CN105343132B - Composition, the drug and preparation method thereof for treating colitis - Google Patents
Composition, the drug and preparation method thereof for treating colitis Download PDFInfo
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- CN105343132B CN105343132B CN201510895295.8A CN201510895295A CN105343132B CN 105343132 B CN105343132 B CN 105343132B CN 201510895295 A CN201510895295 A CN 201510895295A CN 105343132 B CN105343132 B CN 105343132B
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- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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Abstract
The invention discloses a kind of compositions for treating colitis, drug and preparation method thereof, wherein the composition i.e. probiotics and colostrum growth factor for treating colitis are used in combination.The present invention establishes colitis model using Balb/c mouse as subjects, from the apparent state of test mice, changes of weight, colon lengths, pathological conditions angularly, studies the resistive connection enteritis effect of drug of the present invention；Test result shows, drug of the present invention is got well than the effect of probiotics or colostrum growth factor is used alone, and the two has the function of synergy, has good resistive connection enteritis effect, and will not recur in a short time, it can be good at intestinal mucosa injury caused by repairing colitis.
Description
Technical field
The present invention relates to a kind of composition for treating colitis, drug and preparation method thereof, i.e. at the beginning of two probiotics and ox
The drug that newborn growth factor is used in combination, belongs to pharmaceutical technology field.
Background technique
Colitis is also known as nonspecific ulcerative colitis, many kinds of, causes reason multiplicity, and onset is mostly slow
Slowly, the state of an illness differs in weight, main clinical manifestation diarrhea, abdominal pain, Mucous Stool and pus and blood stool, tenesmus, very then constipation, number
Cannot in a few days lead to stool, when and constipation when diarrhea, be often accompanied by thin out of strength etc., more recurrent exerbations.Abdominal pain is generally mostly secret anguish
Or colic pain, it is frequently located in left lower quadrant or underbelly.Other to be presented with loss of appetite, abdominal distension, Nausea and vomiting and hepatomegaly etc., left lower quadrant can
There is tenderness, the colon of spasm can be touched sometimes.Common constitutional symptom has thin, out of strength, fever, anaemia etc..There is small part sick
People is in the chronic course of disease, and the state of an illness deteriorates suddenly or first attack is just in fulminant, shows severe diarrhea, brings to human or animal
Greatly pain.
The method for the treatment of colitis mainly includes drug therapy, nutrition treatment, psychotherapy and operative treatment at present, wherein
Drug therapy is most important method.The drug for being used clinically for treatment colitis mainly has minosalicylates, drugs, kidney
Upper gland glucocorticoids and immunosuppressant etc..Minosalicylates, drugs can inhibit prostaglandin to synthesize well, clearly
Except oxygen radical to mitigate inflammatory reaction；However it can not eradicate colitis, and when patient's medication dose is bigger than normal, be also easy to produce all
More adverse reactions (such as: headache, nausea, leucocyte decline).Glucocorticoid is the head of severe or explosive colitis disease
Select medicine；However such drug easily leads to the side effects such as organism metabolic disorder, water retention, can be used as emergency drug, can not make for a long time
With.Immunosupress class medication medication dependence is big, and treatment phase is longer, thereby increases and it is possible to which intoxicating is carcinogenic, and side effect is larger, so general
It is used as adjuvant treatment.On the whole, said medicine is responded well to treatment fast, good effect, can be relieved the hardship of colitis disease's matter as a fire singeing one's eyebrows,
But there are still many problems.Firstly, patient's colitis can not be eradicated, it is palliative；Secondly, ill-effect is big, easily cause to suffer from
Person's headache, nausea cause body digestive system, hematological abnormalities；Then, potential intoxicating carcinogenicity；Finally, antibiotics
Drug (such as most widely used SASP) easily leads to body drug resistance drug resistance.
