder prepared in example 1 is dissolved in 10mL of physiological saline to obtain litchi polysaccharide solution with the concentration of 99mg/mL for later use; 1g of the Lactobacillus casei lyophilized powder prepared in example 1 was dissolved in 10mL of physiological saline to obtain 100mg/mL of Lactobacillus casei solution, i.e., 1X 10 9 CFU/mL lactobacillus casei solution for later use; 0.99g of fructo-oligosaccharide powder purchased in the market is dissolved in 10mL of physiological saline to obtain fructo-oligosaccharide solution with the concentration of 99mg/mL for later use.
Mixing litchi polysaccharide solution with the concentration of 99mg/mL with normal saline in an equal volume to obtain a reagent 1; mixing litchi polysaccharide solution with concentration of 99mg/mL and litchi polysaccharide solution with concentration of 1×10 9 CFU/mL lactobacillus casei bacteria liquid is mixed in equal volume to obtain a reagent 2; fructooligosaccharide solution with concentration of 99mg/mL and fructooligosaccharide solution with concentration of 1×10 9 Equal volume mixing of CFU/mL lactobacillus casei bacteria liquid to obtain a reagent 3; the concentration was 1X 10 9 CFU/mL lactobacillus casei bacterial liquid and normal saline are mixed in equal volume to obtain a reagent 4 for later use.
The test animals were C57BL/6J male mice (6 weeks old, 21+ -2 g) purchased from Vetong Lihua laboratory (Buddha, china). All mice were kept in a sterile environment with a 12h alternate light and dark cycle at 25.+ -. 2 ℃ and a relative humidity of 65%. After one week of adaptive feeding, the experimental mice were randomly divided into 6 treatment groups: normal control group (NC), antibiotic treatment group (MD), litchi polysaccharide treatment group (PLC), litchi polysaccharide-lactobacillus casei treatment group (PLC-LC), fructo-oligosaccharide-lactobacillus casei treatment group (FOS-LC), lactobacillus casei treatment group (LC), 10 each. Normal Control (NC) and antibiotic treated (MD) mice were perfused with 0.1mL/kg of saline, litchi polysaccharide treated (PLC) mice were perfused with 0.1mL/kg of reagent 1, litchi polysaccharide-lactobacillus casei treated (PLC-LC) mice were perfused with 0.1mL/kg of reagent 2, fructo-oligosaccharide-lactobacillus casei treated (FOS-LC) mice were perfused with 0.1mL/kg of reagent 3, lactobacillus casei treated (LC) mice were perfused with 0.1mL/kg of reagent 4, normal Control (NC) mice were perfused with 0.2mL of saline at 14:00 pm daily, and the remaining 5 treated mice were perfused with 0.2mL of antibiotic solution having concentrations of 12.5g/L cefuroxime acid and 10g/L of levofloxacin throughout the test period of 20 days, wherein the treated group reagent was perfused with 20 days and the antibiotic was perfused with 10 days. All mice can drink and eat freely during the experiment, and feed standard feed.
1.2 cecal index determination
The mice were weighed prior to dissection, ceca were removed during dissection and their weights were weighed and data recorded. The cecal index of the mice was calculated as follows:
the cecal index of 6 treated mice was measured and the results are shown in figure 1.
1.3 fecal moisture content determination
Fresh feces from mice on the day prior to dissection were collected to determine their fecal moisture content. Fresh faeces were weighed on an analytical balance and the wet faeces weight G was recorded 0 Then drying in an oven at 105 ℃ for 2h to constant weight, cooling to room temperature, and recording dry weight G of the feces after the first drying 1 Then the second drying is carried outDrying until the difference of the mass of the two times before and after is not more than 0.002G, and finally recording the dry weight G of the feces of the mice 2 . Units: g.
the calculation formula is as follows:
fresh feces from 6 treatment group mice on the day before dissection were collected, and the feces water content was measured, and the results are shown in FIG. 2.
1.4 fecal pH assay
50mg of fresh feces of each treatment group of mice are accurately weighed, and diluted 15 times by pre-cooled deionized water. After homogenization with a fully automatic tissue grinder, centrifugation at 8000rpm at 4℃for 5min gave supernatant, which was measured for pH with a microphotometer, the results of which are shown in FIG. 3.
1.5 determination of the peristaltic Capacity of the intestine
After feeding for 20 days, mice were fasted for 16 hours, given 5mg/mL of phenol red solution by gavage, 0.2mL of gavage, and sacrificed under anesthesia after 20min of gavage. The small intestine portion was removed, the distance from the pylorus to the furthest end of the colored stool was accurately measured, and the distance traveled by the colored dye in the small intestine was recorded. Intestinal motility was measured by the ratio of the distance traveled by the colored dye to the length of the small intestine. The calculation formula of the intestinal thrust rate of the mice is as follows:
the intestinal thrust of the mice in the 6 treatment groups was measured and the results are shown in FIG. 4.
