Polypeptide capable of promoting hepatocyte proliferation and/or inhibiting hepatocyte apoptosis and application thereof

A liver cell and apoptosis technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of complicated purification methods and high production costs, and achieve the effects of low immunogenicity, small molecular weight, and easy synthesis

Active Publication Date: 2019-03-08
LINK HEALTH GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Now human HIP/PAP protein is generally expressed through recombinant expression system (such

Method used

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  • Polypeptide capable of promoting hepatocyte proliferation and/or inhibiting hepatocyte apoptosis and application thereof
  • Polypeptide capable of promoting hepatocyte proliferation and/or inhibiting hepatocyte apoptosis and application thereof
  • Polypeptide capable of promoting hepatocyte proliferation and/or inhibiting hepatocyte apoptosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0031] Example 2 Detecting the Activity of Promoting Hepatocyte Proliferation in Vitro

[0032] In this example, the human liver cell line HL-7702 was purchased from ATCC.

Embodiment 1

[0033] In this embodiment, the method for detecting the activity of the 6 polypeptides described in Example 1 and HIP / PAP in promoting the proliferation of liver cells in vitro is as follows:

[0034] Take out the frozen HL-7702 cells and thaw them in a water bath at 37°C. Centrifuge at 800 rpm for 5 minutes on a low-speed centrifuge; aspirate the supernatant and resuspend the cells in 1 ml of medium.

[0035] The resuspended cells were transferred to a new culture flask, and an appropriate amount of MEM medium was added. As for CO 2 Incubator culture (cultivation condition 5% CO 2 , saturated humidity, 37°C).

[0036] After the cells grow to 80%, the cells are digested and scattered by blowing and counting, and the cell concentration is adjusted to 1×10 5 cells / ml, added to a 96-well plate, 100 μl per well, that is, 1×10 cells per well 4 indivual.

[0037]After the cells adhered to the wall, different drugs were added for treatment, namely IGL9, PTQ9, LHD9, SLS27, AYG25,...

Embodiment 3

[0041] Example 3 Detecting the Activity of Inhibiting Hepatocyte Apoptosis in Vitro

[0042] In this embodiment, the method for detecting the activity of the 6 polypeptides described in Example 1 and HIP / PAP in vitro inhibiting liver cell apoptosis is as follows:

[0043] Human hepatocytes HL-7702 were spread in 96-well plates, and the cell concentration was 1×10 5 cells / ml, 100 μl per well, that is, 1×10 cells per well 4 indivual.

[0044] Each well was treated with 20ng / mL Act D2 for 30min and 80ng / ml TNF-α for 48h to establish the cell apoptosis model.

[0045] Add corresponding drugs for treatment, respectively IGL9, PTQ9, LHD9, SLS27, AYG25, VSV19, HIP / PAP, the volume of drug addition is 10 μl, and two concentrations of each drug (final concentrations are 1 μg / ml and 10 μg / ml respectively) are added The volume was 10 μl, and the same volume of PBS was used as a negative control.

[0046] After 24 hours of drug treatment, add 100 μl to each well Reagent ( 3 / 7 Assay...

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Abstract

The invention relates to polypeptide and application thereof, in particular to polypeptide capable of promoting hepatocyte proliferation and/or inhibiting hepatocyte apoptosis and application thereof.Amino acid sequence of the polypeptide is selected from at least one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6. The polypeptide has activity equivalent to orbetter than that of HIP/PAP and has the advantages of being small in molecular weight, easy in synthesis and low in immunogenicity, thereby having wide application prospect.

Description

technical field [0001] The present invention relates to polypeptides and uses thereof, in particular to polypeptides promoting liver cell proliferation and / or inhibiting liver cell apoptosis and uses thereof. Background technique [0002] The liver is the largest internal organ in the human body (accounting for about 2.5% of the total body mass), and it is also the central metabolic organ. Parenchymal cells are mainly responsible for liver function, accounting for 80% of the total liver cells. Hepatocytes play a series of roles, including producing plasma, synthesizing carrier proteins, detoxifying digests, conjugating and secreting hormones, regulating the synthesis and metabolism of blood lipids and amino acids, etc. [0003] Apoptosis is an active suicide process of cells under specific circumstances. Apoptosis is two different patterns of apoptosis and death observed by Kerr et al. in 1972 when studying liver ischemia. Apoptosis is characterized by cell shrinkage, cel...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/067A61P1/16A61P31/12A61P37/02A61K38/00C07K7/06C07K7/08C07K14/00
Inventor 宋燕许元生张时群王晔陈伟柱
Owner LINK HEALTH GRP
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