CN114145459A - Lactic acid bacteria-containing composition, preparation method and application thereof - Google Patents
Lactic acid bacteria-containing composition, preparation method and application thereof Download PDFInfo
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- CN114145459A CN114145459A CN202010934231.5A CN202010934231A CN114145459A CN 114145459 A CN114145459 A CN 114145459A CN 202010934231 A CN202010934231 A CN 202010934231A CN 114145459 A CN114145459 A CN 114145459A
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- lactobacillus
- oligosaccharide
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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Abstract
The invention relates to the field of food, in particular to a lactic acid bacteria-containing composition, a preparation process and application thereof. The composition contains at least one of Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium bifidum, Bifidobacterium lactis and Lactobacillus rhamnosus. After animal models and human body experiments, the composition disclosed by the invention can repair disordered intestinal tracts, improve the functions of the intestines and the stomach, regulate intestinal flora and enable the body metabolism to tend to be normal; can also relieve discomfort caused by the medicines such as the Yuhui bowel-relaxing capsule, the new compound aloe capsule and the like, synergistically improve constipation and improve the bioavailability of the medicines.
Description
Technical Field
The invention relates to the field of food, and mainly relates to a lactic acid bacteria-containing composition, a preparation method and application thereof.
Background
Visceral hypersensitivity is defined as an increase in the sensory intensity and a decrease in the threshold value of a patient for visceral pain. Persistent VH is associated with neuronal sensitization, which is manifested as an increase in neuronal activation. Neurotransmitters, such as serotonin (5-hydroxytryptamine, 5-HT), play an important role in neuronal sensitization. Serotonin is a monoamine neuronal transmitter. Previous studies have shown that subcutaneous injection of 5-hydroxytryptophan (5-HTP, a precursor to serotonin) into conscious rats triggers VH. The major site of serotonin synthesis and storage is the enterochromaffin cells of the intestinal mucosa. Serotonin released from enterochromaffin cells activates neuroreflexes associated with intestinal secretion, fluidity, and sensation. According to clinical studies, patients with IBS are often associated with abnormal serotonin metabolism. In addition, serotonin receptor antagonists have been widely used as therapeutic agents, indicating that the pathology of IBS is serotonin-related.
Functional gastrointestinal disorders (FGID) are the most common problem in gastroenterological medicine. These occur as a result of gastrointestinal tract abnormalities and are defined as chronic abdominal complex syndromes such as abdominal pain, diarrhea, constipation, and abdominal distension. According to roman guidelines III, over 20 functional gastrointestinal disorders have been identified. Common FGIDs include, but are not limited to, functional abdominal pain, Irritable Bowel Syndrome (IBS), constipation, functional diarrhea, and functional dyspepsia.
Irritable bowel syndrome is chronic functional gastrointestinal disorder. About 4% to 30% of people worldwide are afflicted with IBS. IBS is characterized by two major symptoms, abdominal pain (chronic pain) and altered bowel habits. Based on these major signs, IBS can be classified into four subgroups: diarrhea-predominant IBS (IBS-D), constipation-predominant IBS (IBS-C), mixed IBS (IBS-M), and non-typeable IBS (IBSU). Of the IBS patients, about 30% of patients suffer from IBS after gastrointestinal inflammation, and those patients are suffering from post-infection/inflammation IBS. Furthermore, a major factor encouraging patients to seek health care and cause significant quality of life to decline is abdominal pain associated with Visceral Hypersensitivity (VH). Visceral hypersensitivity is considered to be one of the major mechanisms responsible for functional gastrointestinal disorders. In addition, recent studies have shown that visceral hypersensitivity is highly specific for IBS.
Lactic acid bacteria are the most important species for use as probiotics and have been considered as an alternative to prevent or treat gastrointestinal health due to their ability to modulate the host intestinal flora. In addition, there is growing evidence in recent years that lactic acid bacteria can also alter the psychological and physiological response of the host to psychological stress (Zareie et al, 2006).
Lactic acid bacteria, a probiotic existing in human bodies, are a general term for a class of bacteria capable of producing a large amount of lactic acid by utilizing fermentable carbohydrates, can help digestion and are beneficial to the health of human intestines. A large number of researches show that the probiotic lactic acid bacteria can regulate normal flora of gastrointestinal tracts of organisms, keep micro-ecological balance, improve the digestibility and the biovalue of food, reduce serum cholesterol, control endotoxin, inhibit growth and propagation of putrefying bacteria and generation of putrefying products in intestinal tracts, produce nutrient substances and stimulate tissue development, thereby having effects on the nutritional state, the physiological function, cell infection, drug effect, toxic reaction, immune reaction, tumorigenesis, aging process, sudden emergency reaction and the like of the organisms. The probiotic lactic acid bacteria not only can improve the nutritive value of food, improve the flavor of food and improve the preservation property and added value of food, but also increasingly attract attention from people on the special physiological activity and nutritional function of the probiotic lactic acid bacteria.
Disclosure of Invention
Accordingly, the present invention provides a composition containing lactic acid bacteria, which can be prepared into ready-to-eat lactic acid bacteria, can be taken alone to repair disordered intestinal tracts and enable the body to have metabolic tendency, and can also be taken together with other medicines, such as aloe vera laxative capsules and compound aloe vera capsules to improve intestinal tract microecology, and other medicines such as orlistat and cetilistat, so as to improve the bioavailability of the medicines and make the synergistic improvement mechanism more obvious.
Specifically, the invention is realized by the following steps:
a lactic acid bacteria-containing strain, wherein said composition comprises:
(1) at least one of Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium bifidum, Bifidobacterium lactis, and Lactobacillus rhamnosus;
(2) inulin, oligosaccharides and optionally food grade or pharmaceutically acceptable adjuvants.
