CN105327338A - Medicine for treating ulcerative colitis and preparation method of medicine - Google Patents

Abstract

The invention discloses a medicine for treating ulcerative colitis and a preparation method of the medicine. The medicine for treating ulcerative colitis is prepared from bovine coloctrum growth factors and probiotics in combination. A Balb/c mouse is taken as a test object, an ulcerative colitis model is established, and the ulcerative colitis resistant effect of the medicine is researched in the aspects about the appearance state, change of weight, the length of the colon, the pathological condition and the like. A test result shows that the medicine has a good ulcerative colitis resistant effect, the ulcerative colitis cannot relapse soon, intestinal mucosa caused by ulcerative colitis can be recovered well, and the medicine can be widely applied to the field of foundation medicine.

Description

A kind of medicine for the treatment of ulcerative colitis and preparation method thereof

Technical field

The present invention relates to a kind of medicine for the treatment of ulcerative colitis and preparation method thereof, namely the medicine of three probiotics and cattle colostrums somatomedin conbined usage, belongs to medical art.

Background technology

Ulcerative colitis (ulcerativecolitis, UC) is a main Types of inflammatory bowel (inflammatoryboweldisease, IBD), is a kind of agnogenic chronic nonspecific enteritis.Data shows, and annual U.S. UC prevalence is 2,00/,100,000 people, and treatment UC spends in about 1 to 1.5 hundred million dollar, and by the end of 2012, UC patient populations was 2.5 times of 2000, and relapse rate reaches 72%; The number of patients of China UC is obvious ascendant trend in recent years.

Ulcerative colitis pathological changes is mainly positioned at mucous membrane of colon and tela submucosa; Clinical manifestation is diarrhoea, mucus bloody purulent stool, stomachache, tenesmus etc., many chronic courses of disease in recurrent exerbation.The generation of ulcerative colitis has badly influenced the healthy of patient and quality of life, is classified as one of modern difficult treatment by World Health Organization (WHO).The cause and onset of disease mechanism of ulcerative colitis is still not fully aware of.Think that the cause of disease of UC may be relevant with environmental factors, inherited genetic factors, immune factor etc. more; Wherein, heredity and immune factor play vital effect in UC causes a disease.Though UC pathogenesis is not also illustrated completely, can affirm that function of intestinal mucosa barrier in patient imbalance take part in the generation of UC.Gut barrier mainly comprises mechanical barrier, immunologic barrier and biological barrier.Mechanical barrier is made up of enterocyte, intestinal mucus; Immunologic barrier (i.e. intestinal immune system of defense), is formed primarily of secreted immunoglobulin and gut associated lymphoid tissue; Biological barrier is made up of normal intestinal flora.Gut epithelium barrier breakdown, mucosa permeability increases, intestinal tissue is exposed in a large amount of antigen for a long time, cause intestinal tract immune system overreaction and wrong identification, cause macrophage and lymphocytic activation, discharge a series of cytokine and inflammatory mediator, activate the immunne response of body, inflammatory reaction is amplified step by step, finally causes tissue injury, occurs pathological change and the clinical manifestation of UC.

The method of current treatment ulcerative 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 UC 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 UC, 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 UC patient; 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 UC patient's matter as a fire singeing one's eyebrows, but still there is a lot of problem.First, patient UC 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.

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.Probiotic bacteria is by carrying out regulating intestinal canal colony balance with harmful bacteria competing binding sites, and Lactobacillus plantarum can produce bacteriocin, suppresses the propagation of harmful bacteria; Bacillus acidophilus has good prebiotic effect.Therefore, we suppose that cattle colostrums somatomedin associating probiotic bacteria has good anti-ulcerative colitis purposes.Therefore, develop a kind of efficient, safety and the medicine of the treatment UC of the less safety of side effect, be of great practical significance.

Summary of the invention

An object of the present invention is to provide a kind of medicine for the treatment of ulcerative colitis, effectively can treat ulcerative colitis, and toxic and side effects is less, not easily produces resistance.

