CN103142656A - Application of bacteroides fragilis in preparing composition for preventing and treating colon cancer - Google Patents
Application of bacteroides fragilis in preparing composition for preventing and treating colon cancer Download PDFInfo
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- CN103142656A CN103142656A CN2013100857015A CN201310085701A CN103142656A CN 103142656 A CN103142656 A CN 103142656A CN 2013100857015 A CN2013100857015 A CN 2013100857015A CN 201310085701 A CN201310085701 A CN 201310085701A CN 103142656 A CN103142656 A CN 103142656A
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Abstract
The invention relates to the technical field of application of bacteroides fragilis, and in particular relates to application of bacteroides fragilis in preparing a composition for preventing and treating colon cancer. Experiments show that bacteroides fragilis induces an organism to generate the anti-tumor effect in vivo, inhibit the tumor growth, accelerate tumor cell apoptosis and prolong the lifetime, so that bacteroides fragilis has a good effect in preventing and treating colon cancer. The invention explores novel use of bacteroides fragilis and develops a novel application field, thereby indicating that the bacteroides fragilis has good edible and officinal prospect. Bacteroides fragilis serving as probiotic can be used for preparing foods or medical compositions for preventing and treating colon cancer so as to provide a clinically health-care and preventing and treating food suitable for human to take.
Description
Technical field
The present invention relates to the applied technical field of bacteroides fragilis, be specifically related to bacteroides fragilis and prevent and treat the application in the colon cancer compositions in preparation.
Background technology
Malignant tumor is the class disease that harm humans health is the most serious, and in more American-European countries, the mortality rate of cancer is only second to cardiovascular system diseases and occupies second.China is along with aged tendency of population, and the M & M of tumor all increases to some extent, and the mortality rate of urbanite's malignant tumor has occupied first of the cause of the death.20 beginnings of the century, it is found that Bulgarian long lived elder ratio is the highest in same area, and the sickness rate of tumor is also relatively low.Originally, the Bulgarian had the custom of drinking Yoghourt every day.The research discovery, the probiotic bacteria contained in matzoon can promote decomposition and the absorption of error residue in stomach, can also decompose lactoprotein, suppresses the growth of pathogenic bacterium in the intestines and stomach.In recent years, increasing research shows that probiotic bacteria has certain antagonism for tumor, and discloses gradually its mechanism of action.Research finds that probiotic bacteria antineoplastic mechanism short of money mainly contains: optimize the combination of intestinal microbial population, suppress to produce the putrefaction bacteria growth of cancerigenic factor; Produce antimutagenic material and cancer-resisting substance; Strengthen host's immune system, preventing chronic recurrent inflammation; In conjunction with albumen mutagenic matter and carcinogen; Affect the expression of tumor cell gene and enzyme.
Colon cancer is one of common pernicious digestive tract tumor, according to the statistics of 2012, shows, the new cases of the annual colon cancer in the whole world, up to 1,200,000, approximately has 60.87 ten thousand patients to die from colon cancer every year.And, in China, the M & M of colon cancer also is continuous ascendant trend.So, the prevention work of how to do well colon cancer and reduce its M & M and become one of current medical circle research direction.
Bacteroides fragilis (
bacteroides fragilis) as a kind of Gram-negative, shaft-like, blunt round and dense the dying in two ends, pod membrane is arranged, without brood cell, motorless obligate anaerobes, minute produce enterotoxin type and non-product enterotoxin type.Bacteroides fragilis, as the part of people and animal intestinal normal flora, mainly is stored in colon, in addition, but the also field planting growth of respiratory tract, gastrointestinal tract and urogenital tract mucosa.What research was more at present is that bacteroides fragilis is as a kind of conditioned pathogen, when host's mucosa is impaired, can invade tela submucosa, cause infection, also can be through blood flow, cause that other organ of health is as intestinal, abdominal cavity, liver, lung, cerebral tissue pyogenic infection the abscess and cause acute and chronic diarrhea etc. of occurring together.In addition, its generation to colorectal cancer also has facilitation.
Yet bacteroides fragilis is not only a kind of conditioned pathogen.Within 1991, Zhang Jijie has delivered " isolation identification of a kind of nontoxic bacteroides fragilis ", reported about nineteen eighty-three JIUYUE separate the large and purer antibacterial of output from a well-developed infant faeces, warp is with serial evaluation, basic test and observation on Clinical Application, belong to a strain in the bacteroides fragilis group without toadstool, be the probiotic bacteria of human body, name the bacterial strain into BF839.Thus, study and found, bacteroides fragilis is also a kind of probiotic bacteria.But bacteroides fragilis is as a kind of probiotic bacteria, and prevent and treat the related application of colon cancer compositions in preparation, report is not yet arranged at present.
Summary of the invention
The object of the present invention is to provide bacteroides fragilis to prevent and treat the application in the colon cancer compositions in preparation, described bacteroides fragilis depositary institution name is called China Committee for Culture Collection of Microorganisms's common micro-organisms center, the depositary institution address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, preservation date is on March 12nd, 2013, deposit number is CGMCC NO.7280, and Classification And Nomenclature is
Bacteroides fragilis.
