CN105343132A - Composition for treating colitis, medicine and preparing method thereof - Google Patents
Composition for treating colitis, medicine and preparing method thereof Download PDFInfo
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- CN105343132A CN105343132A CN201510895295.8A CN201510895295A CN105343132A CN 105343132 A CN105343132 A CN 105343132A CN 201510895295 A CN201510895295 A CN 201510895295A CN 105343132 A CN105343132 A CN 105343132A
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Abstract
The invention discloses a composition for treating colitis, medicine and a preparing method thereof. According to the composition for treating colitis, probiotics and bovine coloctrum growth factors are used in cooperation. Balb/c mice are used as test objects, a colitis model is set up, and the colitis resisting effect of the medicine is studied from the aspects of the apparent state, the body weight change, the colon length, the pathology state and the like of the test mice. The test results show that the effect of the medicine is better than that of medicine singly using the probiotics or the bovine coloctrum growth factors; the the probiotics and the bovine coloctrum growth factors have a cooperative synergism effect, the medicine has a good colitis resistant effect, colitis will not relapse within a short term, and intestinal mucosa injuries caused by colitis can be well repaired.
Description
Technical field
The present invention relates to a kind of compositions, medicine and preparation method thereof for the treatment of colitis, namely the medicine of two probiotics and cattle colostrums somatomedin conbined usage, belongs to medical art.
Background technology
Colitis is also known as nonspecific ulcerative colitis, it is of a great variety, cause reason various, and how slow onset is, state of an illness weight differs, and main clinical manifestation diarrhoea, stomachache, Mucous Stool and bloody purulent stool, tenesmus, very then constipation, can not lead to stool in a few days, time and the time of diarrhoea and constipation, often weak etc. with becoming thin, many recurrent exerbation.Stomachache mostly generally is dull pain or angor, is often positioned at left lower quadrant or lower abdomen.Other performance has inappetence, abdominal distention, Nausea and vomiting and hepatomegaly etc., and left lower quadrant can have tenderness, sometimes can touch the colon of spasm.Common General Symptoms is become thin, weak, heating, anemia etc.Have small part patient in the chronic course of disease, the state of an illness worsens suddenly or first attack is just fulminant, and performance severe diarrhea, brings great misery to human or animal.
The method of current treatment colitis mainly comprises Drug therapy, nutrition treatment, psychotherapy and operative treatment, and its drug treatment is topmost method.The medicine being used for the treatment of colitis clinically mainly contains minosalicylates, drugs, Adrenal Glucocorticoid class and immunosuppressant class etc.Minosalicylates, drugs can well suppress prostaglandin to synthesize, scavenging activated oxygen thus the reaction that reduces inflammation; But it cannot effect a radical cure colitis, and when patient consumes's dosage is bigger than normal, easily produce many untoward reaction (as: headache, feel sick, leukopenia etc.).Glucocorticoid is the drug of first choice of severe or explosive colitis disease; But such medicine easily causes the side effect such as organism metabolic disorder, water retention, can be used as emergent medicine, cannot life-time service.Immunosuppressant class medication medication dependency is large, and the treatment phase is longer, and possibility intoxicating is carcinogenic, and side effect is comparatively large, only uses as auxiliary treatment so general.On the whole, said medicine is responded well to treatment soon, good effect, can alleviate the hardship of colitis disease's matter as a fire singeing one's eyebrows, but still there is a lot of problem.First, patient's colitis cannot be effected a radical cure, cure the symptoms, not the disease; Secondly, ill effect is large, easily causes patient's headache, feels sick, cause body digestive system, hematological abnormalities; Then, potential intoxicating carcinogenecity; Finally, antibiotics (SASP as most widely used) easily causes body drug resistance Drug resistance.
Also patent report is had to adopt Chinese medicine composition to treat colitis, as the patent of invention " medicine for the treatment of chronic colitis and ulcerative colitis and preparation method " that application number is 200710066007.3, application number is the patent of invention " a kind of medicine for the treatment of ulcerative colitis and preparation method thereof " of 200610167659.1, due to the complicated component of these Chinese medicines, curative effect is slow, the course for the treatment of is long, therapeutic effect is also difficult to ensure, the medicine that therefore these Chinese medicine compositions are made is not applied so far on a large scale.So for this disease being difficult to cure for the treatment of colitis, research and develop a kind of new medicine tool and be of great significance.Though there is bibliographical information probiotic bacteria to treat ulcerative colitis, but at present also not about the report adopting probiotic combination cattle colostrums somatomedin conbined usage treatment colitis.
