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Lactobacillus rhamnosus NX-2 and application thereof in preparation of uric acid reducing drugs

A technology of Lactobacillus rhamnosus and NX-2, applied in the field of microorganisms, can solve the problems of relying on import of probiotic strains, lack of functional strains, affecting the promotion of probiotics and their products, etc.

Active Publication Date: 2020-10-27
广东南芯医疗科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many types of probiotic products in the domestic market, there is a lack of functional strains with independent intellectual property rights in my country
The probiotic strains used by domestic production enterprises have long relied on imports, and foreign strains may not be suitable for the intestinal physiological conditions of Chinese residents
In addition, the function of probiotics lacks strong scientific research evidence, which seriously affects the promotion of probiotics and their products

Method used

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  • Lactobacillus rhamnosus NX-2 and application thereof in preparation of uric acid reducing drugs
  • Lactobacillus rhamnosus NX-2 and application thereof in preparation of uric acid reducing drugs
  • Lactobacillus rhamnosus NX-2 and application thereof in preparation of uric acid reducing drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Isolation, Identification and Preservation of Lactobacillus rhamnosus NX-2

[0025] (1) Separation: Inoculate the feces of infants and young children into TPY solid medium, MRS solid medium and BDS solid medium respectively after gradient dilution, culture anaerobically at 37°C for 48 hours, pick single colonies on the plate and separate them by streaking to obtain pure colonies. The pure colonies on the plate were inoculated into MRS liquid medium, cultured anaerobically at 37°C for 12-16h, added 20% glycerol, and stored in a -80°C refrigerator.

[0026] (2) Morphological identification of bacterial strains: Gram-stained strains were observed under a microscope, Gram-positive bacteria were purple, and Gram-negative bacteria were red.

[0027] (3) Molecular biological identification of the strain: Genomic DNA was extracted from the obtained strain, and the full-length 16S rDNA fragment was amplified by PCR using 16S rDNA universal primers 27F and 1492R, follo...

Embodiment 2

[0033] Preparation of Example 2 Lactobacillus rhamnosus NX-2 fermentation supernatant (extracellular secretion), bacterial suspension (thalline) and cell disruption supernatant (intracellular substance)

[0034] Lactobacillus rhamnosus NX-2 was activated and cultured and inoculated in MRS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermentation bacteria was adjusted to 10 9 CFU / mL, centrifuged at 4°C, 6000r / min for 10min to obtain the culture supernatant and cell pellet, the supernatant was filtered through a 0.22μm filter membrane to obtain the fermentation supernatant (extracellular secretion); the cell pellet was filtered through PBS twice After the first wash, the cells were resuspended with PBS, and the cell concentration was adjusted to 10 9 CFU / mL to obtain bacterial suspension (bacteria); process the bacterial suspension in an ice bath with an ultrasonic breaker, work for 3s, interval 8s, ultrasonic break for 15min, centrifuge at 12000×g f...

Embodiment 3

[0035] Example 3 Effect of Lactobacillus rhamnosus NX-2 on uric acid content in high uric acid zebrafish

[0036] Potassium oxalate and allopurinol were purchased from Shanghai Luyuan Biotechnology Co., Ltd., and xanthine sodium salt was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0037] Model construction: the wild-type AB line zebrafish (5dpf) was placed in a six-well plate. The experiment set blank control group, model group, positive control group (allopurinol), sample (bacterial suspension, fermentation supernatant, cell crush supernatant) intervention group, each group set up 3 duplicate holes, each hole 50 Zebrafish (5dpf) were treated with 10mmoL / L potassium oxalate and 0.5mmoL / L xanthine sodium salt for 24h to construct a hyperuricemia model.

[0038] Intervention: add 5mL culture water to each hole of blank control group and model group; add 5mL100mg / L allopurinol solution to each hole of positive control group; add 5mL 10% (v / v, diluted in cu...

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Abstract

The invention discloses lactobacillus rhamnosus NX-2 and an application thereof in preparation of uric acid reducing drugs, and belongs to the technical field of microorganisms. The lactobacillus rhamnosus NX-2 disclosed by the invention has a preservation number of CGMCC No.20110. Non-inactivated and inactivated fermentation supernate, bacterial suspension and cell disruption material supernate of the lactobacillus rhamnosus NX-2 have uric acid reducing effects on a zebrafish high-uric-acid model, and can inhibit the activity of xanthine oxidase in the body of a high-uric-acid zebrafish. Thelactobacillus rhamnosus NX-2 disclosed by the invention has a huge potential application prospect in the aspect of preparing medicines for treating and / or preventing hyperuricemia.

Description

technical field [0001] The invention relates to the technical field of microbes, and more specifically relates to a strain of Lactobacillus rhamnosus NX-2 and its application in the preparation of uric acid-lowering drugs. Background technique [0002] Hyperuricemia (HUA) is a metabolic disease caused by abnormal purine metabolism in the human body that increases the production of uric acid (Uric Acid, UA) and / or reduces the renal excretion of UA. High levels of uric acid in the blood are known to lead to the formation of uric acid crystals in the joints, which can lead to pain, heat, and swelling, often referred to as gout or arthritis. In recent years, hyperuricemia is also considered to be closely related to the occurrence of insulin-resistant diabetes, hyperlipidemia, obesity, kidney disease and other diseases. Xanthine oxidase (Xanthine Oxidase, XO) has been recognized as one of the promising targets for the treatment of hyperuricemia, inhibition of XO activity can eff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/747A61P19/06C12R1/225
CPCA61K35/747A61P19/06C12R2001/225C12N1/205
Inventor 张召郑康帝赵笑笑陈伙带雷春燕田甜陈涛
Owner 广东南芯医疗科技有限公司
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