CN114145461B - Probiotic composition for relieving nonalcoholic fatty liver - Google Patents

Probiotic composition for relieving nonalcoholic fatty liver Download PDF

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CN114145461B
CN114145461B CN202111436490.6A CN202111436490A CN114145461B CN 114145461 B CN114145461 B CN 114145461B CN 202111436490 A CN202111436490 A CN 202111436490A CN 114145461 B CN114145461 B CN 114145461B
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probiotic composition
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lactobacillus
rosell
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郭兆锋
莫景至
郑政东
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Guangzhou Narnia Biotechnology Co ltd
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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
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/125—Casei
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    • A23V2400/11—Lactobacillus
    • A23V2400/147—Helveticus
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    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum
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Abstract

The invention relates to the field of functional foods, in particular to a probiotic composition for relieving nonalcoholic fatty liver. The probiotic composition comprises 30-60 parts of lactobacillus plantarum HA-119, 48-50 parts of lactobacillus casei Rosell-0215 20 and 25-30 parts of lactobacillus helveticus Rosell-0052 10. The invention also provides raw material compositions of solid beverages, tablets and capsules comprising the probiotic composition. The probiotic composition provided by the invention has the advantages of simple selected bacterial strain, excellent efficacy, good stability and activity after compounding, and obvious cooperativity among bacterial strains.

Description

Probiotic composition for relieving nonalcoholic fatty liver
Technical Field
The invention relates to the field of functional foods, in particular to a probiotic composition for relieving nonalcoholic fatty liver.
Background
Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases, which can lead to the occurrence of cirrhosis and liver cancer, and is one of the main causes of end-stage liver disease and liver transplantation. NAFLD pathogenesis is complex, and no effective medicine exists at present.
Common drug therapies include vitamin E, pioglitazone, metformin, pentoxifylline, cholanic acid, statins, and the like. For example, patients with NAFLD with diabetes mellitus may use first-line drugs such as metformin and the like which reduce insulin resistance, patients with hyperlipidemia may be treated with statin lipid-lowering drugs, and lipid peroxidation may be treated with antioxidant drugs such as probucol and the like. In addition, there are some common anti-inflammatory liver-protecting plant extracts such as silybum marianum, glycyrrhizin, etc. To date, NAFLD is considered the only effective therapeutic route to be intervention in unhealthy lifestyles, but patient compliance is often poor.
In recent years, it has been found that dysbacteriosis in the intestinal tract may lead to accumulation of fat in the liver. In the case of dysregulation of intestinal flora, the tight intercellular junctions of intestinal epithelial cells cause an impaired intestinal barrier, which leads to microbial-host interactions, involving release of cell-associated ligands such as lipopolysaccharide into the portal circulation communicating with the liver, affecting the microbial sensing and response system and thus causing inflammation.
Probiotics are able to improve the protective mechanisms of the sexual intestinal barrier and regulate the fibronectin. These effects limit overgrowth of small intestine bacteria and translocation of bacteria, reducing endotoxemia. In addition to improving intestinal mucosa permeability, the mechanisms of action of probiotics include: inhibiting inflammatory response; probiotics regulate the immune system, etc.
Thus, modulation of the interaction of the intestinal flora with the liver has become a major target for NAFLD management in recent years. The beneficial effects of probiotics, prebiotics and synbiotics on the intestinal flora have been demonstrated in earlier experiments and in human research. But the specific effects of the probiotics are different from strain to strain, so that it is very important to screen out the probiotics which have a synergistic effect and can more specifically play a role in relieving the nonalcoholic fatty liver.
