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Lactobacillus curvatus and pharmaceutical application thereof

A technology of Lactobacillus flexi, strains, applied in the direction of antibacterial drugs, bacteria, drug combinations, etc., can solve problems such as poor effect, toxic side effects, etc.

Active Publication Date: 2015-06-03
北京博园新里科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the ineffectiveness of antibiotics in the treatment of bacterial vaginosis and the side effects of liver and kidney damage caused by antibiotics, efforts have been made to explore microecological therapies based on probiotic Lactobacillus preparations at home and abroad.

Method used

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  • Lactobacillus curvatus and pharmaceutical application thereof
  • Lactobacillus curvatus and pharmaceutical application thereof
  • Lactobacillus curvatus and pharmaceutical application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, isolation and identification of Lactobacillus flexus strain

[0039] 1. Isolation of Lactobacillus flexus strains

[0040] MRS medium: peptone 10.0g; beef extract 10.0g; yeast extract 5.0g; diammonium hydrogen citrate 2.0g; glucose 20.0g; Tween 80, 1.0mL; sodium acetate (CH 3 COONa·3H 2 O) 5.0g; Dipotassium hydrogen phosphate (K 2 HPO4·3H 2 O) 2.0g; Magnesium sulfate (MgSO 4 ·7H 2 O) 0.58g; manganese sulfate (MnSO 4 ·H 2 O) 0.25g; distilled water 1000mL; adjust to pH 6.2-6.6.

[0041] More than 200 samples of vaginal secretions from healthy women without symptoms and signs of vaginitis were collected. The samples were numbered sequentially, inoculated in MRS liquid medium, and incubated anaerobically at 37°C for 24 hours. For each sample, 5 single colonies were randomly selected and cultured in 2 mL MRS liquid medium for 24 hours, and the colonies were numbered sequentially. Bacteria are amplified for bacterial conservation and DNA preparation. H...

Embodiment 2

[0051] Embodiment 2, functional analysis of Lactobacillus jensenii (Lactobacillus jensenii) IMC-79

[0052] 1. Lactobacillus jensenii (Lactobacillus jensenii) IMC-79 produces H 2 o 2 detection

[0053] h 2 o 2 Preparation of standard curve: firstly, piperazine-N, N'-di-ethanesulfonic acid (Piperazine-N, N-Bis 2-ethanesulfonic acid, PIPES) with a concentration of 100mmol / L was used to dissolve 30% (v / v) H 2 o 2 (equivalent to 9.128mol / L) stock solution diluted to 1mol / L, and then 1mol / L H with 100mmol / L PIPES 2 o 2 Dilute to 0μmol / L, 20μmol / L, 40μmol / L, 60μmol / L, 80μmol / L, 100μmol / L working solution; then take 100μL of the above H 2 o 2 Mix the working solution with 100 μL of TMB (tetramethylbenzidine) at a concentration of 20 mmol / L and 2 μL of horseradish peroxidase (1 mg / mL), incubate at 16°C for 10 min, and measure the OD 630 value. end up with H 2 o 2 Concentration standard curve: y=0.0033x-0.0496, R 2 = 0.992.

[0054] Lactobacillus jensenii (Lactobacillus j...

Embodiment 3

[0058] Example 3, the adhesion of Lactobacillus flexus IMC-79 to vaginal epithelial cells

[0059] Vaginal epithelial exfoliated cells were obtained from more than 20 healthy women of childbearing age (clinical tests showed no reproductive system infection). Each cell sample was labeled and added to MEM medium (pH 4.0), centrifuged at 120g for 10 minutes, and then repeated centrifuged twice. Wash off the surface bacteria, adjust the concentration of exfoliated vaginal epithelial cells to 79 cells / mL with MEM medium; 8 CFU / mL). The vaginal epithelial exfoliated cells and the Lactobacillus flexus IMC-79 were mixed in equal volumes of 500 μL each, and then placed on a shaker at 37°C (rotational speed: 100r / min, radius of rotation: 2.54cm) and incubated for 1h, and then filtered through a 0.8μm filter membrane , remove the free Lactobacillus flexure IMC-79, and lightly press the filter membrane on the cover glass, fix the cells for Gram staining and observe with an oil microscope...

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Abstract

The invention discloses lactobacillus curvatus and pharmaceutical application thereof. The lactobacillus curvatus provided by the invention specifically is lactobacillus curvatus IMC-79 which has a relatively strong capability of generating H2O2 and can effectively inhibit the growth of external pathogenic bacteria and other conditioned pathogens; and meanwhile, the lactobacillus curvatus IMC-79 also has a relatively strong effect of adhering vaginal epithelial cells to ensure that a strain is fixedly planted on the vaginal epithelium. The lactobacillus curvatus IMC-79 provided by the invention has a certain application prospect in the aspect of preventing and treating bacterial vaginal diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a strain of Lactobacillus crispatus isolated from a human body which can be used to prevent and treat bacterial vaginosis Background technique [0002] Microorganisms inhabit almost every part of the human body, such as the skin, oral cavity, intestinal tract, reproductive tract, etc. There are a large number of bacteria, including many different types of bacteria, which can be roughly divided into "beneficial bacteria" and "conditional pathogenic bacteria". There are two types of germs, and the number and proportion of these bacteria in a healthy human body maintain a stable balance. A variety of bacteria can exist in women's vaginas, among which Lactobacillus is the most common and important beneficial bacteria to maintain a healthy vaginal microecological environment, and Lactobacillus accounts for more than 90% of the total vaginal bacteria in healthy women. They mainly inhibit ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P31/04A61P15/02C12R1/225
CPCA61K35/747C12N1/205C12R2001/225
Inventor 张素华雷天箫陈晏李博宇
Owner 北京博园新里科技有限公司
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