CN109985069B - Probiotic compositions and uses thereof - Google Patents
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Abstract
The present invention provides a probiotic composition, wherein said composition comprises lactobacillus acidophilus, enterococcus faecalis and bacillus cereus. In another aspect, the present invention provides a medicament for the treatment of vaginal diseases, said medicament comprising a probiotic composition as described above. In a further aspect, the present invention provides the use of a probiotic composition of the present invention in the manufacture of a medicament for the prevention and/or treatment of vaginal diseases. The use according to the invention, wherein the vaginal disease is gynaecological vaginal inflammation. By using the probiotic compound, the bacteriostasis rate to escherichia coli is 86.5%, and the bacteriostasis rate to staphylococcus aureus and candida albicans reaches 100%; animal experiments also verified that the probiotic composition of the present invention has no irritating effect on vaginal mucosa.
Description
Technical Field
The invention belongs to the field of biological medicines, and relates to a probiotic composition and application thereof in preparing a product for preventing and/or treating gynecological vaginal diseases. More specifically, the invention relates to a probiotic composition comprising lactobacillus acidophilus, enterococcus faecalis and bacillus cereus and its use in the preparation of a product for preventing and/or treating gynecological inflammations.
Background
The female lower genital tract is an important micro-ecological area of the human body, and in vaginal discharges from healthy women, the ratio of anaerobic to aerobic bacteria is 5: 1, viable count is 102～109cfu/mL, total bacteria count can reach 8 multiplied by 107cfu/mL. More than 200 microorganisms are parasitized in the vagina of women and grow on the surface of the vaginal mucosa of normal people in a special form, and the main persistent bacteria comprise lactobacillus, staphylococcus epidermidis, escherichia coli, clostridium, streptococcus faecalis and the like. In the normal vaginal flora, lactobacilli predominate. The lactobacillus is gram-positive bacillus megaterium, and has microaerophilic property, but better growth in anaerobic environment, and optimal growth temperature35-38 deg.C, per gram of vaginal secretion 107～108Lactobacillus CFU. More than 20 kinds of lactobacillus can be separated from vagina of healthy women. The normal presence of lactobacilli in the vagina plays a key role in maintaining a normal flora in the vagina. Glycogen in the vaginal squamous epithelial cells is decomposed into lactic acid under the action of lactobacillus, so that a weak acid environment (the pH is less than or equal to 4.5 and more than 3.8-4.4) is locally formed in the vagina, and the overgrowth of other parasitic bacteria can be inhibited. In addition, the lactobacillus prevents pathogenic microorganisms from adhering to vaginal epithelial cells through a substitution and competitive exclusion mechanism, and simultaneously secretes hydrogen peroxide, bacteriocin-like substances, biosurfactant and the like to inhibit the growth of the pathogenic microorganisms, so that the balance of the vaginal micro-ecological environment is maintained.
WHO's research on the health status of women worldwide indicates that: the incidence of gynecological diseases reaches 65%, while the prevalence rate of women in the childbearing age reaches 70%. In China, the disease condition of women is not optimistic, the total disease rate of common diseases of women in the whole country in 2010 is 28.8%, wherein vaginitis accounts for the first place, and the disease rate is 13.2%. The unique physiological structure of women, together with the pressure of work and life and environmental factors, causes the incidence of gynecological diseases to be high. In 2015, the number of female population in China is increased to 6.7 hundred million, which accounts for 48.8% of the total population in China. The last half year of 2016, the marketing data of gynecological medicines show that the gynecological inflammation medicines occupy 59.8% of the total amount and are stably positioned at the first place. The total disease rate of common diseases of the national women in 2010 is 28.8%, wherein vaginitis accounts for the first place, and the disease rate is 13.2%. The demand of gynecological anti-inflammatory drugs will keep increasing in the future.