Chinese patent application CN108641988A discloses Lactobacillus plantarum NA136 and application thereof in relieving non-alcoholic fatty liver disease, chinese patent application CN109182191A discloses space Lactobacillus plantarum FH-SS18- -86365L with liver protecting effect and application thereof, the above patents all disclose the liver protecting effect of Lactobacillus plantarum, but a probiotic has very limited therapeutic effect, chinese patent application CN 112244299A discloses a probiotic composition with the function of relieving non-alcoholic fatty liver disease and a preparation method thereof, the invention adopts the combination of Lactobacillus plantarum 1701 and other 4 strains of bacteria, chinese patent application CN 113265361A discloses a compound probiotic preparation capable of relieving non-alcoholic fatty liver disease, a preparation method and application thereof, adopts the combination of Lactobacillus sub-species BLa80, lactobacillus rhamnosus LRa05 and Lactobacillus casei Lp90 of the animal bifidobacterium, and Chinese patent application CN201711402942 discloses a fruit juice beverage for preventing the non-alcoholic fatty liver disease, wherein probiotic powder is Lactobacillus bifidus powder, lactobacillus acidophilus powder, lactobacillus delbrueckii powder and Lactobacillus bifidus powder or Lactobacillus bifidus powder. The patent discloses the use of probiotics for relieving the nonalcoholic fatty liver, but most of the probiotics are either probiotics or excessive strains are adopted, so that dominant strains and ratios are excavated, and a better treatment effect is achieved.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention provides the probiotic composition for relieving the nonalcoholic fatty liver, and the selected strain of the composition is excellent and simple, has good stability and activity after compounding, and has obvious cooperativity.
The invention aims at realizing the following technical scheme:
a probiotic composition with a function of relieving non-alcoholic fatty liver, comprising the following components in parts by weight: 30-60 parts of lactobacillus plantarum HA-119, 30-50 parts of lactobacillus casei Rosell-0215 20 and 20-30 parts of lactobacillus helveticus Rosell-0059.
Preferably, the Lactobacillus plantarum HA119 microorganism is classified as Lactobacillus plantarum Lactobacillus plantarum, deposited with the Belgium joint microorganism deposit center under accession number LMG S-24119.
Preferably, the Lactobacillus casei Rosell-0215 microorganism is classified as Lactobacillus casei Lactobacillus casei and deposited with the national institute of Pasteur, france, microorganism accession number I-3429.
Preferably, the Lactobacillus helveticus Rosell-0052 microorganism classification is named Lactobacillus helveticus Lactobacillus helveticus, deposited with the national center for microbiological deposit of Pasteur institute, france, accession number I-1722.
Preferably, the probiotic composition comprises the following components in parts by weight: 50-60 parts of lactobacillus plantarum HA-119, 0215 20-30 parts of lactobacillus casei Rosell-0215 20-30 parts of lactobacillus helveticus Rosell-0052 20-30 parts.
Preferably, the mass ratio of the lactobacillus plantarum HA-119 to the lactobacillus casei Rosell-0215 is 1.3-3:1.
Preferably, the probiotic composition has a viable bacteria concentration of 1×10 11 cfu/g-2×10 11 cfu/g。
Preferably, the probiotic composition further comprises one or more of bifidobacterium animalis Lafti B94, lactobacillus rhamnosus Rosell-0011, bifidobacterium longum Rosell-0175, lactobacillus plantarum Rosell-1012, bifidobacterium bifidum Rosell-0071 and Saccharomyces boulardii.
The second object of the invention is to provide a preparation method of the probiotic composition, which comprises the steps of weighing the components of the probiotic composition, mixing and stirring uniformly.
Preferably, the weighing is carried out at a temperature of 20.+ -. 2 ℃ and an air relative humidity of less than 40%. The speed of the mixing and stirring is 18-22r/min, and the time of the mixing and stirring is 15-20min.
The invention also aims to provide a solid beverage A, which comprises the following raw materials in parts by weight: 5-10 parts of the probiotic composition, 30-60 parts of prebiotics, 30-60 parts of sugar alcohol, 0-20 parts of flavor substances and 0-10 parts of antioxidants.
Preferably, the total viable count of the solid beverage A is 140-150 hundred million CFU, the prebiotics are one or more of inulin, fructo-oligosaccharide, xylo-oligosaccharide, galacto-oligosaccharide, isomalto-oligosaccharide and stachyose, the sugar alcohol is one or more of erythritol, xylitol and sorbitol, the flavor substances comprise at least one of fruit powder and essence, and the antioxidant is vitamin C.
Still another object of the present invention is to provide a solid beverage B comprising 3 to 10 parts of the above-mentioned probiotic composition, 20 to 30 parts of inulin, 10 to 15 parts of inulin, 5 to 15 parts of honeysuckle powder, 5 to 10 parts of green tea powder, and 30 to 50 parts of xylitol; the total viable count of the solid beverage B is 90-110 hundred million CFU.
