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Application of vitamin C in preparation of medicines for treating and preventing tuberculosis

A technology for vitamins, tuberculosis, applied in the field of biomedicine

Pending Publication Date: 2018-07-27
SHANGHAI PUBLIC HEALTH CLINICAL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have found that oxidative stress has an important impact on cell apoptosis, and excessive oxidative stress levels will induce cell apoptosis, but so far there has been no research on how Mycobacterium tuberculosis can regulate the state of oxidative stress in cells after infection of macrophages. It further affects cell apoptosis, so that latent tuberculosis is released from infected macrophages and killed by VC

Method used

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  • Application of vitamin C in preparation of medicines for treating and preventing tuberculosis
  • Application of vitamin C in preparation of medicines for treating and preventing tuberculosis
  • Application of vitamin C in preparation of medicines for treating and preventing tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Vitamin C (VC) cytotoxicity test:

[0062] The median lethal dose (LD) to infected cells was analyzed by cytotoxicity test (MTT). 50 ), first inoculate RAW264.7 cells in a 96-well plate, about 4,000-10,000 cells per well, and the cells adhered to the wall overnight. The next day, the cells were infected with BCG (MOI=5) for 3 hours, and then different drug action conditions such as 5mM VC were used , 5mM VC+5mM NAC, and 5mM VC+100U / ml of catalase (Catalase) after 24h treatment, discard the supernatant, and then wash with PBS 3 times, and then add 100μl MTT (5mg / mL ) in the DMEM medium, and continue culturing in the cell culture box for 4 hours, discard the medium in the well, add 150 μl DMSO to each well, place it on a microplate reader to read the absorbance at 570 nm, calculate the half-lethal dose of VC, and combine it with clinical VC The amount of treatment, obtain the pharmaceutical dose of VC, and further determine the amount of intravenous infusion of...

Embodiment 2

[0064] Example 2 VC and H 2 o 2 Experiment on molecular mechanism of RAW264.7 macrophages infected with Bacillus Calmette-Guerin (BCG)

[0065] RAW264.7 macrophages were infected with the virulent strain H37Rv and the BCG strain BCG cultured in Michaelis-Menten 7H9 liquid medium respectively, and the infected cells were divided into VC infection group, H 2 o 2 Induction group and VC induction group were tested to detect H 2 o 2 The apoptosis of macrophages after infection was induced by VC and VC, stained by AnnexinV-FITC / PI method, and the change of cell apoptosis rate was detected by flow cytometry; in addition, ROS content was analyzed, cells were stained by DCFH-DA, flow cytometry Type cytometer or laser confocal microscope to detect changes in the content of reactive oxygen species (ROS) in cells, long-term induction of JLP[S] cells by VC to produce drug-resistant JLP[R] cell lines, using O 2 Electrode method to compare the two cell lines cultured for different time ...

Embodiment 3

[0066] Example 3 VC and H 2 o 2 Molecular experiments on treating primary mouse peritoneal macrophages infected with Bacillus Calmette-Guerin (BCG)

[0067]BALB / c female mice were stimulated with broth, and mouse peritoneal macrophages were isolated and cultured. The primary macrophages were respectively infected with the virulent strain H37Rv and BCG strain cultured in Michaelis 7H9 liquid medium, and the post-infection cells according to the infection group, H 2 o 2 Induction group and VC induction group were used for experiments, that is, H 2 o 2 Macrophage apoptosis was induced by VC and VC, the above three groups of cells were stained with Annexin V-FITC / PI, and the change of cell apoptosis rate was detected by flow cytometry; at the same time, the content of ROS was analyzed, and the cells were treated with DCFH-DA Perform staining to detect changes in the active oxygen content in the cells; use an enzyme detection kit to detect changes in the expression of metabolo...

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Abstract

The invention belongs to the field of biomedicines, and relates to an application of vitamin C (ascorbic acid), as a single component, in preparation of medicines for treating and preventing tuberculosis. Through cytobiology and pharmacology experiments with combination of animal experiments, anti-tuberculosis pharmaceutical concentration of the vitamin C is determined; and on the basis of regulatory mechanism on oxidative stress status of mycobacterium tuberculosis after infection by macrophages, interference test is carried out, wherein a result proves that high-dose vitamin C can kill the mycobacterium tuberculosis, so that inhibition effect is achieved through NAC and catalase; the vitamin C treats the macrophages to generate H2O2, wherein functional effect is achieved with the signalchannel being same as H2O2; large-dose vitamin C kills mycobacteria, wherein inhibition function is achieved through a glutathione precursor NAC and CAT; high-dose vitamin C can infect the TNF-alpha signal pathway induced by RAW 264.7 cells via mediation of Bacillus Calmette-Guerin vaccine and H37Rv, thus achieving sterilization effect and influence on body immunity. The invention provides effective treatment strategy for clinical therapy and drug resistance of tuberculosis and tuberculosis in incubation period.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the new application of vitamin C (ascorbic acid) in pharmacy, in particular to the application of vitamin C (ascorbic acid) in preparing medicines for treating and preventing tuberculosis. Background technique [0002] The information disclosed that tuberculosis (Tuberculosis, TB) is currently one of the infectious diseases with the highest morbidity and mortality in the world. According to estimates by the World Health Organization, nearly 1 / 3 of the world's people are infected with Mycobacterium tuberculosis (Mtb) , 10% of them developed into active tuberculosis, and most of them turned into latent infection. According to the assessment of the World Health Organization, the current annual incidence of tuberculosis in my country is about 1.3 million, accounting for 14% of the global incidence, ranking second in the world, second only to India. In recent years, clinical studies have foun...

Claims

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Application Information

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IPC IPC(8): A61K31/375A61K45/06A61P31/06
CPCA61K45/06A61K31/375
Inventor 汪进裴增林张璇纪春霞
Owner SHANGHAI PUBLIC HEALTH CLINICAL CENT
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