Studies currently believe that clinically well-diagnosed and unexplained vaginal inflammation presents different degrees of vaginal dysbiosis. Vaginal microenvironment balance plays an important role in maintaining self-cleaning and host health. Factors affecting vaginal microecological balance include: vaginal pH, migration of flora in different age groups of women, pregnancy, parturition, use of antibacterial/anti-bacterial drugs, anemia, infection, etc. The number of the vaginal lactobacilli of healthy women accounts for more than 95 percent of vaginal microorganisms, and the vaginal lactobacilli are the dominant bacteria in normal flora of healthy women, and the main effects of the lactobacilli in the vagina comprise: firstly, the lactic acid bacteria convert glycogen into lactic acid, so that the vagina is kept in an acidic environment, the pH value is maintained at 3.8-4.2, and the propagation of certain pathogenic bacteria is inhibited; secondly, the vaginal mucosa antibacterial agent has competitive adhesion to bacteria of the vaginal mucosa and can prevent and reduce the adhesion of other vaginal pathogenic bacteria to the vagina; ③ can generate hydrogen peroxide and lactein to inhibit or kill some pathogenic bacteria; has the functions of stimulating immune system and regulating body's immune reaction. Therefore, the lactobacillus plays an important role in maintaining the vaginal ecological flora balance and is a microbial defense line of the female genital tract.
The main treatment means of vaginitis is vaginal medication. The data show that 37-66% of patients are treated by conventional vaginal irrigation, which is liable to destroy the vaginal ecological environment, possibly causes ascending infection and increases the occurrence of pelvic infectious diseases, so that the method is not advocated. Antibiotics are currently the most widely used drugs in clinical treatment of infectious diseases. The antibiotic can inhibit pathogenic bacteria and destroy normal flora in vagina, so that the dominant microorganism is changed, a great amount of pathogenic microorganisms with drug resistance appear, and once the pathogenic microorganisms are infected again, the curative effect of the antibiotic is seriously influenced. Therefore, the treatment of genital tract infection should also pay attention to the regulation and control of the genital tract micro-ecological environment. The efficacy of probiotics in regulating flora is more and more well known, products are already on the market for treating or regulating vaginal inflammation, and along with the popularization of the efficacy of probiotics, the effect of probiotics in treating vaginal inflammation is gradually paid attention.
The Chinese patent application with the application number of 200910024226.4 and the invention name of 'a lactobacillus vaginal capsule and a preparation method thereof' discloses a lactobacillus vaginal capsule which is a powder capsule consisting of an excipient and live streptococcus enterobacteria, wherein each capsule contains at least 2500 ten thousand strains per gram of live streptococcus enterobacteria, and the excipient consists of calcium carbonate powder and medicinal starch with the dosage of 1: 1. Experiments prove that the lactic acid bacteria vaginal capsule can fully supplement the most important lactic acid bacteria in the vagina, thereby inhibiting the growth and the reproduction of pathogenic bacteria. However, the lactic acid bacteria vaginal capsule uses too few probiotics, which is not favorable for later recovery of vaginal flora balance.
Based on the above, the research on the probiotic products for gynecological vaginal diseases in the prior art mainly focuses on the development and application of vaginal lactobacillus, and the research on the compound probiotics is less. Therefore, there is currently a need for probiotic compositions for the treatment of gynaecological vaginal diseases.
Disclosure of Invention
Based on the prior art, the probiotic composition provided by the invention can be used for preventing and/or treating gynecological vaginal inflammation caused by pathogenic bacteria such as escherichia coli, staphylococcus aureus, candida albicans and the like, further regulating vaginal flora, treating vaginal microecological imbalance caused by repeated outbreaks of vaginal inflammation caused by abuse of antibiotics and other products, recovering vaginal flora balance during menstruation, pregnancy and abortion, and playing a role in preventing and/or treating gynecological vaginal inflammation.
In one aspect, the present invention provides a probiotic composition, wherein said composition comprises lactobacillus acidophilus, enterococcus faecalis and bacillus cereus.
Wherein the content of active probiotic bacteria in the probiotic composition is at least 1 × 106cfu/g。
The probiotic composition provided by the invention is characterized in that the preservation number of the lactobacillus acidophilus is CGMCC No. 0460.2;
the preservation number of the enterococcus faecalis is CGMCC No. 0460.3;
the preservation number of the bacillus cereus is CGMCC No. 0460.4.
The lactobacillus acidophilus with the preservation number of CGMCC No.0460.2, the enterococcus faecalis with the preservation number of CGMCC No.0460.3 and the bacillus cereus with the preservation number of CGMCC No.0460.4 are described in the Chinese patent application with the application number of CN01108353.0 and the invention name of "bifidobacterium quadruple viable tablet", and the whole is incorporated by reference.