Also patent report treats using Chinese medicine composition to colitis, as application No. is 200710066007.3
Patent of invention " drug and preparation method for the treatment of chronic colitis and ulcerative colitis ", application No. is 200610167659.1
Patent of invention " a kind of drug and preparation method thereof for treating ulcerative colitis " treated due to the complicated component of these Chinese medicines
Effect is slow, and the course for the treatment of is long, and therapeutic effect is also difficult to ensure, therefore drug made of these Chinese medicine compositions does not obtain on a large scale so far
It promotes and applies.So researching and developing the new drug of one kind with highly important for treating this disease for being difficult to cure of colitis
Meaning.Though there is document report probiotics to can treat ulcerative colitis, there is presently no about using probiotic combination
The report for the treatment of colitis is used in combination in colostrum growth factor.
Summary of the invention
An object of the present invention is to provide a kind of composition for treating colitis, can effectively treat colitis, and
And toxic side effect is smaller, is not easy to produce resistance.
It is a further object to provide a kind of drugs and preparation method thereof for treating colitis, use treatment colitis
Composition, the effect that equally can achieve.
To achieve the goals above, the invention provides the following technical scheme:
A kind of composition for treating colitis, including probiotics and colostrum growth factor, the probiotics are acidophilus
Lactobacillus (Lactobacillus acidophilus) CGMCC10436 and lactobacillus plantarum (Lactobacillus
Plantarum) CGMCC9961, the lactobacillus acidophilus thalline quantity account for the 20~80% of probio thalline total quantity.
Preferably, the thalline quantity of the lactobacillus acidophilus and lactobacillus plantarum ratio is 1:1.
The colostrum growth factor is the growth factor crude extract extracted from colostrum；The probiotics is
The viable bacteria thallus of viable bacteria thallus form, the probiotics is centrifuged respectively by the lactobacillus liquid culture for being in stable initial stage
The bacterium mud of collection.
The probio thalline dosage is 2 × 1010CFU/kg, the colostrum growth factor dosage be 220~
320mg/kg.Lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum i.e. in drug used in every kilogram of animal, human body etc.
The total quantity of CGMCC9961 is 2 × 1010CFU, colostrum growth factor dosage are that 220~320mg specifically can be according to reality
Border situation is calculated, for example, weight is the rat of 20g, contains lactobacillus acidophilus CGMCC10436 and plant in medication
The total quantity of object lactobacillus CGMCC9961 is 2 × 109CFU, colostrum growth factor dosage are 4.4~6.4mg；One weight
For the people of 50kg, in medication the total quantity containing lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum CGMCC9961 be 1 ×
1012CFU, colostrum growth factor dosage are 11~16g.
Wherein, all, which can be made, in the composition can retain its active article, such as dairy products, drug, food.
The invention also discloses a kind of drugs for treating colitis, including composition above-mentioned.
The colitis includes ulcerative colitis, infectious colitis, hemorrhagic colitis, and without being limited thereto.
The preparation method of the drug of above-mentioned treatment colitis, includes the following steps:
(1) respectively by lactobacillus acidophilus and lactobacillus plantarum in 12~16h of MRS culture medium culture；
(2) 9~11 parts by volume of lactobacillus acidophilus bacterium solution for taking step (1) to obtain respectively, 3~5 body of lactobacillus plantarum bacterium solution
Then they are dissolved in the sterile PBS of 1 parts by volume respectively, obtain mixing bacterium mud by product part, centrifuging and taking bacterium mud；
(3) to colostrum centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and dry obtained colostrum growth factor
Crude extract；
(4) 44~64 mass parts of colostrum growth factor crude extract are added in above-mentioned mixing bacterium mud, are stirred evenly, so
After suitable auxiliary material is added clinically-acceptable dosage form is made.Here parts by volume and mass parts understands are as follows: if in step (2)
With the lactobacillus acidophilus bacterium solution of 9~11ml, 3~5ml lactobacillus plantarum bacterium solution, then to the colostrum growth of 44~64mg of application because
Sub- crude extract, i.e. parts by volume/mass parts=ml/mg.
Preferably, the preparation method of the drug of the treatment colitis, includes the following steps:
(1) respectively by lactobacillus acidophilus, lactobacillus plantarum in MRS culture medium culture 14h；
(2) 10 parts by volume of lactobacillus acidophilus bacterium solution for taking step (1) to obtain respectively, 4 parts by volume of lactobacillus plantarum bacterium solution, from
The heart takes bacterium mud, and then they are dissolved in the sterile PBS of 1 parts by volume respectively, obtains mixing bacterium mud；
(3) to colostrum centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and dry obtained colostrum growth factor
Crude extract；Preferably, colostrum is centrifuged at 4 DEG C, it is dry using freeze-drying.