Summary of the invention
An object of the present invention is to provide a kind of compositions for the treatment of colitis, effectively can treat colitis, and toxic and side effects is less, not easily produces resistance.
Another object of the present invention is to provide a kind of medicine for the treatment of colitis and preparation method thereof, uses the compositions treating colitis, the effect that can reach equally.
To achieve these goals, the invention provides following technical scheme:
A kind of compositions for the treatment of colitis, comprise probiotic bacteria and cattle colostrums somatomedin, described probiotic bacteria is bacillus acidophilus (Lactobacillusacidophilus) CGMCC10436 and Lactobacillus plantarum (Lactobacillusplantarum) CGMCC9961, and described bacillus acidophilus's thalline quantity accounts for 20 ~ 80% of probiotic bacteria thalline total quantity.
Preferably, the thalline number ratio of described bacillus acidophilus and Lactobacillus plantarum is 1:1.
Described cattle colostrums somatomedin is the somatomedin crude extract extracted from cattle colostrums; Described probiotic bacteria is viable bacteria thalline form, and the viable bacteria thalline of described probiotic bacteria is distinguished the bacterium mud of collected by centrifugation by the lactobacillus liquid culture being all in the stable initial stage.
Described probiotic bacteria thalline consumption is 2 × 10
10cFU/kg, described cattle colostrums somatomedin consumption is 220 ~ 320mg/kg.Namely in the medicine that the animal, human body etc. of every kilogram are used, the total quantity of bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961 is 2 × 10
10cFU, cattle colostrums somatomedin consumption is 220 ~ 320mg, particularly, can calculate according to practical situation, such as, a weight is the rat of 20g, and the total quantity containing bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961 in its medication is 2 × 10
9cFU, cattle colostrums somatomedin consumption is 4.4 ~ 6.4mg; A weight is the people of 50kg, and the total quantity containing bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961 in its medication is 1 × 10
12cFU, cattle colostrums somatomedin consumption is 11 ~ 16g.
Wherein, described compositions can make all article that can retain its activity, as milk, medicine, food etc.
The invention also discloses a kind of treat colitis medicine, comprise aforesaid compositions.
Described colitis comprises ulcerative colitis, infectious colitis, hemorrhagic colitis, and is not limited thereto.
The preparation method of the medicine of above-mentioned treatment colitis, comprises the steps:
(1) respectively by bacillus acidophilus and Lactobacillus plantarum at MRS culture medium culturing 12 ~ 16h;
(2) get bacillus acidophilus bacterium liquid 9 ~ 11 parts by volume that step (1) obtains respectively, Lactobacillus plantarum bacterium liquid 3 ~ 5 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 1 parts by volume, obtain mixed vaccine mud;
(3) to cattle colostrums centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and dry obtained cattle colostrums somatomedin crude extract;
(4) cattle colostrums somatomedin crude extract 44 ~ 64 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.Here parts by volume and mass parts are interpreted as: if with bacillus acidophilus bacterium liquid, 3 ~ 5ml Lactobacillus plantarum bacterium liquid of 9 ~ 11ml in step (2), then to the cattle colostrums somatomedin crude extract of application 44 ~ 64mg, i.e. parts by volume/mass parts=ml/mg.
Preferably, the preparation method of the medicine of described treatment colitis, comprises the steps:
(1) respectively by bacillus acidophilus, Lactobacillus plantarum at MRS culture medium culturing 14h;
(2) get bacillus acidophilus bacterium liquid 10 parts by volume that step (1) obtains respectively, Lactobacillus plantarum bacterium liquid 4 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 1 parts by volume, obtain mixed vaccine mud;
(3) to cattle colostrums centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and dry obtained cattle colostrums somatomedin crude extract; Preferably, centrifugal at 4 DEG C to cattle colostrums, dry employing lyophilization.
(4) cattle colostrums somatomedin crude extract 54 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.