Preferably, the viable count of Lactobacillus acidophilus in the probiotic composition is at least 1 × 106cfu/g; the viable count of enterococcus faecalis is at least 1 × 106cfu/g; and the viable count of Bacillus cereusAt least 1 × 105cfu/g。
The probiotic composition disclosed by the invention contains lactobacillus acidophilus which can generate an acidic substance to help recover the pH value of the vagina; enterococcus faecalis is a gram-positive and hydrogen peroxide-negative coccus, can produce natural antibiotics and antibacterial substances such as bacteriocin and the like, inhibits the growth of pathogenic bacteria such as escherichia coli and the like, and improves the ecological microenvironment of the vagina; the bacillus cereus can generate antibacterial substances, inhibit the propagation of harmful microorganisms, and consume oxygen in the vagina, so that aerobic pathogenic bacteria such as staphylococcus aureus and the like cannot survive, and the bacillus cereus cannot be fixedly planted in the vagina of a human body. Further, the inventor surprisingly found that, as for the intestinal flora, the three probiotics have a synergistic effect, the lactobacillus acidophilus generates an acidic substance to change the pH of the environment, effectively kill a part of harmful bacteria, and provide a better environment for the growth of enterococcus faecalis and bacillus cereus; the growth of the bacillus cereus can consume a large amount of oxygen, and the lactobacillus acidophilus which is facultative anaerobic can be better propagated and grown. Researches show that the three probiotics provided by the invention can inhibit vaginal pathogenic bacteria more obviously, help to restore vaginal ecological balance in later period, and overcome the defect that gynecological inflammation is recurrent after treatment with antibiotics, wherein the gynecological inflammation is caused by vaginal flora disorder.
In another aspect, the present invention provides a pharmaceutical composition for treating vaginal diseases, comprising the above probiotic composition;
the pharmaceutical composition of the invention comprises pharmaceutically acceptable auxiliary materials;
preferably, the pharmaceutically acceptable excipients are selected from excipients and fillers;
preferably, the pharmaceutically acceptable auxiliary material is selected from one or more of microcrystalline cellulose, corn starch, pregelatinized starch, chitosan, bromelain and sodium carboxymethyl starch.
The pharmaceutical composition provided by the invention is characterized in that the addition amount of microcrystalline cellulose is 10-50wt%, the addition amount of sodium hydroxymethyl starch is 5-30wt%, and the balance of auxiliary materials is medicinal starch, based on the total weight of the pharmaceutical composition.
In a preferred embodiment, the pharmaceutical composition of the invention consists of:
1×106～9×106cfu/g of Lactobacillus acidophilus;
1×106～9×106cfu/g enterococcus faecalis;
1×105～9×105cfu/g of Bacillus cereus;
10-50wt% of microcrystalline cellulose;
5-30wt% sodium starch glycolate; and
the balance being medicinal starch.
The pharmaceutical composition is a solid preparation, a liquid preparation or an enteric preparation;
preferably, the pharmaceutical composition is capsule, granule, powder, tablet or powder in capsule;
the pharmaceutical composition according to the present invention, wherein the route of administration of the pharmaceutical composition is vaginal route.
The pharmaceutical composition according to the present invention, wherein the vaginal disease is gynecological vaginal inflammation;
the pharmaceutical composition according to the present invention, wherein the vaginal disease is a disorder of vaginal flora; wherein the vaginal flora disturbance is caused by fungi, bacteria and bactericidal anti-inflammatory agents, or is caused by vaginal flora imbalance during menstruation, pregnancy and abortion.
In a further aspect, the present invention provides the use of a probiotic composition of the invention in the manufacture of a medicament for the prevention and/or treatment of vaginal diseases;
the use according to the invention, wherein the vaginal disease is gynecological vaginal inflammation;
the use according to the invention, wherein the vaginal disorder is a disorder of vaginal flora; wherein the vaginal flora disturbance is caused by fungi, bacteria and bactericidal anti-inflammatory agents, or by vaginal flora imbalance during menstruation, pregnancy and abortion.
Compared with the prior art, the probiotic compound has the following advantages:
the probiotic composition can obviously inhibit vaginal flora, and the antagonism observed by a test tube method shows that the probiotic compound has the bacteriostasis rate of 86.5 percent on escherichia coli and the inhibition rate of 100 percent on staphylococcus aureus and candida albicans; animal experiments also verified that the probiotic composition had no irritating effect on vaginal mucosa.
