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Application of dihydromyricetin in preparing drugs for treating iron overload diseases

A technology for dihydromyricetin and disease treatment, applied in metabolic diseases, drug combinations, pharmaceutical formulations, etc., can solve the problem that polyphenols do not have the expected effect and other problems, and achieve the effect of protecting cells from damage

Inactive Publication Date: 2019-04-19
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, most polyphenols did not have the expected effect

Method used

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  • Application of dihydromyricetin in preparing drugs for treating iron overload diseases
  • Application of dihydromyricetin in preparing drugs for treating iron overload diseases
  • Application of dihydromyricetin in preparing drugs for treating iron overload diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Cell Viability Detection

[0038] 1) Digest and centrifuge the HT1080 cells and AML12 cells grown to 80%-90% confluence, and dilute them at 1×10 per well 4 The number of cells was inoculated into a 96-well cell culture plate and cultivated for 24 hours;

[0039]2) Add control and 10 μM dihydromyricetin to the 96-well plate containing the cells; then add Fe-8HQ 10 μM to each well plate for 4 hours, and set 6 replicates in each group;

[0040] 3) At 37°C, 5% CO 2 Conditioned cell culture incubator;

[0041] 4) Carefully pipette the treatment solution in the 96-well plate, and add the cell culture medium containing 10% CCK-8 cell viability detection reagent prepared in advance, and continue to store at 37°C, 5% CO 2 Cultivate in a conditioned cell culture incubator for 1 to 4 hours;

[0042] 5) Cell activity was detected on the SpectraMax M5 microplate detection system, and the detection parameter of CCK-8 was 450nm.

[0043] 6) Calculate the cell viability...

Embodiment 2

[0045] Example 2: Determination of cytoplasmic reactive oxygen species ROS.

[0046] There are various ROS in cells, which can be stained by specific reagents, among which cytoplasmic ROS can be stained with CM-H2DCFDA.

[0047] 1) The day before the test, human fibroblastosarcoma carcinoma HT1080 cells grown to 80%-90% confluence were digested and centrifuged, and 2×10 5 The number of cells, inoculated into 6-well cell culture plate, and the cells were grown for 24 hours;

[0048] 2) Treat cells with DMSO, 10 μM Fe-8HQ, 10 μM Fe-8HQ+10 μM dihydromyricetin, 10 μM dihydromyricetin for 4 hours;

[0049] 3) After the treatment time is up, discard the culture medium in the six-well plate, gently wash twice with PBS, add 600 μL of Accutase cell digestion solution to each well to digest the cells, then add the medium to stop the digestion, gently pipette and collect the cells, and rotate at 2500 rpm Centrifuge for 5 minutes;

[0050] 4) After the supernatant was discarded, 500 μL...

Embodiment 3

[0052] Example 3: Determination of cellular mitochondrial reactive oxygen species ROS.

[0053] There are various ROS in cells, which can be stained by specific reagents, among which mitochondrial ROS can be stained by MitoSOX Red.

[0054] 1) The day before the test, human fibroblastosarcoma carcinoma HT1080 cells grown to 80%-90% confluence were digested and centrifuged, and 2×10 5 The number of cells, inoculated into 6-well cell culture plate, and the cells were grown for 24 hours;

[0055] 2) Treat the cells with DMSO, 10 μM Fe-8HQ, 10 μM Fe-8HQ+10 μM dihydromyricetin, 10 μM dihydromyricetin for 4 hours;

[0056] 3) After the treatment time is up, discard the culture medium in the six-well plate, gently wash twice with PBS, add 600 μL of Accutase cell digestion solution to each well to digest the cells, then add the medium to stop the digestion, gently pipette and collect the cells, and rotate at 2500 rpm Centrifuge for 5 minutes;

[0057] 4) After discarding the supern...

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Abstract

The invention discloses application of dihydromyricetin in preparing drugs for treating iron overload diseases. It is shown through studies that the compound can prevent cells from damage by eliminating the generation of reactive oxygen, chelating free iron and keeping the level of intracellular ferritin. A new technical scheme is provided for preparing the drugs for treating the iron overload diseases.

Description

technical field [0001] The invention belongs to the field of pharmacy, and in particular relates to the application of dihydromyricetin in the preparation of medicines for treating iron overload diseases. Background technique [0002] Iron overload is primarily caused by mutations in genes including the gene encoding Hepcidin, HAMP, various genes that regulate Hepcidin expression, and the gene for Hepcidin's target protein, FPN. The lack of Hepcidin or the resistance of FPN to Hepcidin can lead to hemochromatosis. The typical feature of hereditary hemochromatosis is excessive iron absorption in the intestine, iron deposition in body tissues and organs accompanied by damage. According to the age and severity of hemochromatosis onset, different pathological symptoms will appear. Cirrhosis and liver cancer can develop in the liver, iron overload in the heart can lead to heart failure, and iron overload in some endocrine glands can lead to stunted growth, delayed sexual matura...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P3/12
CPCA61K31/352A61P3/12
Inventor 冯杰丁浩轩方升林
Owner ZHEJIANG UNIV
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