The preserving number of bacillus acidophilus CGMCC10436 of the present invention is CGMCCNo.10436, preservation date be 2015 01 month No. 26, the preserving number of described Lactobacillus plantarum CGMCC9961 is CGMCCNo.9961, preservation date is on November 13rd, 2014, depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
Beneficial effect of the present invention:
Cattle colostrums somatomedin by being combined with protection of intestinal mucosal barrier cells tyrosine kinase receptor, thus impels protection of intestinal mucosal barrier cells to grow and propagation, increases villi height, promotes intestinal growth; And promote immunity of organisms by regulation and control inflammatory factor level.The probiotic bacteria that the present invention adopts is by carrying out regulating intestinal canal colony balance with harmful bacteria competing binding sites, and Lactobacillus plantarum CGMCC9961 can produce bacteriocin, suppresses the propagation of harmful bacteria; Bacillus acidophilus CGMCC10436 has good prebiotic effect.Result shows, and the medicine that both conbined usage are made has the effect of good resistive connection enteritis.And there is the effect of Synergistic.Its effect than being used alone two kinds of bacterium, to be used alone cattle colostrums somatomedin effect good.Be in particular in the apparent state significantly improving colitis mice, increase body weight, the colon lengths of colitis mice, and significantly improve the integrity of mouse Colon tissue.
Accompanying drawing explanation
Fig. 1 is the scoring of separate groups of mice Gross pathology.
Detailed description of the invention
When the present inventor feels a delicacy about for the treatment of current global field of medicaments to colitis, can enhancing human body immunity power according to probiotic bacteria and somatomedin, good therapeutic effect is had to human body intestinal canal disease (as diarrhoea etc.), the good living preparation of lactobacillus thalline of two strain probiotic effects is combined with cattle colostrums somatomedin crude extract and is made medicine by the present invention, for animal experiment research, it is below detailed description of the present invention.
Embodiment is only exemplary, does not form any restriction to scope of the present invention.It will be understood by those skilled in the art that and can modify to the details of technical solution of the present invention and form or replace down without departing from the spirit and scope of the present invention, but these amendments and replacement all fall within the scope of protection of the present invention.Various material of the present invention, reagent or apparatus if no special instructions, all can obtain from market.
In order to prove effectiveness of the present invention, applicant carried out following test.
(1) material and reagent
1, culture medium: MRS culture medium.
MRS culture medium is prepared: peptone 5g/L, Carnis Bovis seu Bubali cream 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 0.25g/L, tween 80 1ml/L, pH6.3 ± 0.1, sterilizing in 121 DEG C, 15 minutes.
2, main agents: cattle colostrums is from Wanda Mountain, Heilongjiang Song Bei pasture; Dextran sulfate sodium salt wins company of biotechnology Co., Ltd from Beijing bit; Sulfasalazine is from Shanghai Xinyi Tianping Pharmaceutical Co., Ltd.; Formalin is from Bo Di chemical inc, Tianjin; Potassium dihydrogen phosphate is from the huge chemical apparatuses factory in Dongli District, Tianjin; Sodium hydrogen phosphate is from Tianjin BASF Chemical Co., Ltd..
3, key instrument: ultramicroscope is from Motic company; Ultraviolet-uisible spectrophotometer is from BECKMAN company of the DU800 U.S.; Ophthalmology anatomy box is from Jiu Ming aluminum products factory of Shenyang City; Syringe is from Jiangsu Zhi Yu Medical Devices Co., Ltd..
(2), being separated of bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961
1, from the herdsman family in the areas such as In Xilingol League In Inner Mongolia, Hailar City and Ordos City, the rare butter of tradition or the acidify butter of traditional method making is gathered, for the screening of lactobacillus.Specific as follows:
(1) 25g sample is aseptically shredded, be dissolved in 225mL physiological saline solution, after gradient dilution, select suitable dilution factor, get 1mL diluent and add sterilized petri dishes, be poured on 20mLMRS solid medium, cultivate 72h for 30 DEG C.
(2) choose colony counts 15 ~ 150 flat board, random selecting meets the bacterium colony of lactobacillus colony morphology characteristic, and therefrom a small amount of antibacterial of picking, is dissolved in MRS fluid medium, and carry out 30 DEG C cultivate 48h.
(3) by the bacterial strain be separated on MRS solid medium repeatedly line carry out purification.
(4) bacterial strain good for purification is all carried out Gram’s staining and catalase experiment.