In order to address the above problem, the technical solution adopted in the present invention is:
Provide bacteroides fragilis to prevent and treat the application in the colon cancer compositions in preparation.
Wherein, food or the pharmaceutical composition of described compositions at least containing the bacteroides fragilis that prevents and treats colon cancer.
Wherein, described compositions is selected from any one in milk powder, cheese, curdled milk, Yoghourt, ice cream, milk base fermented food, fermented cereal food, baby food or pet food.
Wherein, described compositions is pill, tablet, granular preparation, capsule, oral liquid or tube feed preparation.
Wherein, described compositions is the liquid bacterial suspension that contains described bacteroides fragilis.
Wherein, the content of described bacteroides fragilis in described compositions is 10
8~ 10
10cfu/mL.
the invention has the beneficial effects as follows:
The present invention has excavated the new purposes of bacteroides fragilis, has opened up a new application.The present invention proves by experiment, and bacteroides fragilis induces body to produce Graft Versus Tumor in vivo, suppresses tumor growth, promotes apoptosis of tumor cells, extends life cycle, for preventing and treating colon cancer, has good effect; And the bacteroides fragilis safety non-toxic, pharmacological action is strong, thereby has indicated that bacteroides fragilis has well edible and prospect in medicine.Bacteroides fragilis, as a kind of probiotic bacteria, can be used for food or pharmaceutical composition that preparation prevents and treats colon cancer, is clinical health care and the prevention non-defective unit that provides a kind of applicable human body to take.
The specific embodiment
Below by embodiment, the present invention is described in further details, these embodiment only are used for illustrating the present invention, do not limit the scope of the invention.
the preparation of embodiment 1 bacteroides fragilis bacterium liquid
(1) separate bacteroides fragilis in infant faeces
Take 0.5 gram stool by " normal flora inspection technique ", be placed in the test tube that the 4.5ml normal saline first is housed, with this serial dilution, become 10
-1~ 10
-6, by the rare long-pending degree of height, to low rare long-pending degree, dripped on the Bd culture medium flat plate, be placed in the anaerobic gas generation box and add the anaerobic gas generation bag and carry out the anaerobism cultivation.Put 37 ℃, can opening inspection after 2 ~ 7 days.Choose single bacterium colony and carry out pure culture, dyeing microscopic examination, cultural character, Physiology and biochemistry detection.
(2) bacterium that increases of bacteroides fragilis is cultivated and bacterium solution preparation
Choose single colony inoculation after purification is increased bacterium and is cultivated in the tryptone meat soup of improvement, the centrifugation of gained bacterium liquid, rotating speed 3000r/min, centrifugal 15min, remove supernatant, after precipitate washs with normal saline, the precipitate normal saline dilution, do the bacterium number with the Maxwell opacity tube and measure, be diluted to 10
8cfu/ml, 10
9cfu/ml, 10
10cfu/ml, save backup.
embodiment 2 bacteroides fragilis virulence experiments
Adopt the bacteroides fragilis bacterium liquid of embodiment 1 preparation to carry out following experiment:
(1) chmice acute virulence experiment
Get 60 of body weight 22 ± 2 gram C57BR/C mices (male and female half and half), be divided into high dose group, low dose group and matched group, each organizes 20.Every mice gavage every day 10 of high dose group
12cfu/ml bacterium liquid 0.5ml, low dose group gavage every day 10
9cfu/ml bacterium liquid 0.5ml, matched group gavage every day 0.5ml normal saline.Continuous Observation 14 days.
Experimental result: the mice normal growth, have no dead, LD50 does not measure.
(2) rat virulence experiment
Get 30 of 150 ± 20 gram SD rats (male and female half and half), be divided into 3 groups: high low dose group and matched group.Every rat gavage every day 10 of high dose group
12cfu/ml bacterium liquid 2ml, low dose group gavage every day 10
9cfu/ml bacterium liquid 2ml, matched group gavage every day 2ml normal saline.Continuous Observation 14 days, phlebotomize after 14 days and survey hemogram, Liver and kidney merit.Get gastrointestinal liver spleen tissue and do the pathological section electron microscopic observation.
Experimental result: be showed no obvious abnormalities.
(3) Rhesus Macacus virulence experiment
Get 10 of Rhesus Macacus (male and female half and half), be divided into 2 groups: high low dose group and matched group.Every of high dose group Rhesus Macacus every day oral 10
12cfu/ml bacterium liquid 100ml, low dose group every day oral 10
9cfu/ml bacterium liquid 100ml.Continuous Observation 60 days.Phlebotomize respectively after 14 days, 30 days, 45 days, 60 days and survey hemogram, Liver and kidney merit, thought-read electrograph.Get gastrointestinal liver spleen tissue and do the pathological section electron microscopic observation.
