阅读说明：本技术 一种乳杆菌组合物及其用途 (Lactobacillus composition and application thereof ) 是由 赵湘江 潘立 于 2018-05-28 设计创作，主要内容包括：本发明公开了一种乳杆菌组合物,含鼠李糖乳杆菌菌株和格氏乳杆菌菌株,可以抵抗细菌性阴道病和各种阴道感染。本发明组合物对细菌性阴道疾病效果显著,且安全无毒、稳定性好、可长期保存。较目前其他市售制剂,本组合物中菌株均能有效定植,植入阴道后可协同发挥作用,可形成阴道微生态,相对于单菌株的作用更为明显。(The invention discloses a lactobacillus composition which contains lactobacillus rhamnosus strain and lactobacillus gasseri strain and can resist bacterial vaginosis and various vaginal infections. The composition has obvious effect on bacterial vaginosis, is safe and nontoxic, has good stability and can be stored for a long time. Compared with other current commercial preparations, the bacterial strains in the composition can be effectively planted, can synergistically play a role after being implanted into a vagina, can form vaginal microecology, and has more obvious effect compared with a single bacterial strain.)

1. A lactobacillus composition comprises a lactobacillus rhamnosus strain and a lactobacillus gasseri strain, and is characterized in that the lactobacillus rhamnosus (RD-0060) has the preservation number of CGMCC No.14184, and is preserved in the China general microbiological culture Collection center; the preservation number of the Lactobacillus gasseri (RD-0046) is CGMCC No.14109, and the Lactobacillus gasseri is preserved in the China general microbiological culture Collection center.

2. A pharmaceutical composition comprising the lactobacillus composition of claim 1 and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition of claim 2, wherein the pharmaceutically acceptable excipient comprises lactose, microcrystalline cellulose, starch, pre-crosslinked starch, silicon dioxide, magnesium stearate.

4. use of a lactobacillus composition according to any of claims 1 to 3 for the preparation of a medicament for the prevention and/or treatment of gynaecological disorders.

5. The lactobacillus composition for use according to claim 4, wherein the gynecological disease medicament comprises vaginal diseases.

6. Use according to claim 5, characterized in that the pathogenic bacteria of said vaginal diseases comprise Denax bacteria, Candida albicans, Atomobacter, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella.

7. Use of a lactobacillus composition according to any of claims 1 to 3 for the preparation of a medicament with the effect of modulating vaginal flora balance.

Technical Field

the invention belongs to the technical field of medicines, and particularly relates to a lactobacillus composition and application thereof, in particular to a medicine for treating gynecological diseases.

background

a plurality of microorganisms exist in the vagina of healthy women, and the microorganisms, a host and the environment form a vaginal microecosystem which is mutually restricted, coordinated and dynamically balanced. The vaginal flora of healthy women is mainly composed of lactobacillus, including lactobacillus rhamnosus, lactobacillus jensenii, lactobacillus gasseri, lactobacillus crispatus, lactobacillus vaginalis, etc. Under normal conditions, the lactobacillus can protect the vagina, and when the vaginal microecological lactobacillus disorder can cause pathogenic bacteria to invade to generate vaginitis.

Bacterial Vaginosis (BV) occurs because of imbalance of vaginal flora and reduction of host lactobacillus, which leads to mass propagation of other conditional pathogenic microorganisms such as Gardner bacteria, various anaerobes, Vibrio flexuosus and the like, and BV is usually a mixed infection mainly comprising Gardner bacteria. The use of antibiotics to treat BV temporarily alleviates the symptoms of BV, but also causes the already reduced lactobacilli to be further reduced, aggravates vaginal dysbiosis, and thus allows BV to recur repeatedly. How to control relapse and radically cure bacterial vaginosis is a delicate problem which needs to be solved urgently by gynecologists.

The existing products on the market at present are not the dominant bacterial flora in the vagina of women in China, have poor planting capability, can not maintain stable viable bacterial count and can not completely meet the clinical requirements of gynecology. The vagina of a healthy woman contains a plurality of lactobacilli, and a single strain is difficult to play a good role when being used as a vaginal flora for regulation, and even if the single strain has good acid production capacity, H2O2 production capacity and the capability of adhering to vaginal epithelial cells, the single strain is difficult to re-fix and enter the vaginal flora, and is difficult to play a role for a long time. When a plurality of strains are successfully planted in the vagina, the synergistic effect is an important basis for the continuous effect of the lactobacillus, is also a key factor for the curative effect of the lactobacillus, and can effectively form a vaginal microecosystem.

Disclosure of Invention

The invention aims to solve the technical problem of providing a lactobacillus composition which can effectively and synergistically play a role, wherein the strain of the composition is from a healthy human body, has active and stable biological characteristics and is a combination of lactobacillus rhamnosus and lactobacillus gasseri with strong bacteriostatic ability.

The technical scheme of the invention is as follows:

A lactobacillus composition comprises a lactobacillus rhamnosus strain and a lactobacillus gasseri strain, wherein the lactobacillus rhamnosus (RD-0060) has the preservation number of CGMCC No.14184, and is preserved in the China general microbiological culture Collection center; the preservation number of the Lactobacillus gasseri (RD-0046) is CGMCC No.14109, and the Lactobacillus gasseri is preserved in the China general microbiological culture Collection center of the culture Collection of microorganisms.