Another object of the present invention is to provide a kind of preparation method for the treatment of the medicine of ulcerative colitis.

To achieve these goals, the invention provides following technical scheme:

Treat a medicine for ulcerative colitis, comprise 132 ~ 198 parts by volume probiotic bacterias and 432 ~ 684 mass parts cattle colostrums somatomedin, wherein parts by volume/mass parts=ml/mg.

Preferably, the medicine of above-mentioned treatment ulcerative colitis, comprises 165 parts by volume probiotic bacterias and 540 mass parts cattle colostrums somatomedin.Described cattle colostrums somatomedin is the somatomedin crude extract extracted from cattle colostrums.Described probiotic bacteria is bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus; And be viable bacteria thalline form.Described probiotics viable bacteria thalline is by the bacterium mud of the lactobacillus liquid culture difference collected by centrifugation being all in the stable initial stage.

Preferably, the number ratio of described bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus is 1: 1: 1.

In the medicine of above-mentioned treatment ulcerative colitis, described probiotic bacteria consumption is 1.5 × 10 ¹⁰cFU/kg, described cattle colostrums somatomedin consumption is 216 ~ 342mg/kg.The total quantity of bacillus acidophilus in the medicine that namely animal, human body etc. of every kilogram are used, Lactobacillus plantarum and lactobacillus helveticus is 1.5 × 10 ¹⁰cFU, cattle colostrums somatomedin consumption is 216 ~ 342mg, particularly, can calculate according to practical situation, and such as, a weight is the rat of 20g, and the total quantity containing bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus in its medication is 3 × 10 ⁸cFU, cattle colostrums somatomedin consumption is 4.32 ~ 6.84mg; A weight is the people of 50kg, and the total quantity of bacillus acidophilus in its medication, Lactobacillus plantarum and lactobacillus helveticus is 7.5 × 10 ¹¹cFU, cattle colostrums somatomedin consumption is 10.8 ~ 17.1g.

The preparation method of the medicine of aforementioned therapies ulcerative colitis, comprises the steps:

(1) respectively by bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus at MRS culture medium culturing;

(2) choose the lactobacillus helveticus of cultivation 14 ~ 18h respectively, cultivate the Lactobacillus plantarum of the bacillus acidophilus of 12 ~ 16h and cultivation 12 ~ 16h;

(3) lactobacillus helveticus bacterium liquid 77 ~ 115 parts by volume that step (2) obtains is got respectively, bacillus acidophilus bacterium liquid 40 ~ 60 parts by volume, Lactobacillus plantarum bacterium liquid 15 ~ 23 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 10 parts by volume, obtain mixed vaccine mud;

(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 432 ~ 684 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.

Preferably, the preparation method of the medicine for the treatment of ulcerative colitis, comprises the steps:

(1) respectively by bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus at MRS culture medium culturing;

(2) choose logarithm end of term stable phase probiotic bacteria just, namely choose the lactobacillus helveticus of cultivation 16h, the bacillus acidophilus of cultivation 14h and the Lactobacillus plantarum of cultivation 14h respectively;

(3) lactobacillus helveticus bacterium liquid 96 parts by volume that step (2) obtains is got respectively, bacillus acidophilus bacterium liquid 50 parts by volume, Lactobacillus plantarum bacterium liquid 19 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 10 parts by volume, obtain mixed vaccine mud;

(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 540 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.

Beneficial effect of the present invention:

The medicine that the present invention makes with probiotic bacteria and cattle colostrums somatomedin crude extract has good anti-ulcerative colitis effect.Be in particular in the apparent state significantly improving ulcerative colitis mice, increase body weight, the colon lengths of ulcerative colitis mice, and significantly improve the integrity of mouse Colon tissue.

