CN117085042A - Bifidobacterium bifidum BL002 metagen and application thereof - Google Patents
Bifidobacterium bifidum BL002 metagen and application thereof Download PDFInfo
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- CN117085042A CN117085042A CN202311365371.5A CN202311365371A CN117085042A CN 117085042 A CN117085042 A CN 117085042A CN 202311365371 A CN202311365371 A CN 202311365371A CN 117085042 A CN117085042 A CN 117085042A
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- 241000186016 Bifidobacterium bifidum Species 0.000 title claims abstract description 34
- 229940002008 bifidobacterium bifidum Drugs 0.000 title claims abstract description 34
- 239000003814 drug Substances 0.000 claims abstract description 16
- 238000004321 preservation Methods 0.000 claims abstract description 12
- 241000186000 Bifidobacterium Species 0.000 claims abstract description 10
- 238000009629 microbiological culture Methods 0.000 claims abstract description 6
- 230000002040 relaxant effect Effects 0.000 claims abstract description 6
- 108020004465 16S ribosomal RNA Proteins 0.000 claims abstract description 4
- 238000002360 preparation method Methods 0.000 claims description 10
- 230000001580 bacterial effect Effects 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 5
- 239000008187 granular material Substances 0.000 claims description 2
- 206010010774 Constipation Diseases 0.000 abstract description 15
- 229940079593 drug Drugs 0.000 abstract description 7
- 244000005700 microbiome Species 0.000 abstract description 3
- 241000699670 Mus sp. Species 0.000 description 25
- 230000000968 intestinal effect Effects 0.000 description 13
- -1 compound diphenoxylate Chemical class 0.000 description 11
- 229960004192 diphenoxylate Drugs 0.000 description 11
- 230000002496 gastric effect Effects 0.000 description 11
- 241001465754 Metazoa Species 0.000 description 10
- 239000002609 medium Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 210000000813 small intestine Anatomy 0.000 description 8
- 230000008855 peristalsis Effects 0.000 description 7
- 230000013872 defecation Effects 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical class [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 206010016100 Faeces discoloured Diseases 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 244000215068 Acacia senegal Species 0.000 description 3
- 229920000084 Gum arabic Polymers 0.000 description 3
- 239000000205 acacia gum Substances 0.000 description 3
- 235000010489 acacia gum Nutrition 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 235000013325 dietary fiber Nutrition 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 229960001571 loperamide Drugs 0.000 description 3
- RDOIQAHITMMDAJ-UHFFFAOYSA-N loperamide Chemical compound C=1C=CC=CC=1C(C=1C=CC=CC=1)(C(=O)N(C)C)CCN(CC1)CCC1(O)C1=CC=C(Cl)C=C1 RDOIQAHITMMDAJ-UHFFFAOYSA-N 0.000 description 3
- 238000009835 boiling Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 235000012631 food intake Nutrition 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 238000003304 gavage Methods 0.000 description 2
- 210000002429 large intestine Anatomy 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 210000001187 pylorus Anatomy 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 206010053155 Epigastric discomfort Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010054949 Metaplasia Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 208000019790 abdominal distention Diseases 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 210000004534 cecum Anatomy 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 208000010643 digestive system disease Diseases 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- 230000030136 gastric emptying Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 230000003870 intestinal permeability Effects 0.000 description 1
- 239000008141 laxative Substances 0.000 description 1
- 229940125722 laxative agent Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000015689 metaplastic ossification Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 150000004666 short chain fatty acids Chemical class 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 231100000033 toxigenic Toxicity 0.000 description 1
- 230000001551 toxigenic effect Effects 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/10—Laxatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses bifidobacterium bifidum BL002 metagen and application thereof, and relates to the technical field of microorganisms. The invention provides a bifidobacterium bifidum BL002 metagen, which is obtained by processing bifidobacterium bifidum BL002, wherein the bifidobacterium bifidum BL002 is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms, the preservation address is North Silu No.1, 3 in the Korean region of Beijing city, the preservation date is 2021, 10 months and 25 days, the preservation number is CGMCC No.23664, and the classification name is bifidobacterium bifidumBifidobacterium bifidumThe 16S rDNA sequence is shown as SEQ ID NO. 1. The bifidobacterium bifidum BL002 metagen can be used for preparing medicines for relaxing bowel and effectively relieving constipation.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to bifidobacterium bifidum BL002 metagen and application thereof.