(4) 54 mass parts of colostrum growth factor crude extract are added in above-mentioned mixing bacterium mud, are stirred evenly, then plus
Enter suitable auxiliary material and clinically-acceptable dosage form is made.
The deposit number of lactobacillus acidophilus CGMCC10436 of the present invention is CGMCC No.10436, the deposit date is
01 month 2015 No. 26, the deposit number of the lactobacillus plantarum CGMCC9961 is CGMCC No.9961, and the deposit date is 2014
In on November 13, in, depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center, and address is Beijing
The institute 3 of city, North Star West Road, Chaoyang District 1, Institute of Microorganism, Academia Sinica.
Beneficial effects of the present invention:
Colostrum growth factor can be by conjunction with protection of intestinal mucosal barrier cells tyrosine kinase receptor, to promote protection of intestinal mucosal barrier cells
Growth and proliferation, increase villi height, promote intestinal growth；And immunity of organism can be promoted by regulation inflammatory factor level
Power.The probiotics that the present invention uses can be by adjusting intestinal flora balance, and lactobacillus plantarum with harmful bacteria competing binding sites
CGMCC9961 can produce bacteriocin, inhibit the proliferation of harmful bacteria；Lactobacillus acidophilus CGMCC10436 has good prebiotic work
With.The results show that manufactured drug, which is used in combination, in the two has preferable resistive connection enteritis effect.And there is synergistic work
With.Its effect is got well than two kinds of bacterium are used alone, colostrum growth factor effect is used alone.It is in particular in and significantly improves knot
The apparent state of enteritis mouse, increases weight, the colon lengths of colitis mice, and significantly improves the complete of mouse Colon tissue
Property.
Detailed description of the invention
Fig. 1 is the scoring of separate groups of mice Gross pathology.
Specific embodiment
In the case that the present inventor feels a delicacy about to the treatment of colitis for current global field of medicaments, according to prebiotic
Immunity of organisms can be enhanced in bacterium and growth factor, has good therapeutic effect to human body intestinal canal sick (such as diarrhea), the present invention
The preferable living preparation of lactobacillus thallus of two plants of probiotic effects is combined with colostrum growth factor crude extract, drug is made, is used for animal
Experimental study is detailed description of the present invention below.
Examples are merely exemplary, and it is not intended to limit the scope of the present invention in any way.Those skilled in the art should
Understand, without departing from the spirit and scope of the invention can details to technical solution of the present invention and form modify
Or replacement, but these modifications and replacement are fallen within the protection scope of the present invention.A variety of materials of the present invention, reagent or
Apparatus unless otherwise specified, can be obtained from market.
In order to prove effectiveness of the invention, applicant carried out following tests.
(1) material and reagent
1, culture medium: MRS culture medium.
MRS culture medium is prepared: peptone 5g/L, beef extract 5g/L, tryptone 10g/L, yeast powder 5g/L, glucose
20g/L, anhydrous sodium acetate 5g/L, citric acid hydrogen diamine 2g/L, dipotassium hydrogen phosphate 2g/L, magnesium sulfate 0.58g/L, manganese sulfate
6.3 ± 0.1,121 DEG C of 0.25g/L, Tween-80 1ml/L, pH, it sterilizes within 15 minutes.
2, main agents: colostrum comes from the pasture Wanda Mountain, Heilongjiang Song Bei；Dextran sulfate sodium salt comes from Beijing bit
Rich company, biotechnology Co., Ltd；Sulfasalazine comes from Shanghai Xinyi Tianping Pharmaceutical Co., Ltd.；Formaldehyde is molten
Liquid comes from the chemical inc Tianjin Bo Di；Potassium dihydrogen phosphate comes from the extremely big chemical apparatuses factory in Tianjin Dongli District；Phosphoric acid hydrogen
Disodium comes from Tianjin BASF Chemical Co., Ltd..