(5) all Gram-positives, negative catalase, the bacterial strain (being tentatively judged as lactobacillus) that do not form spore are elected, glycerol is added in its culture fluid, make the ultimate density of glycerol be 30% (v/v), be stored in-80 DEG C stand-by.
(6) above-mentioned being separated to after the bacterial strain of fixing tentatively as lactobacillus activated for 3 generations respectively in MRS fluid medium is carried out identification experiment.
2, the qualification of bacillus acidophilus CGMCC10436
(1) Physiology and biochemistry qualification
With reference culture bacillus acidophilus bacterial strain in contrast, carry out following physiological and biochemical test: H
2s test, methyl red test, litmus milk tests, gelatin liquefaction test, Starch Hydrolysis test, citrate utilize test, urea decomposition test, nitrate Degrading experiment and acetoin to produce test, choose the lactobacillus identical with reference culture bacillus acidophilus physiological and biochemical test.
(2) Molecular Identification
Extract strain gene group DNA, carry out 16SrDNA order-checking, its nucleotide sequence is as shown in sequence table SeqIDNo1, the 16SrDNA sequence of bacterial strain of the present invention and the sequence of NCBI bacillus acidophilus are compared, 16SrDNA sequence and the Lactobacillusacidophilus sequence similarity of bacterial strain of the present invention reach more than 95%, according to 16SrDNA sequence analysis, this bacterial strain is accredited as bacillus acidophilus (Lactobacillusacidophilus), and be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 01 26th, 2015, microbial preservation number is CGMCCNo.10436, preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
3, the qualification of Lactobacillus plantarum CGMCC9961
(1) Physiology and biochemistry qualification
With standard plant lactobacillus bacterial strain in contrast, carry out following physiological and biochemical test: H
2s test, methyl red test, litmus milk tests, gelatin liquefaction test, Starch Hydrolysis test, citrate utilize test, urea decomposition test, nitrate Degrading experiment and acetoin to produce test, choose the lactobacillus identical with Lactobacillus plantarum physiological and biochemical test.
(2) Molecular Identification
Extract strain gene group DNA, carry out 16SrDNA order-checking, its nucleotide sequence is as shown in sequence table SeqIDNo2, the 16SrDNA sequence of bacterial strain of the present invention and the sequence of NCBI Lactobacillus plantarum are compared, 16SrDNA sequence and the Lactobacillusplantarum sequence similarity of bacterial strain of the present invention reach more than 95%, according to 16SrDNA sequence analysis, identify, this bacterial strain is bacillus acidophilus (Lactobacillusplantarum).Deposit number is CGMCCNO.9961, preservation date is on November 13rd, 2014, depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), and address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
(3) preparation of gavage probiotic bacteria
Bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961 is cultivated 14h respectively, carries out gradient plate counting, the bacterium number needed for bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961 every mice is 2 × 10
8cFU, get the bacillus acidophilus CGMCC10436 bacterium liquid 1mL that equal stable phase goes out respectively, Lactobacillus plantarum CGMCC9961 bacterium liquid 0.4mL, centrifuging and taking bacterium mud, is then refitted in 0.1mLPBS respectively, for the probiotic bacteria given low of every mice.
(4) preparation of cattle colostrums somatomedin crude extract
By the cattle colostrums somatomedin crude extract obtained to cattle colostrums centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and lyophilization; The processing method such as centrifugal, ultrafiltration, dialysis, drying of indication of the present invention, can use any method known to a person of ordinary skill in the art, as long as it can keep somatomedin effect, preferably talk about, centrifugal pass through 4 DEG C centrifugal, dryly adopt lyophilization.
(5) preparation of the medicine of colitis is treated
Cattle colostrums somatomedin crude extract, given low is 5.4mg/20g (cattle colostrums somatomedin crude extract quality/mice quality); The gavage form of cattle colostrums somatomedin crude extract is, takes out the bacillus acidophilus be dissolved in the aseptic PBS solution of 0.1mL, Lactobacillus plantarum bacterium mud tubule, mixing, then adds the cattle colostrums somatomedin crude extract of described dosage.
(6) gavage of other group mices prepares
Antibiotic group mouse stomach medicine: antibiotic given low is 9.2mg/20g (willow nitrogen thiamine pyridine opaque amount/mice weights); The gavage form of willow nitrogen thiamine pyridine is, takes out and is dissolved in the aseptic PBS solution of 0.3mL, and mixing, then 4 DEG C of preservations, in order to gavage.
