CN112618580A - Lactobacillus rhamnosus Probio-M9 and application of product thereof in preparing medicament for treating autism - Google Patents
Lactobacillus rhamnosus Probio-M9 and application of product thereof in preparing medicament for treating autism Download PDFInfo
- Publication number
- CN112618580A CN112618580A CN202011600715.2A CN202011600715A CN112618580A CN 112618580 A CN112618580 A CN 112618580A CN 202011600715 A CN202011600715 A CN 202011600715A CN 112618580 A CN112618580 A CN 112618580A
- Authority
- CN
- China
- Prior art keywords
- lactobacillus rhamnosus
- rhamnosus probio
- probio
- btbr
- mice
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000218588 Lactobacillus rhamnosus Species 0.000 title claims abstract description 83
- 206010003805 Autism Diseases 0.000 title claims abstract description 19
- 208000020706 Autistic disease Diseases 0.000 title claims abstract description 19
- 239000003814 drug Substances 0.000 title claims abstract description 17
- 230000011273 social behavior Effects 0.000 claims abstract description 10
- 208000029560 autism spectrum disease Diseases 0.000 claims description 20
- 238000002360 preparation method Methods 0.000 claims description 10
- 230000005856 abnormality Effects 0.000 claims description 6
- 239000008187 granular material Substances 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 6
- 239000006041 probiotic Substances 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 5
- 235000018291 probiotics Nutrition 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 4
- 230000000529 probiotic effect Effects 0.000 claims description 4
- 239000002775 capsule Substances 0.000 claims description 3
- 239000002552 dosage form Substances 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 239000006187 pill Substances 0.000 claims description 3
- 235000013361 beverage Nutrition 0.000 claims description 2
- 235000009508 confectionery Nutrition 0.000 claims description 2
- 230000036541 health Effects 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 4
- 239000002671 adjuvant Substances 0.000 claims 2
- 230000001580 bacterial effect Effects 0.000 claims 1
- 235000010855 food raising agent Nutrition 0.000 claims 1
- 239000000546 pharmaceutical excipient Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 64
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 8
- 230000002159 abnormal effect Effects 0.000 abstract description 5
- 238000010171 animal model Methods 0.000 abstract description 5
- 230000006872 improvement Effects 0.000 abstract description 5
- 230000006400 anxiety behaviour Effects 0.000 abstract description 4
- 238000011160 research Methods 0.000 abstract description 4
- 235000013305 food Nutrition 0.000 abstract description 2
- 229960000074 biopharmaceutical Drugs 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 69
- 241000699666 Mus <mouse, genus> Species 0.000 description 35
- 238000012360 testing method Methods 0.000 description 32
- 238000003556 assay Methods 0.000 description 12
- 238000012346 open field test Methods 0.000 description 9
- 206010020112 Hirsutism Diseases 0.000 description 8
- 230000006399 behavior Effects 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 238000009960 carding Methods 0.000 description 6
- 230000000968 intestinal effect Effects 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 4
- 230000006978 adaptation Effects 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 238000010998 test method Methods 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 208000035475 disorder Diseases 0.000 description 3
- 230000002496 gastric effect Effects 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 230000003997 social interaction Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 239000003826 tablet Substances 0.000 description 3
- 208000019901 Anxiety disease Diseases 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 230000036506 anxiety Effects 0.000 description 2
- 239000012752 auxiliary agent Substances 0.000 description 2
- 230000003542 behavioural effect Effects 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 238000010411 cooking Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 230000003252 repetitive effect Effects 0.000 description 2
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010058314 Dysplasia Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010022998 Irritability Diseases 0.000 description 1
- 208000019022 Mood disease Diseases 0.000 description 1
- 208000029726 Neurodevelopmental disease Diseases 0.000 description 1
- 206010041243 Social avoidant behaviour Diseases 0.000 description 1
- 206010042008 Stereotypy Diseases 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- 208000013404 behavioral symptom Diseases 0.000 description 1
- 238000009227 behaviour therapy Methods 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000004641 brain development Effects 0.000 description 1
- 230000001149 cognitive effect Effects 0.000 description 1
- 230000003931 cognitive performance Effects 0.000 description 1
- 208000030251 communication disease Diseases 0.000 description 1