Further, the composition can also comprise pharmaceutically acceptable pharmaceutic adjuvants such as lactose, microcrystalline cellulose, starch, pre-crosslinked starch, silicon dioxide, magnesium stearate and the like, and can also not contain pharmaceutic adjuvants.

The lactobacillus composition is used for preparing medicines for preventing and/or treating gynecological diseases.

The application of the lactobacillus composition in preparing a medicament for preventing and/or treating vaginal diseases.

Furthermore, the pathogenic bacteria of the vaginal diseases comprise one or more of Gardnerella, Clostridium, Candida albicans, atrophaera, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Salmonella.

the application of the lactobacillus composition in preparing a medicament with the function of adhering vaginal epithelial cells.

The lactobacillus composition is used for preparing a medicament for regulating the balance of vaginal flora.

Two Lactobacillus rhamnosus RD-060 and Lactobacillus gasseri RD-0046 of the invention are screened from vaginal secretions of healthy women of China, and a large number of experiments prove that the Lactobacillus rhamnosus RD-060 and the Lactobacillus gasseri RD-0046 of the invention have stronger acid-producing, H2O 2-producing and vaginal epithelial cell adhesion capabilities. The medicine for preventing and/or treating the gynecological diseases, in particular to the medicine for preventing and/or treating vaginal pathogenic bacteria, the medicine for preventing and/or treating vaginal diseases, the medicine for regulating vaginal flora balance and the medicine with the function of adhering vaginal epithelial cells can play a good medicinal effect due to the characteristics of the medicine.

The beneficial effect that adopts above-mentioned technical scheme to produce lies in: (1) the Lactobacillus rhamnosus RD-060 and Lactobacillus gasseri RD-0046 strains of the invention can independently and synergistically resist bacterial vaginosis and various vaginal infections, including Candida albicans vaginitis, gonorrhea, viral vaginitis, urinary tract infection and the like. (2) The strain is directly collected from a healthy human body, has active and stable biological characteristics, does not need domestication and rejuvenation processes, and can be directly prepared by a preparation process. (3) The strain has the effects of inhibiting Gardnerella, clostridium, Candida albicans, atrophaera, staphylococcus aureus, Escherichia coli, pseudomonas aeruginosa, salmonella and the like, has obvious advantages compared with the current commercially available preparation, has strong field planting capability when implanted, and can effectively play a role. (4) Can effectively form vaginal microecology, which is obviously different from the planting effect of single strain.

Preservation information

The Lactobacillus rhamnosus RD-060 is screened from vaginal secretions of women of healthy child-bearing age in China, wherein the Lactobacillus rhamnosus RD-0060 is deposited in China general microbiological culture collection center of China microbiological culture collection management Committee, the preservation place is Beijing in China, the preservation number is CGMCC No.14184, and the preservation date is 2017, 05 and 24 days; wherein the Lactobacillus gasseri RD-0046 preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, the preservation place is Beijing, China, the preservation number is CGMCC No.14109, and the preservation date is 2017, 05 and 10 days.

Drawings

FIG. 1: composition bacteriostatic cake pattern-staphylococcus aureus;

FIG. 2: the composition is bacteriostatic, namely clostridium;

FIG. 3: a result graph of extraction of the genomic DNA of Lactobacillus gasseri RD-0046;

FIG. 4: a PCR product amplification result chart of the Lactobacillus gasseri RD-0046;

FIG. 5: a genomic DNA extraction result chart of lactobacillus rhamnosus RD-0060;

FIG. 6: and (3) a PCR product amplification result chart of the lactobacillus rhamnosus RD-0060.

Detailed Description

The test reagent consumables used in the following examples are all conventional biochemical reagents unless otherwise specified; the experimental methods are conventional methods unless otherwise specified; in the following examples,% represents mass%.

The bacterial culture media used in the following examples can be prepared as follows:

1. broth solid Medium (MRS) preparation:

(1) preparing agar powder into a solution, and adding 1.5g/100ml deionized water;

(2) Adding 17.91g/100ml agar solution of MRS culture medium, and mixing;

(3) Placing into a sterilizing pot, and sterilizing for 20 minutes by 1.0MPa steam;

(4) Pouring the culture medium into a culture dish after the temperature of the culture medium is reduced to room temperature, wherein the culture medium is about 10 ml/piece or 20 ml/piece according to the size of the culture dish;

(5) operating in a clean bench. Cooling to become agar, marking the name of the culture medium and the preparation date, and placing in a refrigerator at 4 ℃ for later use.

2. Broth liquid Medium (MRS) preparation:

(1) Adding the MRS culture medium into deionized water in a ratio of 17.9l g/100 ml;

(2) Placing into a sterilizing pot, and sterilizing for 20 minutes by 1.0MPa steam;

(3) Taking out the sterilized pan when the pressure of the sterilized pan is not high, and subpackaging the sterilized pan into EP tubes with each 1.0 ml. Marking the name of the culture medium and the preparation date, and placing the culture medium in a refrigerator at 4 ℃ for later use.