3, key instrument: electron microscope comes from Motic company；Ultraviolet-uisible spectrophotometer comes from the U.S. DU800
BECKMAN company；Ophthalmology anatomy box comes from Shenyang City Jiu Ming aluminum products factory；Syringe controls the limited public affairs of space medical instrument from Jiangsu
Department.
(2), the separation of lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum CGMCC9961
1, conventional method is acquired from the herdsman family in the area such as In Xilingol League In Inner Mongolia, Hailar City and Ordos City
The traditional dilute cream or acidification cream of production, the screening for lactic acid bacteria.It is specific as follows:
(1) 25g sample is aseptically shredded, is dissolved in 225mL sterile saline, select to close after gradient dilution
Suitable dilution takes 1mL dilution that sterilized petri dishes are added, is poured on 20mL MRS solid medium, 30 DEG C of culture 72h.
(2) plate of the colony counts 15~150 is chosen, the bacterium colony for meeting lactic acid bacteria colony morphology characteristic is randomly selected,
The therefrom a small amount of bacterium of picking is dissolved in MRS fluid nutrient medium, and carries out 30 DEG C of culture 48h.
(3) by the bacterial strain being separated to, scribing line is purified repeatedly on MRS solid medium.
(4) purified bacterial strain is carried out to Gram's staining and catalase experiment.
(5) by all Gram-positives, negative catalase, the bacterial strain (being tentatively judged as lactic acid bacteria) for not forming gemma
It elects, glycerol is added in its culture solution, make the ultimate density 30% (v/v) of glycerol, be stored in -80 DEG C for use.
(6) above-mentioned be separated to after the bacterial strain for fixing tentatively as lactic acid bacteria activated for 3 generations respectively in MRS fluid nutrient medium is reflected
Fixed experiment.
2, the identification of lactobacillus acidophilus CGMCC10436
(1) Physiology and biochemistry is identified
Using reference culture lactobacillus acidophilus as control strain, following physiological and biochemical test: H is carried out2S test, methyl red
Test, litmus milk tests, gelatin liquefaction test, Starch Hydrolysis test, citrate utilize test, urea decomposition test, nitre
Hydrochlorate Degrading experiment and 3-hydroxy-2-butanone generate test, choose lactic acid identical with reference culture lactobacillus acidophilus physiological and biochemical test
Bacterium.
(2) Molecular Identification
Strain gene group DNA is extracted, 16SrDNA sequencing, nucleotide sequence such as 1 institute of sequence table Seq ID No are carried out
Show, the 16SrDNA sequence of bacterial strain of the present invention is compared with the sequence of NCBI lactobacillus acidophilus, bacterial strain of the present invention
16SrDNA sequence and Lactobacillus acidophilus sequence similarity are up to 95% or more, according to 16SrDNA sequence point
Analysis, the bacterial strain are accredited as lactobacillus acidophilus (Lactobacillus acidophilus), and are protected on 01 26th, 2015
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, microbial preservation number is CGMCC No.10436,
Preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica.
3, the identification of lactobacillus plantarum CGMCC9961
(1) Physiology and biochemistry is identified
Using standard plant lactobacillus as control strain, following physiological and biochemical test: H is carried out2S test, methyl red test,
Litmus milk tests, gelatin liquefaction test, Starch Hydrolysis test, citrate are dropped using test, urea decomposition test, nitrate
Solution test and 3-hydroxy-2-butanone generate test, choose lactic acid bacteria identical with lactobacillus plantarum physiological and biochemical test.
(2) Molecular Identification
Strain gene group DNA is extracted, 16SrDNA sequencing, nucleotide sequence such as 2 institute of sequence table Seq ID No are carried out
Show, the 16SrDNA sequence of bacterial strain of the present invention is compared with the sequence of NCBI lactobacillus plantarum, bacterial strain of the present invention
16SrDNA sequence and Lactobacillus plantarum sequence similarity are analyzed up to 95% or more according to 16SrDNA sequence,
It identifies, this bacterial strain is lactobacillus acidophilus (Lactobacillus plantarum).Deposit number is CGMCC NO.9961, is protected
The hiding date is on November 13rd, 2014, and depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center
(CGMCC), address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica.