The invention also aims to provide a probiotic capsule for relieving the non-alcoholic fatty liver, which comprises the following raw materials in parts by weight: 5-15 parts of the probiotic composition, 0-10 parts of the Bradyyeast, 5-20 parts of the prebiotics, 10-15 parts of the dandelion extract, 20-30 parts of curcumin, 10-15 parts of the antioxidant and 20-30 parts of microcrystalline cellulose, wherein the specification of the probiotic capsule is 0.5 g/granule, and the total viable count of the probiotic capsule is 80-110 hundred million CFU.
Preferably, the antioxidant is vitamin C.
The invention further aims to provide a probiotic tablet for relieving non-alcoholic fatty liver, which comprises the following raw materials in parts by weight: 5-25 parts of the probiotic composition, 10-15 parts of plant extract, 10-15 parts of phyllanthus emblica powder, 10-20 parts of antioxidant, 20-35 parts of sugar alcohol and 15-25 parts of microcrystalline cellulose.
Preferably, the plant extract is a wolfberry extract.
Preferably, the antioxidant is vitamin C, the sugar alcohol is sorbitol, the specification is 0.6 g/tablet, and the total viable count of the probiotic tablet is 50-70 hundred million CFU.
It is also an object of the present invention to provide the use of the probiotic composition described above for the preparation of a product having a relief of non-alcoholic fatty liver disease.
Compared with the prior art, the invention has the beneficial effects that:
(1) Through repeated researches, the invention discovers that the combination of lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 and lactobacillus helveticus Rosell-0052 HAs stronger effects of relieving the nonalcoholic fatty liver and improving the lipid metabolism in vivo. Compared with other strains and strains, the three strains have excellent synergistic effect, and the strain has better stability and activity after being compounded and better effect of relieving the non-alcoholic fatty liver disease than that of a single strain.
(2) The invention also discovers that lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 and lactobacillus helveticus Rosell-0052 have the optimal proportion, and the lactobacillus powder formed by the lactobacillus plantarum HA-119 and lactobacillus helveticus Rosell-0052 HAs stronger fat absorption inhibition effect, HAs smaller difference between the effect of a high-dose group and the effect of a positive group, and HAs the advantage of no toxic or side effect.
(3) The probiotic composition has simple strain, simple preparation method and no toxic or side effect, and provides a new preferred scheme for clinically treating the non-alcoholic fatty liver.
Drawings
FIG. 1 is a graph showing the intensity of staining of intestinal to tail vascular fat of zebra fish after treatment with a lactic acid bacteria composite powder; wherein the dotted line frame is the analysis part intestinal canal to tail blood vessel.
Fig. 2 shows the intestinal to tail vascular fat staining intensity of zebra fish after lactobacillus complex (probiotic composition) treatment. Wherein p <0.01 and p <0.001 compared to the model control group.
FIG. 3 is a graph of the HE staining results of rat liver tissue, wherein the control group, model group and example 1 group were followed from left to right.
Detailed Description
The technical scheme of the invention is further described below in connection with specific embodiments.
In the following embodiments, lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215, lactobacillus helveticus Rosell-0052 were obtained from Lallemannd. Wherein the lactobacillus plantarum HA119 microorganism is classified as lactobacillus plantarum Lactobacillus plantarum and is deposited in the Belgium joint microorganism deposit center with the deposit number of LMG S-24119; lactobacillus casei Rosell-0215 microorganism classification name is Lactobacillus casei Lactobacillus casei, which is deposited in the national center for microbiological deposit of Pasteur institute, france, accession number I-3429; lactobacillus helveticus Rosell-0052 microorganism classification is named Lactobacillus helveticus Lactobacillus helveticus and is deposited with the national collection of microorganisms at the national institute of Pasteur, france under the accession number I-1722.
Example 1
A probiotic composition for relieving non-alcoholic fatty liver comprises the following raw materials: lactobacillus plantarum HA-119 50 parts, lactobacillus casei Rosell-0215 30 parts, lactobacillus helveticus Rosell-0052 20 parts. The live bacteria specification of the probiotic composition is 1500 hundred million CFU/g.
The preparation method comprises weighing Lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 and Lactobacillus helveticus Rosell-0052 respectively, and mixing and stirring at 20r/min for 15 min. The whole preparation is carried out at a temperature of 20+/-2 ℃ and an air relative humidity of less than 40%.
