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Application of luteolin in preparation of medicine for inhibiting functional activity of pyogenic streptococcus hemolysin

A technology of streptolysin and luteolin, applied in the field of medicine, can solve the problems of immune system dysfunction, exacerbation of drug resistance of pathogenic bacteria, and reduction of clinical treatment effect

Pending Publication Date: 2022-05-13
ACADEMY OF MILITARY MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Western medicine mainly uses antibiotics for the treatment of pathogen-related diseases, but the widespread and irregular application of antibiotics also aggravates the emergence of drug resistance of pathogenic bacteria
This also reduces the effect of clinical treatment, and the single use of antibiotics gradually fails to achieve more satisfactory results. At the same time, it also causes dysfunction of the immune system to a certain extent.

Method used

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  • Application of luteolin in preparation of medicine for inhibiting functional activity of pyogenic streptococcus hemolysin
  • Application of luteolin in preparation of medicine for inhibiting functional activity of pyogenic streptococcus hemolysin
  • Application of luteolin in preparation of medicine for inhibiting functional activity of pyogenic streptococcus hemolysin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, antibacterial activity and time killing test

[0027] GAS (ATCC, USA; SF370) was cultured at 37°C until OD 600=0.8, and the bacterial solution was diluted to 5×10 5 Density in CFU / mL. The minimum inhibitory concentration (MIC) test was performed using the broth microdilution method according to the procedures in the CLSI guidelines (M31-A2). MIC is recorded as the lowest drug concentration at which the culture system becomes turbid after 24 hours.

[0028] Another 18 15mL glass test tubes were taken and divided into 3 groups, 6 in each group. Prepare luteolin (Shanghai Yuanye Biotechnology Co., Ltd.; B20888) into 5 concentrations of 64, 128, 256, 512, and 1024 μg / mL, with a total of 6 mL for each concentration, and add them to the test tubes in sequence, and add 2mL, add THY medium without luteolin to the last test tube, and dilute the bacterial solution to 2×10 5 The density of CFU / mL was added to 2.5 mL of the above test tubes respectively. The fin...

Embodiment 2

[0032] Embodiment 2, hemolytic activity test

[0033] Refer to the previous experimental protocol of red blood cell hemolysis [Dong J, Liu Y, Xu N, Yang Q, Ai X. MorinProtects Channel Catfish From Aeromonas hydrophila Infection by Blocking Aerolysin Activity. Front Microbiol. 2018; 9:2828, Dong J, Zhang L, Liu Y, Xu N, Zhou S, Yang Y, et al. Luteolin decreases the pathogenicity of Aeromonas hydrophilavia inhibiting the activity of aerolysin. Virulence. 2021; 12:165-76] with modifications. Briefly, recombinant SLO (0.1 g / mL) was added to the reaction system with or without luteolin, and sheep red blood cells (RBCs) were collected and added to the above reaction system to the final concentration 2% (v / v). The entire mixed system was incubated at 37°C for 30 minutes, then centrifuged at 5000 rpm for 5 minutes, and the OD of the supernatant was obtained 540 . The completely hemolyzed system treated with 0.1% Triton-X100 was used as a positive control. Final hemolysis percentag...

Embodiment 3

[0035] Embodiment 3, surface plasmon resonance analysis (Surface plasmon resonance, SPR)

[0036] The affinity and kinetics of luteolin for SLO were measured by SPR at 25 °C on a REICHERT 4SPR using a carboxymethyl dextran hydrogel surface sensor chip. Firstly, to measure the binding of protein to small molecules, the protein was dissolved in 10 mM sodium acetate (pH 4.5) and immobilized on a chip with 6000 response units (RU) at a flow rate of 10 μL / min. Luteolin was serially diluted in PBST buffer (PBS containing 0.005% Tween 20) to a concentration of 0.125g / mL, 0.25g / mL, 0.5g / mL, 1g / mL, 2g / mL, 4g / mL, The infusion was performed at a flow rate of 25 μL / min for 3 min; for dissociation, the flow rate was set at 25 μL / min for 5 min. Reference values ​​and signals from buffer-injected controls were subtracted and sensor traces were fitted to a 1:1 Langmuir binding model using the data analysis program Scriber 2.0 to calculate ka, kd and KD values.

[0037] In order to verify th...

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Abstract

The invention discloses an application of luteolin in preparation of a medicine for inhibiting functional activity of pyogenic streptococcus hemolysin. Experiments show that luteolin can bind to SLO protein with high affinity to inhibit the functional activity of pyogenic streptococcus hemolysin, and it is proved that luteolin may become a potential inhibitor for effectively treating GAS infection.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to the application of luteolin in the preparation of medicines for inhibiting the functional activity of streptolysin pyogenes. Background technique [0002] Group A streptococcus (GAS), also known as Streptococcus pyogenes, is a Gram-positive bacterium that can cause a variety of illnesses ranging from pharyngitis, erysipelas, and cellulitis to more serious life-threatening diseases such as streptococcal toxic shock syndrome and necrotizing fasciitis. In addition, persistent GAS infection can lead to severe immune-mediated diseases, such as acute glomerulonephritis, rheumatic fever, and rheumatic heart disease [Martin JM, Green M. Group Astreptococcus. Semin Pediatr Infect Dis. 2006; 17:140 -8; Steer AC, Danchin MH, Carapetis JR. Group A streptococcal infections in children. J Paediatr ChildHealth. 2007; 43:203-13; Walker MJ, Barnett TC, McArthur JD, Cole JN, Gillen CM, Henning...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/352A61P31/04
CPCA61K31/352A61P31/04
Inventor 刘鹏姜永强郭婷婷黄文华江华孔德聪律清宇
Owner ACADEMY OF MILITARY MEDICAL SCI
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