- 231100000867 compulsive behavior Toxicity 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 238000013480 data collection Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000000358 effect on autism Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 208000021302 gastroesophageal reflux disease Diseases 0.000 description 1
- 238000002575 gastroscopy Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 210000004392 genitalia Anatomy 0.000 description 1
- 210000004251 human milk Anatomy 0.000 description 1
- 235000020256 human milk Nutrition 0.000 description 1
- 208000013403 hyperactivity Diseases 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 210000005027 intestinal barrier Anatomy 0.000 description 1
- 230000007358 intestinal barrier function Effects 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 208000019116 sleep disease Diseases 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Images
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
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G3/00—Sweetmeats; Confectionery; Marzipan; Coated or filled products
- A23G3/34—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
- A23G3/36—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds
- A23G3/364—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
- A23G3/366—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
Abstract
The application provides Lactobacillus rhamnosus Probio-M9 and application of a product thereof in preparing a medicine for treating autism, and the application shows that the autism research using a mouse as an animal model shows that the strain has an improvement effect on abnormal social behaviors and preference disorder of novelty things of a BTBR mouse, can effectively relieve anxiety behaviors of the BTBR mouse, has common food attributes and biopharmaceutical strain potential, provides a new strategy and rational application basis for relieving and treating autism, is relatively low in cost, and has wide application prospect and popularization value.
Description
Technical Field
The application belongs to the technical field of biology, and particularly relates to lactobacillus rhamnosus Probio-M9 and application of a product thereof in preparing a medicament for treating autism.
Background
Autism, the disease name Autism Spectrum Disorder (ASD), is a neurodevelopmental disorder, which originates in early childhood, and has a tendency of a global sharply rising incidence in recent years, mainly due to social interaction communication disorder and repetitive stereotypy and interest, but the etiology and pathogenesis of the disease are not clear yet. Research shows that ASD has genetic basis and can be induced to cause diseases by some factors, the factors which can induce diseases comprise maternal immunity factors, autoimmune disorder factors, natural environment factors and the like, and the diseased individuals often have abnormal brain development. However, the etiology and pathogenesis of ASD are not completely clear.
Most ASD patients suffer from other mental and physical ailments such as: hyperactivity disorders, mood disorders, sleep problems, digestive tract dysfunction, immune problems, and nutritional and metabolic problems, among others. Studies have shown that the digestive tract problems of ASD patients are significant, some ASD patients have significant manifestations, such as constipation, acid reflux or diarrhea, and some ASD patients have no significant symptoms, but when they are examined by gastroscopy or enteroscopy, they find that the digestive tract is abnormal.
Autistic patients with gastrointestinal problems exhibit more severe symptoms of irritability, anxiety, and social withdrawal than autistic patients without gastrointestinal problems. Through genome detection, the composition of intestinal flora, including the type and the number of flora, of the ASD population is greatly different from that of the healthy population.
Disclosure of Invention
One of the purposes of the application is to provide the application of lactobacillus rhamnosus Probio-M9 for preparing the medicine for treating autism, wherein the microorganism preservation number of the lactobacillus rhamnosus Probio-M9 is CGMCC No. 18639.
The lactobacillus rhamnosus Probio-M9 is separated from breast milk of healthy women, the microorganism preservation number of the lactobacillus rhamnosus Probio-M9 is CGMCC No.18639, and the classification name is as follows: lactobacillus rhamnosus (Lactobacillus rhamnosus); the preservation unit is China general microbiological culture Collection center; the preservation time is 2019, 10 and 8 days; and (4) storage address: western road No.1 institute 3, institute of microbiology, china academy of sciences, north chen, chaoyang, china.
Research shows that the lactobacillus rhamnosus Probio-M9 has excellent gastrointestinal fluid tolerance and good bile salt tolerance and has basic conditions as probiotics.
The application also aims to provide a product for treating autism, wherein the product comprises lactobacillus rhamnosus Probio-M9, and the microorganism preservation number of the lactobacillus rhamnosus Probio-M9 is CGMCC No. 18639.
In an achievable manner, the content of Lactobacillus rhamnosus Probio-M9 in the preparation is 1X 10 based on the total weight or volume of the preparation6CFU/g or 1X 106CFU/mL or more.