The lactobacillus is selected from at least two of Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium bifidum, Bifidobacterium lactis and Lactobacillus rhamnosus.
Further, the lactic acid bacteria are selected from at least three of lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium bifidum, bifidobacterium lactis and lactobacillus rhamnosus.
Further, the lactic acid bacteria are selected from at least four of lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium bifidum, bifidobacterium lactis and lactobacillus rhamnosus.
Further, the lactobacillus is a mixed bacterium of lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium bifidum, bifidobacterium lactis and lactobacillus rhamnosus.
The food-grade or pharmaceutically acceptable auxiliary materials are adhesive and filling agent; further, the food grade or pharmaceutically acceptable excipients include, but are not limited to, polyols, starches.
The composition comprises the following components in parts by weight:
the oligosaccharide is at least one of fructo-oligosaccharide, isomaltooligosaccharide, xylooligosaccharide and stachyose; the polyhydric alcohol is at least one of lactitol, inositol, galactitol, sorbitol, xylitol, maltitol and mannitol; the starch is selected from at least one of corn starch and wheat starch.
Calculated by weight ratio, the oligosaccharide is selected from 50-200 parts of fructo-oligosaccharide, 150-400 parts of isomaltooligosaccharide, 1-15 parts of xylo-oligosaccharide and 5-15 parts of stachyose; the polyol is selected from 500 parts of lactitol 250-; the starch is selected from 5-20 parts of corn starch.
The lactobacillus is selected from Lactobacillus plantarum LP 4515-45 parts, Bifidobacterium lactis BAL 5311-10 parts, Bifidobacterium bifidum TMC 31151-10 parts, Lactobacillus acidophilus La 281-10 parts, and Lactobacillus rhamnosus LR 5191-10 parts.
The Lactobacillus plantarum LP45 (namely Lactobacillus plantarum, YMC1005) is preserved in China general microbiological culture Collection center (CGMCC), and the strain is preserved in Beijing West Lu No.1 Hospital, Chaoyang district, Beijing, and the preservation number is 8 months and 26 days in 2013: CGMCC No. 8072.
Lactobacillus acidophilus La28 (Lactobacillus acidophilus) which is preserved in China general microbiological culture Collection center at the preservation address of No. 3 Xilu-Chen-Xilu-1 of the area of the south-facing-the-Yang, Beijing, the institute of microbiology, China academy of sciences, with the preservation date of 2015, 10 months and 15 days, and the preservation number of CGMCC No. 11506.
Bifidobacterium bifidum TMC3115 (Bifidobacterium bifidum) is preserved in China general microbiological culture Collection center (CGMCC), with the preservation date of 11 months and 11 days in 2013 and the preservation number of CGMCC No.8462 at Hodgkin, West Lu No.1 institute of North American district, China institute of microbiology, Beijing.
Bifidobacterium lactis BAL531 (Bifidobacterium lactis) is preserved in China general microbiological culture Collection Center (CCM) with the preservation date of 2019, 03 and 13 days, and the preservation number of CGMCC No.17329
Lactobacillus rhamnosus Lr-G14 (Lactobacillus rhamnosus) is preserved in China general microorganism culture Collection, with the address of Beijing Corp-Yangzhou region of Suzuku-Xilu No.1, Zhongke institute of microbiology, the preservation date of 2018, 06 and 20 days, and the preservation number of GMCCNo.15969
The composition comprises, by weight, 20-40 parts of Lactobacillus plantarum LP 4520-40 parts, Bifidobacterium bifidum TMC 31154-8 parts, Bifidobacterium lactis BAL 5314-8 parts, Lactobacillus acidophilus La 281-10 parts, Lactobacillus rhamnosus LR 5191-10 parts, 300 parts of inulin 100-; more preferably, the feed comprises, by weight, 4530 parts of lactobacillus plantarum LP, 5315 parts of bifidobacterium lactis BAL, 31155 parts of bifidobacterium bifidum TMC, 285 parts of lactobacillus acidophilus La, 5194 parts of lactobacillus rhamnosus LR, 200 parts of inulin, 100 parts of fructo-oligosaccharide, 270 parts of isomalto-oligosaccharide, 10 parts of xylo-oligosaccharide, 10 parts of stachyose, 320 parts of lactitol and 10 parts of corn starch. The composition comprises the following steps:
(1) mixing inulin and oligosaccharide, adding food-grade or pharmaceutically-acceptable adjuvants, wet granulating, sterilizing, and sieving to obtain granule;
(2) mixing lactic acid bacteria with the granules obtained in step (1), and preparing according to a conventional process.
The composition of the invention contains at least 10 per g of the composition8To 1012CFU lactic acid bacteria.
The composition of the invention contains at least 10 per g of the composition9To 1011CFU lactic acid bacteria.
The composition can be made into any one of pill, tablet, capsule, powder, granule or oral liquid.
The invention also provides a medicine, food or health product containing the composition; preferably, the food or health product includes but is not limited to probiotics, prebiotics, biscuits, snacks.
The invention also provides an application of the traditional Chinese medicine composition in preparing medicines, foods or health-care products for regulating intestinal functions; and the application in preparing the medicine or food for assisting the efficacy of the aloe capsule for expelling toxin, beautifying, relaxing the bowels and moistening the intestines.
The invention also provides instant lactic acid bacteria, and the instant lactic acid bacteria have the composition.
The invention also provides a pharmaceutical composition which contains the instant lactobacillus and any one of orlistat, aloe vera capsules, compound aloe capsules, cistanche cathartica oral liquid, edestan pills, qingning pills, cathartic capsules, cistanche cathartica capsules, eden intestine moistening pills, eden spleen nourishing pills, constipate, tongle granules, neolistat, amoxicillin, irinotecan, isoniazid, rifampicin, rifapentine and fluorouracil antineoplastic drugs.