1.6 determination of gastrointestinal motility
5 mice were randomly selected for each group, and the mice were perfused with 5mg/mL of phenol red solution at a rate of 0.2mL and allowed free access to food and water after one hour. The time from the end of the stomach lavage to the time of obtaining the first colored stool particles, i.e., the gastrointestinal transit time in minutes, was recorded as the time from colored diet to colored stool discharge for each group of mice, and the results are shown in fig. 5.
1.7 determination of ROS
Molecular probe dichloro-dihydro-fluorescein (H) 2 DCF-DA) to determine the accumulation of ROS in fecal and intestinal tissues. Collecting feces and intestinal tissue of 20mg of 6 mice in treatment group, diluting with pre-cooled physiological saline 15 times, homogenizing with full-automatic tissue grinder, centrifuging at 4deg.C and 8000rpm for 5min, collecting 50ul supernatant, adding into 96-well plate, and adding 50ul H into the plate 2 DCF-DA (concentration 0.5 mg/mL) was incubated at 37℃for 1h and the fluorescence intensity was measured using an EnSpire multimode plate reader (Perkinelmer), the results of which are shown in FIGS. 6 and 7.
1.8 data analysis
All experiments were repeated at least three times and the results were expressed as mean ± standard deviation (mean ± SD), and the p-value was less than 0.05 as a significant difference using a single factor anova using GraphPad prism9.0.0 edition (GraphPad Software, inc., san Diego, CA) software. * Indicating significant differences (< 0.05, < p <0.01, < p <0.001 and < p < 0.0001).
(II) results of experiments
PLC-LC capable of alleviating side effects caused by antibiotics
The protective effect of PLC-LC on antibiotic-exposed mice was evaluated by analyzing the cecal index, fecal moisture content, and fecal pH changes of the mice. As can be seen from fig. 1, exposure of the antibiotics caused the cecum enlargement of the mice, and the cecum index increased significantly, while the cecum enlargement phenomenon of the mice was alleviated after the PLC-LC intervention, and the cecum index was lowered significantly; as can be seen from fig. 2 and 3, exposure to antibiotics significantly increased the fecal moisture content and pH of the mice. After the PLC-LC dry prognosis, the water content and the pH value of the feces of the mice are obviously reduced (p is less than 0.001), wherein, the water content of the feces is reduced by 11.26 percent compared with the MD of a model group, and the pH value of the feces is recovered to be equivalent to that of a normal mouse.
PLC-LC having an alleviation effect on gastrointestinal tract function caused by antibiotics
The small intestine is one of the most important digestive organs of the human body, and is not only responsible for the digestion and absorption of food, but also has a crucial influence on the intestinal flora and on the metabolism of unabsorbed food. The mechanical peristalsis of the small intestine can promote adequate contact of the food chyme with the small intestine villi to absorb proteins, fats and other nutrients. Thus, the invention evaluates the physiological state of the small intestine by measuring the intestinal peristalsis function and the gastrointestinal tract operation capability of the mice in each treatment group. As can be seen from fig. 4, exposure to antibiotics significantly reduced the intestinal motility of mice, while PLC-LC was able to alleviate the weakening of the intestinal motility caused by antibiotics.
As can be seen from fig. 5, exposure to antibiotics causes an increase in the gastrointestinal transit time in mice, which may be associated with a cecum enlargement, while PLC-LC can alleviate the gastrointestinal function impairment caused by antibiotics, reduce the transit time in the gastrointestinal tract, and promote defecation in mice.
PLC-LC can relieve intestinal oxidative stress injury caused by antibiotics
Oxidative stress refers to an imbalance between the production of excess free radicals and low antioxidant levels in cells when the body is stimulated either endogenously or exogenously. Oxidative stress increases to cause intestinal mucosa barrier injury, intestinal permeability increase, intestinal flora disorder, etc., thereby affecting the balance system of the organism. The present invention evaluates the effect of PLC-LC on oxidative stress damage to mice exposed to antigen by measuring free radical ROS levels in the feces and intestinal tissues of each treatment group of mice. As can be seen from fig. 6, exposure to antibiotics significantly destroyed the oxidative stress level of mice, increased the ROS level of their feces, while oxidative stress injury was alleviated and the ROS level of their feces was significantly reduced with PLC-LC intervention. As can be seen from fig. 7, the exposure to antibiotics also increased the level of ROS in the intestinal tract of mice, whereas the level of ROS in the intestinal tract was significantly reduced after PLC-LC intervention, and the oxidative stress damage in the intestinal tract was alleviated. As can be seen from fig. 6 and 7, the intervention of PLC-LC significantly alleviates oxidative stress damage caused by antibiotics.