Furthermore, the product comprises the viable bacteria of the lactobacillus rhamnosus Probio-M9.
Further, the product comprises functional health products such as direct vat set starter, probiotic solid beverage, tablet candy, functional medicine auxiliary materials and the like.
It is another object of the present application to provide the use of said preparation for the preparation of at least one of the following medicaments:
(A) alleviating or treating autism spectrum disorders;
(B) improving social behavior abnormalities;
(C) improving the preference obstacle to novelty.
In one implementation, the dosage form of the medicament includes granules, capsules, tablets, granules, oral liquids, injections, powders or pills.
Optionally, the medicament further comprises pharmaceutically acceptable auxiliary materials and/or auxiliary agents.
Another object of the present application is to provide the use of lactobacillus rhamnosus Probio-M9 for the preparation of at least one of the following drugs, wherein the lactobacillus rhamnosus Probio-M9 has the microbial collection number CGMCC No. 18639:
(A) alleviating or treating autism spectrum disorders;
(B) improving social behavior abnormalities;
(C) improving the preference obstacle to novelty.
In one implementation, the dosage form of the medicament includes granules, capsules, tablets, granules, oral liquids, injections, powders or pills.
Optionally, the medicament further comprises pharmaceutically acceptable auxiliary materials and/or auxiliary agents.
The study on autism with a mouse as an animal model shows that lactobacillus rhamnosus Probio-M9 has an improvement effect on abnormal social behaviors and novelty preference disorder of BTBR mice, and can effectively relieve the anxiety behavior of the BTBR mice, and the lactobacillus rhamnosus Probio-M9 and the products thereof provided by the application have important significance on relieving and treating autism spectrum disorder. The strain has common food attributes and biological pharmacy strain potentials, provides a new strategy and a rational application basis for relieving and treating the autism, has relatively low cost, and has wide application prospects and popularization values.
Drawings
FIG. 1A shows the effect of Lactobacillus rhamnosus Probio-M9 on the sociability of BTBR mice in a three-box social test (D22-23);
FIG. 1B shows the effect of Lactobacillus rhamnosus Probio-M9 on the social freshness of BTBR mice in a three-box social test (D22-23);
FIG. 2A shows the effect of Lactobacillus rhamnosus Probio-M9 on the sociability of BTBR mice in a three-box social test (D40-41);
FIG. 2B shows the effect of Lactobacillus rhamnosus Probio-M9 on the social freshness of BTBR mice in a three-box social test (D40-41);
FIG. 3A shows the effect of Lactobacillus rhamnosus Probio-M9 on the mean hair management time in BTBR mice in a hair management trial (D22-23);
FIG. 3B shows the effect of Lactobacillus rhamnosus Probio-M9 on the number of false teasing in BTBR mice in a pilatory assay (D22-23);
FIG. 4A shows the effect of Lactobacillus rhamnosus Probio-M9 on the mean hairiness time of BTBR mice in a hairiness test (D39-40) the effect of Lactobacillus rhamnosus Probio-M9 on the mean hairiness time of BTBR mice in a hairiness test (D39-40);
FIG. 4B shows the effect of Lactobacillus rhamnosus Probio-M9 on the number of false teasing in BTBR mice in a pilatory assay (D39-40);
FIG. 5A shows the effect of Lactobacillus rhamnosus Probio-M9 on the search time of BTBR mice in a novel object identification assay (D28-31);
FIG. 5B shows the effect of Lactobacillus rhamnosus Probio-M9 on BTBR mouse recognition index in a novel object recognition assay (D28-31);
FIG. 6A shows the effect of Lactobacillus rhamnosus Probio-M9 on the search time of BTBR mice in a new object identification assay (D45-48);
FIG. 6B shows the effect of Lactobacillus rhamnosus Probio-M9 on BTBR mouse recognition index in a novel object recognition assay (D45-48);
FIG. 7A shows the effect of Lactobacillus rhamnosus Probio-M9 on the total distance of activity of BTBR mice in an open field trial (D24);
FIG. 7B shows the effect of Lactobacillus rhamnosus Probio-M9 on the central regional course of activity of BTBR mice in an open field trial (D24);
FIG. 7C shows the effect of Lactobacillus rhamnosus Probio-M9 on the central region time of activity of BTBR mice in the open field trial (D24);
FIG. 7D shows the effect of Lactobacillus rhamnosus Probio-M9 on the number of BTBR mouse standings in the open field test (D24);
FIG. 7E shows the effect of Lactobacillus rhamnosus Probio-M9 on standing time of BTBR mice in the open field test (D24);
FIG. 8A shows the effect of Lactobacillus rhamnosus Probio-M9 on the total distance of activity of BTBR mice in an open field trial (D42);
FIG. 8B shows the effect of Lactobacillus rhamnosus Probio-M9 on the central regional distance of activity of BTBR mice in an open field trial (D42);
FIG. 8C shows the effect of Lactobacillus rhamnosus Probio-M9 on the central region time of activity of BTBR mice in the open field trial (D42);
FIG. 8D shows the effect of Lactobacillus rhamnosus Probio-M9 on the number of BTBR mouse standings in the open field test (D42);
FIG. 8E shows the effect of Lactobacillus rhamnosus Probio-M9 on standing time of BTBR mice in the open field test (D42).