Accompanying drawing explanation

Fig. 1 is bacillus acidophilus's microscopy picture (100 times);

Fig. 2 is bacillus acidophilus's bacterium colony figure;

Fig. 3 is lactobacillus helveticus microscopy picture (100 times);

Fig. 4 is lactobacillus helveticus bacterium colony figure;

Fig. 5 is Lactobacillus plantarum microscopy picture (100 times);

Fig. 6 is Lactobacillus plantarum bacterium colony figure;

Fig. 7 is three strain lactobacillus growth curves;

Fig. 8 is the apparent state of enteritis group mice;

Fig. 9 is the apparent state feature of enteritis group mice;

Figure 10 is enteritis group mice internal anatomy;

Figure 11 is enteritis group mouse Colon length representative figure;

Figure 12 is complete naive mice colon lengths representative graph;

Figure 13 is enteritis group mice HE colored graph;

Figure 14 is plan design figure;

Figure 15 is for combining the apparent state diagram of group mice;

Figure 16 is for combining group mice internal anatomy;

Figure 17 is that later observation combines the apparent state of group mice;

Figure 18 is the apparent state of antibiotic group mice;

Figure 19 is antibiotic group mice internal anatomy;

Figure 20 is the apparent state of later observation antibiotic group mice;

Figure 21 is the apparent state of complete naive mice;

Figure 22 is naive mice internal anatomy;

Figure 23 is the apparent state of later observation naive mice;

Figure 24 is for combining group mouse Colon length representative figure;

Figure 25 is antibiotic group mouse Colon length representative figure;

Figure 26 is complete naive mice colon lengths representative graph;

Figure 27 is for combining group mice HE colored graph;

Figure 28 is antibiotic group mice HE colored graph;

Figure 29 is complete naive mice HE colored graph;

Figure 30 is for combining group mice later observation HE colored graph;

Figure 31 is antibiotic group mice later observation HE colored graph;

Figure 32 is complete naive mice later observation HE colored graph.

Detailed description of the invention

When the present inventor feels a delicacy about for the treatment of current global field of medicaments to ulcerative 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.), therefore the good living preparation of lactobacillus thalline of three strain probiotic effects is combined with cattle colostrums somatomedin crude extract make medicine, 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) preparation of gavage probiotic bacteria

1, three probiotics colony morphological observation

The bacillus acidophilus of preservation, lactobacillus helveticus and Lactobacillus plantarum are taken out respectively, repeatedly activated for 2 generations carries out the setting-out of 3rd district on solid medium, cultivates about 24h, carries out Gram’s staining and oily sem observation for 37 DEG C.

The microscopy picture of bacillus acidophilus, lactobacillus helveticus and Lactobacillus plantarum sees Fig. 1, Fig. 3 and Fig. 5 respectively, its colonial morphology is shown in Fig. 2, Fig. 4 and Fig. 6 respectively, three probiotics thalli morphologies and microscopy result are basically identical with the description on uncle Jie Shi Bacteria Identification handbook, are typical bacillus acidophilus, lactobacillus helveticus and Lactobacillus plantarum.

2, the choosing of probiotic bacteria gavage time

Picking half bacterium colony from the solid medium cultivated, after an activation generation, pass into 0h, 1h, 2h, 3h, 4h, 6h, 8h, 10h, 12h, 14h, 16h, 18h, 20h, 22h, 24h, 26h, 28h, 30h, the fluid medium of totally 18 time periods, when arriving the above-mentioned time, taking-up ultraviolet spectrophotometer, measures OD600, makes growth curve; Take incubation time as abscissa, OD600 value is vertical coordinate, draws the growth curve of 3 strain lactobacilluss, sees Fig. 7.According to the growth curve of three strain lactobacilluss, the time that known lactobacillus helveticus, bacillus acidophilus and Lactobacillus plantarum enter stable phase is respectively 16h, 14h and 14h.

Because the activity of probiotic of logarithmic (log) phase is the highest, and the probiotics bacterial number of stable phase reaches maximum, so choose logarithm end of term stable phase probiotic bacteria just, ulcerative colitis mice is treated, namely cultivate the lactobacillus helveticus of 16h, 14h and 14h, bacillus acidophilus and Lactobacillus plantarum respectively.

