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Bifidobacterium breve strain capable of relieving rheumatoid arthritis, and application of bifidobacterium breve strain

A bifidobacterium breve, rheumatoid technology, applied in the field of microorganisms, can solve the problems of lack, research, etc., and achieve the effect of low cost

Active Publication Date: 2020-12-22
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, some progress has been made in the prevention and / or treatment of rheumatoid arthritis. Inflammation, however, the Bifidobacterium bifidum KCTC13474BP relieves rheumatoid arthritis, mainly focusing on the anti-inflammatory effect in vitro cells and the inhibition of rheumatoid arthritis autoantibodies, and there is still a lack of research on other indicators

Method used

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  • Bifidobacterium breve strain capable of relieving rheumatoid arthritis, and application of bifidobacterium breve strain
  • Bifidobacterium breve strain capable of relieving rheumatoid arthritis, and application of bifidobacterium breve strain
  • Bifidobacterium breve strain capable of relieving rheumatoid arthritis, and application of bifidobacterium breve strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1: Screening and strain identification of Bifidobacterium breve CCFM1078

[0061] 1. Screening

[0062] Stool samples from healthy infants in Wuxi were stored in 30% glycerol in a -80°C freezer. After taking out the sample and thawing it at low temperature, mix the sample evenly, draw 0.5mL sample and add it to 4.5mL, carry out gradient dilution with 0.9g / 100mL physiological saline containing 0.05g / 100mL cysteine, and select the appropriate gradient dilution solution to apply on the On the MRS solid medium containing 0.05g / 100mL cysteine, cultured at 37°C for 48h, picked typical colonies to streak and purify on the MRS solid medium, picked a single colony and transferred to MRS liquid medium (containing 0.05 g / 100mL cysteine), the cultured thalli were preserved in 30% glycerol to obtain bacterial strain CCFM1078.

[0063] 2. Identification

[0064] The genome of CCFM1078 was extracted, and the 16S rDNA of CCFM1078 was amplified and sequenced (by Huada Gene Te...

Embodiment 2

[0067] Embodiment 2: the cultivation of Bifidobacterium breve CCFM1078

[0068] The Bifidobacterium breve (Bifidobacterium breve) CCFM1078 obtained in Example 1 was inserted into the MRS solid medium (containing 0.05g / 100mL cysteine) and cultivated in an anaerobic incubator at 37°C for 48h, and its colonies were observed, and it was found that Its colonies are milky white, smooth, and rounded.

[0069] The Bifidobacterium breve (Bifidobacterium breve) CCFM1078 obtained in Example 1 was inserted into MRS liquid medium (containing 0.05g / 100mL cysteine) and after anaerobic culture at 37°C for 24h, it was transferred to fresh MRS liquid medium (containing 0.05g / 100mL cysteine), cultivated under the same conditions for 24 hours, centrifuged the bacteria at 6000g for 15min, washed the bacteria with 0.9g / 100mL normal saline, centrifuged again at 6000g for 10min to obtain the bacteria, and resuspended with 30% sucrose solution , and stored at -80°C until use.

Embodiment 3

[0070] Example 3: Tolerance of Bifidobacterium breve CCFM1078 to simulated gastrointestinal fluid

[0071] 1. Tolerance of Bifidobacterium breve CCFM1078 to simulated gastric juice

[0072]Insert the Bifidobacterium breve (Bifidobacterium breve) CCFM1078 obtained in Example 1 into the MRS liquid medium and culture it anaerobically at 37°C for 18 hours, then collect the cells by centrifugation, wash the collected cells with physiological saline, and centrifuge again after the washing is completed. Collect the cells, resuspend the collected cells in physiological saline containing 3g / L pepsin at a pH of 3 (pH is adjusted by HCl), take 0.1mL of the bacterial liquid to count viable bacteria on the plate by the pouring method as the bacterial liquid The original number of viable bacteria of Bifidobacterium breve (Bifidobacterium breve) CCFM1078, put the remaining bacterial liquid at 37°C for anaerobic culture for 3 hours, take 0.1mL of the bacterial liquid to count the viable bacte...

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Abstract

The invention discloses a bifidobacterium breve strain capable of relieving rheumatoid arthritis, and application of the bifidobacterium breve strain, and belongs to the technical field of microorganisms. According to the invention, the bifidobacterium breve strain CCFM1078 is screened out; the bifidobacterium breve strain CCFM1078 has the effect of relieving rheumatoid arthritis and is specifically embodied in that: RAW264.7 cells are remarkably promoted to secrete an anti-inflammatory factor IL-10; the joint thickness, clinical score and morbidity of a rat suffering from rheumatoid arthritisare remarkably reduced; the content of pro-inflammatory cytokines TNF alpha and IL-1 beta in serum of the rat suffering from rheumatoid arthritis is remarkably reduced; the content of short-chain fatty acid in excrement of the rat suffering from rheumatoid arthritis is remarkably adjusted; the proportion of Th17 cells in mesenteric lymph nodes of the rat suffering from rheumatoid arthritis is remarkably reduced; and, a pro-inflammatory factor IL-6 secreted by synovial cells is remarkably reduced.

Description

technical field [0001] The invention relates to a strain of bifidobacterium breve capable of alleviating rheumatoid arthritis and its application, belonging to the technical field of microorganisms. Background technique [0002] Rheumatoid Arthritis (RheumatoidArthritis, RA) is a complex autoimmune disease, the pathological features are mainly joint synovitis, deterioration to cartilage and bone tissue destruction, eventually leading to joint stiffness and dysfunction. The incidence of rheumatoid arthritis (RheumatoidArthritis, RA) is higher in women than in men, and higher in middle-aged and elderly people than in children, and it shows a certain degree of heredity. [0003] At present, in the treatment of rheumatoid arthritis, traditional drugs such as antirheumatic drugs, glucocorticoids, non-steroidal anti-inflammatory drugs, and some new biological agents are often used. These drugs have limitations. For example, in the initial stage of the disease, traditional drugs a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23C9/12A23C13/16A23C15/06A23C15/12A23C15/16A23C17/02A23C19/032A23C19/06A23L11/00A23L13/40A23L29/00A23L33/135A61K35/745A61P19/02A61P29/00C12R1/01
CPCC12N1/20A23L33/135A23L13/45A23L29/065A23C9/1203A23C13/16A23C15/065A23C15/123A23C15/165A23C17/02A23C19/0323A23C19/062A61K35/745A61P19/02A61P29/00A23V2002/00C12R2001/01C12N1/205A23V2400/519A23V2200/30A23L2/02A23L2/382A23L2/52A23L33/40
Inventor 杨波陈卫樊哲新赵建新张灏
Owner JIANGNAN UNIV
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