Two bacterium combination group mouse stomach medicines:
By bacillus acidophilus on the market and Lactobacillus plantarum the same carry out microorganism counting, the process such as centrifugal, get respectively and be in the first bacillus acidophilus of stable phase and Lactobacillus plantarum each 2 × 10
8cFU, both mixing, are then dissolved in the aseptic PBS solution of 0.3mL, and mixing, then 4 DEG C of preservations, in order to gavage.
Below in conjunction with zoopery, further research is done to medicine of the present invention, so that effectiveness of the present invention to be described.
(1) making of mouse models of colitis and EXPERIMENTAL DESIGN
Dextran sulfate sodium (DSS) is dissolved in water bottle, Balb/c mice is allowed freely to drink 8 days, with provocative test mouse models of colitis, then with Mouse Weight change, apparent situation, colon lengths, pathology etc. for index, checking mouse colitis model whether be successfully established.
During modeling, Mouse Weight declines, and enteritis group mice is apparent is not in good state, and is in particular in that mice is One's eyesight is restrained, hair is obscure curling, has Qiang's hair phenomenon; Further, there is ecchymosis at enteritis group mice root of the tail place, illustrates that mice has dominant phenomenon of having blood in stool; There is phenomenon of festering at enteritis group mouse Colon place after dissection, and intestinal tissue has sends out pus, turbidity phenomenon; And during modeling, enteritis group mouse Colon length naive mice colon significantly shortens; Further, from HE colored graph, model group mouse Colon tissue, intestinal mucosa atrophy, mucous membrane surface passivation, crypts destroys.
In sum, acute colitis model is successfully established, and can carry out next step Drug therapy test.
(2) experiment conclusion
(1) explanation is tested
Treatments period, only carries out PBS gavage to enteritis naive mice, does not treat, dead 11 of enteritis naive mice during result gavage; Illustrate that colitis model is successfully established on the one hand, illustrate that colitis mice can not be fully recovered by self-regeneration in addition on the one hand; So follow-up contrast, is across comparison, the contrast namely before and after same group, the contrast after mice treatment and before treatment.
(2) medicine of the present invention obviously improves the apparent state of mice
After the gavage of two courses for the treatment of terminates, the poly-god of medicine group mice sight of the present invention, action are active, and mouse hair is smooth, brilliant white, compared with apparent state during enteritis, difference is extremely obvious, and compared with two bacterium combination groups and independent cattle colostrums somatomedin group, the apparent state of mice obviously improves; And compared with the apparent state of antibiotic group mice, difference is not remarkable.Illustrate that medicine of the present invention (probiotic combination cattle colostrums somatomedin) has good effect to treatment colitis, and in a short time without recurrence sign.
(3) medicine of the present invention increases mouse Colon length
Table 1 treatments period mouse Colon length variations table
As can be seen from Table 1, after two courses for the treatment of, medicine of the present invention significantly can increase the colon of mice, significantly be better than two bacterium combination groups and cattle colostrums somatomedin group, and little with antibiotic group mouse Colon length of tissue difference, illustrate that the colon's atrophy caused because of colitis has been returned to normal level by the Drug therapy of the present invention of two courses for the treatment of, therefore, from mouse Colon length angle, medicine of the present invention can rise appreciably mouse Colon length of tissue, strengthens gut barrier effect.
(4) medicine of the present invention obviously improves mice pathology
By enteritis group, medicine group (combining group) of the present invention, two bacterium combination groups, cattle colostrums somatomedin group, antibiotic group contrasts, as shown in Figure 1, enteritis group mice histological scores is higher, enteritis group mice intestinal mucosa ulcer is described, there is serious hyperemia, suppuration phenomenon, after medicine group (the combining group) treatment of the present invention of two courses for the treatment of, the colon Gross pathology scoring of combining group mice significantly reduces, and be significantly better than two bacterium combination groups, cattle colostrums somatomedin group, and with antibiotic group mice without significant difference, medicine of the present invention is described, two probiotics associating cattle colostrums somatomedin have good synergism, the intestinal mucosa injury of mouse Colon can be repaired significantly, it is made to become neat, rule.