3, the mensuration of gavage bacterium number

Lactobacillus helveticus, bacillus acidophilus and Lactobacillus plantarum are cultured to 16h, 14h and 14h respectively, and carried out gradient plate counting, result shows, lactobacillus helveticus, bacillus acidophilus and Lactobacillus plantarum arrive stable phase bacterium number just and be respectively 1.25 × 10 ⁹cFU/mL, 2.42 × 10 ⁹cFU/mL, 6.3 × 10 ⁹cFU/mL.

Research obtains, and the gavage amount of mice is 3 × 10 ⁸time CFU/20g (bacterium number/Mouse Weight), have and well repair intestinal mucosa injury effect; This experiment mice average weight calculates with 20g, and according to 1: 1: 1 ratio, lactobacillus helveticus, bacillus acidophilus, Lactobacillus plantarum should be respectively 1 × 10 ⁸cFU, therefore, get the lactobacillus helveticus bacterium liquid 0.96mL that equal stable phase goes out, bacillus acidophilus bacterium liquid 0.5mL, Lactobacillus plantarum bacterium liquid 0.19mL, centrifuging and taking bacterium mud, is then refitted in 0.1mLPBS respectively, for the probiotic bacteria given low of every mice.

(3) 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.

(4) preparation of the medicine of ulcerative colitis is treated

Cattle colostrums somatomedin crude extract, given low is 5.4mg/20g (cattle colostrums somatomedin crude extract quality/mice weights); 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, lactobacillus helveticus bacterium mud tubule, mixing, then adds the cattle colostrums somatomedin crude extract of described dosage.

(5) 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.

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 ulcerative colitis mouse model 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 Ulcerative Colitis mouse model, then with Mouse Weight change, apparent situation, colon lengths, pathology etc. for index, checking Mouse Ulcerative Colitis Model whether be successfully established.

During modeling, Mouse Weight rate of descent is 13.76%; And enteritis group mice is apparent is not in good state, is in particular in that mice is One's eyesight is restrained, hair is obscure curling, has Qiang's hair phenomenon, see Fig. 8; Further, there is ecchymosis at enteritis group mice root of the tail place, illustrates that mice has dominant phenomenon of having blood in stool, sees Fig. 9; There is phenomenon of festering at enteritis group mouse Colon place after dissection, and intestinal tissue has sends out pus, turbidity phenomenon, sees Figure 10; And during modeling, enteritis group mouse Colon length is about 8.64cm, comparatively in naive mice colon 12.82cm, significantly shortens, see Figure 11, Figure 12; Further, from HE colored graph (see Figure 13), model group mouse Colon tissue, intestinal mucosa atrophy, mucous membrane surface passivation, crypts destroys.

In sum, acute ulcer Colitis Model is successfully established, and can carry out next step Drug therapy test, this experimental test design is shown in Figure 14.

(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 Ulcerative Colitis Model is successfully established on the one hand, illustrate that ulcerative 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, sees Figure 15, and compared with apparent state during enteritis, difference is extremely obvious; And compared with the apparent state of complete naive mice (Figure 21) and the apparent state of antibiotic group mice (Figure 18), difference is not remarkable.From mice internal anatomy, medicine of the present invention significantly improves the festering of the colon of enteritis mice, ecchymosis phenomenon, sees Figure 16; And medicine group mice of the present invention compared with complete naive mice (Figure 22), antibiotic group mouse Colon tissue (Figure 19) without significant difference.From later observation figure, compared with the apparent state of antibiotic group mice (see Figure 20) and the complete apparent state of naive mice (23), combine the apparent state of group mice normal, have no adverse reaction, see Figure 17, illustrate that medicine of the present invention has good effect to treatment ulcerative colitis, and in a short time without recurrence sign.

(3) medicine of the present invention (combining group) increases mouse Colon length

Table 1 treatments period mouse Colon length variations table

Associating group, the mouse Colon length of antibiotic group and completely blank group, see Figure 24 respectively, Figure 25, Figure 26, and by table 1, Figure 24, Figure 25, Figure 26 is known, after two courses for the treatment of, medicine of the present invention (combining group) significantly can increase the colon of mice, by the 8.64cm of mouse Colon tissue after modeling, rise to about 12.93, and difference is not remarkable compared with complete blank group, and be significantly better than antibiotic group, illustrate that the colon's atrophy caused because of ulcerative colitis is 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, strengthen gut barrier effect.