Detailed Description
Reference will now be made in detail to the exemplary embodiments, examples of which are illustrated in the accompanying drawings. When the following description refers to the accompanying drawings, like numbers in different drawings represent the same or similar elements unless otherwise indicated. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present invention. Rather, they are merely examples of methods consistent with certain aspects of the invention, as detailed in the appended claims.
The details provided herein are set forth in the following examples.
First, a brief description will be given of a model employed in this example. The model used in this example is a BTBR T- (+) tf/J (abbreviated as "BTBR") mouse model, which is an inbred mouse that has not only the core symptoms of ASD: social contact is reduced, fewer ultrasonic waves are emitted in social occasions, repeated hairing behaviors are severe, and the brain dysplasia and the immune biochemical index abnormality similar to ASD are also provided, so that the model is a main application model for researching autism. The BTBR mouse model is used for research, and can reflect whether the drug to be tested can play a role in treating the ASD patient.
In the following examples of the present application, it was demonstrated by animal experiments whether or not lactobacillus rhamnosus Probio-M9 and its products have a relieving effect or a treating effect on autism in both of BTBR mice and normal mice of 6 months of age.
(I) Experimental method
1.1 preparation of Experimental Strain
Lactobacillus rhamnosus Probio-M9 strain powder, wherein the viable bacteria content is 2 × 1011CFU/g, available from Beijing Ke Tuhengtong Biotechnology GmbH.
1.2 Experimental animals
The experiment adopts 6-month-old BTBR mice and 6-month-old normal mice, which are provided by Beijing Witongliwa laboratory animal technology Limited company and are used for establishing an autism spectrum disorder model.
1.3 design of the experiment
The two mice are respectively randomly divided into 3 groups, each group comprises 13 mice, and the treatment of the autism spectrum disorder model is respectively carried out, wherein the experimental method comprises the following steps:
the first group was a B6 control group consisting of normal mice: feeding sterile normal saline;
the second group is the BTBR-Vehicle group consisting of BTBR mice: feeding sterile normal saline;
the third group is a BTBR-M9 group consisting of BTBR mice: feeding sterile normal saline, and performing intragastric administration of Lactobacillus rhamnosus Probio-M9 daily at a total daily intragastric dose of 5 × 109CFU/time until the end of day 21 dosing.
Behavioral test time points: before grouping (D-4), after the 21 st dose (D22-31), before the end of the experiment (D39-48).
Examples
Example 1 social behavior test-three-box social test
The test method comprises the following steps:
(1) an adaptation stage: the mice to be tested were placed in the apparatus (40cm x 60cm x 22cm) and allowed to move freely in 3 communicating chambers for 10min to acclimate.
(2) And (3) a testing stage:
well-adapted mice to be tested were taken out and randomly placed in an empty cage (designated as "empty cage") and a cage with strange mice (designated as "mouse 1") in the left and right compartments, respectively.
Gently placing the mouse from the middle chamber into the left and right chambers for 10min, and recording the stay times (n)Air conditionerAnd nA method for preparing a catalyst) And the time of interaction with the empty cage and the strange mouse, respectively (T)Air conditionerAnd TA method for preparing a catalyst) After completion, the test mice were gently removed.
