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Application of orychophragmine D in preparation of medicine for inhibiting small intestine crypt epithelial cell ferroptosis

A technology of epithelial cells and ferroptosis, applied in the field of biomedicine, can solve problems such as narrow treatment time window, poor drug effect, and inhibition of spermatogenic cells, and achieve the effect of protecting radiation damage

Pending Publication Date: 2022-02-11
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Clinically, the existing anti-radiation drugs such as aminothiol-based anti-radiation drugs, the representative drug is amifostine, which has a narrow treatment time window and serious adverse reactions such as hypotension, dizziness, and vomiting, which limit its clinical application Hormonal radiation damage prevention and treatment drugs such as nylestriol have good curative effect, but hormone drugs have a huge impact on sexual organs. Long-term repeated use can cause female menstrual disorders, male testicular atrophy, and side effects such as spermatogenic cell inhibition, which limits its clinical application
It can be seen that for radiation injury, the effect of conventional treatment drugs is not good, and it is urgent to study the new mechanism of nuclear radiation intestinal injury and develop new and more effective treatment techniques and products

Method used

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  • Application of orychophragmine D in preparation of medicine for inhibiting small intestine crypt epithelial cell ferroptosis
  • Application of orychophragmine D in preparation of medicine for inhibiting small intestine crypt epithelial cell ferroptosis
  • Application of orychophragmine D in preparation of medicine for inhibiting small intestine crypt epithelial cell ferroptosis

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preparation example Construction

[0024] The present invention has no special limitation on the preparation method of the zhugraine D, and the conventional preparation method in this field can be adopted. In a specific embodiment of the present invention, the preparation of the zhugraine D OV16 is preferably prepared according to the application number Prepared for the patent document of 201910175859.9.

[0025] In the present invention, the Erastin is a classical ferroptosis activator, acting on the mitochondrial voltage-dependent anion channel (VDAC) of various cell types. In order to establish the ferroptosis model of the rat intestinal crypt epithelial cell line IEC6, the present invention uses different concentrations of Erastin to treat the IEC6 cells, and observes the cell death induced by Erastin.

[0026] In the specific embodiment of the present invention, data analysis uses Graphpad Prism version 5.0 software (GraphPad software, Inc., La Jolla, CA, USA). All results are expressed as mean ± standard...

Embodiment 1

[0033] Erastin induces ferroptosis in rat intestinal crypt epithelial cell line IEC6

[0034] Cell viability was evaluated using CCK-8 (Dojindo Laboratories, KμMamoto, Japan) according to the kit instructions. IEC6 cells at 5 x 10 3 The concentration of cells (100 μL / well per well) was inoculated in a 96-well plate, and they were divided into two groups, Erastin treatment group and Fer-1 treatment group, with an average of 4 replicate wells in each group. Among them, the Erastin treatment group used Erastin (Tocris, Minneapolis, MN, USA) to treat the cells, and the final concentration of the medium was 0 μM, 0.25 μM, 0.5 μM, 1 μM, 2 μM, 5 μM, 10 μM; At the same time, the cells were treated with ferrostatin-1 (Fer-1) (Sigma-Aldrich, St Louis, MO, USA), and the final concentration of Fer-1 in the medium was 1 μM. Cells in each group were stored at 37°C, 5% CO 2 Culture in an incubator, detect cell viability after 24 hours, then add CCK-8 10 μL / well in each group, at 37°C, 5% ...

Embodiment 2

[0037] Zhugraine D (OV16) as an inhibitor of Erastin-induced ferroptosis in IEC6 cells

[0038] After digesting the IEC6 cells in the logarithmic growth phase, 5×10 3 The concentration of cells (100 μL per well) was inoculated in 96-well plates, and they were divided into three groups, namely Erastin treatment group, Fer-1 treatment group and OV16 treatment group. Among them, the Erastin treatment group was only treated with 2.5 μM Erastin, and the final concentration of the medium was 2.5 μM; the Fer-1 treatment group was treated with 2.5 μM Erastin plus 1 μM Fer-1; the OV16 treatment group was treated with 2.5 μM Erastin and 1, 5, Treatment with 10 μM zhugraine D (OV16) natural product compound. After each group was treated and reacted for 24 hours, the cell viability was evaluated using Cell Counting Kit-8 (CCK-8) according to the instructions (Dojindo Laboratories, Tokyo, Japan), and the results were as follows: figure 2 . All data are expressed as mean ± standard devi...

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Abstract

The invention belongs to the technical field of biological medicine. The invention provides an application of orychophragmus violaceine D, namely an OV16 monomeric compound, in preparation of drugs for inhibiting small intestine crypt epithelial cell ferroptosis, the orychophragmus violaceine D OV16 can inhibit rat small intestine crypt epithelial cell ferroptosis induced by Erastin, and the orychophragmus violaceine D OV16 is an effective inhibitor for rat small intestine crypt epithelial cell ferroptosis induced by Erastin. Experiments prove that when the OV16 is used for treating 14.5 Gy abdominal irradiation cobalt-source mouse C57 strain, radiation injury of mouse intestinal crypt tissues can be protected, and a theoretical basis is provided for research on treatment and prevention of intestinal radiation injury by the orychophragine DOV16 in the future.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of zhurinaine D in the preparation of drugs for inhibiting ferroptosis of small intestinal crypt epithelial cells. Background technique [0002] In today's society, with the wide application of ionizing radiation technology in the fields of national economy, medical treatment and military affairs, while benefiting mankind, they also bring potential nuclear and radiation dangers to the world today, and various nuclear accidents and radiation accidents occur frequently How to prevent and treat body damage caused by ionizing radiation has become a major concern worldwide. [0003] Radiation-induced intestinal injury (RII) is an important type of tissue injury in acute radiation sickness, and it is also a major scientific problem faced by radiation medicine. Clinically, the existing anti-radiation drugs such as aminothiol-based anti-radiation drugs, the represent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/53A61P1/00
CPCA61K31/53A61P1/00
Inventor 肖凤君张广杰杜丽卢育新程晓晨董俊兴王立生
Owner ACADEMY OF MILITARY MEDICAL SCI
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