(4) medicine of the present invention increases Mouse Weight

Table 2 treatments period Mouse Weight change list

As shown in Table 2, after gavage one course for the treatment of, drug effect of the present invention is remarkable, not only it is dead to show that medicine group mice of the present invention does not have because of the reason weight loss of enteritis, and body weight increases 11.02% on the contrary, after two courses for the treatment of terminate, medicine group mice of the present invention increases 25.42% relative to modeling initial stage body weight, and compared with antibiotherapy, difference is not remarkable, the method utilizing medicine of the present invention and Medication in Ulcerative Colitis traditional is on the market described, without significant difference; The later observation result of 2 weeks shows, medicine group Mouse Weight of the present invention sustainable growth, does not occur the sign recurred.And completely after two courses for the treatment of of naive mice body weight rate of increase be 11.01%, may be do not carry out Modling model process due to this group mice, body weight reduce, and is in normal level, then Mouse Weight growth space is less; In addition because this group Mouse Weight radix is comparatively large, then mice rate of increase is also relatively little.Therefore, from body weight angle, medicine of the present invention has good anti-ulcerative colitis purposes, and does not have the sign of recurrence in a short time.

(5) mice intestinal mucosa injury repaired by medicine of the present invention

After utilizing medicine of the present invention to carry out the treatment of two courses for the treatment of to mice, dissect mice, get its colon, carry out Hematoxylin-eosin dyeing (HE dyeing), the pathology figure of enteritis group, medicine group of the present invention (combining group), antibiotic group, completely blank group is shown in Figure 13, Figure 27, Figure 28, Figure 29 respectively; After the later observation of two weeks, repeat the same process, medicine group of the present invention, complete blank group, the pathology figure of antibiotic group is shown in Figure 30, Figure 31, Figure 32 respectively.

As seen from the figure, enteritis group mice, intestinal mucosa atrophy, mucous membrane surface passivation, crypts destroys, and is substantially in gut barrier imbalance state; After utilizing medicine of the present invention to carry out the treatment of two courses for the treatment of to mice, protection of intestinal mucosal barrier cells arrangement is substantially neat, intestinal mucosa structural integrity, inorganization necrosis phenomena, illustrates that medicine of the present invention can well the intestinal mucosa injury of repairing ulcer colitis mice; And compared with antibiotic group, completely naive mice, without significant difference; The pathology figure of later observation shows, medicine of the present invention short-term and long-term in all there is good anti-ulcerative colitis effect.

Embodiment 1 one kinds treats the preparation method of the medicine of ulcerative colitis, comprises the steps:

(1) respectively by bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus at MRS culture medium culturing;

(2) lactobacillus helveticus cultivating 14h, the Lactobacillus plantarum cultivating 12h bacillus acidophilus and cultivation 12h is chosen respectively;

(3) lactobacillus helveticus bacterium liquid 77 parts by volume that step (2) obtains is got respectively, bacillus acidophilus bacterium liquid 40 parts by volume, Lactobacillus plantarum bacterium liquid 15 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 10 parts by volume, obtain mixed vaccine mud;

(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 432 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.

Embodiment 2 one kinds treats the preparation method of the medicine of ulcerative colitis, comprises the steps:

(1) respectively by bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus at MRS culture medium culturing;

(2) lactobacillus helveticus cultivating 18h, the Lactobacillus plantarum cultivating 16h bacillus acidophilus and cultivation 16h is chosen respectively;

(3) lactobacillus helveticus bacterium liquid 115 parts by volume that step (2) obtains is got respectively, bacillus acidophilus bacterium liquid 60 parts by volume, Lactobacillus plantarum bacterium liquid 23 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 10 parts by volume, obtain mixed vaccine mud;

(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 684 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.