Experimental result: be showed no obvious abnormalities.
embodiment 3 bacteroides fragiliss are for the preventive effect of colon cancer
Adopt the bacteroides fragilis bacterium liquid of embodiment 1 preparation to carry out following experiment:
Animal Model: 60 ICR mices are divided into to 2 large groups at random, that is: normal saline group (10) and model group (50).1,2-Dimethylhydrazine (DMH) is dissolved in normal saline, and making its final concentration is 20mg/kg, with 0.25mol/L NaOH, adjusts pH to 6.5, and lumbar injection gives mice.After 1 week, add Dextran sodium sulfate (DSS) in the drinking-water of model group, concentration is 20g/L, gives mice and drinks continuously 7 days.When hemafecia appears for the second time in mice, and give the 9th week after DMH, model mice is divided into to 4 groups at random by weight, that is: model group (20) and bacteroides fragilis group: high dose group (10
9cfu/ml), middle dosage group (10
8cfu/ml), low dose group (10
7cfu/ml).Bacteroides fragilis bacterium liquid gavage, every day 1 time, each 0.5ml, totally 11 weeks.
Evaluation index: test the etherization mice the 20th week.Cut off mouse peritoneal, in ice bath, from anus, upwards intercept colon 8cm, with after normal saline flushing, filter paper blots, and claims weight.Record the colon tumor number, get the about 1cm of colon and do pathologic finding.
Result: the model group mice was at the 20th week, and proctoptosis appears in 50% mice, and three dosage groups of bacteroides fragilis have no the proctoptosis mice.Model group has the mice colon that surpasses 80% one or more tumor to occur, and colon hyperemia thickens.And the bacteroides fragilis low dose group only has 3 colon tumor to occur, senior middle school's dosage group only has 1 appearance.Pathological examination shows, the disorder of model group cell arrangement, infiltrative growth, and obscurity boundary, be diagnosed as adenomatoid hyperplasia; Three dosage group mucosa disappearances of bacteroides fragilis, inflammatory cell extensively infiltrates, and is typical inflammation and changes.
Experimental result shows, bacteroides fragilis successfully rests on inflammatory conditions by the mice colon, has stoped the transformation of colitis to colon cancer.As can be seen here, bacteroides fragilis has good preventive effect to colon cancer.
embodiment 4 bacteroides fragiliss are for the therapeutical effect of colon cancer
Adopt the bacteroides fragilis bacterium liquid of embodiment 1 preparation to carry out following experiment:
30 of BALB/c mouse, every Mus back subcutaneous vaccination CT26 cell (1*10
5cell/only), inoculation is divided into 2 groups by mice, 15 every group next day at random.First group is the normal saline group, every day gavage 0.5ml normal saline; Second group is the bacteroides fragilis group, gavage bacteroides fragilis bacterium liquid, every gavage amount 0.5ml*10
9cfu/ml, totally 30 days.Observe medication tumor size in the time of 30 days, mice with tumor life cycle, the part mice is put to death and carries out In situ cell apoptosis analysis (TU NEL).
Experimental result: bacteroides fragilis group colon lump is significantly less than the normal saline group, and the mean survival time obviously is longer than the normal saline group.Bacteroides fragilis group mouse tumor cell (CT26) apoptosis number positive is apparently higher than the normal saline matched group.
Experimental result shows, bacteroides fragilis induces body to produce Graft Versus Tumor in vivo, suppresses tumor growth, promotes apoptosis of tumor cells, extends life cycle.Illustrated that bacteroides fragilis has good therapeutical effect for colon cancer.
Claims (6)
1. bacteroides fragilis prevents and treats the application in the colon cancer compositions in preparation.
2. bacteroides fragilis according to claim 1 prevents and treats the application in the colon cancer compositions in preparation, it is characterized in that: food or the pharmaceutical composition of described compositions at least containing the bacteroides fragilis that can prevent and treat colon cancer.
3. bacteroides fragilis according to claim 2 prevents and treats the application in the colon cancer compositions in preparation, it is characterized in that: described compositions is selected from any one in milk powder, cheese, curdled milk, Yoghourt, ice cream, milk base fermented food, fermented cereal food, baby food or pet food.
4. bacteroides fragilis according to claim 2 prevents and treats the application in the colon cancer compositions in preparation, it is characterized in that: described compositions is pill, tablet, granular preparation, capsule, oral liquid or tube feed preparation.
5. bacteroides fragilis according to claim 1 prevents and treats the application in the colon cancer compositions in preparation, it is characterized in that: described compositions is the liquid bacterial suspension that contains described bacteroides fragilis.
6. bacteroides fragilis according to claim 1 prevents and treats the application in the colon cancer compositions in preparation, it is characterized in that: the content of described bacteroides fragilis in described compositions is 10
8~ 10
10cfu/mL.
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| CN114306615A (en) * | 2022-01-12 | 2022-04-12 | 广州知易生物科技有限公司 | Novel application of bacteroides fragilis capsular polysaccharide A and immune checkpoint inhibitor |
| WO2023134195A1 (en) * | 2022-01-12 | 2023-07-20 | 广州知易生物科技有限公司 | Use of bacteroides fragilis combined with immune checkpoint inhibitor in treatment of skin tumors |
| CN114306615B (en) * | 2022-01-12 | 2023-11-17 | 广州知易生物科技有限公司 | Novel application of bacteroides fragilis capsular polysaccharide A and immune checkpoint inhibitor |
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Application publication date: 20130612 |