CN114146101B - Application of bifidobacterium animalis subspecies lactis BLa80 in preparation of medicines or foods for regulating intestinal motility - Google Patents
Application of bifidobacterium animalis subspecies lactis BLa80 in preparation of medicines or foods for regulating intestinal motility Download PDFInfo
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- CN114146101B CN114146101B CN202111389232.7A CN202111389232A CN114146101B CN 114146101 B CN114146101 B CN 114146101B CN 202111389232 A CN202111389232 A CN 202111389232A CN 114146101 B CN114146101 B CN 114146101B
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- bla80
- bifidobacterium animalis
- animalis subspecies
- bifidobacterium
- subspecies
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Abstract
The invention provides an application of bifidobacterium animalis subspecies lactis BLa80 in preparing medicines or foods for regulating intestinal motility. The medicine or food for regulating intestinal motility comprises a single microbial agent of bifidobacterium animalis subspecies gla 80 microbial agent. The bifidobacterium animalis subspecies of lactic acid bacteria BLa80 microbial inoculum is a simple bifidobacterium animalis subspecies of lactic acid bacteria BLa80 or a combination of bifidobacterium animalis subspecies of lactic acid bacteria BLa80 and a protective agent, wherein the protective agent comprises an emulsifying agent, polysaccharide and glycerin. The medicine or food for regulating intestinal motility can increase intestinal motility contraction frequency, improve intestinal osmotic pressure, promote water absorption, reduce pH value in intestinal tract, increase content of beneficial bacteria in intestinal tract, and promote intestinal motility recovery.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to application of bifidobacterium animalis subspecies lactis BLa80 in preparation of medicines or foods for regulating intestinal motility.
Background
Constipation, a common chronic gastrointestinal disorder, is often manifested by a reduced number of bowel movements and difficulty in bowel movements, has become one of the important factors affecting the quality of life of modern people. Constipation is classified into organic constipation and functional constipation, and functional constipation refers to chronic constipation without structural abnormality or metabolic disorder due to lack of organic etiology, except for irritable bowel syndrome, mainly due to intestinal dysfunction. In daily life, constipation is caused by many factors, such as age, living habit, mental condition or intestinal tract disease. The research shows that the incidence rate of adult chronic constipation is 4-6%, the incidence rate of constipation in the aged population can reach 15-20%, and the incidence rate tends to increase year by year.
Currently, the classes of drugs for treating constipation are bulk laxatives, osmotic laxatives, and stimulant laxatives, which are effective in relieving constipation. CN105435231a discloses a composition for treating constipation, which contains an oligosaccharide-based bifidus factor and an osmotic purgative agent, the content of the osmotic purgative agent being 0.5-10 parts by weight relative to 1 part by weight of the oligosaccharide-based bifidus factor. The composition can achieve a significantly enhanced effect of treating constipation relative to a single xylooligosaccharide or osmotic laxative. However, osmotic purgatives are not suitable for long-term administration and tend to cause adverse reactions.
CN108030847a discloses a pharmaceutical composition for external use for treating constipation, which comprises: almond oil, honey, hemp seed oil, sesame oil, betel nut oil and longan juice. The pharmaceutical composition has no toxic or side effect, has good curative effect on constipation, can obviously improve symptoms such as abdominal distension and constipation of constipation patients, is suitable for long-term use, is a safe and effective medicament for treating constipation, and has good application prospect. However, the pharmaceutical composition comprises a large variety of raw materials and has high economic cost.
CN109965280a discloses an intestinal microecological preparation for regulating constipation in children and adolescents, said intestinal microecological preparation comprising: hawthorn fruit, orange peel, malt, lutein, cassia seed, inulin, kudzuvine root, turmeric and probiotics; the viable count of the probiotics is 10 8 -10 10 cfu/g. The intestinal microecological preparation can effectively improve the regulating effect of the intestinal microecological preparation for constipation of children and teenagers by controlling the mass fraction and the viable count of probiotics.
Based on the above study, it can be seen that the probiotics as microorganisms have the functions of enhancing immunity and regulating intestinal tracts, and compared with the traditional laxative, the probiotics has less side effects and obvious constipation relieving effect. Therefore, further searching for strains that can relieve constipation is a hotspot and an important point of current research.
