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Application of polypeptide HDP-1 of Haliotis discus hannai to preparation of drugs used for preventing and treating tumors and diseases related to oxidation

A technology of HDP-1 and wrinkle plate abalone, applied in the field of peptides, can solve problems such as oxidative damage of cells, tissues or organs, instability of free radicals, etc.

Active Publication Date: 2018-11-16
SHENZHEN INST OF GUANGDONG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ROS and free radicals are usually unstable and easily react with substances in organisms, causing oxidative damage to cells, tissues or organs
However, there is no research on the antioxidant and anti-tumor effects and mechanisms of the polypeptide (HDP-1) isolated from the boiled by-product of Abalone wrinkle.

Method used

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  • Application of polypeptide HDP-1 of Haliotis discus hannai to preparation of drugs used for preventing and treating tumors and diseases related to oxidation
  • Application of polypeptide HDP-1 of Haliotis discus hannai to preparation of drugs used for preventing and treating tumors and diseases related to oxidation
  • Application of polypeptide HDP-1 of Haliotis discus hannai to preparation of drugs used for preventing and treating tumors and diseases related to oxidation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: HDP-1 free radical scavenging activity

[0029] Biological oxidation has an inseparable relationship with free radicals and reactive oxygen species. 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) has the characteristics of stable properties and easy control of the reaction, and is usually used as the first choice reagent for the determination of antioxidant free radical scavenging ability in vitro. Among active oxygen species, superoxide Anion radicals (O 2- ), hydroxyl radicals (·OH), hydrogen peroxide (H 2 o 2 ) has a certain representativeness and is a commonly used experimental object in antioxidant research. Therefore, this experiment detects the in vitro scavenging ability of HDP-1 on DPPH free radicals, superoxide anion free radicals, hydroxyl free radicals and peroxyl free radicals. In order to better evaluate the free radical scavenging ability of HDP-1, in addition to in vitro experiments, this study also carried out the measurement of HDP-1's in...

Embodiment 2

[0043] Example 2: Cytotoxic effect of HDP-1 on HT1080 cells

[0044] Determination of Cytotoxicity by MTT Assay

[0045] Take out the HT1080 cells from the incubator and digest them with an appropriate amount of trypsin. After the cells have detached from the wall, add complete medium, blow and blow to make the cells evenly dispersed, inoculate them into 96-well plates at a volume of 100 μL per well, and place them in a carbon dioxide incubator. Incubate for 24 hours. The original medium was aspirated, replaced with fresh medium, and HDP-1 samples were added respectively so that the final concentrations of the samples were 0, 10, 20, 50, and 100 μM (there were 3 parallel groups for the same concentration), and incubated for 24 hours. Aspirate the medium, add 100 μL of 5 mg / mL MTT to each well, and incubate for 4 hours. Aspirate the MTT, add 100 μL DMSO to each well to dissolve the crystals, measure the absorbance value with a microplate reader at a wavelength of 570 nm, and ...

Embodiment 3

[0047] Example 3: Protective effect of HDP-1 on DNA

[0048] 3.1 DNA extraction

[0049] Several flasks of HT1080 cells (obtained from Example 2) were taken and digested with trypsin. After the cells were detached, an appropriate amount of medium was added, transferred to a centrifuge tube, and centrifuged at 1000 rpm for 10 min in a centrifuge. Remove the supernatant, wash with PBS, add 10 mL of PBS containing 5 mM EDTA, and centrifuge at 1000 rpm for 10 min. Remove the supernatant, add 350 μL 0.2M sodium acetate, 25 μL 10% SDS, 10 μL proteinase K and 10 μL 0.5 mg / mL RNase, mix thoroughly, and after standing at 54 °C for 1 hour, add phenol at a ratio of 1:1: Chloroform: isoamyl alcohol (25:24:1) solution, centrifuged at 12000 rpm for 10 min. The supernatant was mixed with 100% cold ethanol at a ratio of 1:1.5, and centrifuged at 12000 rpm for 5 min to remove ethanol. After the precipitate was fully dried, it was dissolved in TE buffer and stored at low temperature.

[005...

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Abstract

The invention relates to application of the polypeptide HDP-1 of Haliotis discus hannai to preparation of drugs, health products or dietary supplements used for preventing and treating tumors and diseases related to oxidation. HDP-1 has strong free radical removing capability and can inhibit free radical-mediated DNA oxidation damage. HDP-1 can also inhibit the migration of HT1080 cells and the expression of MMP-2 and MMP-9, and the inhibitory intensity of HDP-1 is dose-dependent; that is, HDP-1 has antioxidant and antitumor activities.

Description

technical field [0001] The present invention relates to the field of polypeptides, and in particular to the use of abalone wrinkled polypeptide HDP-1 in the preparation of medicines, health products or dietary supplements for preventing and treating tumors and oxidation-related diseases. Background technique [0002] Tumor is a new biological mass formed by excessive proliferation and abnormal differentiation of human tissue cells under the action of tumorigenic factors, and is the enemy of human health. According to reports, in 2012, there were about 3.586 million new cases of malignant tumors nationwide, and 2.187 million deaths, with a mortality-incidence ratio as high as 62%. Tumor cell metastasis is the key cause of death in malignant tumors, and matrix metalloproteinases (MMPs) play a key role in the process of tumor cell metastasis [Shen Peiliang, Liu Zhaoguo, Wang Xu, et al. Research progress in tumor ECM fibrogenesis and tumor metastasis. China Bulletin of Physiolo...

Claims

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Application Information

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IPC IPC(8): A61K38/08A61P35/00A61P35/04A61P39/06A23L33/18
CPCA61K38/08A61P35/00A61P35/04A61P39/06A23L33/18A23V2002/00A23V2200/308A23V2200/30
Inventor 千忠吉巩芳李承勇周春霞洪鹏志孙省利
Owner SHENZHEN INST OF GUANGDONG OCEAN UNIV
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