Detailed Description
Example 1 preparation of probiotic composition capsules of the invention
Freeze-drying probiotics: after lactobacillus acidophilus, enterococcus faecalis and bacillus cereus are subjected to secondary activation, probiotic powder is prepared into freeze-dried powder, and the viable bacteria content of the obtained bacterium powder is respectively as follows: the viable count of the lactobacillus acidophilus powder is 1011cfu/g, viable count of enterococcus faecalis 1010cfu/g, the viable count of the bacillus cereus is 109cfu/g。
② capsule preparation: mixing Lactobacillus acidophilus, enterococcus faecalis and Bacillus cereus powder with microcrystalline cellulose, medicinal starch and sodium carboxymethyl starch according to calculated amount, and preparing into pharmaceutical composition capsule, wherein:
high dose capsules: the viable count of Lactobacillus acidophilus is 1x107cfu/g, viable count of enterococcus faecalis 1x107cfu/g, viable count of Bacillus cereus 1x106cfu/g, total viable count of 107cfu/g；
And (3) a medium-dose capsule: the viable count of Lactobacillus acidophilus is 1x106cfu/g, viable count of enterococcus faecalis 1x106cfu/g, viable count of Bacillus cereus 1x105cfu/g, total viable count of 106cfu/g；
Low dose capsules: the viable count of Lactobacillus acidophilus is 1x105cfu/g, viable count of enterococcus faecalis 1x105cfu/g, viable count of Bacillus cereus 1x104cfu/g, total viable count of 105cfu/g。
In the capsule, the addition amount of microcrystalline cellulose is 12% by weight, the addition amount of sodium hydroxymethyl starch is 6.7% by weight, and the rest is filled with medicinal starch, wherein the medicinal starch is used as a dispersing agent and a protective agent to ensure that the strain is in a semi-anaerobic and semi-sealed state for a long time, so that the number of viable bacteria in the specified effective period of the medicine is realized, and the safety and effectiveness of the medicine are ensured. Encapsulating the medicinal powder to obtain probiotic compound capsule, wherein the weight of each capsule is about 0.5 g.
Example 2 in vitro bacteriostatic experiments on probiotic composition capsules of example 1
2.1 Observation of the bacteriostatic action by Oxford cup method
2.1.1 g of the freeze-dried powder obtained in example 1 was put into 100mL of seed solution for activation culture, wherein the preparation method of the lactobacillus acidophilus seed solution comprises the following steps: weighing 10g of casein peptone, 10g of beef extract, 5g of yeast extract, 20g of glucose, 5g of dipotassium hydrogen phosphate trihydrate, 2g of triammonium citrate, 10g of sodium acetate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and 1g of polysorbate, adding 1L of purified water for dissolving, and adjusting the pH value to 5.8;
the enterococcus faecalis seed liquid formula comprises: weighing 10g of peptone, 5g of yeast extract, 5g of beef extract, 5g of glucose, 5g of sodium chloride and 1g of polysorbate, adding 1L of purified water for dissolving, and adjusting the pH value to 7.4;
the bacillus cereus seed liquid comprises the following components in percentage by weight: weighing 10g of peptone, 5g of beef extract and 5g of sodium chloride, adding 1L of purified water for dissolving, and adjusting the pH value to 7.4.
2.1.2 preparation of Single-bacterium culture solutions of Lactobacillus acidophilus, enterococcus faecalis and Bacillus cereus (bacterial solutions were diluted to 1X106cfu/mL、1×106cfu/mL and 1X105cfu/mL), and preparing two-by-two mixed bacterial liquids of lactobacillus acidophilus, enterococcus faecalis and bacillus cereus (the concentration of each bacterial liquid is 1 multiplied by 10 respectively)6cfu/mL、1×106cfu/mL and 1X105cfu/mL, total viable count of 106cfu/mL)；
2.1.3 preparation of a sample of the capsules of example 1, in which each granule isThe weight of the capsule is 0.5g, each capsule contains about 0.48g of powder, 10g of powder in the capsule with high, low and medium dosage is respectively dissolved in 10mL of water to be used as sample liquid, and the viable count content of the sample liquid is about 105cfu/mL、106cfu/mL、107cfu/mL of high, low and medium dosage sample solution.