Example 2
A probiotic composition for relieving non-alcoholic fatty liver comprises the following raw materials: lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 20, lactobacillus helveticus Rosell-0052 10. The live bacteria specification of the probiotic composition is 1500 hundred million CFU/g.
The preparation method comprises weighing Lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 and Lactobacillus helveticus Rosell-0052 respectively, and mixing and stirring at 18r/min for 25 min. The whole preparation is carried out at a temperature of 20+/-2 ℃ and an air relative humidity of less than 40%.
Example 3
A probiotic composition for relieving non-alcoholic fatty liver comprises the following raw materials: lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 45, lactobacillus helveticus Rosell-0052 30. The live bacteria specification of the probiotic composition is 1500 hundred million CFU/g.
The preparation method comprises weighing Lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 and Lactobacillus helveticus Rosell-0052 respectively, and mixing and stirring at 22r/min for 20min. The whole preparation is carried out at a temperature of 20+/-2 ℃ and an air relative humidity of less than 40%.
Example 4
A probiotic composition for relieving non-alcoholic fatty liver comprises the following raw materials: lactobacillus plantarum HA-119 50 parts, lactobacillus casei Rosell-0215 30 parts, lactobacillus helveticus Rosell-0052 20 parts, bifidobacterium animalis Lafti B94 3 parts, and Saccharomyces boulardii 3 parts. The live bacteria specification of the probiotic composition is 1500 hundred million CFU/g.
The preparation method comprises the steps of weighing each probiotic component respectively, and then mixing and stirring for 15min at 20 r/min. The whole preparation is carried out at a temperature of 20+/-2 ℃ and an air relative humidity of less than 40%.
Example 5
A probiotic composition for relieving non-alcoholic fatty liver comprises the following raw materials: lactobacillus plantarum HA-119 50 parts, lactobacillus casei Rosell-0215 30 parts, lactobacillus helveticus Rosell-0052 20 parts, bifidobacterium longum Rosell-0175 parts, bifidobacterium bifidum Rosell-0071 parts. The live bacteria specification of the probiotic composition is 1500 hundred million CFU/g.
The preparation method comprises the steps of weighing each probiotic component respectively, and then mixing and stirring for 15min at 20 r/min. The whole preparation is carried out at a temperature of 20+/-2 ℃ and an air relative humidity of less than 40%.
Example 6
A solid beverage A was prepared by mixing with a specification of 2 g/bag and a viable bacteria specification of 150 hundred million CFU, the composition of which is shown in Table 1 below. Wherein the probiotic composition was prepared using the raw materials and methods of example 1.
Table 1 solid beverage a formulation
Raw materials Parts by weight
Probiotic compositions 5
Inulin 40
Erythritol 40
Apple fruit powder 10
Vitamin C 5
Example 7
A solid beverage B was prepared by mixing, with a specification of 2 g/bag and a viable bacteria specification of 100 hundred million CFU, with the composition shown in Table 2 below. Wherein the probiotic composition was prepared using the raw materials and methods of example 2.
Table 2 solid beverage B formulation
Raw materials Parts by weight
Probiotic compositions 3.33
Inulin 26.00
Xylitol 40.67
Chrysanthemum flower powder 12.00
Honeysuckle powder 10.00
Green tea powder 8.00
Example 8
A capsule, the composition of which is shown in Table 3 below, was obtained by mixing, dry granulating, and encapsulating to a specification of 0.5 g/granule and a viable bacteria specification of 100 hundred million CFU. Wherein the probiotic composition was prepared using the raw materials and methods of example 1.
Table 3 capsule formulation
Raw materials Parts by weight
Probiotic compositions 12.0
Saccharomyces boulardii 10.0
Inulin 16.0
Taraxacum mongolicum extract 10.0
Curcumin 20.0
Vitamin C 12.0
Microcrystalline cellulose 20.0
Example 9
A tablet having the composition shown in Table 4 below was obtained by dry granulation of the components, and the tablet had a specification of 0.6 g/tablet and a viable bacteria specification of 60 hundred million CFU. Wherein the probiotic composition was prepared using the raw materials and methods of example 3.
Table 4 tablet formulation
Comparative example 1
The comparative example differs from example 1 in the composition ratio, specifically: lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 75, lactobacillus helveticus Rosell-0052, 5. The live bacteria specification of the probiotic composition is 1500 hundred million CFU/g.