The medicinal composition can be prepared into granules by the conventional process from the contents of instant lactic acid bacteria and the aloe constipation relieving capsule (Chinese standard character Z20150041), and the proportion of the contents of the instant lactic acid bacteria and the aloe constipation relieving capsule is 1-10:1 by weight; preferably, the ratio of the content of the instant lactobacillus and the aloe cathartic capsule is 4.26: 1;
the pharmaceutical composition can be prepared from instant lactobacillus and orlistat, wherein the ratio of the instant lactobacillus to the orlistat is 1-50: 1; preferably, the ratio of the instant lactic acid bacteria to the orlistat is 10-30:1 in terms of weight and dosage ratio; more preferably, the ratio of the instant lactic acid bacteria to orlistat is 27:1 by weight.
The pharmaceutical composition can also be prepared into granules by the conventional process of the content of the instant lactic acid bacteria and the compound aloe capsule (Chinese medicine standard Z13020306), and the proportion of the content of the instant lactic acid bacteria and the compound aloe capsule is 1-20:1 by weight; preferably, the proportion of the content of the instant lactobacillus and the compound aloe capsule is 1-10:1 by weight; further preferably, the proportion of the content of the instant lactic acid bacteria to the content of the compound aloe capsule is 5.4:1 by weight.
The invention also aims to provide a preparation method of the instant lactic acid bacteria, and the technological process of the method is shown in figure 1 in the attached drawing of the specification.
Compared with the prior art, the invention has the advantages that:
1) in animal model experiments, the pharmaceutical composition can effectively change intestinal flora, regulate the intestinal flora structure, improve the microenvironment and further improve the intestinal health, has good regulation effect on intestinal flora disturbance caused by various reasons, and obtains good technical effect.
2) The results of comparative experiments show that the specific proportion of the composition provided by the invention to orlistat, aloe vera laxative capsules, compound aloe capsules and the like is more excellent in the effects of regulating weight loss, constipation and intestinal flora structure than that of a single composition or the orlistat, the aloe vera laxative capsules and the compound aloe capsules, and the composition has better synergistic effect after being combined with the specific weight proportion of the orlistat, the aloe vera laxative capsules, the compound aloe capsules and the like.
Drawings
FIG. 1 is a flow chart of the preparation process of the instant lactic acid bacteria.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
The present invention is further illustrated by the following examples, which are intended to be purely exemplary and are not intended to limit the scope of the invention, as various equivalent modifications of the invention will occur to those skilled in the art upon reading the present disclosure and fall within the scope of the appended claims.
Example 1: an instant lactic acid bacteria, the used raw materials and the preparation process are as follows:
the preparation process comprises the following steps:
(1) mixing inulin and fructo-oligosaccharide, adding lactitol and corn starch, granulating by wet method, sterilizing, and sieving to obtain granules;
(2) mixing Lactobacillus plantarum LP45, Bifidobacterium bifidum TMC3115 and the granules in step (1), and preparing according to conventional process.
Example 2: an instant lactic acid bacteria, the used raw materials and the preparation process are as follows:
the preparation process is the same as in example 1.
Example 3: an instant lactic acid bacteria, the used raw materials and the preparation process are as follows:
100 g of inulin, 50 g of fructo-oligosaccharide, 150 g of isomalto-oligosaccharide, 5g of xylo-oligosaccharide, 5g of stachyose, 250 g of lactitol and 5g of corn starch.
The preparation process is the same as in example 1.
Example 4: an instant lactic acid bacteria, the used raw materials and the preparation process are as follows:
300 g of inulin, 150 g of fructo-oligosaccharide, 400 g of isomalto-oligosaccharide, 15g of xylo-oligosaccharide, 15g of stachyose, 500 g of lactitol and 15g of corn starch.
The preparation process is the same as in example 1.
Example 5: an instant lactic acid bacteria, the used raw materials and the preparation process are as follows:
200 g of inulin, 100 g of fructo-oligosaccharide, 270 g of isomalto-oligosaccharide, 10 g of xylo-oligosaccharide, 10 g of stachyose, 320 g of lactitol and 10 g of corn starch.
The preparation process is the same as in example 1.
Example 6: an instant lactic acid bacteria, the used raw materials and the preparation process are as follows:
lactobacillus plantarum LP 4530 g
Bifidobacterium lactis BAL 5315 g
Bifidobacterium bifidum TMC 31155 g
200 g of inulin, 100 g of fructo-oligosaccharide, 270 g of isomalto-oligosaccharide, 10 g of xylo-oligosaccharide, 10 g of stachyose, 320 g of lactitol and 10 g of corn starch.
The preparation process is the same as in example 1.
Example 7: an instant lactic acid bacteria, the used raw materials and the preparation process are as follows:
200 g of inulin, 100 g of fructo-oligosaccharide, 270 g of isomalto-oligosaccharide, 10 g of xylo-oligosaccharide, 10 g of stachyose, 320 g of lactitol and 10 g of corn starch.
The preparation process is the same as in example 1.
Example 8: a medicine containing instant lactobacillus comprises the following raw materials and preparation process:
200 g of inulin, 100 g of fructo-oligosaccharide, 270 g of isomalto-oligosaccharide, 10 g of xylo-oligosaccharide, 10 g of stachyose, 320 g of lactitol and 10 g of corn starch.
The above raw materials were prepared into a pharmaceutical composition according to the preparation process described in example 1, and then the contents of the aloe vera laxative capsule were added in a weight ratio of 4.26:1, and prepared into granules according to a conventional process, and then the granules were bagged, 3g per bag.