(3) preparation of stomach-filling probiotics
Lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum CGMCC9961 are cultivated into 14h respectively, carry out gradient plate meter
Number, according to bacterium number needed for every mouse of lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum CGMCC9961 be 2 ×
108CFU, the lactobacillus acidophilus CGMCC10436 bacterium solution 1mL for taking equal stationary phase to go out respectively, lactobacillus plantarum CGMCC9961 bacterium solution
0.4mL, centrifuging and taking bacterium mud, is then refitted in 0.1mLPBS respectively, the probiotics given low for every mouse.
(4) preparation of colostrum growth factor crude extract
As raw to colostrum made from colostrum centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and freeze-drying
Long factor crude extract；The processing methods such as signified centrifugation, ultrafiltration, dialysis, drying of the invention, can be used ordinary skill people
Any method well known to member, as long as it is able to maintain growth factor effect, preferably, centrifugation is dry using cold by 4 DEG C of centrifugations
It is lyophilized dry.
(5) preparation of the drug of colitis is treated
Colostrum growth factor crude extract, given low are 5.4mg/20g (colostrum growth factor crude extract quality/small
Mouse quality)；The stomach-filling form of colostrum growth factor crude extract is to take out the acidophilus cream bar being dissolved in the sterile PBS solution of 0.1mL
Bacterium, lactobacillus plantarum bacterium mud tubule, mixing, are then added the colostrum growth factor crude extract of the dosage.
(6) stomach-filling of other group of mouse prepares
Antibiotic group intragastric administration on mice drug: the given low of antibiotic be 9.2mg/20g (willow nitrogen thiamines pyridine silty amount/
Mice weights)；The stomach-filling form of willow nitrogen thiamines pyridine is that taking-up is dissolved in the sterile PBS solution of 0.3mL, is mixed, then 4 DEG C of guarantors
It deposits, in case stomach-filling.
Two bacterium groups are combined intragastric administration on mice drug:
By on the market lactobacillus acidophilus and lactobacillus plantarum be same as above and carry out the processing such as counting, centrifugation of microorganism, distinguish
Take the lactobacillus acidophilus at the beginning of being in stationary phase and lactobacillus plantarum each 2 × 108CFU, the two mixing, is then dissolved in the nothing of 0.3mL
In bacterium PBS solution, mixing, then 4 DEG C of preservations, in case stomach-filling.
It is studied below with reference to zoopery is further to drug of the invention, to illustrate effectiveness of the invention.
(1) production and experimental design of mouse models of colitis
Dextran sulfate sodium (DSS) is dissolved in water bottle, allows Balb/c mouse freely to drink 8 days, with provocative test knot
Enteritis mouse model verifies mouse Colon then with mouse weight variation, apparent situation, colon lengths, pathology etc. for index
Whether scorching model is successfully established.
Mouse weight declines during modeling, and enteritis group mouse is apparently not in good state, be in particular in mouse it is One's eyesight is restrained,
The obscure curling of hair has Qiang's hair phenomenon；Also, there is blood cake at enteritis group mouse root of the tail, illustrate that mouse has dominant hematochezia phenomenon；Solution
There is phenomenon of festering at enteritis group mouse Colon after cuing open, intestinal tissue has hair purulence, turbidity phenomenon；Moreover, during modeling, enteritis group
Mouse Colon length naive mice colon significantly shortens；Also, by HE colored graph it is found that model group mouse Colon tissue, intestines
Mucosal atrophy, mucous membrane surface passivation, crypts destroy.
In conclusion the success of acute colitis model foundation, can carry out next step drug therapy test.
(2) experiment conclusion
(1) explanation is tested
During treatment, PBS stomach-filling only is carried out to enteritis naive mice, without treatment, as a result enteritis is empty during stomach-filling
White group dead mouse 11；On the one hand illustrate that colitis model is successfully established, still further aspect illustrates that colitis mice cannot be leaned on
Self-regeneration is fully recovered；So subsequent comparison, is across comparison, i.e., comparison with group front and back, after mouse treatment with control
Comparison before treatment.