2.1.4 respectively intensively inoculating staphylococcus aureus and escherichia coli on a nutrient agar plate, inoculating candida albicans on a Sabouraud agar plate, putting into an oxford cup, and respectively adding 100 mu L of the bacterial liquid into the oxford cup; the cells were allowed to absorb at 4 ℃ for 4 hours, and then incubated at 37 ℃ for 24 hours, each 3 times, and the zone diameter was measured with a vernier caliper. The zone of inhibition is directly shown in table 1 below:
TABLE 1 size of zone of inhibition
The results show that the inhibition effect of lactobacillus acidophilus in the three strains on vaginal pathogenic bacteria escherichia coli, staphylococcus aureus and candida albicans is the best, the inhibition effect of enterococcus faecalis is the next time, and the inhibition effect of wax samples on the three strains of pathogenic bacteria is not obvious. In the two mixed bacteria liquids, the acidophilic bacteria and feces and intestine mixed bacteria culture solution has a good inhibition effect on pathogenic bacteria, but the bacteria liquid mixed by the acidophilic bacteria and the feces and intestine mixed bacteria has the best inhibition effect, so that three probiotic compounds are selected to form the probiotic compound capsule. The research on the in-vitro antibacterial effect of the sample liquid with high, low and medium doses proves that the antibacterial effect of the sample liquid with low concentration is slightly worse than that of the sample liquid with medium dose and the sample liquid with high dose, and the antibacterial effect of the sample liquid with medium dose and the sample liquid with high dose is not much different.
2.2 comparison experiment results with commercial probiotic gynecological disinfection products
Selecting the currently marketed vaginal capsule product 1 (the main effective components are lactobacillus and live streptococcus enterobacteria), and the viable count of the probiotics is about 1x107cfu/g; the commercial freeze-dried active factor bacteriostatic powder product 2 (the main effective component is lactic acid bacteria) has the viable count of about 1x106cfu/g; commercial freeze-dried active factor bacteriostatic powder product 3 (the main effective component is lactobacillus) with probiotic activityThe number of bacteria is about 1x107The results of in vitro bacteriostasis experiments on the three cfu/g products compared with the invention are shown in the following tables 2 and 3:
TABLE 2 comparison of the zones of inhibition
TABLE 3 comparison of bacteriostatic rates
The experimental results show that although the viable bacteria of the probiotic product is one order of magnitude lower than that of the commercial product, the bacteriostatic effect of the probiotic product in vitro in inhibiting the vaginal pathogenic bacteria escherichia coli, staphylococcus aureus and candida albicans is superior to that of the selected three commercial products, so that the probiotic product disclosed by the invention generates a synergistic effect through the combination of the three bacteria, and the bacteriostatic effect is superior to that of the commercial products. .
Example 3 animal experiments with probiotic composition capsules of example 1
Based on example 1, each capsule of the composite probiotic capsule for preventing or treating gynecological inflammation of the invention weighs 0.5g, each capsule contains about 0.48g of powder, 10g of high-dose capsule powder is dissolved in 10mL of water, and the viable count content of the obtained high-dose sample liquid is about 1x107cfu/mL; dissolving 10g of the medium-dose capsule powder in 10mL of water to obtain medium-dose sample solution with viable count content of about 1x106cfu/mL; dissolving low-dose capsule powder l0g in 10mL water to obtain medium-dose sample solution with viable count content of about 1x105cfu/mL. The number of viable bacteria at each dose was checked before the experiment.
3.1 mucosal irritation test
24 healthy and mature New Zealand female white rabbits with the age of about 5 months and the weight of about 2.5Kg are divided into an experimental group and 6 groups of high dose, medium dose and low dose, and are subjected to a vaginal mucosa stimulation experiment. The vaginal opening of the animal was examined for the absence of secretions, congestion, edema and other lesions and then used in the experiment. A blunt hose with the length of about 8cm is connected with a 2.5mL syringe, and the syringe and the catheter are filled with a test object (probiotic compound capsule powder diluent) for later use. Fixing the animal on the back, exposing perineum and vaginal orifice, wetting the head of the catheter with a control solution, inserting the catheter into vagina (4-5 cm) gently, injecting 1mL of probiotic compound capsule diluent slowly with an injector, and withdrawing the catheter. Control animals were treated with saline as well. The animals were sacrificed 24h after contact with the vaginal mucosa, the whole vagina was removed, cut longitudinally and observed for signs of congestion, edema, etc. Then the vagina is put into 10 percent formalin solution for fixation for more than 24 hours, tissues at two ends and the center of the vagina are selected for flaking, and histopathological examination is carried out after HE staining. Histopathological examination results (vaginal mucosa irritation response) were scored according to "Disinfection specifications" and are shown in Table 4.