Example 9: a medicine containing instant lactobacillus comprises the following raw materials and preparation process:
200 g of inulin, 100 g of fructo-oligosaccharide, 270 g of isomalto-oligosaccharide, 10 g of xylo-oligosaccharide, 10 g of stachyose, 320 g of lactitol and 10 g of corn starch.
The raw materials are prepared into a pharmaceutical composition according to the preparation process of example 1, orlistat is added according to the weight ratio of 27:1, and the pharmaceutical composition is prepared into granules according to a conventional process and is bagged, wherein each bag contains 3g of orlistat.
Example 10: a medicine containing instant lactobacillus comprises the following raw materials and preparation process:
200 g of inulin, 100 g of fructo-oligosaccharide, 270 g of isomalto-oligosaccharide, 10 g of xylo-oligosaccharide, 10 g of stachyose, 320 g of lactitol and 10 g of corn starch.
The raw materials are prepared into a pharmaceutical composition according to the preparation process of example 1, then the content of the compound aloe capsule is added according to the weight ratio of 5.4:1, the compound aloe capsule is prepared into granules according to the conventional process, and the granules are packaged, wherein each bag contains 3g of the compound aloe capsule.
Comparative example 1: an instant lactobacillus-containing food, which comprises the following raw materials and preparation process:
49 g of microcrystalline cellulose, 200 g of inulin, 10 g of fructo-oligosaccharide, 27 g of isomalto-oligosaccharide, 10 g of xylo-oligosaccharide, 10 g of stachyose, 320 g of lactitol and 10 g of corn starch.
The preparation process is the same as in example 1.
Comparative example 2: a composition containing instant lactobacillus comprises the following raw materials and preparation process:
80 g of inulin, 300 g of fructo-oligosaccharide, 270 g of isomalto-oligosaccharide, 60 g of xylo-oligosaccharide, 60 g of stachyose, 320 g of lactitol and 10 g of corn starch.
The preparation process is the same as in example 1.
Comparative example 3: a composition containing instant lactobacillus comprises the following raw materials and preparation process:
50 g of inulin, 100 g of fructo-oligosaccharide, 277 g of isomalto-oligosaccharide, 110 g of xylo-oligosaccharide, 15g of stachyose, 120 g of lactitol and 5g of corn starch.
The preparation process is the same as in example 1.
Comparative example 4: a composition containing instant lactobacillus comprises the following raw materials and preparation process:
the preparation process comprises the following steps:
taking lactobacillus plantarum, lactobacillus acidophilus and lactobacillus paracasei according to the prescription amount, uniformly mixing the lactobacillus plantarum, lactobacillus acidophilus and lactobacillus paracasei to form bacterial powder, sterilizing, sieving, mixing the bacterial powder, and bagging, wherein each bag is 3g, so as to obtain the lactobacillus paracasei.
Comparative example 5: a composition containing instant lactobacillus comprises the following raw materials and preparation process:
the preparation process comprises the following steps:
taking lactobacillus plantarum, lactobacillus acidophilus and lactobacillus paracasei according to the prescription amount, uniformly mixing to obtain bacterium powder, adding aspartame and fruit powder, sterilizing, sieving, mixing with the bacterium powder, and bagging, wherein each bag is 3g, thus obtaining the lactobacillus paracasei beverage.
Verification examples
First, the Effect of the composition of the present invention on the intestinal flora
Test materials and test sites
Experimental unit: pharmacological center of Shandong New times pharmaceutical Co., Ltd, Shandong pharmaceutical group, Lunan
Animals: the SD rat is provided by Lunan pharmaceutical group GmbH, and the batch number of the animal qualification certificate is as follows: SYXK (Lu)
20180008, half male and female, the body weight is 180-220 g.
1.1 Molding and grouping
The molded rats were fed with high fat diet (formula 78.8% basal diet, 1% cholesterol, 10% egg yolk powder, 10% lard, 0.2% bile salt) continuously for 7 days.
Taking 110 successfully molded rats, randomly dividing the rats into 11 groups of 10 rats, and respectively taking a model control group, an example 5 group, an example 8 group, an example 9 group, an example 10 group, a compound aloe capsule group, an aloe vera capsule group, an orlistat group, a comparative example 3 group, a comparative example 4 group and a comparative example 5 group; another 10 rats fed with normal feed were used as a blank control group.
1.2 modes of administration and doses were as follows:
example 5 group: intragastrically administering the ready-to-eat lactic acid bacteria of example 5 (0.81 g/kg/d);
example 8 group: the drug of example 8 (equivalent to 0.81g/kg/d lactic acid bacteria composition +0.19g/kg/d aloe vera laxative capsule) was administered by gavage;
example 9 group: the drug of example 9 (equivalent to 0.81g/kg/d lactic acid bacteria composition +0.03g/kg/d orlistat) was administered by gavage;
example 10 group: the drug of example 10 (equivalent to 0.81g/kg/d lactic acid bacteria composition +0.15g/kg/d compound aloe capsule) was administered by gavage;
compound aloe capsule group: compound aloe capsule (0.15g/kg/d) is administrated after intragastric administration;
the aloe capsule group for relaxing bowels: the fleece-flower root, aloe and cathartic capsule (0.19g/kg/d) is given after intragastric administration;
comparative example 3 group: the composition of comparative example 3 (0.81g/kg/d) was gavaged;
comparative example 4 group: gavage the composition of comparative example 4 (0.062 g/kg/d);
comparative example 5 group: the composition of comparative example 5 (0.069g/kg/d) was gavaged;
the normal control group and the model control group are administered with purified water with equal volume by intragastric administration.