(2) drug of the invention is obviously improved the apparent state of mouse
After the stomach-filling of two courses for the treatment of, the poly- mind of medicine group mouse sight of the present invention, action are active, and mouse hair is suitable
Sliding, brilliant white, compared with apparent state when enteritis, difference is extremely obvious, and is combined with two bacterium groups and independent colostrum growth factor
Group is compared, and the apparent state of mouse obviously improves；And compared with the apparent state of antibiotic group mouse, difference is not significant.Illustrate this hair
Bright drug (probiotic combination colostrum growth factor) has good effect to treatment colitis, and in a short time without recurrence mark
As.
(3) drug of the invention increases mouse Colon length
Mouse Colon length changes table during table 1 is treated
As it can be seen from table 1 drug of the invention can significantly increase the colonic tissue of mouse after two courses for the treatment of, it is significant excellent
In two bacterium groups be combined with colostrum growth factor group, and it is little with antibiotic group mouse Colon length of tissue difference, illustrate two
The drug therapy of the present invention of the course for the treatment of will be restored to normal level, therefore, from mouse because of colonic tissue atrophy caused by colitis
From the point of view of colon lengths angle, drug of the invention can rise appreciably mouse Colon length of tissue, enhancing gut barrier effect.
(4) drug of the invention is obviously improved mouse pathology
Enteritis group, medicine group of the present invention (joint group), two bacterium groups are combined, colostrum growth factor group, antibiotic group into
Row comparison, as shown in Figure 1, enteritis group mouse histological scores are higher, illustrate enteritis group mouse intestinal mucosa ulcer, there is serious fill
Blood, suppuration phenomenon, after medicine group of the present invention (joint group) treatment by two courses for the treatment of, the colon macroscopic view disease of joint group mouse
Neo-Confucianism scoring significantly reduces, and be significantly better than two bacterium groups be combined, colostrum growth factor group, and it is aobvious with the nothing of antibiotic group mouse
Difference is write, illustrates drug of the present invention, two probiotics, which combine colostrum growth factor, has synergistic effect well, can be significant
The intestinal mucosa injury for repairing mouse Colon, makes it become neat, regular.
(5) drug of the invention is substantially reduced mouse cytokine inflammatory reaction
Enteritis group, medicine group of the present invention (joint group), two bacterium groups are combined, colostrum growth factor group, complete blank group,
Antibiotic group compares, and after the stomach-filling treatment of two courses for the treatment of, dissection mouse takes its blood, measures four in mice serum after centrifugation
Kind cytokine content, is shown in Table 2.As shown in Table 2, -1 β, IL-10 of enteritis group serum IL and TNF-α content are higher, due to mouse
Inflammatory factor is largely secreted after illness, mediated immunity reaction, and after the joint group drug therapy of two courses for the treatment of, mice serum
In above-mentioned cytokine content significantly reduce, substantially with antibiotic medication on the market without significant difference, and significant
Difference is in individual therapeutic effect, and therefore, drug of the present invention can be reacted by reducing cytokine levels come mediated immunity, thus
Play anti-ulcerative colitis purposes.
2 each group mice serum cytokine content of table compares (X ± SD)
Embodiment 1
A kind of drug for treating colitis, including 12 parts by volume probiotics and 64 mass parts colostrum growth factors.It is described
Colostrum growth factor be the growth factor crude extract extracted from colostrum；The probiotics is lactobacillus acidophilus
CGMCC10436 and lactobacillus plantarum CGMCC9961, and the quantity ratio of the two is 1:1, and is viable bacteria thallus form.It is described
Viable bacteria thallus be the bacterium mud being collected by centrifugation respectively by the lactobacillus liquid culture for being in stable initial stage.Wherein, parts by volume/
Mass parts=ml/mg, and following embodiment is same.