TABLE 4 irritation test results
| Categories | Integration | Stimulation index |
| Control group (viable count) | 5.45 | / |
| High dose (1x 10)7cfu/mL) | 7.51 | 2.06 |
| Middle dose (1x 10)6cfu/mL) | 4.98 | -0.47 |
| Low dose (1X 10)5cfu/mL) | 4.74 | -0.71 |
Tests show that when the content of viable bacteria of the probiotic compound is 106Under the condition of cfu/g (medium dose), the mean value of the integral of an experimental group is 5.45, the integral of a control group is 4.98, the stimulation index is-0.47, and the test method is non-irritant. From the test results, the probiotic compound capsule has no irritation to the vaginal mucosa of rabbits at the using dosage and the low dosage, and has slight irritation to the mucosa at the high dosage. Therefore, the low-dose group and the medium-dose group are selected to be used for the next animal treatment test.
3.2 probiotic bacteria for vaginitis effect research experiment
a. Modeling of test animals
Taking 18 healthy and mature New Zealand female white rabbits with the age of about 5 months and the body weight of about 2.5Kg, checking and confirming that the vaginal orifice of the animal has no secretion, congestion, edema and other damaged conditions, and carrying out the probiotic vaginal bacteriostasis experiment. Mixing 24h cultures of common pathogenic bacteria Escherichia coli, Staphylococcus aureus and Candida albicans in equal proportion, injecting 1mL of bacteria liquid into vagina by using a syringe for each rabbit (the posture of the injection is kept for 15min after the injection, and then the injection liquid is put back into a cage to prevent the bacteria liquid from flowing out immediately), and waiting for 1 time every day until corresponding clinical symptoms appear. The successfully modeled vaginal mucosa of the rabbit is swollen and congested, mucous secretion adheres to the periphery of the vulva, appetite is reduced, and escherichia coli, staphylococcus aureus and candida albicans are detected by a smear microscope.
b. Study of therapeutic Effect
After successful modeling, 18 rabbits were randomly divided into 3 groups of 6 rabbits each, namely a control group, a low-dose treatment group and a medium-dose treatment group.Injecting into vagina with 1mL of liquid by syringe, wherein the control group is injected with normal saline, and the low dose group is injected with viable bacteria with a count of about 105The low-dose probiotic compound of cfu/mL has the live bacteria number of about 10 in the middle-dose group injection6A medium dose of probiotic complex cfu/mL, 1 time per day for 7 days. In the control group, 1mL of sterile physiological saline was injected into the vagina of each rabbit with a syringe 1 time per day. Clinical changes such as body temperature, mental state, activity, eating condition and the like of the experimental group and the control group are observed every day. The animals are sacrificed after stopping the medicine and observing for 5 days, the complete vagina is taken out, the longitudinal incision is carried out, whether the vaginal mucous membrane is smooth or not is observed, and the sampling culture is carried out to determine whether escherichia coli, staphylococcus aureus and candida albicans exist in the vagina or not.
Test results show that the vaginitis clinical symptoms of the rabbits in the medium-dose treatment group completely disappear after 7 days of continuous injection, and pathogenic bacteria escherichia coli, staphylococcus aureus and candida albicans are not found in the vagina collection of the rabbits cultured after 5 days of drug withdrawal. In the low dose treatment group, 4 out of 6 rabbits showed a significant improvement in clinical symptoms. The control group had increased clinical symptoms and even had no food intake, so the dose was set to a medium dose (viable count content of 1X 10)6cfu/mL)。
(1) The results of in vitro experiments by an Oxford cup method show that the inhibition effect of Lactobacillus acidophilus in the three strains on vaginal pathogenic bacteria Escherichia coli, Staphylococcus aureus and Candida albicans is the best, the inhibition effect of a wax sample on the three strains of pathogenic bacteria is not obvious, the inhibition effect of a mixed bacterium culture solution of acidophilic bacteria and fecal intestine in two-two combination on the pathogenic bacteria is better, but the bacterium solution of the mixed bacterium solution of the acidophilic bacteria and the fecal intestine has the best inhibition effect, so that three probiotic compounds are selected to form the probiotic compound capsule. The antagonism observed by a test tube method shows that the probiotic compound has the inhibition rate of 86.5 percent on escherichia coli and the inhibition rate of 100 percent on staphylococcus aureus and candida albicans. Through the research on the in vitro bacteriostatic experiments of three strains of bacteria with different dosages, the result shows that the bacteriostatic effect of the low-dosage sample liquid is inferior to that of the medium-high dosage sample liquid, and the inhibitory effects of the medium-high dosage sample liquid on the gynecological vaginal pathogenic bacteria are not greatly different, so that the medium dosage, namely 1, is selected from the aspect of cost06cfu/g was taken as the viable bacteria concentration of the final capsule.