Another strange mouse (denoted "mouse 2") was placed in the empty cage, and the test mouse was again gently placed in the middle chamber. The mice to be tested are recorded in the left and right chambers within the next 10minNumber of dwells (n)CookingAnd nUnfamiliar 2) And the time of interaction with the previous strange mouse and the new strange mouse, respectively (T)CookingAnd TUnfamiliar 2) The results are shown in fig. 1A, 1B, 2A and 2B, in which,
FIG. 1A shows the effect of Lactobacillus rhamnosus Probio-M9 on the sociability of BTBR mice in a three-box social test (D22-23);
FIG. 1B shows the effect of Lactobacillus rhamnosus Probio-M9 on the social freshness of BTBR mice in a three-box social test (D22-23);
FIG. 2A shows the effect of Lactobacillus rhamnosus Probio-M9 on the sociability of BTBR mice in a three-box social test (D40-41);
FIG. 2B shows the effect of Lactobacillus rhamnosus Probio-M9 on the social freshness of BTBR mice in a three-box social test (D40-41).
As can be seen from FIGS. 1A and 1B, the social interaction behavior of the B6 control group of test mice was normal and had social novelty preference, and the BTBR-Vehicle group of test mice had a significantly greater preference for the familiar mouse ("mouse 1") than for "mouse 2" during the second phase of the test. After 21 days of continuous administration of Lactobacillus rhamnosus Probio-M9, the mice in BTBR-probiotic group had improved social behavior, but all had statistical differences (P > 0.05).
As can be seen from fig. 2A and 2B, the social interaction behavior of the test mice in the B6 control group was normal and social novelty preference was observed, and the contact time of the test mice in the BTBR-Vehicle group to the empty cage in the first stage of the test was significantly longer than that of the strange mice (P < 0.05); lactobacillus rhamnosus Probio-M9 has a remarkable improvement effect on BTBR social behavior abnormality.
EXAMPLE 2 stencil behavior test-Hair Conditioning test
The test method comprises the following steps:
(1) an adaptation stage: the mice were taken from the animal room to the test room 1h before the video recording activity started, and the room was illuminated at about 40Lux for 1h of acclimatization. The testing time of each time is 17: 00-21: 00. For the experiment, the mice were gently transferred into a transparent observation box (specification L.times.WXH: 20 cm. times.20 cm. times.50 cm) and data collection was carried out simultaneously.
(2) And (3) a testing stage: each mouse was placed in a clean observation box with padding on the bottom, and a camera was placed directly above the observation box. Under low light conditions (about 40Lux), after the observation box is pre-adapted for 10min each time, the behavior of the mouse is recorded for 10min, the testers are kept quiet and far away from the tested mouse in the whole process, the observers keep the standards consistent and the groups are unknown, and the repeated self-combing behavior of the mouse within 10min is scored.
Specifically, 5min is used as a unit interval, the accumulated carding time and the number of times of correct/wrong carding actions (rounds) in each interval are recorded, and the average accumulated carding time(s) and the average wrong carding time percentage (%) are calculated, wherein a carding analysis algorithm is used for analyzing the carding actions as rounds, and the sequence of one round from head to tail is as follows: paw, face, body, leg, tail/genitals; percent (%) false combing number-total number of times that the normal cranial combing sequence number/combing behavior was not followed.
The results are shown in fig. 3A, 3B, 4A and 4B, in which,
FIG. 3A shows the effect of Lactobacillus rhamnosus Probio-M9 on the mean hair management time in BTBR mice in a hair management trial (D22-23);
FIG. 3B shows the effect of Lactobacillus rhamnosus Probio-M9 on the number of false teasing in BTBR mice in a pilatory assay (D22-23);
FIG. 4A shows the effect of Lactobacillus rhamnosus Probio-M9 on the mean hairiness time of BTBR mice in a hairiness test (D39-40) the effect of Lactobacillus rhamnosus Probio-M9 on the mean hairiness time of BTBR mice in a hairiness test (D39-40);
FIG. 4B shows the effect of Lactobacillus rhamnosus Probio-M9 on the number of false teasing in BTBR mice in a pilatory assay (D39-40).
