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A strain of Lactobacillus casei that can alleviate rheumatoid arthritis and its application

A technology of Lactobacillus casei and rheumatoid, applied in the field of microorganisms, can solve the problems of high cost, excessive suppression of the immune system, and induction of other diseases, and achieve the effect of low cost

Active Publication Date: 2021-04-13
无锡食生臻选生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, in the treatment of rheumatoid arthritis (Rheumatoid Arthritis, RA), the clinical use is limited to the use of some biological agents targeting inflammatory factors such as IL-6 or TNFα, and these biological agents are effective in short-term clinical application. However, it can only temporarily relieve and improve the condition, and it is difficult to fundamentally control the development of the condition. It is recurrent and expensive. In addition, biological agents are a cascade reaction that forcibly prevents immune inflammation in the body. Long-term use will cause patients The immune system is oversuppressed, and there is a risk of inducing other diseases

Method used

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  • A strain of Lactobacillus casei that can alleviate rheumatoid arthritis and its application
  • A strain of Lactobacillus casei that can alleviate rheumatoid arthritis and its application
  • A strain of Lactobacillus casei that can alleviate rheumatoid arthritis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1: Screening of Lactobacillus casei and identification of bacterial species

[0064] 1. Filter

[0065] Healthy human fecal samples from the Wuxi area were stored in a -80°C freezer in 30% glycerol. After the samples were taken out and thawed at low temperature, 0.5mL samples were mixed and added to 4.5mL, and 0.9% normal saline containing 0.05% cysteine ​​was used for gradient dilution. On the MRS solid medium of cystine, culture at 37°C for 48h, pick typical colonies to streak purification on MRS solid medium, pick a single colony and transfer it to MRS liquid medium (containing 0.05% cysteine) Cultured in 30% glycerol, the cultured cells were stored in 30% glycerol to obtain strains CCFM1074, L1 and L2.

[0066] 2. Identification

[0067] Extract the genome of CCFM1074, L1, L2, the 16S rDNA of CCFM1074, L1, L2 is amplified and sequenced (by Huada Gene Technology Co., Ltd., wherein, the nucleotide sequence of the 16S rDNA amplification of CCFM1074 is suc...

Embodiment 2

[0068] Example 2: Cultivation of Lactobacillus casei

[0069] Lactobacillus casei CCFM1074 was inserted into MRS solid medium (containing 0.05% cysteine) and cultured at 37°C for 48 hours, and the colonies were observed, and the colonies were found to be milky white, with smooth surface and round protrusions.

[0070] Lactobacillus casei CCFM1074 was inserted into MRS liquid medium (containing 0.05% cysteine) and cultured anaerobic at 37°C for 24 h, then transferred to fresh MRS liquid medium (containing 0.05% cysteine). ), cultured under the same conditions for 24h, centrifuged the cells at 6000g for 15min, washed the cells with 0.9% saline, and centrifuged them again at 6000g for 10min to obtain the cells, resuspended in 30% sucrose solution, and frozen at -80°C for later use.

Embodiment 3

[0071] Example 3: Tolerance of different Lactobacillus casei to simulated gastrointestinal fluids

[0072] 1. Tolerance of different Lactobacillus casei to simulated gastric juice

[0073] Lactobacillus casei (Lactobacillus casei) CCFM1074, Lactobacillus casei (Lactobacillus casei) L1, and Lactobacillus casei (Lactobacillus casei) L2 obtained in Example 1 were respectively inserted into the MRS liquid medium and cultured at 37° C. for 18 h, and the cells were collected by centrifugation. , the collected cells were washed with physiological saline, after washing, the cells were collected by centrifugation again, and the collected cells were resuspended in pH 3 (pH adjusted by HCl) containing 3 g / L of pepsin-containing physiological saline, kept The initial concentrations of Lactobacillus casei CCFM1074, Lactobacillus casei L1, and Lactobacillus casei L2 in normal saline were the same, and 0.1 mL of bacterial liquid was taken to count the viable bacteria on the plate by pouring me...

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Abstract

The invention discloses a strain of Lactobacillus casei which can alleviate rheumatoid arthritis and application thereof, belonging to the technical field of microorganisms. The present invention has screened out a strain of Lactobacillus casei CCFM1074, and this Lactobacillus casei CCFM1074 has the function of alleviating rheumatoid arthritis, which is embodied in: significantly alleviating the weight loss of rheumatoid arthritis rats; significantly reducing the rheumatoid arthritis The contents of pro-inflammatory cytokines TNFα and IL-1β in the serum of rats with rheumatoid arthritis; the content of short-chain fatty acids in feces of rats with rheumatoid arthritis was significantly regulated; the content of Th17 and Treg in mesenteric lymph nodes of rats with rheumatoid arthritis was significantly reduced The proportion of cells; improve the intestinal flora of rheumatoid arthritis rats at the genus level; significantly inhibit the growth of rheumatoid arthritis pathogenic bacteria Prevotella in vitro.

Description

technical field [0001] The invention relates to a Lactobacillus casei strain capable of relieving rheumatoid arthritis and its application, belonging to the technical field of microorganisms. Background technique [0002] Rheumatoid Arthritis (RA) is an autoimmune disease with symmetrical polyarthritis as the main clinical manifestation. The disease is characterized by chronic inflammation of the joint synovium and progressive destruction of the joints. The clinical manifestations are varied and the prognosis is often poor. In the advanced stage, it can cause joint stiffness, deformity and serious damage to function. Damage to organs and multiple systems seriously endangers the health and quality of life of patients. [0003] Rheumatoid arthritis (Rheumatoid Arthritis, RA) occurs all over the world, and northern my country is the high incidence area of ​​the disease. Therefore, the prevention and treatment of rheumatoid arthritis (RA) have important theoretical and social ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/747A61P29/00A61P19/02A61P31/04A23L33/135C12R1/245
CPCA61K35/747A61P29/00A61P19/02A61P31/04A23L33/135A23V2002/00C12R2001/245C12N1/205A23V2400/125A23V2200/30
Inventor 陈卫杨波樊哲新赵建新张灏
Owner 无锡食生臻选生物科技有限公司
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