(2) From the results of animal experiments, it can be seen that the probiotic compound capsule is non-irritating to the vaginal mucosa of rabbits at the use concentration. The test results show that the clinical symptoms of vaginitis of the rabbits in a treatment group disappear after 7 days of continuous injection, the clinical symptoms of a control group become worse, and even the situation of no eating appears, and pathogenic bacteria escherichia coli, staphylococcus aureus and candida albicans are not found in the vagina collection of the rabbits cultured after the medicine is stopped for 5 days, so that the probiotic compound capsule has a good treatment effect on vaginitis caused by the pathogenic bacteria.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and are not limited. Although the present invention has been described in detail with reference to the embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the scope of the present invention. Are intended to be covered by the scope of the claims of the present invention.
Claims (13)
1. A probiotic composition for use in the preparation of a medicament for the treatment of vaginal diseases, wherein said composition consists of lactobacillus acidophilus, enterococcus faecalis and bacillus cereus;
the lactobacillus acidophilus has a preservation number of CGMCC number 0460.2, the enterococcus faecalis has a preservation number of CGMCC number 0460.3, and the bacillus cereus has a preservation number of CGMCC number 0460.4;
wherein the viable count of Lactobacillus acidophilus in the probiotic composition is at least 1 × 106cfu/g; the viable count of enterococcus faecalis is at least 1 × 106cfu/g; the viable count of Bacillus cereus is at least 1 × 105 cfu/g。
2. A pharmaceutical composition for treating vaginal diseases comprising the probiotic composition of claim 1.
3. The pharmaceutical composition of claim 2, wherein the medicament comprises a pharmaceutically acceptable excipient.
4. The pharmaceutical composition of claim 3, wherein the pharmaceutically acceptable excipient is selected from excipients and fillers.
5. The pharmaceutical composition according to claim 3, wherein the pharmaceutically acceptable excipients are selected from one or more of microcrystalline cellulose, corn starch, pregelatinized starch, chitosan, bromelain and sodium carboxymethyl starch.
6. The pharmaceutical composition of claim 2, wherein the pharmaceutical composition consists of:
1×106～9×106cfu/g of Lactobacillus acidophilus;
1×106～9×106cfu/g enterococcus faecalis;
1×105～9×105cfu/g of Bacillus cereus;
10-50wt% of microcrystalline cellulose;
5-30wt% sodium starch glycolate; and
the balance being medicinal starch.
7. The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition is a solid formulation, a liquid formulation or an enteric formulation.
8. The pharmaceutical composition of claim 2, wherein the pharmaceutical composition is a capsule, granule, powder, tablet, or powder in a capsule.
9. The pharmaceutical composition according to any one of claims 2-8, wherein the vaginal disease is gynecological vaginal inflammation or vaginal flora disturbance.
10. The pharmaceutical composition of claim 9, wherein the vaginal flora disorder is caused by fungi, bacteria, and bactericidal anti-inflammatory agents; or the vaginal dysbacteriosis is caused by an imbalance in vaginal flora during menstruation, pregnancy and after abortion.
11. Use of a probiotic composition according to claim 1, in the preparation of a medicament for the prevention and/or treatment of vaginal diseases.
12. The use according to claim 11, wherein the vaginal disease is gynecological vaginal inflammation or vaginal flora disturbance.
13. The use according to claim 11 or 12, wherein the disorder of vaginal flora is caused by fungi, bacteria and bactericidal anti-inflammatory agents, or the disorder of vaginal flora is an imbalance of vaginal flora during menstruation, pregnancy and post-abortion.
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