Comparative example 2
The difference between this comparative example and example 1 is the strain, specifically: lactobacillus plantarum Rosell-1012 50 parts, lactobacillus casei Rosell-0215 30 parts, lactobacillus helveticus Lafti L10 parts. The live bacteria specification of the probiotic composition is 1500 hundred million CFU/g.
Comparative example 3
The difference between the comparative example and the example 1 is that the raw material components and the proportions are different, specifically: 25 parts of lactobacillus acidophilus, rosell-0011 25 parts of lactobacillus rhamnosus, rosell-0215 30 parts of lactobacillus casei and Rosell-0052 20 parts of lactobacillus helveticus.
Comparative example 4
The comparative example differs from example 1 in that the probiotic composition was Lactobacillus plantarum HA-119 parts, lactobacillus casei Rosell-0215 40 parts.
Comparative example 5
The difference between this comparative example and example 1 is that the probiotic composition is Lactobacillus casei Rosell-0215 55 parts, lactobacillus helveticus Rosell-0052 45 parts.
Comparative example 6
The difference between this comparative example and example 1 is that the probiotic composition was Lactobacillus plantarum HA-119 parts, lactobacillus helveticus Rosell-0052 35 parts.
Experimental example 1 ability of the probiotic composition of the present invention to inhibit fat absorption
1.1 experimental materials:
zebra fish are all cultivated in water for fish cultivation at 28 deg.C (water quality: 200mg instant sea salt is added into 1L reverse osmosis water, conductivity is 450-550 mu S/cm, pH is 6.5-8.5, hardness is 50-100 mg/L CaCO) 3 ). The melanin allele mutant semitransparent Albino strain zebra fish is carried out in a natural pairing mating propagation mode. Zebra fish aged 5 days after fertilization (5 dpf) are used for measuring the maximum detection concentration (MTC) of the fat absorption inhibition effect of the lactobacillus composite powder and evaluating the effect of the zebra fish.
Experimental samples from the probiotic composition of example 1, a mother liquor of 20.0mg/mL was prepared with standard dilution water, ready to use. The positive control was orlistat capsule (hereinafter abbreviated as orlistat), the capsule content was white powder, lot number 190200901, shandong New age pharmaceutical Co., ltd., cool in shade, and dry storage. Stock solution of 15.0mg/mL was prepared with DMSO and stored at-20 ℃.
1.2 experimental method:
evaluation of fat absorption inhibition efficacy
Randomly selecting 5dpf melanin allele mutant semitransparent Albino strain zebra fish in beakers, and treating 30 zebra fish in each beaker. The lactic acid bacteria complex powder (probiotic composition of example 1, concentrations see table 5 below) was given in water, the positive control orlistat at a concentration of 15.0 μg/mL, while the model control group was set at a capacity of 20mL per beaker. After treatment at 28 ℃ for 1 hour, each experimental group was Bai Tianshui dissolved to give 0.1% and then removed at night using oil red O egg yolk powder solution to establish a zebra fish food fat absorption model. After culturing at 28 ℃ for 30 hours, fixing, dehydrating and dyeing, randomly selecting 10 zebra fish from each experimental group after dyeing, photographing under an dissecting microscope, analyzing and collecting data by NIS-Elements D3.20 advanced image processing software, analyzing the fat dyeing intensity from the intestinal tract to the tail blood vessel of the zebra fish, and evaluating the fat absorption inhibition effect of the lactobacillus composite powder according to the statistical analysis result of the index. Statistical treatment results are expressed in mean+ -SE. Statistical analysis was performed with SPSS 26.0 software, p <0.05 indicated that the differences were statistically significant.
1.3 experimental results:
the evaluation of the fat absorption inhibition effect is shown in Table 5, FIG. 1 and FIG. 2.
Table 5 results of experiment for evaluation of fat absorption inhibition efficacy of probiotic compositions (n=10)
P <0.01, p <0.001 compared to model control group
The result shows that the probiotic composition has stronger fat absorption inhibition effect, compared with a model group, the difference is remarkable, compared with a positive group, a high-dose group, the effect is equivalent, and no toxic or side effect exists.
Experimental example 2 the ability of the probiotic composition of the present invention to alleviate non-alcoholic fatty liver
2.1 experimental materials:
healthy SD male rats were selected and weighing 200+ -20 g. Adopting a feeding environment with the temperature of 22+/-2 ℃ and the humidity of 50% -60% and circularly lighting for 12 hours, and taking food and drinking water freely.
