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Application of anthocyanin in preparation of medicine for treating cell damage caused by p-aminophenylarsenic acid

A technology of p-aminophenylarsenic acid and cell damage, which can be used in drug combinations, pharmaceutical formulations, plant raw materials, etc., can solve the problems of anthocyanin cell apoptosis and other problems that have not yet been seen, and achieve the effect of expanding medical uses.

Active Publication Date: 2019-10-15
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on anthocyanins antagonizing cell apoptosis induced by p-aminophenylarsenic acid

Method used

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  • Application of anthocyanin in preparation of medicine for treating cell damage caused by p-aminophenylarsenic acid
  • Application of anthocyanin in preparation of medicine for treating cell damage caused by p-aminophenylarsenic acid
  • Application of anthocyanin in preparation of medicine for treating cell damage caused by p-aminophenylarsenic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: the preparation of Solanum solanum anthocyanins

[0029] (1) Remove the petiole of the fresh nightshade fruit and wash it, beat it with a juicer to obtain a slurry.

[0030] (2) Use hydrochloric acid aqueous solution with pH=1 as the extraction solvent, and ultrasonically extract the slurry, the quality of the extraction solvent and the slurry is 1:6, the ultrasonic power is 600W, ultrasonic 3 times, each time 30min, after three times, use 200 mesh gauze Filter to obtain the crude extract of solanum anthocyanins.

[0031] (3) Mix the crude extract of Solanum solanum anthocyanin with ethyl acetate at a volume ratio of 1:3, perform the first extraction after fully shaking and mixing, and collect the lower aqueous solution; then mix the lower aqueous solution with ethyl acetate in a volume ratio 1:3 for mixing, fully oscillating and mixing for the second extraction, collecting the aqueous solution of the lower layer to obtain the extract of Solanum solanum a...

Embodiment 2

[0034]Embodiment 2: CCK-8 method detects cell viability

[0035] 1. Test method:

[0036] The cell viability was detected by CCK-8 method to study the effect of p-aminophenylarsenic acid and anthocyanin on the apoptosis of DF-1 cells. The anthocyanins are the solanum anthocyanins prepared in Example 1.

[0037] Culture DF-1, and when the cell fusion degree reaches more than 80%, the cells (5×10 5 cells / well) to a 96-well plate (100 μL per well) and cultured for 24 h. The experiment was divided into three treatments, namely: control group, ASA group, NA and ASA co-incubation group; among them, the control group was a blank control without treatment; the ASA group was treated with 0-100mM / L p-aminobenzene arsenic acid Cells for 24 hours; NA and ASA co-incubation group treated cells with 10mM / L p-aminophenyl arsenic acid and 0-200μg / ml anthocyanin for 24 hours.

[0038] After 24 hours, 10 μL CCK-8 reagent was added to each well. Incubate in the incubator for 2h. Be careful ...

Embodiment 3

[0044] Example 3: Detection of cell apoptosis rate by flow cytometry

[0045] 1. Test method:

[0046] (1) Cell culture: DF-1 cells were cultured until the confluence of the cells reached more than 80%, and then subcultured to a 6-well plate and cultured for 24 hours.

[0047] (2) Cell poisoning: the experiment was divided into four groups, control group, p-aminophenylarsenic acid (ASA) challenge group, ASA (10mM) alone group, anthocyanin (NA) (50μg / mL)+ASA (10mM ) co-incubation group, NA (100μg / mL)+ASA (10mM) co-incubation group, the exposure time was 24h. The anthocyanins are the solanum anthocyanins prepared in Example 1.

[0048] (3) Cell collection: absorb the old culture medium and store it in a centrifuge tube, wash the cells twice with PBS, collect the cleaning solution in a centrifuge tube, digest the cells with EDTA-free trypsin, and control the digestion time at 5-7 minutes. Cells were collected upon completion of digestion. Centrifuge at 1000rpm for 3min, disca...

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Abstract

The invention discloses application of anthocyanin in preparation of drugs for treating cell injury caused by arsanilic acid. An in-vitro chick embryo fibroblast DF-1 cell model is established for studying application of arsanilic acid and anthocyanin in apoptosis of DF-1 cells. The application of arsanilic acid and the anthocyanin in apoptosis of the DF-1 cells firstly discovers that arsanilic acid has cytotoxicity on the DF-1 cells and is capable of inducing the apoptosis. The anthocyanin with a certain concentration being 50-100 micrograms per milliliter can resist the apoptosis caused by arsanilic acid; the medical application of the anthocyanin is expanded; the development of drugs taking the anthocyanin as an effective component and having new indications can be facilitated.

Description

technical field [0001] The invention relates to the field of medical applications of anthocyanins, in particular to the application of anthocyanins in the preparation of medicines for treating cell damage caused by p-aminophenylarsenic acid. Background technique [0002] Arsanilic acid (ASA), commonly known as arsanilic acid, has the molecular formula C 6 h 8 AsNO 3 , is an organic arsenic compound formed by substituting the 4-position of the benzene ring of phenylarsonic acid with an amino group. P-aminophenylarsenic acid has the functions of promoting growth, antibacterial and preventing diseases, so it is often used as a drug feed additive and widely used in livestock and poultry breeding. In the early 1950s, the United States took the lead in developing and producing p-aminophenylarsenic acid that can be used as a feed additive, and then it was popularized and used in various countries. In the 1980s, China began to study the use of p-aminophenyl arsenic acid and it w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/352A61P39/02A61K36/81
Inventor 刘建柱谢宁丁新华李洋刘永夏刘康平
Owner SHANDONG AGRICULTURAL UNIVERSITY
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