Disclosure of Invention
Aiming at the defects and the actual requirements of the prior art, the invention aims to provide the application of bifidobacterium animalis subspecies of milk Bla80 in preparing medicines or foods for regulating intestinal motility. The bifidobacterium animalis subspecies of the Lactobacillus Bla80 has the effects of regulating intestinal tract, promoting the growth of acidic bacteria in the intestinal tract, increasing the content of beneficial bacteria in the intestinal tract, and can be used for preparing medicines or foods for regulating intestinal motility.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
in a first aspect, the invention provides the use of bifidobacterium animalis subspecies lactis BLa80 in the manufacture of a medicament or foodstuff for modulating intestinal motility.
The bifidobacterium animalis subspecies of the invention BLa80 can significantly improve the content level of short-chain fatty acids in individuals, the short-chain fatty acids can stimulate intestinal wall peristalsis, increase intestinal peristalsis contraction frequency, improve intestinal osmotic pressure and promote water absorption, the short-chain fatty acids can also reduce the pH value in the intestinal tract, further promote the growth of acidic bacteria such as lactobacillus or bifidobacterium in the intestinal tract and further increase the content of beneficial bacteria in the intestinal tract. Meanwhile, the bifidobacterium animalis subspecies of milk Bla80 can promote the recovery of individual small intestine power and improve the small intestine propulsion rate. The bifidobacterium animalis subspecies of milk Bla80 can be used for preparing medicines or foods for regulating intestinal power, so that the intestinal power of an individual is ensured to be maintained at a normal level, and constipation can be effectively relieved.
In a second aspect, the invention provides a medicament for modulating intestinal motility comprising a single bacterial agent of bifidobacterium animalis subspecies gla 80 bacterial agent.
The bifidobacterium animalis subspecies (Bifidobacterium animalis subsp.lacti) Bla80 is preserved in China general microbiological culture Collection center (CGMCC), the preservation time is 3 months and 5 days in 2018, the preservation number is CGMCC No.15410, and the addresses are: no.1 and No. 3 of the north cinquefoil of the morning sun area of beijing city.
In the invention, the bifidobacterium animalis subspecies of the Bla80 microbial inoculum is a simple bifidobacterium animalis subspecies of the Bla80 or a combination of the bifidobacterium animalis subspecies of the BLa80 and a protective agent.
Preferably, the protective agent includes an emulsifier, a polysaccharide, and glycerin.
Preferably, the emulsifier comprises skim milk.
Preferably, the polysaccharide comprises trehalose and/or sucrose.
In the present invention, the dosage form of the medicine for regulating intestinal motility includes any one of suppository, powder, capsule, powder, solution, emulsion, enema, gel or tablet.
In the present invention, the medicine for regulating intestinal motility further includes pharmaceutically acceptable auxiliary materials, where the auxiliary materials include any one or a combination of at least two of a carrier, a diluent, an excipient, a filler, an adhesive, a wetting agent, a disintegrating agent, an emulsifying agent, a cosolvent, a solubilizer, an osmotic pressure regulator, a surfactant, a coating material, a colorant, a pH regulator, an antioxidant, a bacteriostatic agent or a buffer, and the combination may be, for example, a combination of a carrier and a diluent or a combination of a diluent and an excipient, and any other combination modes may be selected and will not be repeated here.
In the invention, the viable count of the bifidobacterium animalis subspecies lactis BLa80 microbial inoculum is 1 multiplied by 10 9 cfu/mL, cfu is colony forming unit (colony forming units).
In a third aspect, the present invention provides a food for modulating intestinal motility comprising a single bacterial agent of the bifidobacterium animalis subspecies gla 80 bacterial agent.
The bifidobacterium animalis subspecies (Bifidobacterium animalis subsp.lacti) Bla80 is preserved in China general microbiological culture Collection center (CGMCC), the preservation time is 3 months and 5 days in 2018, the preservation number is CGMCC No.15410, and the addresses are: no.1 and No. 3 of the north cinquefoil of the morning sun area of beijing city.
In the invention, the bifidobacterium animalis subspecies of the Bla80 microbial inoculum is a simple bifidobacterium animalis subspecies of the Bla80 or a combination of the bifidobacterium animalis subspecies of the BLa80 and a protective agent.
Preferably, the protective agent includes an emulsifier, a polysaccharide, and glycerin.
Preferably, the emulsifier comprises skim milk.
Preferably, the polysaccharide comprises trehalose and/or sucrose.
In the invention, the food for regulating intestinal motility comprises any one of solid beverage, fermented milk or fermented fruits and vegetables.
Compared with the prior art, the invention has the following beneficial effects:
according to the invention, the bifidobacterium animalis subspecies BLa80 is adopted to prepare the medicine or the food for regulating the intestinal motility, and the bifidobacterium animalis subspecies BLa80 can obviously improve the content level of short chain fatty acids in an individual, so that the peristalsis of intestinal walls is stimulated, the peristaltic contraction frequency of the intestinal tracts is increased, the osmotic pressure in the intestinal tracts is improved, the water absorption is promoted, the pH value in the intestinal tracts is reduced, the growth of acidic bacteria such as lactobacillus or bifidobacterium in the intestinal tracts is promoted, and the content of beneficial bacteria in the intestinal tracts is further increased. Meanwhile, the bifidobacterium animalis subspecies of milk Bla80 can promote the recovery of individual small intestine power and improve the small intestine propulsion rate.
Drawings
FIG. 1 is a graph showing the results of first example black discharge time of mice of different groups according to the present invention;
wherein, the statistical value P between a and b is less than 0.01, the statistical value P between a and c is less than 0.01, the statistical value P between b and c is less than 0.05, P is less than 0.05 and represents the difference is significant, and P is less than 0.01 and represents the difference is extremely significant;
FIG. 2 is a graph showing the results of the fecal moisture content of mice of different groups according to the present invention;
wherein, the statistical value P <0.01 between a and b, P <0.01 indicates that the difference is very significant;
FIG. 3 is a graph showing the results of total short chain fatty acid content of the feces of different groups of mice according to the present invention;
wherein statistics between a and b are P <0.01, statistics between a and c are P <0.01, statistics between a and d are P <0.01, statistics between b and c are P <0.01, statistics between b and d are P <0.01, statistics between c and d are P <0.01, P <0.05 indicates that the difference is significant, and P <0.01 indicates that the difference is extremely significant;
FIG. 4 is a graph showing the results of intestinal tract propulsion rates of different groups of mice according to the present invention;
wherein, the statistical value P <0.01 between a and b, the statistical value P <0.01 between a and c, the statistical value P <0.05 between b and c, P <0.05 indicates that the difference is significant, and P <0.01 indicates that the difference is extremely significant.
Detailed Description
The technical scheme of the invention is further described by the following specific embodiments. It will be apparent to those skilled in the art that the examples are merely to aid in understanding the invention and are not to be construed as a specific limitation thereof.
The sources of the corresponding materials and raw materials in the following examples are as follows:
wherein SPF-grade BALB/c mice were purchased from Shanghai Laike laboratory animals Co., ltd; gum arabic is purchased from national pharmaceutical group chemical company; compound diphenoxylate is purchased from Jiangsu Huayang pharmaceutical Co., ltd (national medicine standard code H32020933); the bifidobacterium animalis subspecies (Bifidobacterium animalis subsp. Lactis) BLa80 is a strain preserved in China general microbiological culture Collection center (CGMCC) with the preservation time of 2018, 3 months and 5 days and the preservation number of CGMCC No. 15410. The other materials and raw materials, without specific description, are all commercially available.
Preparation example 1
The preparation example provides a bifidobacterium animalis subspecies lactis Bla80 bacterial suspension, and the preparation method comprises the following steps:
inoculating single colony of Lactobacillus bifidus subspecies BLa80 of animal with rejuvenation on plate into 10mL of TPY liquid culture medium, anaerobically culturing at 37deg.C for 12h, inoculating activated bacterial liquid into 100mL of TPY liquid culture medium with 2% inoculum size, anaerobically culturing at 37deg.C for 12h, centrifuging at 4deg.C for 10min at 5000g, collecting centrifuged bacterial precipitate, re-suspending with sterile PBS buffer solution, and adjusting bacterial suspension concentration to 10 9 cfu/mL, obtaining the bifidobacterium animalis subspecies lactis BLa80 bacterial suspension, and preserving at 4 ℃ for later use.
Example 1
This example explores the effect of different doses of bifidobacterium animalis subspecies gla 80 on the time to black bowel movement in mice. 50 healthy SPF-grade BALB/c mice were selected, weighing 18-20g, and kept in animal houses according with animal test protocol: after maintaining room temperature at 25 ℃, relative humidity at 55%,12h light, 12h darkness, free feeding and drinking water, all mice were treated as follows after 7 days of adaptive feeding observations without abnormalities:
(1) Grouping into 5 groups: blank, model, low dose bifidobacterium animalis subspecies gla 80 intervention group, medium dose bifidobacterium animalis subspecies glabra 80 intervention group and high dose bifidobacterium animalis subspecies glabra 80 intervention group, 10 each, wherein the blank is filled with sterile PBS buffer solution, the model is filled with sterile PBS buffer solution, and the low dose bifidobacterium animalis subspecies glabra 80 intervention group is filled with 1 x 10 per kg body weight of mice 7 cfu of Bifidobacterium animalis subspecies BLa80 bacterial suspension, medium dose Bifidobacterium animalis subspecies BLa80 intervention group 1X 10 stomach lavage per kg body weight mice 8 Suspension of cfu Bifidobacterium animalis subspecies BLa80 bacteria, 1X 10 stomach lavage per kg body weight of mice in high dose Bifidobacterium animalis subspecies BLa80 intervention group 9 The cfu animal bifidobacterium lactis subspecies BLa80 bacterial suspension is subjected to gastric lavage once every 9 points a day for 14 days.
(2) The method for establishing the constipation mouse model comprises the following steps: the method comprises the steps of lavaging a mouse with 10mg of compound diphenoxylate per kilogram of body weight, inducing constipation of the mouse, and exploring the influence of different doses of bifidobacterium animalis subspecies BLa80 on the time of first grain urination and defecation of the mouse based on a constructed constipation mouse model. The specific experimental method is as follows:
preparing ink: weighing 100g of gum arabic, adding 800mL of water, boiling until the solution is transparent, weighing 50g of active carbon, adding into the solution, boiling for 3 times, cooling, adding water to 1000mL, preserving in a refrigerator at 4 ℃, and shaking uniformly before use;
experiment: after 14 days of gastric lavage, each group of mice was fasted without water withdrawal overnight. On day 15 blank group was gavaged with 0.9% NaCl solution, model group, low, medium and high dose animal bifidobacterium subspecies of Lactobacillus 80 intervention group were respectively gavaged with compound diphenoxylate (per kg body weight)Mice were perfused with 10mg of Compound diphenoxylate) and after 0.5h of Compound diphenoxylate, the mice of the blank and model groups were perfused with 0.1mL/kg of gastric ink, and the low, medium and high doses of Bifidobacterium animalis subspecies BLa80 were used to intervene in groups with Bifidobacterium animalis subspecies BLa80 of 1X 10 per kg body weight of mice 8 cfu、1×10 9 cfu and 1×10 10 The amount of cfu was 0.1mL/kg of gastric juice. Starting from the stomach-filling ink, the first black stool time was recorded for each mouse and statistically analyzed.
As shown in fig. 1, the mice had a reduced first grain urination and defecation time compared to the model group for the different doses of bifidobacterium lactis BLa 80-infused animals. Statistical analysis shows that the black stool discharge time of the first example of mice in the blank group and the model group is extremely remarkably different (P < 0.01), the black stool discharge time of the first example of mice in the blank group and the low-dose animal bifidobacterium lactis type BLa80 intervention group is remarkably different (P < 0.05), and the black stool discharge time of the first example of mice in the blank group and the medium-dose and high-dose animal bifidobacterium lactis type BLa80 intervention group is not remarkably different. The model group has a significant difference (P < 0.05) in the first black stool discharge time of mice in the low-dose animal bifidobacterium lactis sub-species BLa80 intervention group and has a significant difference (P < 0.01) in the first black stool discharge time of mice in the medium-and high-dose animal bifidobacterium lactis sub-species BLa80 intervention group. For different doses of bifidobacterium animalis subspecies BLa80 intervention groups, the low dose of bifidobacterium animalis subspecies BLa80 intervention groups are quite obvious in the first black stool discharge time difference (P < 0.01) of mice in the middle and high dose of bifidobacterium animalis subspecies BLa80 intervention groups, and the first black stool discharge time difference of mice in the middle and high dose of bifidobacterium animalis subspecies BLa80 intervention groups is not obvious, which indicates that the bifidobacterium animalis subspecies BLa80 has a dose effect in reducing the first grain black stool discharge time of the mice.
Example 2
This example refers to the mouse model constructed in example 1 and explores the effect of different doses of bifidobacterium animalis subspecies gla 80 on the moisture content of the faeces of mice. The specific method comprises the following steps:
using the mice obtained in example 1, on days 16-20, the blank was filled with 0.9% NaCl solution, model groupGastric lavage of Compound diphenoxylate (10 mg of Compound diphenoxylate per kg body weight of mice), 1×10 of Compound diphenoxylate was respectively gastric lavaged per kg body weight of mice in low, medium and high dose bifidobacterium lactis subspecies Bla80 intervention group 8 cfu、1×10 9 cfu and 1×10 10 After the 19 th day of stomach irrigation, a single mouse is put into a cage box filled with water absorbing paper, feces are collected, wet weight is weighed, the animal bifidobacterium lactis subspecies Bla80 bacterial suspension of cfu is divided into two parts, dry weight is recorded after one part is dried, and the water content of the feces is calculated according to the following formula:
moisture content in feces= (wet fecal weight-dry fecal weight)/wet fecal weight x 100%.
As shown in fig. 2, the number of the reduction of the fecal moisture content of the mice in the model group is larger than that of the blank group, and the fecal moisture content of the mice in the intervention group of bifidobacterium lactis BLa80 in different gastric lavage doses is improved. In combination with statistical analysis, there was a very significant difference (P < 0.01) between the blank and model groups, and between the blank and low and medium dose bifidobacterium lactis subspecies BLa80 intervention groups (P < 0.01), there was no significant difference between the blank and medium dose bifidobacterium lactis subspecies BLa80 intervention groups. The model group has no obvious difference from the low and medium dose animal bifidobacterium lactis subspecies BLa80 intervention group, and has extremely obvious difference (P < 0.01) from the high dose animal bifidobacterium lactis subspecies BLa80 intervention group. For the different doses of bifidobacterium animalis subspecies BLa80 intervention groups, there was a very significant difference (P < 0.01) between the low and medium doses of bifidobacterium animalis subspecies BLa80 intervention groups and the high doses of bifidobacterium animalis subspecies BLa80 intervention groups, and the high doses of bifidobacterium animalis subspecies BLa80 intervention groups restored the mouse fecal moisture content to normal levels, indicating that there was a dose effect of bifidobacterium animalis subspecies BLa80 on improving constipation mouse fecal moisture content.
Example 3
This example refers to the mouse model constructed in example 1 and explores the effect of different doses of bifidobacterium animalis subspecies gla 80 on total short chain fatty acid content in mouse faeces. The specific method for detection is as follows:
another stool fraction collected in example 2 was used for determination of the content of SCFAs, and the contents of acetic acid, propionic acid, n-butyric acid and isobutyric acid were determined by gas chromatography-mass spectrometry (Gas Chromatography-Mass Spectrometry, GC-MS), respectively, followed by calculation of the sum to obtain the total fatty acid content in the mouse stool.
As shown in fig. 3, compared with the blank group, the total fatty acid content in the mouse feces of the model group is obviously reduced, and the total fatty acid content in the mouse feces of the animal bifidobacterium lactis sub-species BLa80 intervention group with different intragastric administration doses is obviously improved. In combination with statistical analysis, there was a very significant difference between the blank and model groups (P < 0.01), and between the blank and low, medium and high dose bifidobacterium lactis subspecies BLa80 intervention groups (P < 0.01). The model group was very significantly different from the low, medium and high dose animal bifidobacterium lactis BLa80 intervention group (P < 0.01). For the low, medium and high dose animal bifidobacterium lactis subspecies BLa80 intervention groups, there was a very significant difference (P < 0.01) between the medium dose animal bifidobacterium lactis subspecies BLa80 intervention group and the low and high dose animal bifidobacterium lactis subspecies BLa80 intervention group, and there was no significant difference between the low dose animal bifidobacterium lactis subspecies BLa80 intervention group and the high dose animal bifidobacterium lactis subspecies BLa80 intervention group, indicating that animal bifidobacterium subspecies BLa80 has a dose effect on total fatty acid content in constipation-improving mouse faeces.
