Small peptide and application thereof

A product, functional food technology, applied in the biological field to achieve the effect of relieving diabetic complications and strong antioxidant capacity

Active Publication Date: 2017-05-31
NANJING UNIV OF FINANCE & ECONOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, there is a lack of drugs and functional foods that can regulate the abnormal proliferation of VSMCs induced by high glucose, regulate the apoptosis of s

Method used

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  • Small peptide and application thereof
  • Small peptide and application thereof
  • Small peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Antioxidant properties of small peptides of the present invention

[0022] This example examines the antioxidant properties of the small peptides of the present invention.

[0023] 1. Determination of oxygen radical absorption capacity (ORAC)

[0024] Pipette 20μL of the sample solution to be tested (the final concentration in the reaction system is 50μM) and 80μL of the sodium fluorescein solution (the final concentration of the fluorescein sodium in the reaction system is 175nM) in each well of a black 96-well plate, mix well, After 15min incubation at 37°C. A multichannel pipette was used to add 100 μL of AAPH (azobisisobutamidine hydrochloride, the final concentration of AAPH in the reaction system was 24 mM) to each well of the microplate to start the reaction. Then place the microplate in the microplate reader, shake and mix well, and measure at 37°C, 485nm excitation wavelength, and 538nm emission wavelength. Measure the fluorescence intensity every 2min, s...

Embodiment 2

[0032] Example 2 The protective effect of the small peptide of the present invention on the abnormal proliferation of VSMCs induced by high glucose

[0033] The following experiment was performed using the small peptide of the present invention.

[0034] 1. Thiazole blue (MTT) method to detect cell proliferation

[0035] Take VSMCs in logarithmic growth phase and adjust the cell suspension to 5×10 with low-sugar DMEM medium containing 2% FBS 4 Cells / mL, seeded in a 96-well plate, 100μL per well (the blank control group is added with 100μL low-sugar DMEM medium containing 2% FBS), the number of cells is 5×10 3 Pcs / well, placed at 37℃, 5% CO 2 After culturing in the incubator for 48 hours, aspirate the culture solution and add 100 μL of low-sugar DMEM medium to each well. The blank control group did not add any substance, and the other wells were randomly divided into three groups: the normal group did not add any substance; the high sugar group (G) added an appropriate amount of 45% g...

Embodiment 3

[0048] Example 3 Effect of the small peptide of the present invention on the apoptosis of VSMCs incubated with high glucose

[0049] The following experiment was performed using the small peptide of the present invention.

[0050] Annexin V-FITC / PI double staining method to detect cell apoptosis rate: the apoptosis rate of each group of cells uses Annexin V-FITC cell apoptosis detection kit (Biyuntian biological reagent company), namely Annexin V-FITC / PI double staining law. Take VSMCs in logarithmic growth phase, digest them with 0.05% EDTA-trypsin solution, and adjust the cell suspension to 5×10 with low-sugar DMEM medium containing 10% FBS 4 Cells / mL, seeded in a 6-well plate, 2mL per well, so that the number of cells is 1×10 5 Pcs / well, placed at 37℃, 5% CO 2 After culturing in the incubator for 48 hours, aspirate the culture medium, switch to serum-free low-sugar DMEM medium, and randomly divide it into three groups: the normal group does not add any substances; the high-sugar...

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PUM

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Abstract

The invention provides a small peptide and application thereof, and relates to the technical field of biology. The amino acid sequence of the small peptide is shown as SEQ ID NO:1. The invention also provides the application of the small peptide to preparation of functional food for alleviating diabetic complications, anti-atherosclerosis functional food, antioxidant functional food, a free radical scavenging product and a lipid peroxidation inhibition product. The small peptide not only has relatively high antioxidant capability, but also can be used for regulating high glucose-induced abnormal proliferation of VSMCs (vascular smooth muscle cells) and regulating apoptosis of the VSMCs in a high glucose environment to finally make the VSMCs tending to get normal, and thus has a broad development prospect on the aspects of prevention of atherosclerosis formation in the high glucose environment, alleviation of other diabetic complications and the like.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to a small peptide and its application. Background technique [0002] Diabetes is a group of metabolic diseases characterized by increased chronic blood sugar levels. The auto-oxidation of glucose caused by hyperglycemia, the enhancement of protein glycosylation and abnormal lipid metabolism are the main reasons for the increase of free radicals in diabetic patients, especially reactive oxygen species (ROS). It also leads to a significant decrease in the activity of some antioxidant enzymes. So as to make diabetic patients in a certain degree of oxidative stress. Oxidative stress not only participates in the whole process of the occurrence, development and deterioration of diabetes, but also participates in the important pathological changes of the complications of diabetes-atherosclerosis (AS) of various organs. An important component of the blood vessel wall is vascular smooth muscle cel...

Claims

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Application Information

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IPC IPC(8): C07K7/06A61K38/08A61P39/06A61P3/10
CPCA61K38/00C07K7/06
Inventor 沈新春曹小舟陈思远俞凌汤晓智方勇
Owner NANJING UNIV OF FINANCE & ECONOMICS
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