Also, the preparation method of the drug of above-mentioned treatment colitis, includes the following steps:
(1) respectively by lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum CGMCC9961 in MRS culture medium culture；
(2) the lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum CGMCC9961 for cultivating 12h are chosen respectively；
(3) 9 parts by volume of lactobacillus acidophilus CGMCC10436 bacterium solution for taking step (2) to obtain respectively, lactobacillus plantarum
Then they are dissolved in the sterile PBS of 1 parts by volume respectively, obtain Mixed Microbes by 5 parts by volume of CGMCC9961 bacterium solution, centrifuging and taking bacterium mud
Mud；
(4) colostrum growth is made to colostrum centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and freeze-drying
Factor crude extract；
(5) 64 mass parts of colostrum growth factor crude extract are added in above-mentioned mixing bacterium mud, are stirred evenly, then plus
Enter suitable auxiliary material and clinically-acceptable dosage form is made.
Embodiment 2
A kind of drug for treating colitis, including 16 parts by volume probiotics and 44 mass parts colostrum growth factors.Wherein
The colostrum growth factor is the growth factor crude extract extracted from colostrum；The probiotics is lactobacillus acidophilus
CGMCC10436 and lactobacillus plantarum CGMCC9961, and the quantity ratio of the two is 1:1, and is viable bacteria thallus form.Institute
The viable bacteria thallus stated is the bacterium mud being collected by centrifugation respectively by the lactobacillus liquid culture for being in stable initial stage.
Wherein, the preparation method for treating the drug of colitis, includes the following steps:
(1) respectively by lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum CGMCC9961 in MRS culture medium culture；
(2) the lactobacillus acidophilus CGMCC10436 and lactobacillus plantarum CGMCC9961 for cultivating 16h are chosen respectively；
(3) 11 parts by volume of lactobacillus acidophilus CGMCC10436 bacterium solution for taking step (2) to obtain respectively, lactobacillus plantarum
Then they are dissolved in the sterile PBS of 1 parts by volume respectively, obtain Mixed Microbes by 3 parts by volume of CGMCC9961 bacterium solution, centrifuging and taking bacterium mud
Mud；
(4) colostrum growth is made to colostrum centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and freeze-drying
Factor crude extract；
(5) 44 mass parts of colostrum growth factor crude extract are added in above-mentioned mixing bacterium mud, are stirred evenly, then plus
Enter suitable auxiliary material and clinically-acceptable dosage form is made.
Embodiment 3
A kind of drug for treating colitis, including 14 parts by volume probiotics and 54 mass parts colostrum growth factors.Wherein
The colostrum growth factor is the growth factor crude extract extracted from colostrum；The probiotics is lactobacillus acidophilus
CGMCC10436 and lactobacillus plantarum CGMCC9961, and the quantity ratio of the two is 1:1, and is viable bacteria thallus form.It is described
Viable bacteria thallus be the bacterium mud being collected by centrifugation respectively by the lactobacillus liquid culture for being in stable initial stage.
The preparation method of the drug of above-mentioned treatment colitis, includes the following steps:
(1) respectively by lactobacillus acidophilus CGMCC10436, lactobacillus plantarum CGMCC9961 in MRS culture medium culture；
(2) probiotics at the beginning of stationary phase in the logarithm end of term is chosen, i.e., chooses the lactobacillus acidophilus for cultivating 14h respectively
CGMCC10436 and lactobacillus plantarum CGMCC9961；
(3) 10 parts by volume of lactobacillus acidophilus CGMCC10436 bacterium solution for taking step (2) to obtain respectively, lactobacillus plantarum
Then they are dissolved in the sterile PBS of 1 parts by volume respectively, obtain Mixed Microbes by 4 parts by volume of CGMCC9961 bacterium solution, centrifuging and taking bacterium mud
Mud；
(4) colostrum growth is made to colostrum centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and freeze-drying
Factor crude extract；
(5) 54 mass parts of colostrum growth factor crude extract are added in above-mentioned mixing bacterium mud, are stirred evenly, then plus
Enter suitable auxiliary material and clinically-acceptable dosage form is made.
Claims (9)
1. a kind of composition for treating colitis, which is characterized in that including probiotics and colostrum growth factor, described is prebiotic
Bacterium is lactobacillus acidophilus (Lactobacillus acidophilus) CGMCC10436 and lactobacillus plantarum (Lactobacillus
Plantarum) CGMCC9961, the lactobacillus acidophilus thalline quantity account for the 20~80% of probio thalline total quantity.