Embodiment 3 one kinds treats the preparation method of the medicine of ulcerative colitis, comprises the steps:

(1) respectively by bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus at MRS culture medium culturing;

(2) choose logarithm end of term stable phase probiotic bacteria just, namely choose the lactobacillus helveticus cultivating 16h, the Lactobacillus plantarum cultivating 14h bacillus acidophilus and cultivation 14h respectively;

(3) lactobacillus helveticus bacterium liquid 96 parts by volume that step (2) obtains is got respectively, bacillus acidophilus bacterium liquid 50 parts by volume, Lactobacillus plantarum bacterium liquid 19 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 10 parts by volume, obtain mixed vaccine mud;

(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 540 mass parts joined in above-mentioned mixed vaccine mud, stir, clinical acceptable dosage form made by the adjuvant then adding conjunction alive.

Claims (9)

1. treat a medicine for ulcerative colitis, it is characterized in that, comprise 132 ~ 198 parts by volume probiotic bacterias and 432 ~ 684 mass parts cattle colostrums somatomedin, wherein parts by volume/mass parts=ml/mg.

2. the medicine for the treatment of ulcerative colitis according to claim 1, is characterized in that, comprises 165 parts by volume probiotic bacterias and 540 mass parts cattle colostrums somatomedin.

3. the medicine for the treatment of ulcerative colitis according to claim 1 and 2, is characterized in that, described cattle colostrums somatomedin is the somatomedin crude extract extracted from cattle colostrums.

4. the medicine for the treatment of ulcerative colitis according to claim 1 and 2, is characterized in that, described probiotic bacteria is bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus; And be viable bacteria thalline form.

5. the medicine for the treatment of ulcerative colitis according to claim 4, is characterized in that, the number ratio of described bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus is 1: 1: 1.

6. the medicine for the treatment of ulcerative colitis according to claim 4, is characterized in that, described probiotics viable bacteria thalline is by the bacterium mud of the lactobacillus liquid culture difference collected by centrifugation being all in the stable initial stage.

7. the medicine for the treatment of ulcerative colitis according to claim 1, is characterized in that, described probiotic bacteria consumption is 1.5 × 10 ¹⁰cFU/kg, described cattle colostrums somatomedin consumption is 216 ~ 342mg/kg.

8. the preparation method of the medicine of the treatment ulcerative colitis according to claim 1 ~ 7 any one, is characterized in that, comprise the steps:

(1) respectively by bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus at MRS culture medium culturing;

(2) choose the lactobacillus helveticus of cultivation 14 ~ 18h respectively, cultivate the Lactobacillus plantarum of the bacillus acidophilus of 12 ~ 16h and cultivation 12 ~ 16h;

(3) lactobacillus helveticus bacterium liquid 77 ~ 115 parts by volume that step (2) obtains is got respectively, bacillus acidophilus bacterium liquid 40 ~ 60 parts by volume, Lactobacillus plantarum bacterium liquid 15 ~ 23 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 10 parts by volume, obtain mixed vaccine mud;

(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 432 ~ 684 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 for the treatment of ulcerative colitis according to claim 8, is characterized in that, comprise the steps:

(1) respectively by bacillus acidophilus, Lactobacillus plantarum and lactobacillus helveticus at MRS culture medium culturing;

(2) choose logarithm end of term stable phase probiotic bacteria just, namely choose the lactobacillus helveticus of cultivation 16h, the bacillus acidophilus of cultivation 14h and the Lactobacillus plantarum of cultivation 14h respectively;

(3) lactobacillus helveticus bacterium liquid 96 parts by volume that step (2) obtains is got respectively, bacillus acidophilus bacterium liquid 50 parts by volume, Lactobacillus plantarum bacterium liquid 19 parts by volume, centrifuging and taking bacterium mud, then respectively they are dissolved in the aseptic PBS of 10 parts by volume, obtain mixed vaccine mud;

(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 540 mass parts is joined in above-mentioned mixed vaccine mud, stir, then add suitable adjuvant and make clinical acceptable dosage form.