As can be seen from FIGS. 3A, 3B, 4A and 4B, administration of Lactobacillus rhamnosus Probio-M9 did not significantly improve the mean hair management time and the percentage of false combing times in BTBR mice.
Example 3 cognitive behavioral testing-New object recognition
The test method comprises the following steps:
(1) an adaptation period: on day 1, the test mice were acclimated to the test room for 1h, and then placed in an open field box (no object in the box) for 20 min.
(2) In the familiarity stage: on day 2 (after 24h adaptation), the test mice were adapted to the test room for 1h, then placed in an open field box for at least 5min, then removed from the open field box and placed in a clean temporary holding cage for about 2 min. Two yellow cylindrical bottles with the same size, color and texture are placed at the left end and the right end of the same side in the open field box to serve as familiar objects, the tested mouse is placed in the box with the back facing the familiar objects, the condition that the tested mouse explores the familiar objects within 5min is recorded, and the timing standard is the time for exploring the familiar objects within the range that the nose or mouth of the mouse is 2cm away from the objects.
(3) And (3) testing period: after the familiarity period of 2h, a yellow cylindrical bottle in the open field box is randomly replaced by a green conical bottle with the same size as the yellow cylindrical bottle to serve as a novelty object, then the mouse is placed in the box with the back facing the object, and the exploration time (Tn) of the mouse on the novelty object and the exploration time (Tf) of the familiar object within 5min are recorded. The identification index (RI) is used as a detection index of the recognition capability of the novel object, and is calculated according to the formula RI of Tn/(Tn + Tf).
The results are shown in fig. 5A, 5B, 6A and 6B, wherein,
FIG. 5A shows the effect of Lactobacillus rhamnosus Probio-M9 on the search time of BTBR mice in a novel object identification assay (D28-31);
FIG. 5B shows the effect of Lactobacillus rhamnosus Probio-M9 on BTBR mouse recognition index in a novel object recognition assay (D28-31);
FIG. 6A shows the effect of Lactobacillus rhamnosus Probio-M9 on the search time of BTBR mice in a new object identification assay (D45-48);
FIG. 6B shows the effect of Lactobacillus rhamnosus Probio-M9 on the BTBR mouse recognition index in a novel object recognition assay (D45-48).
As can be seen from FIGS. 5A, 5B, 6A and 6B, administration of Lactobacillus rhamnosus Probio-M9 did not significantly improve cognitive performance in BTBR mice.
Example 4 anxiety behavior test-open field test
The test method comprises the following steps:
under the condition of low light (about 40lux), the tested mouse is placed in an open field analysis box to adapt for 2min, and the standing times, standing time, total area moving distance, central area moving distance and moving time of the mouse in the open field analysis box within 10min are measured by an animal spontaneous movement video analysis system.
Standing standard: the front 2 limbs were all lifted off the ground and kept for 1s and above.
The test data includes: total distance, central area time, standing times and standing time.
The results are shown in fig. 7A, 7B, 7C, 7D, 7E, 8A, 8B, 8C, 8D and 8E, wherein,
FIG. 7A shows the effect of Lactobacillus rhamnosus Probio-M9 on the total distance of activity of BTBR mice in an open field trial (D24);
FIG. 7B shows the effect of Lactobacillus rhamnosus Probio-M9 on the central regional course of activity of BTBR mice in an open field trial (D24);
FIG. 7C shows the effect of Lactobacillus rhamnosus Probio-M9 on the central region time of activity of BTBR mice in the open field trial (D24);
FIG. 7D shows the effect of Lactobacillus rhamnosus Probio-M9 on the number of BTBR mouse standings in the open field test (D24);
FIG. 7E shows the effect of Lactobacillus rhamnosus Probio-M9 on standing time of BTBR mice in the open field test (D24);
FIG. 8A shows the effect of Lactobacillus rhamnosus Probio-M9 on the total distance of activity of BTBR mice in an open field trial (D42);
FIG. 8B shows the effect of Lactobacillus rhamnosus Probio-M9 on the central regional distance of activity of BTBR mice in an open field trial (D42);
FIG. 8C shows the effect of Lactobacillus rhamnosus Probio-M9 on the central region time of activity of BTBR mice in the open field trial (D42);
FIG. 8D shows the effect of Lactobacillus rhamnosus Probio-M9 on the number of BTBR mouse standings in the open field test (D42);
FIG. 8E shows the effect of Lactobacillus rhamnosus Probio-M9 on standing time of BTBR mice in the open field test (D42).