Example 4
This example refers to the mouse model constructed in example 1 and explores the effect of different doses of bifidobacterium animalis subspecies gla 80 on intestinal thrust in mice. The specific method comprises the following steps:
after day 20 of gastric lavage of each group of mice in example 2, each group of mice was fasted without water withdrawal overnight. The weights of the mice in each group were weighed and recorded and the average value was calculated on day 21, morning. Subsequently, the blank and model groups were filled with 0.9% NaCl solution, and the model group and low, medium and high dose animals were filled with 0.1mL/kg of the ink prepared in example 2, and the low, medium and high dose animals were filled with the bifidobacterium milk subspecies BLa80 intervention group filled with Compound diphenoxylate (10 mg of Compound diphenoxylate per kg of body weight of mice) for 30minBifidobacterium lactosub-species BLa80 mice per kg body weight in the intervention group were treated with the animal Bifidobacterium lactosub-species BLa80 at 1X 10, respectively 8 cfu、1×10 9 cfu and 1×10 10 The amount of cfu was 0.1mL/kg of gastric juice. Immediately after 30min, mice were sacrificed, blood from the mice was collected and added with anticoagulant and preserved at 4 ℃ for later use, then immediately the abdominal cavity was opened to separate the mesentery, the intestinal tube from the lower end of the pylorus to the tail of the cecum was cut, placed on a tray, the small intestine was gently pulled into a straight line, the length of the intestinal tube was measured, defined as the "total length of the small intestine", the length of the pylorus to the front of the ink was measured, and the length of the ink propulsion was defined as the "ink propulsion length". The intestinal tract thrust rate of the mice was calculated as follows:
small intestine propulsion = ink propulsion length/small intestine total length x 100%.
As shown in fig. 4, compared with the blank group, the small intestine propulsion rate of the model mice is obviously reduced, and the intestinal propulsion rate of the mice in the intervention group of bifidobacterium lactis BLa80 in different intragastric doses is improved. In combination with statistical analysis, there was a very significant difference (P < 0.01) between the blank and model groups, the blank and low and medium dose bifidobacterium lactis subspecies BLa80 intervention groups were very significant differences (P < 0.01), but there was no significant difference between the blank and high dose bifidobacterium lactis subspecies BLa80 intervention groups. There was no significant difference between the model group and the low dose bifidobacterium animalis subspecies BLa80 intervention group, there was a significant difference between the medium dose bifidobacterium animalis subspecies BLa80 intervention group (P < 0.05), and there was a very significant difference between the high dose bifidobacterium animalis subspecies BLa80 intervention group (P < 0.01). For the low, medium and high dose bifidobacterium animalis subspecies gla 80 intervention groups, there was a significant difference (P < 0.05) between the low dose bifidobacterium animalis subspecies gla 80 intervention group and the medium dose bifidobacterium animalis subspecies gla 80 intervention group, there was a very significant difference (P < 0.01) between the low dose bifidobacterium animalis subspecies glabra 80 intervention group and the high dose bifidobacterium animalis subspecies glabra 80 intervention group, and there was a very significant difference (P < 0.01) between the medium dose bifidobacterium glabra 80 intervention group and the high dose bifidobacterium glabra 80 intervention group, indicating that bifidobacterium animalis gla 80 has a dose effect in improving constipation in mice intestinal thrust.
Example 5
This example explores the effect of bifidobacterium animalis subspecies gla 80 on the treatment of functional constipation groups.
The specific method comprises the following steps:
100 patients who were treated in hospitals and confirmed to be functional constipation were selected as study subjects, and the patients were asked in advance for the weekly defecation frequency and somatosensory score, and the feces of the patients were collected and classified according to bristol stool classification. Under the condition that the patient is fully voluntary, the patient is tested and tested for 4 weeks of feeding test of the bifidobacterium animalis subspecies BLa80 powder, and the dosage of the bifidobacterium animalis subspecies BLa80 powder is 2.0x10 10 cfu/d, the preparation method of the bifidobacterium animalis subspecies lactis BLa80 bacterial powder comprises the following steps:
inoculating the Bifidobacterium animalis subspecies BLa80 bacterial suspension obtained in preparation example 1 into 10mL of TPY liquid culture medium, and culturing at 37 ℃ for 12h to obtain first-stage seed liquid; inoculating the first-level seed liquid into 10mL of TPY liquid culture medium according to the inoculum size of 2% (v/v), and culturing for 12h at 37 ℃ to obtain a second-level seed liquid; inoculating the secondary seed solution into 10mL of TPY liquid culture medium according to the inoculum size of 1% (v/v), culturing at 37 ℃ for 12h to obtain bacterial solution, centrifuging 6000g of bacterial solution for 15min, and collecting precipitate; washing the precipitate with PBS buffer solution with pH of 7.4 twice, and centrifuging 6000g for 10min again to obtain thallus; the strain was resuspended to a cell concentration of 1X 10 with a protectant solution comprising 80g/L skim milk, 20g/L trehalose, 2 g/L sucrose and 10g/L glycerol 10 cfu/mL to obtain BLa80 emulsion; and freeze-drying the emulsion to obtain the bifidobacterium animalis subspecies BLa80 bacterial powder.