The basic feed consists of corn, bean cake, fish meal and other raw materials. The high-fat and high-sugar feed is prepared by adding 10% of lard, 5% of sucrose and 2% of cholesterol into basic feed.
2.2 experimental method:
the experimental rats were randomly divided into 13 groups of 10 rats each. The groups were classified into a control group, a model group, an example 1 group, an example 2 group, an example 3 group, an example 4 group, an example 5 group, a comparative example 1 group, a comparative example 2 group, a comparative example 3 group, a comparative example 4 group, a comparative example 5 group, and a comparative example 6 group. The control group is fed according to basic feed; the other groups are fed with high-fat and high-sugar feed, wherein each of the groups of examples 1-5 and comparative examples 1-6 is fed with high-fat and high-sugar feed simultaneously with each otherProbiotic compositions prepared from the corresponding group of the Tianchang, once a day, 1×10 per day 8 The CFU dose was filled with physiological saline of the same volume as the control and model groups.
2.3 sample collection and processing
After 10 weeks of the experiment, each group was anesthetized with 2% sodium pentobarbital (45 mg/kg), weighed and sacrificed by cervical spine removal after blood collection. Dissecting and taking out the liver, and then placing the liver into 4% paraformaldehyde solution for preservation. Paraffin-embedded sections were used for detection, stained by HE method, and histopathological changes were observed under a microscope. Blood samples were centrifuged at 4000rpm and serum was taken for blood lipid testing.
2.4 experimental results
(1) General status of animals: rats in the control group are full of energy, flexible and good, normal in diet, clean in skin and hair and slow in weight increase; the model group has large food intake, sleepiness, yellow hair color and quicker weight increase; the rats in the other experimental groups have mild conditions compared with the model groups, and certain differences exist between the groups.
(2) Example 1, model and control rat liver tissue HE staining is shown in figure 3. The liver lobule structure of the control group is clear and distinguishable, the sizes of liver cells are basically consistent, and the cell nucleus is complete and cytoplasmic red staining is performed. The liver cells of the model group have increased volume, fat blebs and vacuoles are mostly seen, the cell nucleus is slightly atrophic, and the liver sinus is slightly narrowed. Example 1 the liver tissue of the rats showed a degree of steatosis, but the degree of cellular degeneration was improved relative to the model group, as evidenced by a reduction in the number and volume of intracellular lipid droplets.
(3) The body weight and liver index changes after each group of rats are shown in Table 6 below.
Table 6 body weight and liver index results after each group of rats were tested
Group of Liver weight (g) Liver index (liver/body weight,%)
Control group 9.51±1.32 ** 2.93±0.30 **
Model group 14.12±0.91 3.86±0.18
Example 1 group 10.14±0.97 **a 2.98±0.32 **a
Example 2 group 10.16±0.65 **a 3.00±0.16 **a
Example 3 group 10.16±0.39 **a 2.99±0.18 **a
Example 4 group 10.24±0.67 **a 3.08±0.24 **a
Example 5 group 10.27±0.87 **a 3.12±0.37 **a
Comparative example 1 group 10.83±0.16 **b 3.29±0.85 **b
Comparative example 2 group 11.21±0.65 **b 3.35±0.21 **b
Comparative example 3 group 12.15±1.14 *c 3.48±0.19 *c
Comparative example 4 group 11.87±1.18 **b 3.34±0.38 *b
Comparative example 5 group 12.96±1.09 *c 3.67±0.26 *c
Comparative example 6 group 12.41±1.21 *c 3.54±0.15 *c
Differences from the model group are shown below with P <0.05; * P <0.01. The different english alphabets of the same column for each example and comparative example group indicate that the differences between the groups are significant, with P <0.05.
As can be seen from the above table, the liver of the model rats was significantly heavier than that of the blank control group, suggesting that the model rats had developed fatty liver. And under the intervention of probiotics, the weight and index of the liver of rats in the experimental group are lower than those of rats in the model group, which indicates that the occurrence of fatty liver can be relieved by the intervention of the probiotics. The intervention effect of 3 groups of the combination of lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 and lactobacillus helveticus Rosell-0052 is very remarkable compared with that of the model group and the rest of the comparative example groups, and is superior to that of other single bacteria and the rest of probiotics, so that the probiotic formula HAs excellent synergistic effect.