From the indicators of standing time shown in fig. 7E and fig. 8E, the standing time of B6 control mice was significantly greater than that of BTBR model mice (P <0.05) regardless of the vehicle group or the probiotic group. No obvious difference is seen in the total regional movement distance, the central regional movement distance and the time index of B6 control group mice and BTBR mice at the end of dosing D24/D42 when taking Lactobacillus rhamnosus Probio-M9 (P > 0.05).
The study on autism with a mouse as an animal model shows that the lactobacillus rhamnosus Probio-M9 has a certain improvement effect on abnormal social behaviors and novelty preference disorder of a BTBR mouse, can effectively relieve the anxiety behavior of the BTBR mouse, and has important significance on relieving and treating autism spectrum disorder.
Furthermore, the applicant thinks that the autism is closely related to the unbalance of intestinal flora and the abnormity of the intestinal-cerebral axis, the probiotic lactobacillus rhamnosus Probio-M9 and the product thereof provided by the application can regulate the composition of the intestinal flora by taking the intestinal flora as a target spot, improve the integrity of the intestinal barrier, further reduce the leakage of specific intestinal metabolites, rebuild the metabolites in serum, effectively reduce repetitive and compulsive behaviors, reduce anxiety and impulsion, improve sleeping and social abilities and other behavioral habits related to the autism, recover the normal intestinal flora, reduce inflammation and improve the behavioral symptoms related to the autism, or can become an effective adjuvant therapy scheme for the autism.
The present application has been described in detail with reference to specific embodiments and illustrative examples, but the description is not intended to limit the application. Those skilled in the art will appreciate that various equivalent substitutions, modifications or improvements may be made to the presently disclosed embodiments and implementations thereof without departing from the spirit and scope of the present disclosure, and these fall within the scope of the present disclosure. The protection scope of this application is subject to the appended claims.
Claims (9)
1. The application of lactobacillus rhamnosus Probio-M9 in preparing a medicament for treating autism, wherein the microorganism preservation number of the lactobacillus rhamnosus Probio-M9 is CGMCC No. 18639.
2. A product for treating autism, which comprises Lactobacillus rhamnosus Probio-M9, wherein the microorganism accession number of the Lactobacillus rhamnosus Probio-M9 is CGMCC No. 18639.
3. The preparation according to claim 2, wherein the lactobacillus rhamnosus Probio-M9 is present in the preparation in an amount of 1 x 10 based on the total weight or volume of the preparation6CFU/g or 1X 106CFU/mL or more.
4. The product according to claim 2 or 3, characterized in that it comprises viable bacterial cells of Lactobacillus rhamnosus Probio-M9.
5. The product according to any one of claims 2 to 4, wherein the product comprises functional health products such as ready-to-use leavening agents, probiotic solid beverages, tabletted confectioneries and functional pharmaceutical aids.
6. Use of the article of manufacture of any one of claims 2 to 5 for the manufacture of at least one of the following medicaments:
(A) alleviating or treating autism spectrum disorders;
(B) improving social behavior abnormalities;
(C) improving the preference obstacle to novelty.
7. The article of manufacture of claim 6, wherein the pharmaceutical dosage form comprises a granule, a capsule, a tablet, a granule, an oral liquid, an injection, a powder, or a pill.
8. The article of manufacture of claim 6 or 7, wherein the medicament further comprises pharmaceutically acceptable adjuvants and/or adjuvants.