(5) medicine of the present invention obviously reduces mouse cytokine inflammatory reaction
By enteritis group, medicine group of the present invention (combining group), two bacterium combination groups, cattle colostrums somatomedin group, complete blank group, antibiotic group contrasts, after the gavage treatment of two courses for the treatment of, dissect mice and get its blood, four kinds of cytokine content in centrifugal rear mensuration mice serum, in table 2.As shown in Table 2, enteritis group serum IL-1 β, IL-10 and TNF-alpha content is higher, because the ill rear inflammatory factor of mice is secreted in a large number, mediated immunity reacts, and through two courses for the treatment of combine group Drug therapy after, above-mentioned cytokine content in mice serum significantly reduces, substantially with antibiotic medication on the market without significant difference, and significant difference is in independent therapeutic effect, therefore, medicine of the present invention carrys out mediated immunity reaction by reducing cytokine levels, thus plays anti-ulcerative colitis purposes.
Table 2 respectively group mice serum cytokine content compares (X ± SD)
Embodiment 1
Treat a medicine for colitis, comprise 12 parts by volume probiotic bacterias and 64 mass parts cattle colostrums somatomedin.Described cattle colostrums somatomedin is the somatomedin crude extract extracted from cattle colostrums; Described probiotic bacteria is bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961, and both number ratio are 1:1, and is viable bacteria thalline form.Described viable bacteria thalline is by the bacterium mud of the lactobacillus liquid culture difference collected by centrifugation being all in the stable initial stage.Wherein, parts by volume/mass parts=ml/mg, and following examples are same.
Further, the preparation method of the medicine of above-mentioned treatment colitis, comprises the steps:
(1) respectively by bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961 at MRS culture medium culturing;
(2) the bacillus acidophilus CGMCC10436 all cultivating 12h and Lactobacillus plantarum CGMCC9961 is chosen respectively;
(3) get bacillus acidophilus CGMCC10436 bacterium liquid 9 parts by volume that step (2) obtains respectively, Lactobacillus plantarum CGMCC9961 bacterium liquid 5 parts by volume, then they are dissolved in the aseptic PBS of 1 parts by volume, obtain mixed vaccine mud by centrifuging and taking bacterium mud respectively;
(4) cattle colostrums somatomedin crude extract is obtained to cattle colostrums centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and lyophilization;
(5) cattle colostrums somatomedin crude extract 64 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.
Embodiment 2
Treat a medicine for colitis, comprise 16 parts by volume probiotic bacterias and 44 mass parts cattle colostrums somatomedin.Wherein said cattle colostrums somatomedin is the somatomedin crude extract extracted from cattle colostrums; Described probiotic bacteria is bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961, and both number ratio are 1:1, and is viable bacteria thalline form.Described viable bacteria thalline is by the bacterium mud of the lactobacillus liquid culture difference collected by centrifugation being all in the stable initial stage.
Wherein, the preparation method of the medicine for the treatment of colitis, comprises the steps:
(1) respectively by bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961 at MRS culture medium culturing;
(2) the bacillus acidophilus CGMCC10436 all cultivating 16h and Lactobacillus plantarum CGMCC9961 is chosen respectively;
(3) get bacillus acidophilus CGMCC10436 bacterium liquid 11 parts by volume that step (2) obtains respectively, Lactobacillus plantarum CGMCC9961 bacterium liquid 3 parts by volume, then they are dissolved in the aseptic PBS of 1 parts by volume, obtain mixed vaccine mud by centrifuging and taking bacterium mud respectively;
(4) cattle colostrums somatomedin crude extract is obtained to cattle colostrums centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and lyophilization;
(5) cattle colostrums somatomedin crude extract 44 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.
Embodiment 3
Treat a medicine for colitis, comprise 14 parts by volume probiotic bacterias and 54 mass parts cattle colostrums somatomedin.Wherein said cattle colostrums somatomedin is the somatomedin crude extract extracted from cattle colostrums; Described probiotic bacteria is bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961, and both number ratio are 1:1, and is viable bacteria thalline form.Described viable bacteria thalline is by the bacterium mud of the lactobacillus liquid culture difference collected by centrifugation being all in the stable initial stage.