The results show that compared with other treatment groups, the litchi polysaccharide-lactobacillus casei treatment group can obviously relieve cecum swelling caused by antibiotics and increase the water content of excrement; reducing the pH value of the feces of the mice exposed by the antibiotics to keep the health of intestinal tracts and relieving the side effects of the exposure of the antibiotics to organisms. Further experiments show that the substance can also relieve the inhibition of antibiotics on gastrointestinal functions, and simultaneously, the substance can also relieve intestinal oxidative stress injury caused by antibiotics by reducing the level of free radical ROS. Indicating that it has great potential to alleviate antibiotic side effects.
According to the embodiment, the litchi polysaccharide-lactobacillus casei synbiotic and explosive bead, the preparation method and the application thereof are provided, and the litchi polysaccharide-lactobacillus casei synbiotic and explosive bead prepared by the method can obviously relieve the cecum swelling caused by antibiotics and increase the water content of excrement; reducing the pH value of the feces of the mice exposed by the antibiotics to keep the health of intestinal tracts, relieving the side effect of the exposure of the antibiotics to organisms, also relieving the inhibition effect of the antibiotics to the gastrointestinal tract functions, and relieving the oxidative stress injury of the intestinal tracts caused by the antibiotics by reducing the level of free radical ROS.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A synbiotics with antibiotic side effects alleviation, characterized by comprising the following components:
litchi polysaccharide and lactobacillus casei;
the mass ratio of the litchi polysaccharide to the lactobacillus casei is 1-2: 1 to 2.
2. The synbiotics according to claim 1, wherein the viable count of Lactobacillus casei is 0.5-1.5X10 10 CFU/g。
3. Use of a synbiotics according to any one of claims 1-2 for the preparation of a health product for alleviating side effects of antibiotics.
4. The litchi polysaccharide-lactobacillus casei gel is characterized by comprising the following components:
litchi polysaccharide, lactobacillus casei, sodium alginate, calcium chloride and water;
the viable count of the lactobacillus casei is 0.5 to 1.5X10 10 CFU/g;
The litchi polysaccharide, lactobacillus casei, sodium alginate and calcium chloride are in a mass ratio of 1-2: 1-2: 0.6 to 2:2 to 6;
the volume ratio of the total mass of the litchi polysaccharide, the lactobacillus casei, the sodium alginate and the calcium chloride to the water is 4.6-12.0 g:1100mL.
5. The method for preparing the litchi polysaccharide-lactobacillus casei gel as claimed in claim 4, comprising the steps of:
(1) Mixing sodium alginate, litchi polysaccharide and lactobacillus casei with 1/11-1/10 of water to obtain a mixed solution;
(2) Mixing calcium chloride with the rest of water to obtain a calcium chloride aqueous solution;
(3) And (3) mixing the mixed solution obtained in the step (1) with a calcium chloride aqueous solution to obtain the litchi polysaccharide-lactobacillus casei gel.
6. Use of the litchi polysaccharide-lactobacillus casei gel as claimed in claim 4 or the litchi polysaccharide-lactobacillus casei gel as claimed in claim 5 in the preparation of a health product for alleviating side effects of antibiotics.
7. The litchi polysaccharide-lactobacillus casei explosive bead is characterized by comprising the following components:
litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide, sodium alginate, calcium lactate and water;
the litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide, sodium alginate and calcium lactate are in a mass ratio of 1-2: 1-2: 0.5 to 1:4 to 9;
the total volume ratio of the total mass of the litchi polysaccharide-lactobacillus casei gel, the litchi polysaccharide, the sodium alginate and the calcium lactate to the water is 6.5-14 g:1100mL;
the litchi polysaccharide-lactobacillus casei gel is the litchi polysaccharide-lactobacillus casei gel in claim 4 or the litchi polysaccharide-lactobacillus casei gel prepared by the preparation method in claim 5.
8. The method for preparing litchi polysaccharide-lactobacillus casei explosive beads as claimed in claim 7, comprising the following steps:
(1) Mixing litchi polysaccharide-lactobacillus casei gel, litchi polysaccharide and sodium alginate with 1/11-1/10 of water to obtain mixed solution;
(2) Mixing calcium lactate with the rest of water to obtain a calcium lactate aqueous solution;
(3) And (3) mixing the mixed solution obtained in the step (1) with a calcium lactate aqueous solution to obtain the explosion beads.
9. The method according to claim 8, wherein the mixing time in the step (3) is 10 to 30 seconds.
10. Use of the litchi polysaccharide-lactobacillus casei explosive bead of claim 7 or the litchi polysaccharide-lactobacillus casei explosive bead prepared by the preparation method of any one of claims 8-9 in the preparation of health care products for relieving side effects of antibiotics.
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