The volume of the composition administered by gavage to rats was 10mL/kg, and the composition was gavage 1 time per day for 4 days. The number of ingested viable bacteria was the same for each group by calculation.
During the test period, the blank control group was continuously given normal diet, and the model control group and each administration group were continuously given high fat diet.
1.3 detection of intestinal flora in rats
Before the last administration, taking a proper amount of excrement aseptically (0.1 +/-0.01 g), placing the excrement in a dry sterilized test tube, weighing again, calculating the weight of the excrement of the rat, adding 0.9% sodium chloride solution, shaking for dissolution, preparing 10mg/mL suspension, sequentially diluting by 10 times, selecting proper dilution, respectively inoculating to the following selective culture media by adopting a spiral inoculator, and culturing according to the following requirements.
And (3) lactobacillus: LBS agar medium, at 37 deg.C for 48h, aerobically culturing;
bifidobacteria: BBL agar medium, 37 ℃, 48h, anaerobic culture.
After completion of the culture, the cells were formed into colony forming units (cfu. g)-1) Counts were made and results are expressed as log of bacterial colonies per gram of feces. The changes of lactobacillus and bifidobacterium among groups are compared.
1.4 data processing
Statistical analysis is carried out by SPSS19.0 software in statistical processing, and the mean value plus or minus standard deviation is used for experimental data
Showing that the two-sided t test is used for significance analysis among groups, and P <0.05 is difference with statistical significance.
2. Results
2.1 Effect of the compositions of the invention on the intestinal flora of rats
The test results are shown in table 1, and compared with the normal control group, the numbers of the bacteria and the bifidobacteria in the rat manure sample of the model control group are obviously reduced (P is less than 0.01).
After the composition of the invention is used, the numbers of lactobacillus and bifidobacterium in rat manure samples of the groups of example 2, example 5, example 8, example 9 and example 10 are all obviously increased, and the difference has significance (P is less than 0.01) compared with the model control group.
Compared with the groups of the comparative examples 3 to 5, the numbers of the lactic acid bacteria and the bifidobacteria in the groups of the examples 1 to 3 are obviously increased, and the difference has significance (P <0.05 or P < 0.01).
TABLE 1 Effect of the compositions of the present invention on the amount of Lactobacillus and Bifidobacterium in rats
Compared with the normal control group, the composition has the advantages that,*P<0.05，**P<0.01；
compared with the model control group,&P<0.05，&&P<0.01。
as can be seen from table 1: the composition of the embodiment of the invention can remarkably increase the number of lactobacillus and bifidobacterium in the intestinal tract and regulate the intestinal flora.
2. Treatment evaluation of the composition of the invention on the enteritis experiment of mice
Experimental unit: pharmacological center of Shandong New times pharmaceutical Co., Ltd, Shandong pharmaceutical group, Lunan
Animals: BALB/C mice were provided by the Lunan pharmaceutical group, Inc., and the certification batch numbers of the experimental animals were:
SYXK 20180008, male and female halves, 3-4 weeks old, with a weight of 18-22 g.
2.1 Molding and grouping
The mice establish a mouse enteritis model according to the establishment and evaluation modes of a mouse enteritis model induced by enterotoxigenic escherichia coli K88 such as Qiyu and the like. Taking 140 mice which are successfully molded, and randomly dividing the mice into a model control group, a positive control group, example groups 2, 3, 4, 5 and 6 and comparative example groups 1, 2, 3, 4 and 5, wherein each group comprises 10 mice; another 10 normal mice were used as a blank control group.
Each administration group was administered with the corresponding drug in the following manner:
example 2 group: gavage the composition of example 2 (1.00 g/kg/d);
example 3 group: gavage the composition of example 3 (1.00 g/kg/d);
example 4 group: gavage the composition of example 4 (1.00 g/kg/d);
example 5 group: gavage the composition of example 5 (1.00 g/kg/d);
example 6 group: gavage the composition of example 6 (1.00 g/kg/d);
example 7 group: gavage the composition of example 7 (1.00 g/kg/d);
comparative example 1 group: the composition of comparative example 1 (1.00g/kg/d) was gavaged;
comparative example 2 group: the composition of comparative example 2 (1.00g/kg/d) was gavaged;
comparative example 3 group: the composition of comparative example 3 (1.00g/kg/d) was gavaged;
comparative example 4 group: the composition of comparative example 4 (1.00g/kg/d) was gavaged;
comparative example 5 group: the composition of comparative example 5 (1.00/kg/d) was gavaged;
positive control group: tablets for enteritis (administration dose: 0.8736g/kg/d) were taken.
The blank and model mice were administered starch (dose 1.0 mg/day/gavage) by gavage for 4 days. During the experiment, mice in each group were fed normally.
2.2 detection of indicators
Diarrhea and pain. The most obvious manifestation of enteritis is diarrhea, namely the times of defecation, which is the result most capable of visually reflecting colitis, and the experiments of the invention count the times of defecation of mice within 24 hours; another characterization of colitis is pain, measured by the inventors by observing the number of body torsions in the mice over 8h, and the results of each group of measurements are shown in Table 2.
2.3 statistical treatment
All experimental data are expressed as means ± standard deviation, and a t-test for significance between groups was performed using the sps 19.0 software.