During the test, the defecation frequency and the somatosensory score of the tested patient are recorded once a week, the patient's faeces are collected after the test and classified according to the bristo stool classification method, the bristo stool classification method refers to table 1, and the somatosensory score is selected from part of indexes in the chronic constipation severity score scale.
TABLE 1
The chronic constipation severity score scale is shown in table 2 below:
TABLE 2
After the 31 null patients were removed, the frequency of bowel movement, bristo stool typing, and bowel movement somatosensory scores were statistically analyzed for the remaining 69 patients before and after intervention, as shown in table 3 below (wherein the different letters on the right-hand top of the data in the table indicate significant differences between the groups, and the same letters indicate no significant differences):
TABLE 3 Table 3
| Group of | Average defecation frequency | Bristo stool type | Stool feeling score |
| Before taking the powder | 3.42±0.89 a | 1.86±0.35 a | 7.23±0.86 a |
| After taking the medicine powder | 5.54±1.36 b | 3.47±0.82 b | 5.12±0.44 b |
From the above table data, the average defecation frequency, bristo stool type and defecation somatosensory score of the patients all have significant differences (P < 0.05) before and after taking the medicine powder, the average defecation frequency of the patients is significantly improved after taking the medicine powder, the bristo stool type is recovered to be normal from constipation, and the defecation somatosensory is significantly improved, which indicates that the bifidobacterium lactis sub-species BLa80 of animals can effectively relieve constipation symptoms.
In summary, the invention adopts the bifidobacterium animalis subspecies of the Lactobacillus Bla80 to prepare the medicine or the food for regulating the intestinal motility, and the bifidobacterium animalis subspecies of the Lactobacillus Bla80 can obviously improve the content level of short-chain fatty acid in an individual, stimulate the peristalsis of intestinal walls, increase the peristaltic contraction frequency of intestinal tracts, improve the osmotic pressure in intestinal tracts, reduce the pH value in intestinal tracts and increase the content of beneficial bacteria in intestinal tracts. Meanwhile, the bifidobacterium animalis subspecies of milk Bla80 can promote the recovery of individual small intestine power, improve the small intestine propulsion rate and effectively relieve constipation.
The applicant declares that the above is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be apparent to those skilled in the art that any changes or substitutions that are easily conceivable within the technical scope of the present invention disclosed by the present invention fall within the scope of the present invention and the disclosure.
Claims (4)
1. Bifidobacterium animalis subspecies lactisBifidobacterium animalis subsp. lactis) Use of Bla80 as single active ingredient for the preparation of a medicament for the treatment of constipation, characterized in that said bifidobacterium animalis subspecies lactis（Bifidobacterium animalis subsp. lactis) BLa80 is preserved in China general microbiological culture Collection center (CGMCC), the preservation time is 2018, 3 months and 5 days, the preservation number is CGMCC No.15410, and the address is: beijing, chaoyang area, north Chenxi Lu No.1, 3;
the medicament further comprises a protective agent, wherein the protective agent comprises an emulsifying agent, polysaccharide and glycerin;
the emulsifier comprises skim milk.
2. Use according to claim 1, wherein the polysaccharide comprises trehalose and/or sucrose.
3. The use according to claim 1, wherein the pharmaceutical dosage form comprises any one of suppositories, powders, capsules, powders, solutions, emulsions, enemas, gels or tablets.
4. The use according to claim 1, wherein the medicament further comprises pharmaceutically acceptable excipients comprising any one or a combination of at least two of diluents, fillers, binders, wetting agents, disintegrants, emulsifiers, co-solvents, solubilisers, tonicity adjusting agents, surfactants, coating materials, colorants, pH adjusting agents, antioxidants, bacteriostats or buffers.
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| CN114480229B (en) * | 2022-04-15 | 2022-08-09 | 微康益生菌(苏州)股份有限公司 | Bifidobacterium animalis subsp lactis strain WKB148 and product and application thereof |
| CN115747121A (en) * | 2022-12-12 | 2023-03-07 | 威凯海思(山东)生物工程有限公司 | Bifidobacterium lactis HSLA-009, microbial inoculum and preparation method and application thereof |
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