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A clam enzymatic oligopeptide capable of repairing non-alcoholic fatty liver disease cell model and its preparation method

A fatty liver disease and cell model technology, applied in the fields of biochemical equipment and methods, peptide, and microorganism determination/inspection, etc., can solve the problems of lack of specific methods and poor results in conservative treatment, and achieve the ability of cells to scavenge oxygen free radicals Enhances, reduces the degree of damage, and alleviates the degree of lipid peroxidation

Active Publication Date: 2020-12-18
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current treatment methods mainly adopt conservative treatments such as weight loss treatment, lipid-lowering treatment and angiotensin-converting enzyme inhibitors, but there is still a lack of specific methods for conservative treatment, and the effect is not good

Method used

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  • A clam enzymatic oligopeptide capable of repairing non-alcoholic fatty liver disease cell model and its preparation method
  • A clam enzymatic oligopeptide capable of repairing non-alcoholic fatty liver disease cell model and its preparation method
  • A clam enzymatic oligopeptide capable of repairing non-alcoholic fatty liver disease cell model and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Establishment of NAFLD model

[0025] Normal liver Chang liver cells (hereinafter referred to as normal group cells) were cultured in DMEM culture medium, 37°C, 5% CO2 incubator, and the culture medium was discarded after the cells adhered to 80%, digested with 0.25% trypsin, and passaged to cultivate. Select cells with good growth status, induce Chang liver cells with 15 μg / mL palmitic acid, collect cells after treatment for 12h, 24h, 48h, and 72h, and measure triglyceride (TG) content. The TG detection method follows the kit instructions conduct. The induction time was selected to significantly increase the TG content of normal liver Chang liver cells and the cells grew well, and an in vitro NAFLD cell model (hereinafter referred to as model group cells) was established.

[0026] The TG content of normal liver Chang liver cells induced by 15 μg / mL palmitic acid showed that the TG content increased significantly after the treatment with palmitic acid, but after 48...

Embodiment 2

[0047] Example 2 The oligopeptide obtained by enzymolysis of clams with alkaline protease has obvious repairing effect on NAFLD cell model

[0048] Inoculate the cells of the normal group and the model group on the cover glass in the six-well culture plate, discard the nutrient solution after 24 hours and divide them into three groups of cells, namely, the cells of the normal group, the cells of the model group and the clam hydrolyzate prepared in Example 1 The cells in the model group (hereinafter referred to as the drug group) under the action were routinely cultured in the normal group and the model group, and the cells in the drug group were treated with a drug concentration of 10 mg / mL, and the experiment was ended after 24 hours of culture. Fix the cells with 10% neutral formaldehyde for 10 minutes, incubate with oil red O staining solution for 15 minutes, discard the staining solution, wash with PBS, wash with PBS after hematoxylin staining for 5 minutes, and observe red...

Embodiment 3

[0050] Embodiment 3 Determination of liver cell function index

[0051] Alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), glutathione S-transferase (GSH-ST), γ-glutamyltransferase (GSH-ST), and The content of aminoacyltranspeptidase (γ-GT), superoxide dismutase (SOD) and other indicators, the detection method is carried out according to the kit instructions. Among them, the test results of liver cell function indicators are shown in Table 6. It can be seen that the contents of MDA, GSH-ST, ALT, AST, and γ-GT in the model group were significantly higher than those in the normal group, and the levels of the drug group after being treated with clam oligopeptides decreased significantly. , which was statistically significant compared with the model group (P<0.05). The content of SOD decreased in the model group and increased in the drug group, but there was no statistical significance compared with the model group. ALT mainly exists in the ...

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Abstract

The invention relates to an enzymatically decomposed clam oligopeptide having a recovery effect on a non-alcoholic fatty liver disease (NAFLD) cell model. The clam oligopeptide is characterized by comprising an amino acid sequence of Gln Leu Asn Trp Asp. The invention also relates to a preparation method of the enzymatically decomposed clam oligopeptide having the recovery effect on the NAFLD cell model. Compared with the prior art, the following advantages can be achieved: the NAFLD cell model is established by virtue of induction of palmitic acid and the damaged liver cells in a body are simulated sufficiently, and the result indicates that the TG content of the in-vitro NAFLD cell model established 48 hours after induction by 15 micrograms / milliliter palmitic acid increased remarkably in comparison with that of normal liver cells; oil red O staining shows that the number of intracellular lipid droplets of the model group is increased in contrast with that of the cells of a normal group, and the cell mortality rate is low and the repeatability is good, and therefore, the modeling method is simple, convenient and feasible; meanwhile, the enzymatically decomposed clam oligopeptide having the obvious recovery effect on the NAFLD cell model can be separated out from clams.

Description

technical field [0001] The invention relates to an extract obtained from an animal and a preparation method thereof, in particular to a clam extract capable of repairing a cell model of non-alcoholic fatty liver disease and a preparation method thereof. Background technique [0002] Meretrix meretrix L. belongs to the phylum Molluscs, Clamidae, and the genus Meretrix. Its shells are hard and thick, and the two shells are equal in size. It likes to grow in inner bays with fresh water and fine sandy beaches near estuaries. It is one of the four major species in my country. One of the farmed shellfish. Meretrix meretrix is ​​delicious and contains protein, fat, trace elements, etc. necessary for the human body. my country has a long history of using clams to treat diseases. It is recorded in "Compendium of Materia Medica" that it can cure diseases such as "sores, boils, swollen poison, disperse accumulation, and hangover poison". [0003] Recent studies have shown that clam ex...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C12P21/06C12Q1/02
Inventor 杨最素赵莎莎丁国芳黄芳芳赵玉勤余方苗
Owner ZHEJIANG OCEAN UNIV
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