TABLE 2 measurement results of various indexes of examples
| Group of | Number of stools | Number of body torsions (times/8 h) |
| Blank control group | 2.1±0.56 | 3.52±0.88 |
| Model control group | 7.2±1.71** | 19.72±4.01** |
| Tablets for treating enteritis | 2.8±0.32&&## | 5.25±1.73&&##@ |
| EXAMPLE 2 group | 3.0±0.40&&## | 5.19±1.14&&##@ |
| EXAMPLE 3 group | 2.3±0.96&&## | 4.25±1.27&&##@ |
| EXAMPLE 4 group | 2.5±0.29&&## | 3.99±0.73&&##@ |
| EXAMPLE 5 group | 2.1±0.55&&##@ | 3.25±1.09&&##@ |
| EXAMPLE 6 group | 2.8±0.44&&## | 4.15±1.13&&##@ |
| EXAMPLE 7 group | 2.7±0.63&&## | 4.09±0.87&&##@ |
| Comparative example 1 group | 7.0±1.22 | 18.65±3.22 |
| Comparative example 2 group | 5.8±1.30 | 10.25±2.77&& |
| Comparative example 3 group | 4.4±1.91& | 9.69±3.12&& |
| Comparative example 4 group | 2.4±1.5&&## | 4.56±1.27&&## |
| Comparative example 5 group | 2.3±0.85&&## | 4.44±0.92&&## |
Note: compared with the blank control group, the composition of the composition,*P<0.05，**P<0.01；
compared with the model control group,&P<0.05，&&P<0.01；
in comparison with the group of comparative example 1,#P<0.05，##P<0.01；
in comparison with the group of comparative example 2,@P<0.05，@@P<0.01。
the results in table 2 show the stool frequency and body torsion frequency of the mice, and the blank control group is compared with the model control group, so that the modeling of the invention is successful, and each group of the examples has improvement effect on the stool frequency and the body torsion frequency of the mice, but the treatment effect of the comparative example is obviously lower than that of the group of the examples of the invention (P <0.05 and P < 0.01).
3. Research on function of outlet obstruction type rat constipation model
3.1 materials and methods
Experimental animals: SD rat, sex is half respectively, and the weight is 180 ~ 220g, is provided by Lunan pharmaceutical group member company Limited, and experimental animals qualification batch number is: SYXK (lu) 20180008; one week of adaptive feeding before the experiment.
3.2 drugs and reagents
Example 5 group: gavage the composition of example 5 (0.81 g/kg/d);
example 8 group: the drug of example 8 (equivalent to 0.81g/kg/d lactic acid bacteria composition +0.19g/kg/d aloe vera laxative capsule) was administered by gavage;
example 10 group: the drug of example 10 (equivalent to 0.81g/kg/d lactic acid bacteria composition +0.15g/kg/d compound aloe capsule) was administered by gavage;
compound aloe capsule group: compound aloe capsule (0.15g/kg/d) is administrated after intragastric administration;
the aloe capsule group for relaxing bowels: the fleece-flower root, aloe and cathartic capsule (0.19g/kg/d) is given after intragastric administration;
comparative example 1: the composition of comparative example 1 (0.81g/kg/d) was gavaged;
comparative example 3: the composition of comparative example 3 (0.81g/kg/d) was gavaged;
comparative example 5: the composition of comparative example 5 (0.81g/kg/d) was administered by gavage.
3.3 Molding method
The rectum incompetence ligation method is adopted for molding. Rats were randomly divided into 10 groups of 10 rats each with hermaphrodite halves, each group consisting of blank group, example 5, example 8, example 10, compound aloe capsule group, aloe vera laxative capsule group, comparative example 1, comparative example 3, and comparative example 5. The blank groups were not processed. The rats in other groups were anesthetized with sodium pentobarbital at a dose of 30mg/kg, opened, the rectum exposed, and 10 gauge silk was passed through the abdominal wall from 1-1.5cm from the anus, passed under the rectum in a "8" pattern, and then led out through the abdominal wall, with the distance between the silk introduction and extraction points being about 0.5 cm. And (3) sleeving an iron wire with the diameter of 0.5cm in the silk thread, knotting, and keeping the rectum partially narrowed and suspended on the abdominal wall and the intestinal wall in a normal blood circulation state. The abdominal cavity was closed and the rats were given a normal diet after awakening.
And (3) postoperative administration, wherein the administration is carried out for 3 days continuously, once a day, 1h after the last administration, the administration of the medicines is carried out by using ink according to the weight for intragastric administration, after the intragastric administration is finished for 30min, cervical vertebrae are taken off immediately to kill rats, the abdominal cavity is opened, mesentery is separated, the length from the upper end of a large intestine to a pylorus is taken as the length of a small intestine, and the length from the pylorus to the front edge of the ink is taken as the length of ink propulsion.
Calculating the ink propelling rate: the ink propulsion rate (%) < ink propulsion length/small intestine length x 100%
Weighing the wet weight of the small intestine, drying the small intestine to constant weight at 80 ℃, and weighing.
Calculating the water content of the intestinal cavity: water content (%) of small intestine (wet weight of small intestine-dry weight of small intestine)/wet weight of small intestine.
Data results were analyzed using SPSS19.0 software.
3.4 Experimental results:
the results are shown in Table 3.