(4) Serum index concentrations after each group of rats are shown in Table 7 below.
TABLE 7 changes in serum TC, TG, AST, ALT, AST, ALT concentration in rats
Note that: the same column of different english alphabets indicates that the differences are significant, P <0.05, in comparison between the corresponding groups.
According to the table, compared with the rats in the control group, the rats in the model group have a certain degree of increase in serum TC and LDL, TG is obviously increased, and HDL is obviously reduced; furthermore, AST and ALT, which are markers of liver injury, both appear to increase significantly in the model group. In the probiotic intervention group, the intervention effects of the example group using the combination of lactobacillus plantarum HA-119, lactobacillus casei Rosell-0215 and lactobacillus helveticus Rosell-0052 were significantly different from those of the model group, and the effects of the probiotic combination were more significant than those of the comparative examples 1-6, so that the synergistic effect of the probiotic combination was further demonstrated.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.

Claims (9)

1. The probiotic composition for relieving the non-alcoholic fatty liver is characterized by comprising the following raw materials in parts by weight: 30-60 parts of lactobacillus plantarum HA-119, 0215 20-50 parts of lactobacillus casei Rosell-0215 20 and 0052 10-30 parts of lactobacillus helveticus;
the lactobacillus plantarumThe mass ratio of HA-119 to Lactobacillus casei Rosell-0215 is 1.3-3:1; the probiotic composition has a viable bacteria concentration of 1×10 11 cfu/g-2×10 11 cfu/g。
2. A method for preparing the probiotic composition according to claim 1, which comprises weighing and mixing the components of the probiotic composition, and stirring uniformly.
3. The solid beverage A is characterized by comprising the following raw materials in parts by weight: the probiotic composition of claim 1, wherein the probiotic composition comprises 5-10 parts of prebiotics, 30-60 parts of sugar alcohol, 30-60 parts of flavoring substances, 0-20 parts of antioxidants and 0-10 parts of antioxidants.
4. The solid beverage a according to claim 3, wherein the solid beverage a has a specification of 1-3 g/bag, a total viable count of 140-150 hundred million CFU, and the prebiotic is one or more of inulin, fructo-oligosaccharide, xylo-oligosaccharide, galacto-oligosaccharide, isomaltooligosaccharide, stachyose; the sugar alcohol is one or more of erythritol, xylitol and sorbitol; the flavor substance is at least one of fruit powder and essence; the antioxidant is vitamin C.
5. The solid beverage B is characterized by comprising the following raw materials in parts by weight: 3-10 parts of the probiotic composition of claim 1, 20-30 parts of inulin, 10-15 parts of chrysanthemum powder, 5-15 parts of gold-silver pollen and 5-10 parts of green tea powder, and 30-50 parts of xylitol; the specification of the solid beverage B is 2 g/bag, and the total viable count is 90-110 hundred million CFU.
6. The probiotic capsule for relieving the non-alcoholic fatty liver is characterized by comprising the following raw materials in parts by weight: the probiotic composition of claim 1, wherein the probiotic capsule has a specification of 0.5 g/granule and a total viable count of 80-110 hundred million CFU, and the probiotic composition comprises 5-15 parts of saccharomyces boulardii, 0-10 parts of saccharomyces boulardii, 5-20 parts of prebiotics, 10-15 parts of dandelion extract, 20-30 parts of curcumin, 10-15 parts of antioxidant and 20-30 parts of microcrystalline cellulose.
7. The probiotic tablet for relieving the non-alcoholic fatty liver is characterized by comprising the following raw materials in parts by weight: the probiotic composition of claim 1, wherein the probiotic composition comprises 5-25 parts of plant extract 10-15 parts, emblic powder 10-15 parts, antioxidant 10-20 parts, sugar alcohol 20-35 parts, and microcrystalline cellulose 15-25 parts.
8. The probiotic tablet of claim 7, wherein the plant extract is a wolfberry extract, the antioxidant is vitamin C, the sugar alcohol is sorbitol, the probiotic tablet has a specification of 0.6 g/tablet and a total viable count of 50-70 hundred million CFU.
9. Use of a probiotic composition according to claim 1 for the preparation of a product for alleviating non-alcoholic fatty liver disease.
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