2. the composition for the treatment of colitis according to claim 1, which is characterized in that the lactobacillus acidophilus and plant
The thalline quantity ratio of lactobacillus is 1:1.
3. the composition for the treatment of colitis according to claim 1, which is characterized in that the colostrum growth factor is
The growth factor crude extract extracted from colostrum；The probiotics is viable bacteria thallus form, the work of the probiotics
Bacterium thallus is the bacterium mud being collected by centrifugation respectively by the lactobacillus liquid culture for being in stable initial stage.
4. the composition for the treatment of colitis according to claim 1, which is characterized in that the probio thalline dosage is
2×1010CFU/kg, the colostrum growth factor dosage are 220~320mg/kg.
5. a kind of drug for treating colitis, which is characterized in that including composition described in Claims 1 to 4 any one.
6. the drug for the treatment of colitis according to claim 5, which is characterized in that the colitis includes exedens knot
Enteritis, infectious colitis, hemorrhagic colitis.
7. the preparation method of the drug for the treatment of colitis according to claim 5, which comprises the steps of:
(1) respectively by lactobacillus acidophilus and lactobacillus plantarum in 12~16h of MRS culture medium culture；
(2) 9~11 parts by volume of lactobacillus acidophilus bacterium solution for taking step (1) to obtain respectively, 3~5 parts by volume of lactobacillus plantarum bacterium solution,
Then they are dissolved in the sterile PBS of 1 parts by volume respectively, obtain mixing bacterium mud by centrifuging and taking bacterium mud；
(3) colostrum centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and dry obtained colostrum growth factor are slightly mentioned
Object；
(4) 44~64 mass parts of colostrum growth factor crude extract are added in above-mentioned mixing bacterium mud, are stirred evenly, then plus
Enter suitable auxiliary material and clinically-acceptable dosage form is made.
8. the preparation method of the drug for the treatment of colitis according to claim 7, which comprises the steps of:
(1) respectively by lactobacillus acidophilus, lactobacillus plantarum in MRS culture medium culture 14h；
(2) 10 parts by volume of lactobacillus acidophilus bacterium solution for taking step (1) to obtain respectively, 4 parts by volume of lactobacillus plantarum bacterium solution, centrifuging and taking
Then they are dissolved in the sterile PBS of 1 parts by volume respectively, obtain mixing bacterium mud by bacterium mud；
(3) colostrum centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and dry obtained colostrum growth factor are slightly mentioned
Object；
(4) 54 mass parts of colostrum growth factor crude extract are added in above-mentioned mixing bacterium mud, are stirred evenly, be then added and close
Clinically-acceptable dosage form is made in suitable auxiliary material.
9. the preparation method of the drug for the treatment of colitis according to claim 7 or 8, which is characterized in that step (3) is to ox
Colostrum is centrifuged at 4 DEG C, dry using freeze-drying.
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| CN112236155A (en) * | 2018-05-31 | 2021-01-15 | 深圳华大生命科学研究院 | Composition and application thereof |
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| WO2013008039A2 (en) * | 2011-07-14 | 2013-01-17 | Gt Biologics Ltd | Bacterial strains isolated from pigs |
| CN103705543A (en) * | 2013-12-25 | 2014-04-09 | 广州华农大实验兽药有限公司 | Manure-borne compound probiotics soluble powder for animals and preparation method thereof |
| CN105106246A (en) * | 2015-08-20 | 2015-12-02 | 江南大学 | Lactobacillus plantarum ZS2058 and application thereof |
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| WO2013008039A2 (en) * | 2011-07-14 | 2013-01-17 | Gt Biologics Ltd | Bacterial strains isolated from pigs |
| CN103705543A (en) * | 2013-12-25 | 2014-04-09 | 广州华农大实验兽药有限公司 | Manure-borne compound probiotics soluble powder for animals and preparation method thereof |
| CN105106246A (en) * | 2015-08-20 | 2015-12-02 | 江南大学 | Lactobacillus plantarum ZS2058 and application thereof |
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