TABLE 3 comparison of carbon dust propulsion rate and Small intestine Water content
| Grouping | Carbon dust advancing Rate (%) | Moisture content of small intestine (%) |
| Normal group | 42.7±5.7 | 58.1±10.5 |
| Model set | 30.3±6.1** | 67.2±11.3** |
| EXAMPLE 5 group | 36.9±7.7&&#@ | 60.9±9.6&&# |
| EXAMPLE 8 group | 44.6±8.5&&##@@ | 55.3±8.1&&## |
| EXAMPLE 10 group | 41.5±9.4&&##@@ | 58.6±11.9&& |
| Compound aloe capsule group | 38.6±7.5&&##@@ | 50.3±6.9&&## |
| Aloe capsule for relieving constipation | 40.36±9.4&&##@@ | 53.2±11.9&& |
| Comparative example 1 group | 31.0±5.3 | 65.7±12.2 |
| Comparative example 3 group | 34.9±4.8& | 64.1±9.3 |
| Comparative example 5 group | 33.8±6.6 | 65.3±10.8 |
Note: compared with the blank control group, the composition of the composition,*P<0.05，**P<0.01；
compared with the model control group,&P<0.05，&&P<0.01；
in comparison with the group of comparative example 1,#P<0.05，##P<0.01；
in comparison with the group of comparative example 2,@P<0.05，@@P<0.01。
as can be seen from table 3, the model group had the worst and significant differences in the voluntary defecation function (P <0.01) relative to the normal group. The carbon powder propulsion rate and the small intestine water content of each group of the examples and the group of the comparative examples are improved to different degrees. However, as can be seen from the statistical data, the groups of the examples and the group of the comparative examples have significant difference in promoting the intestinal peristalsis of the rats and reducing the water content of the intestinal cavity (P <0.05 and P < 0.01).
Second, the staff of the Lunan pharmaceutical group takes the pharmaceutical composition of the invention to investigate
Medicine preparation: aloe laxative capsules, manufacturer: southwestern, pacho, pharmaceuticals, inc.
The experimenter: randomly selecting 120 volunteers from the crowd with the desire to relieve functional constipation
Grouping and taking methods:
experimental group (ii): 2 hui tong capsules/time + 5g of the composition of example 5/time, once in the morning, in the middle and evening, three times a day;
experiment group two: the aloe cathartic capsule is 2 granules per time, and is taken once in the morning, in the middle and at night, three times per day;
experimental group iii group: the composition of example 5 was administered at 5 g/dose, once in the morning, in the middle of the evening, three times per day.
Experimental group iv group: 2 hui laxative capsules/time + 5g of the composition of comparative example 5/time, once in the morning, in the middle and in the evening, three times a day;
experimental group v group: 2 hui laxative capsules/time + 5g of the composition of comparative example 1/time, once in the morning, in the middle and in the evening, three times a day;
observation group: the compound aloe capsule is 2 granules/time, once in the morning and at night.
Taking the medicine for 14 days without interruption, observing and recording results of each group by a statistical experimental group, and counting constipation and gastrointestinal tract condition information again after 14 days.
The main indexes are as follows:
1) stool characteristics: number of bowel movements and satisfaction, number of bowel movements within 24 hours after taking the relevant example drug/composition, statistics were taken for 14 consecutive days. Ease of defecation: very satisfactory 10 points, satisfactory 7 points, generally 3 points, and 1 point difference.
2) Abdominal characteristics: the degrees of alleviating abdominal pain, borborborygmus, diarrhea and abdominal distension are respectively 10 points of satisfaction, 7 points of satisfaction, 3 points of general satisfaction and 1 point of difference.
3) After 14 days, the defecation frequency and the abdominal pain, the borborygmus, the diarrhea and the abdominal distension are counted.
Secondary indexes are as follows:
1) degree of mouthfeel: satisfaction of mouthfeel of the composition taken: very satisfactory 10 points, satisfactory 7 points, generally 3 points, and 1 point difference.
2) Diet: appetite following administration of the composition: very satisfactory 10 points, satisfactory 7 points, generally 3 points, and 1 point difference.
3) Odor: the faint scent degree of the composition is as follows: very satisfactory 10 points, satisfactory 7 points, generally 3 points, and 1 point difference.
4) And (3) halitosis improvement: degree of change in halitosis by taking the composition: very satisfactory 10 points, satisfactory 7 points, generally 3 points, and 1 point difference.
As a result: the final scores were pooled and statistically analyzed using a sps 19.0.
TABLE 4 Abdominal characteristics statistics
Note: compared with the second experimental group,#p is less than 0.05, compared with the experimental group IV@P＜0.05；
Compared with the experimental group (v),￥p is less than 0.05; in comparison with the observation group,▲P＜0.05，▲▲P＜0.01。
TABLE 5 statistical results of stool characteristics
Note: compared with the experimental group III,▲P＜0.05，▲▲P＜0.01。
as can be seen from tables 4 and 5, the abdominal cavity characteristics and the stool characteristics of each experimental group were improved to different degrees, but the most significant improvement effect was found to be that the first experimental group was superior to other experimental groups in that the constipation problem of patients could be improved by using only the aloea cathartic capsule, but the composition of example 5 of the present invention had a significant effect in improving abdominal characteristics, such as diarrhea and abdominal pain, and improving symptoms of abdominal pain and diarrhea.
TABLE 6 measurement results of various indexes of examples
| Group of | Degree of mouthfeel | Smell(s) | Diet | Halitosis |
| Experimental group I group | 95.8±11.3@￥ | 96.5±8.6@@￥ | 88.5±6.6##@@▲▲ | 87.9±6.6##@@￥￥▲▲ |
| Experiment group 2 | -- | -- | 57.5±7.3 | 65.5±7.3▲ |
| Experimental group iii group | 96.3±9.6@￥ | 96.8±6.5@@￥ | 87.2±8.5##@@￥￥▲▲ | 84.8±8.2##@@￥￥▲▲ |
| Experimental group iv | 86.2±8.5 | 60.2±5.9 | 54.3±5.2 | 70.1±7.5▲ |
| Experimental group v group | 82.9±9.4 | 87.2±7.2 | 54.2±7.7 | 66.2±5.6▲ |
| Observation group | -- | -- | 53.8±9.9 | 60.8±9.9 |
Note: compared with the second experimental group,#P＜0.05，##p is less than 0.01; compared with the experimental group IV@P＜0.05，@@P＜0.01；
Compared with the experimental group (v),￥P＜0.05，￥￥p is less than 0.01; in comparison with the observation group,▲P＜0.05，▲▲p is less than 0.01. As can be seen from Table 3, additionThe degree of mouthfeel and the smell of the fruit powder and the aspartame are more easily accepted, but the problems of halitosis and appetite cannot be solved; the experimental group (iv) and the experimental group (v) have a great difference from the composition of the present invention in the aspects of taste, diet, etc.