Background
Bifidobacteria are a bacterial genus. Bifidobacterium genus ]Bifidobacterium) Is a gram positive, motionless, rod-shaped cell, and branch-shaped cell with one end being strictly anaerobic, and widely exists in the digestive tract, vagina, oral cavity and other habitats of human and animals. Bacteria of the genus bifidobacterium are one of the important constituent members of the intestinal flora of humans and animals. Some strains of bifidobacteria may be used as probiotics in food, pharmaceutical and feed applications.
Constipation is a clinical manifestation of digestive system diseases, and is a group of symptoms of defecation times, difficult or uncomfortable defecation, dry and hard feces, and is common in children and the elderly. The time that the stool remains in the large intestine after constipation occurs is prolonged, and moisture in the stool is then absorbed, resulting in dryness of the stool, thereby inhibiting intestinal peristalsis and causing intestinal imbalance and intestinal pain. At present, the common clinical treatment method of constipation is taking laxatives and prokinetic medicines, but long-term taking of anti-constipation medicines has great side effects, and can cause medicine dependence, and constipation is aggravated after stopping taking the medicines.
The dietary fiber is not digested and absorbed generally after entering a human body, can increase the water content of the excrement and keep the intestinal canal lubricated by absorbing the water, can stimulate the intestinal wall and increase the intestinal peristalsis, can generate short-chain fatty acid through fermentation, change the pH of the intestinal canal, and improve the reproduction environment of beneficial flora, thereby accelerating the intestinal peristalsis and smoothly discharging the excrement. And a large amount of dietary fiber is supplemented from the diet, so that a considerable amount of food containing dietary fiber is needed to be eaten, the gastrointestinal burden of a patient can be increased, and the gastric emptying delay is caused, so that symptoms such as abdominal distention, epigastric discomfort, belch, appetite reduction, dyspepsia and the like are caused. Microbial preparations for improving constipation are limited in the prior art, and the effect of the preparations for improving constipation is still required to be further improved. The invention thus provides bifidobacterium bifidum BL002 metagen and uses thereof.
Disclosure of Invention
The invention aims to provide bifidobacterium bifidum BL002 metagen and application thereof, which can be used for preparing medicines for relaxing bowel and effectively relieving constipation.
In order to achieve the above purpose, the invention provides the application of the bifidobacterium bifidum BL002 metazoan in preparing the medicine for relaxing bowel, the bifidobacterium bifidum BL002 metazoan is obtained by processing the bifidobacterium bifidum BL002, the bifidobacterium bifidum BL002 is preserved in the China general microbiological culture Collection center of the China general microbiological culture Collection center, the preservation address is North Chen West road No.1 in the Korean region of Beijing, the preservation date is 2021, 10 months and 25 days, the preservation number is CGMCC No.23664, and the classification name is bifidobacterium bifidumBifidobacterium bifidumThe 16S rDNA sequence is shown as SEQ ID NO. 1.
Preferably, the method is applied to the preparation of medicaments.
Preferably, the medicament comprises a bacterial powder preparation, a tablet and a granule.
The bifidobacterium bifidum BL002 metagen and the application thereof have the advantages and positive effects that:
1. in the invention, bifidobacterium bifidum BL002 shortens normal transportation time and increases defecation frequency by increasing intestinal peristalsis and softening the consistency of feces, and improves intestinal mucosa.
2. After intestinal tract colonization, bifidobacterium bifidum BL002 of the invention can metabolize to produce organic acid, can reduce the pH value in the intestinal cavity, and can directly or indirectly act on large intestine and other organs by influencing intestinal flora, adjusting intestinal permeability and immunological parameters and producing regulatory or bioactive metabolites, thereby promoting gastric digestion and intestinal peristalsis.
The technical scheme of the invention is further described in detail through examples.
Detailed Description
The technical scheme of the invention is further described below by examples.
Unless defined otherwise, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this invention belongs.
Example 1
Bifidobacterium bifidum BL002, which is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) with a preservation address of 1 # 3 of North Silu, chaoyang district of Beijing, a preservation date of 2021, 10 months and 25 days, a preservation number of 23664 CGMCC No. and classified and named bifidobacterium bifidumBifidobacterium bifidumThe 16S rDNA sequence is shown as SEQ ID NO. 1.