The preparation method of the medicine of above-mentioned treatment colitis, comprises the steps:
(1) respectively by bacillus acidophilus CGMCC10436, Lactobacillus plantarum CGMCC9961 at MRS culture medium culturing;
(2) choose logarithm end of term stable phase probiotic bacteria just, namely choose the bacillus acidophilus CGMCC10436 and Lactobacillus plantarum CGMCC9961 that all cultivate 14h respectively;
(3) get bacillus acidophilus CGMCC10436 bacterium liquid 10 parts by volume that step (2) obtains respectively, Lactobacillus plantarum CGMCC9961 bacterium liquid 4 parts by volume, then they are dissolved in the aseptic PBS of 1 parts by volume, obtain mixed vaccine mud by centrifuging and taking bacterium mud respectively;
(4) cattle colostrums somatomedin crude extract is obtained to cattle colostrums centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and lyophilization;
(5) cattle colostrums somatomedin crude extract 54 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.
Claims (9)
1. treat the compositions of colitis for one kind, it is characterized in that, comprise probiotic bacteria and cattle colostrums somatomedin, described probiotic bacteria is bacillus acidophilus (Lactobacillusacidophilus) CGMCC10436 and Lactobacillus plantarum (Lactobacillusplantarum) CGMCC9961, and described bacillus acidophilus's thalline quantity accounts for 20 ~ 80% of probiotic bacteria thalline total quantity.
2. the compositions for the treatment of colitis according to claim 1, is characterized in that, the thalline number ratio of described bacillus acidophilus and Lactobacillus plantarum is 1:1.
3. the compositions for the treatment of colitis according to claim 1, is characterized in that, described cattle colostrums somatomedin is the somatomedin crude extract extracted from cattle colostrums; Described probiotic bacteria is viable bacteria thalline form, and the viable bacteria thalline of described probiotic bacteria is distinguished the bacterium mud of collected by centrifugation by the lactobacillus liquid culture being all in the stable initial stage.
4. the compositions for the treatment of colitis according to claim 1, is characterized in that, described probiotic bacteria thalline consumption is 2 × 10
10cFU/kg, described cattle colostrums somatomedin consumption is 220 ~ 320mg/kg.
5. treat a medicine for colitis, it is characterized in that, comprise the compositions described in Claims 1 to 4 any one.
6. the medicine for the treatment of colitis according to claim 5, is characterized in that, described colitis comprises ulcerative colitis, infectious colitis, hemorrhagic colitis.
7. the preparation method of the medicine for the treatment of colitis according to claim 5, is characterized in that, comprise the steps:
(1) respectively by bacillus acidophilus and Lactobacillus plantarum at MRS culture medium culturing 12 ~ 16h;
(2) get bacillus acidophilus bacterium liquid 9 ~ 11 parts by volume that step (1) obtains respectively, Lactobacillus plantarum bacterium liquid 3 ~ 5 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 1 parts by volume, obtain mixed vaccine mud;
(3) to cattle colostrums centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and dry obtained cattle colostrums somatomedin crude extract;
(4) cattle colostrums somatomedin crude extract 44 ~ 64 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.
8. the preparation method of the medicine for the treatment of colitis according to claim 7, is characterized in that, comprise the steps:
(1) respectively by bacillus acidophilus, Lactobacillus plantarum at MRS culture medium culturing 14h;
(2) get bacillus acidophilus bacterium liquid 10 parts by volume that step (1) obtains respectively, Lactobacillus plantarum bacterium liquid 4 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 1 parts by volume, obtain mixed vaccine mud;
(3) to cattle colostrums centrifugal degreasing, the heavy casein of acid, second ultrafiltration, dialysis and dry obtained cattle colostrums somatomedin crude extract;
(4) cattle colostrums somatomedin crude extract 54 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.
9. the preparation method of the medicine of the treatment colitis according to claim 7 or 8, is characterized in that, step (3) is centrifugal at 4 DEG C to cattle colostrums, dry employing lyophilization.
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Effective date of registration: 20220114 Address after: 166500 Manjianghong street, Zhaoyuan County, Daqing City, Heilongjiang Province (east of Heilongjiang Tianyuan Food Co., Ltd.) Patentee after: Daqing Shi Shi Yuan Dairy Co.,Ltd. Address before: 150030 Northeast Agricultural University, Wood Street, Xiangfang District, Harbin City, Heilongjiang Province Patentee before: Northeast Agricultural University |