Claims (16)
1. A lactic acid bacteria-containing composition, comprising the following components:
(1) at least one of Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium bifidum, Bifidobacterium lactis, and Lactobacillus rhamnosus;
(2) inulin, oligosaccharides and optionally food grade or pharmaceutically acceptable adjuvants.
2. The composition of claim 1, wherein the composition comprises at least two of lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium bifidum, bifidobacterium lactis, lactobacillus rhamnosus; further, the composition contains at least three of lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium bifidum, bifidobacterium lactis and lactobacillus rhamnosus; further, the composition contains at least four of lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium bifidum, bifidobacterium lactis and lactobacillus rhamnosus; further, the composition contains lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium bifidum, bifidobacterium lactis and lactobacillus rhamnosus.
3. The composition of claim 1, wherein the food-grade or pharmaceutically acceptable excipients are binders, fillers; further, the food grade or pharmaceutically acceptable excipients include, but are not limited to, polyols, starches.
4. The composition of claim 3, wherein the composition comprises the following components in parts by weight:
5. the composition of claim 4, wherein the oligosaccharide is at least one member selected from the group consisting of fructooligosaccharide, isomaltooligosaccharide, xylooligosaccharide, stachyose; the polyhydric alcohol is at least one of lactitol, inositol, galactitol, sorbitol, xylitol, maltitol and mannitol; the starch is selected from at least one of rice starch and wheat starch.
6. The composition as claimed in claim 5, wherein the oligosaccharide comprises 50-200 parts of fructo-oligosaccharide, 150-400 parts of isomalto-oligosaccharide, 1-15 parts of xylo-oligosaccharide and 5-15 parts of stachyose; the polyol is 500 parts of lactitol 250-; the starch is 5-20 parts of corn starch.
7. The composition according to claim 2, wherein the composition comprises, by weight, lactobacillus plantarum LP 4515-45 parts, bifidobacterium lactis BAL 5311-10 parts, bifidobacterium bifidum TMC 31151-10 parts, lactobacillus acidophilus La 281-10 parts, and lactobacillus rhamnosus LR 5191-10 parts.
8. The composition as claimed in claim 7, wherein the composition comprises, by weight, Lactobacillus plantarum LP 4520-40 parts, Bifidobacterium bifidum TMC 31154-8 parts, Bifidobacterium lactis BAL 5314-8 parts, Lactobacillus acidophilus La 281-10 parts, Lactobacillus rhamnosus LR 5191-10 parts, inulin 100-300 parts, fructo-oligosaccharide 50-150 parts, isomalto-oligosaccharide 150-400 parts, xylo-oligosaccharide 1-15 parts, stachyose 5-15 parts, lactitol 250-500 parts, and corn starch 5-15 parts; more preferably, the feed comprises, by weight, 4530 parts of lactobacillus plantarum LP, 5315 parts of bifidobacterium lactis BAL, 31155 parts of bifidobacterium bifidum TMC, 285 parts of lactobacillus acidophilus La, 5194 parts of lactobacillus rhamnosus LR, 200 parts of inulin, 100 parts of fructo-oligosaccharide, 270 parts of isomalto-oligosaccharide, 10 parts of xylo-oligosaccharide, 10 parts of stachyose, 320 parts of lactitol and 10 parts of corn starch.
9. A method of preparing the composition of claim 1, comprising the steps of:
(1) mixing inulin and oligosaccharide, adding food-grade or pharmaceutically-acceptable adjuvants, wet granulating, sterilizing, and sieving to obtain granule;
(2) mixing lactic acid bacteria with the granules obtained in step (1), and preparing according to a conventional process.
10. The composition of claim 9, comprising at least 10 per gram of the composition8To 1012A CFU lactic acid bacterium; further preferably, it comprises at least 10 per g of the composition9To 1011CFU lactic acid bacteria.
11. The composition of claim 10, wherein the composition is formulated as any one of a pill, a tablet, a capsule, a powder, a granule, or an oral liquid.
12. A pharmaceutical, food or health product comprising the composition of any one of claims 1 to 10.
13. Use of a composition according to any one of claims 1 to 10 for the preparation of a medicament, food or health product for regulating intestinal function.
14. Use of a composition according to any one of claims 1 to 10 for the preparation of a medicament, health product or food for assisting the efficacy of hui constipation relieving capsules in expelling toxins, nourishing the skin, relieving constipation and moistening the intestines.
15. A ready-to-eat lactic acid bacterium comprising the composition of any one of claims 1 to 10.
16. A pharmaceutical composition, which comprises the ready-to-eat lactic acid bacteria of claim 15, orlistat, hui tong capsule, fu fang lu shi ye, ma ren shi wan, qing ning wan, tong ling jiao nang, cong tong fu li wan, ma ren zi pi wan, kou tong li ke li, xinli shi stan, amoxicillin, irinotecan, isoniazid, rifampin, rifapentine, fluorouracil antineoplastic; preferably, the pharmaceutical composition comprises any one of the instant lactobacillus and aloe cathartic capsule, compound aloe capsule, desert cistanche cathartic oral liquid, edestan pill, qingning pill, cathartic capsule, desert cistanche cathartic capsule, edestan intestine moistening pill, edestan spleen nourishing pill, constipated catharsis and tongle granule as claimed in claim 15.
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