EXAMPLE 2 preparation of Bifidobacterium bifidum BL002 metazoan
S1, preparation of seed liquid
Activating strain BL002 in MRS liquid culture medium, inoculating 5% of the strain into 100mL of MRS liquid culture medium, and performing amplification culture at 40deg.C for 12 hr to obtain Bifidobacterium bifidum BL002 seed solution with viable count of 6X10 9 cfu/mL；
S2, strain fermentation
Inoculating the seed solution into a fermentation culture medium according to 10% of inoculum size, and fermenting at 40 ℃ for 20h to obtain fermentation liquor;
s3, preparation of metazoan
Centrifuging the fermentation liquor at 6000rpm for 30min at normal temperature, discarding the supernatant, taking wet precipitate, adding equal amount of sterilized corn starch, uniformly stirring, sieving with 40 mesh sieve, and vacuumizing at room temperature until the bacterial powder is completely dried to obtain bifidobacterium bifidum BL002 metagen.
Example 3 application of Bifidobacterium bifidum BL002 metagen in relaxing bowel
The model making medicine compound diphenoxylate is administrated by oral lavage, and a small intestine peristalsis inhibition model of the mice is established.
Instrument and reagent
Surgical scissors, ophthalmic forceps, ruler, syringe, balance, activated carbon powder, gum arabic, compound diphenoxylate or loperamide.
Preparing ink: accurately weighing 100g of gum arabic, adding 800mL of water, boiling until the solution is transparent, weighing 50g of activated carbon (powder) and adding into the solution, boiling for three times, cooling the solution, adding water to 1000mL, keeping the volume at 4 ℃ in a refrigerator, and shaking uniformly before use.
Preparing a compound diphenoxylate suspension: the concentration was 0.025%.
The compound diphenoxylate tablet contains 2.5mg of compound diphenoxylate, 25mg (10 tablets) of compound diphenoxylate tablet is taken, ground into powder by a mortar, added with water to 100mL and prepared just before use.
Experimental method
S1, selecting adult male mice, wherein the weight of the adult male mice is 18-22g, and the total number of the adult male mice is 5, and the adult male mice are divided into a control group, a model group, a low-dose group, a medium-dose group and a high-dose group. The normal person (60 kg) eats 6g (total bacterial count 2×10) of Bifidobacterium bifidum BL002 metazoan daily 11 Based on each gram), the food consumption is respectively 5, 10, 20 times of the low, medium and high dose groups, namely the low dose group of the total bacterial count of the bifidobacterium bifidum BL002 is 2 multiplied by 10 9 The dosage group is 4×10 9 Individual/g, high dose group 8X 10 9 The mice had food during the test, and the weight and mass changes before and after each group of mice were recorded, and the dose slightly changed with the changes.
S2, all mice are normally bred for 7 days to adapt to the environment. The test substance is administered by means of gastric lavage, the tube trace is 0.2mL, the gastric lavage is continued for one week, and after the 7 th day of gastric lavage, no water is forbidden after continuous 16h of fasted. The model group, the low dose group, the medium dose group and the high dose group were respectively given compound diphenoxylate (10 mg/kg) by gavage, and the blank group was given the same dose of distilled water. After 30min of gastric lavage, the low, medium and high test groups respectively lavage the ink containing the corresponding test object, and the control group and the model group lavage the same dosage of ink. After the stomach is irrigated for 25min, each group of mice is killed through cervical dislocation, the mice are dissected, the whole section of small intestine from the pylorus to the cecum is cut off and taken out, the total length of the small intestine and the toxigenic from the pylorus to the front of the ink are measured after the small intestine is tiled and lightly pulled to a straight state, and the small intestine propulsion rate of the mice is calculated according to the following formula.