9. Use of lactobacillus rhamnosus Probio-M9 for the preparation of at least one of the following medicaments, wherein the lactobacillus rhamnosus Probio-M9 has the microbial deposit number of CGMCC No. 18639:
(A) alleviating or treating autism spectrum disorders;
(B) improving social behavior abnormalities;
(C) improving the preference obstacle to novelty.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011600715.2A CN112618580A (en) | 2020-12-30 | 2020-12-30 | Lactobacillus rhamnosus Probio-M9 and application of product thereof in preparing medicament for treating autism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011600715.2A CN112618580A (en) | 2020-12-30 | 2020-12-30 | Lactobacillus rhamnosus Probio-M9 and application of product thereof in preparing medicament for treating autism |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112618580A true CN112618580A (en) | 2021-04-09 |
Family
ID=75286391
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011600715.2A Pending CN112618580A (en) | 2020-12-30 | 2020-12-30 | Lactobacillus rhamnosus Probio-M9 and application of product thereof in preparing medicament for treating autism |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112618580A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100216783A1 (en) * | 2009-02-26 | 2010-08-26 | Laxminarayan Bhat | Compositions, synthesis, and methods of utilizing arylpiperazine derivatives |
| CN110982733A (en) * | 2019-11-14 | 2020-04-10 | 北京科拓恒通生物技术股份有限公司 | Lactobacillus rhamnosus for preventing and treating mastitis and application thereof |
-
2020
- 2020-12-30 CN CN202011600715.2A patent/CN112618580A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100216783A1 (en) * | 2009-02-26 | 2010-08-26 | Laxminarayan Bhat | Compositions, synthesis, and methods of utilizing arylpiperazine derivatives |
| CN110982733A (en) * | 2019-11-14 | 2020-04-10 | 北京科拓恒通生物技术股份有限公司 | Lactobacillus rhamnosus for preventing and treating mastitis and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 吴晓丽等: "肠道微生物与自闭症研究进展", 《科学通报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2211879B1 (en) | Probiotic compositions and methods for inducing and supporting weight loss | |
| CN110893195B (en) | Lactobacillus paracasei ET-22 with function of relieving intestinal inflammation | |
| US8092793B2 (en) | Treating inflammatory bowel disease with live bacteria | |
| US20080199444A1 (en) | Composition and method for reducing feces toxins and treating digestive disorders | |
| US20230048705A1 (en) | Use of probiotics in the treatment and/or prevention of atopic dermatitis | |
| EA035949B1 (en) | Use of a composition comprising bacterial blautia hydrogenotrophica strain and method of treating or preventing visceral hypersensitivity | |
| JP2022516983A (en) | Rhodococcus louver products and their pharmaceutical use | |
| WO2022206300A1 (en) | Composition capable of facilitating defecation and use thereof | |
| CN102091099A (en) | Application of bacillus coagulans in preparing medicine for treating inflammatory bowel disease | |
| CN108348558A (en) | The method for treating colitis | |
| CN111575204B (en) | Lactobacillus paracasei capable of relieving or treating allergic diseases and application thereof | |
| WO2005030230A1 (en) | Compositions and methods for treatment or prevention of psoriasis and related disorders | |
| CN101138573A (en) | Application of clostridium butyricum and condensate bacillus in the preparation of medicament for treating inflammatory bowel disease | |
| US20080118482A1 (en) | Treating mouth ulcer with live bacteria | |
| CN103157095A (en) | Kidney bean phytolectin applications in preparation of human drugs and drug composition thereof | |
| CN114681493B (en) | Application of bifidobacterium animalis subspecies lactis | |
| CN106389478A (en) | Applications of Bacteroides fragilis in treatment and/or prevention of obesity or diabetes mellitus | |
| CN116121154B (en) | Leuconostoc lactis and application thereof | |
| KR102636571B1 (en) | Prevention and/or treatment of pneumococcal infection | |
| CN112618580A (en) | Lactobacillus rhamnosus Probio-M9 and application of product thereof in preparing medicament for treating autism | |
| WO2019227414A1 (en) | Composition and uses thereof | |
| CN113512509B (en) | Lactobacillus crispatus and uses thereof | |
| CN113005066A (en) | Compound bifidobacterium preparation with antiallergic, immunity enhancing, blood sugar reducing, blood fat reducing and weight losing functions and preparation method thereof | |
| CN105194658A (en) | Traditional Chinese medicinal preparation for treating obesity and enhancing physique and preparation method of traditional Chinese medicinal preparation | |
| WO2019205506A1 (en) | Egg-shaped bacteroides for relieving endotoxin infection and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |