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Lactobacillus reuteri composition capable of relieving rheumatoid arthritis

A Lactobacillus reuteri, rheumatoid technology, applied in the field of Lactobacillus reuteri composition, to achieve the effect of inhibiting level, relieving rheumatoid arthritis, and improving the composition of intestinal flora

Active Publication Date: 2019-12-27
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no research report on the occurrence and development of Lactobacillus reuteri on rheumatoid arthritis

Method used

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  • Lactobacillus reuteri composition capable of relieving rheumatoid arthritis
  • Lactobacillus reuteri composition capable of relieving rheumatoid arthritis
  • Lactobacillus reuteri composition capable of relieving rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Adhesion performance of Lactobacillus reuteri to human colon adenocarcinoma cell Caco2 in vitro

[0039] Caco2 cells were cultured in DEME (5% fetal bovine serum, 1% antibiotics) medium, and the medium was changed every day until the degree of cell aggregation was 80-90%. After washing three times with PBS, the cells were digested with pre-warmed 0.25% trypsin, and the cell concentration was adjusted to 5×10 5 cells / mL, and inoculated into a six-well culture dish, and placed the culture dish at 37°C, 95% air / 5% CO 2 Cultivate in an incubator, replace the medium once a week or two, and change the medium every day after one week until the growth becomes a dense monolayer.

[0040] Wells with a single cell layer were used for the adhesion experiment. After washing three times with 2 mL of pre-warmed PBS, 1 mL of DEME medium containing 5% fetal bovine serum without antibiotics and 1 mL of DEME culture medium resuspended with end-logarithmic bacteria were added t...

Embodiment 2

[0045] Example 2: Determination of Lactobacillus reuteri tolerance to simulated gastrointestinal fluid in vitro

[0046] Prepare simulated gastric juice: weigh a certain amount of pepsin, dissolve it in 2 / 3 of the required volume of 0.5% (w / v) normal saline, adjust the pH to 3.0 with 6M hydrochloric acid, and make the final concentration of pepsin 3g / L.

[0047] Preparation of simulated intestinal juice: Weigh a certain amount of trypsin and bile salts, dissolve in 2 / 3 of the required volume of 0.5% (w / v) normal saline, adjust to pH 8.0 with 0.1mol / L NaOH, constant volume, make The final concentrations were 1g / L and 0.3g / L, respectively.

[0048] The simulated gastric juice and simulated intestinal juice are prepared and used immediately, and the simulated liquid after constant volume is filtered and sterilized by a 0.22 μm sterile filter membrane before use.

[0049] The activated two-generation Lactobacillus reuteri continued to be subcultured, cultured for 18 hours, coll...

Embodiment 3

[0052] Embodiment 3: the preparation of Lactobacillus reuteri composition preparation

[0053] With Lactobacillus reuteri CCFM14 and CCFM8631, respectively in the MRS medium after continuous activation for three generations, the strains after the activation were transferred to the MRS medium with 5% inoculum (V / V) (adding 0.5% cysteine Hydrochloride) to expand the culture. Centrifuge the expanded cultures of Lactobacillus reuteri CCFM14 and CCFM8631 respectively (5000g, 15min), discard the supernatant, resuspend the sludge with sterile physiological saline, and resuspend the sludge in 30% sterile sucrose solution after cleaning , making the concentration of each strain 5×10 10 CFU / mL, aliquoted, stored in a -80°C refrigerator, thawed before use, and diluted tenfold with sterile normal saline. The diluted cultures of the two strains were uniformly mixed at a ratio of 1:1 of viable bacteria, and placed in a 37°C incubator for 15 minutes to form a Lactobacillus reuteri composit...

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Abstract

The invention discloses a lactobacillus reuteri composition capable of relieving rheumatoid arthritis, and belongs to the technical field of food microorganisms. The concentrated biological preparation is prepared by activating lactobacillus reuteri CCFM8631 and lactobacillus reuteri CCFM14, performing enlarge cultivation, performing centrifugation, and performing re-suspension in a 30% sucrose solution; and the numbers of viable bacteria of the lactobacillus reuteri CCFM8631 and CCFM14 obtained by a plate count process are both greater than 10<10> CFU / mL. The lactobacillus reuteri composition provided by the invention can significantly alleviate rheumatoid arthritis induced by type II collagen, relieve joint redness and swelling symptoms, inhibit the level of antibodies resisting againstthe type II collagen, reduce the content of proinflammatory cytokines and anti-inflammatory cytokines in serum, reduce the levels of Th1 and Th2 cell-related effect characteristic factors, improve anintestinal function, and has broad market application prospects.

Description

technical field [0001] The invention relates to a Lactobacillus reuteri composition capable of alleviating rheumatoid arthritis, and belongs to the technical field of food microorganisms. Background technique [0002] Rheumatoid arthritis (Rheumatoid Arthritis, RA) is a chronic, systemic autoimmune disease, characterized by small joint symmetry and persistent pain. The initial manifestations are the formation of synovitis and pannus, and then joint pain Cartilage destruction, bone tissue inflammation, arthrosclerosis in the later stage, significant narrowing or disappearance of the joint cavity, and eventually develops into osteoarthritis, narrowing or even disappearing of the joint cavity, joint deformity, and functional disability. Epidemiological surveys show that about 0.5%-1% of people worldwide suffer from rheumatoid arthritis, and the prevalence in my country is about 0.32%-0.36%. The occurrence of rheumatoid arthritis is affected by many factors, such as genetics, l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/747A61P19/02A61P29/00A23L33/135
CPCA61K35/747A61P19/02A61P29/00A23L33/135A23V2002/00A23V2400/173A61K2300/00A23V2200/30
Inventor 陈卫杨波樊哲新赵建新张灏
Owner JIANGNAN UNIV
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