；
The results are shown in Table 1:
TABLE 1
；
Note that: p < 0.05, P < 0.01 compared to control; in contrast to the set of models, # P＜0.05， ## P＜0.01。
as can be seen from Table 1, after the compound diphenoxylate drug is adopted to construct a constipation model of mice, compared with a control group, the ink propulsion length and the propulsion rate of the mice in the model group are obviously reduced, the difference is obvious, and the statistical significance (P is less than 0.01) is achieved, which indicates that after the compound diphenoxylate drug is adopted to perfuse the stomach of the mice in the model group, the continuous model establishment of small intestine peristalsis is successful. Under the condition that the model is established, the ink propulsion length of the mice treated by each dose group of bifidobacterium bifidus BL002 metaplasia is obviously increased, which indicates that the samples of each dose group can obviously accelerate the movement of the small intestine, strengthen the small intestine and improve the ink propulsion rate, and the differences are obvious, thereby having statistical significance (P is less than 0.05 or P is less than 0.01).
Example 4 application of Bifidobacterium bifidum BL002 metagen in relieving constipation
The model making medicine loperamide is administrated by oral gastric lavage to establish a model of constipation of mice.
Instrument and reagent
Ophthalmic forceps, syringe, analytical balance, activated carbon powder, gum arabic, compound diphenoxylate, loperamide.
Preparation of reagent preparation ink: same as in example 3.
Experimental method
S1, selecting adult male mice, wherein the weight of the adult male mice is 18-22g, and the total number of the adult male mice is 5, and the adult male mice are divided into a control group, a model group, a low-dose group, a medium-dose group and a high-dose group. The normal person (60 kg) eats 6g (total bacterial count 2×10) of Bifidobacterium bifidum BL002 metazoan daily 11 Based on each gram), the food consumption is respectively 5, 10, 20 times of the low, medium and high dose groups, namely the low dose group of the total bacterial count of the bifidobacterium bifidum BL002 is 2 multiplied by 10 9 The dosage group is 4×10 9 Individual/g, high dose group 8X 10 9 The mice had food during the test, and the weight and mass changes before and after each group of mice were recorded, and the dose slightly changed with the changes.
S2, all mice are normally bred for 7 days to adapt to the environment. The test substance is administered by means of gastric lavage, the tube trace is 0.2ml, the gastric lavage is continued for one week, and after the 7 th day of gastric lavage, no water is forbidden after continuous 16h of fasted. The model group, the low dose group, the medium dose group and the high dose group were respectively given compound diphenoxylate (10 mg/kg) by gavage, and the blank group was given the same dose of distilled water. After 30min of gastric lavage, the low, medium and high test groups respectively lavage the ink containing the corresponding test object, and the control group and the model group lavage the same dosage of ink.
The first black stool discharge time, the number of black stools discharged within 6 hours, and the weight of each mouse (the first black stool discharge stool should be removed in the statistics) were recorded.
The results are shown in Table 2:
TABLE 2
；
Note that: p < 0.05, P < 0.01 compared to control; in contrast to the set of models, # P＜0.05， ## P＜0.01。
as can be seen from table 2, the time to first discharge the black stool was increased, the time exceeded 100min, and the number of stool particles was reduced, the total mass of the stool was reduced, and the variability was significant (P < 0.01) compared to the control group, indicating that the constipation model mice were successfully established. Under the condition that the model is established, the low, medium and high dose groups of bifidobacterium bifidum BL002 metazoan can obviously shorten the first defecation time of the mice, increase the number of defecation grains and the total mass of excrement in 6h of the mice, and show that the bifidobacterium bifidum BL002 metazoan can play a role in promoting defecation of the constipation model mice.
Therefore, the bifidobacterium bifidum BL002 metagen and the application thereof can be used for preparing the medicine for relaxing bowel and effectively relieving constipation.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention and not for limiting it, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that: the technical scheme of the invention can be modified or replaced by the same, and the modified technical scheme cannot deviate from the spirit and scope of the technical scheme of the invention.
Claims (2)
1. The application of bifidobacterium bifidum BL002 metagen in preparing the medicine for relaxing bowel is characterized in that: the post-metazoan of the bifidobacterium bifidum BL002 is obtained by processing the bifidobacterium bifidum BL002, the bifidobacterium bifidum BL002 is preserved in the China general microbiological culture Collection center with the preservation address of North Chen West Lu No.1 of the Korean area of Beijing, the preservation date is 2021, 10 and 25 days, the preservation number is CGMCC No.23664, and the classification is named as the bifidobacterium bifidumBifidobacterium bifidumThe 16S rDNA sequence is shown as SEQ ID NO. 1.
2. The use according to claim 1, characterized in that: the medicine comprises a bacterial powder preparation, a tablet and a granule.
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