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Polypeptide GPR5 and application thereof to promoting of hepatocyte proliferation and inhibiting of hepatocyte apoptosis

A liver cell and apoptosis technology, applied in the field of cell proliferation and apoptosis, to achieve the effect of small molecular weight, easy synthesis, and promotion of liver cell proliferation

Active Publication Date: 2020-01-10
LINK HEALTH GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] GPR5 is a sequence that mimics fibrinogen. There is a polymerization site "A" that can be activated by thrombin in the E domain of fibrinogen. The "A" site is broken from the α-chain of fibrin peptide A to expose GPR. sequence (Gly-Pro-Arg), a synthetic short peptide that mimics the GPR sequence can bind fibrinogen and inhibit its polymerization, thereby being used for related applications of hemostasis and coagulation (see Chinese patent CN105916524A, and non-patent literature 1), but so far There is no report that GPR5 is used to promote liver cell regeneration and inhibit liver cell apoptosis

Method used

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  • Polypeptide GPR5 and application thereof to promoting of hepatocyte proliferation and inhibiting of hepatocyte apoptosis
  • Polypeptide GPR5 and application thereof to promoting of hepatocyte proliferation and inhibiting of hepatocyte apoptosis
  • Polypeptide GPR5 and application thereof to promoting of hepatocyte proliferation and inhibiting of hepatocyte apoptosis

Examples

Experimental program
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Effect test

Embodiment 2

[0040] Example 2 Detecting the Activity of Promoting Hepatocyte Proliferation in Vitro

[0041] In this example, the human liver cell line LO2 was purchased from ATCC.

[0042] In this embodiment, the method for detecting the activity of the polypeptide GPR5 described in Example 1 in promoting the proliferation of liver cells in vitro is as follows:

[0043]After LO2 cells were revived according to conventional methods, they were placed at 37°C and 5% CO 2 In the incubator, culture with DMEM complete medium containing 10% FBS and 1% double antibody. When the cells have grown to about 85% of the bottom of the bottle, digest with trypsin-EDTA, add DMEM complete medium and blow gently to make a cell suspension, adjust the cell concentration, inoculate into a 96-well plate, 100 μL per well, Then place at 37°C, 5% CO 2 cultured in an incubator until use.

[0044] Inoculate LO2 in the logarithmic phase at a density of 5000 / well in a 96-well plate and culture for 24 hours. Divid...

Embodiment 3

[0046] Example 3 Detecting the Activity of Inhibiting Hepatocyte Apoptosis in Vitro

Embodiment 1

[0047] In this example, the method for detecting the activity of the polypeptide GPR5 described in Example 1 in inhibiting liver cell apoptosis in vitro is as follows:

[0048] 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.

[0049] Each well was treated with 20ng / mL actinomycin D2 (Act D2) for 30min and 80ng / ml tumor necrosis factor α (TNF-α) for 48h to establish the apoptosis model.

[0050] The corresponding polypeptide GPR5 was added for treatment, the dosing volume was 10 μl, and the final concentrations were 0.1, 1, and 10 μg / ml respectively, and the same volume of PBS was used as a negative control, and 0.2 μg / ml epidermal growth factor (EGF) was used as a positive control.

[0051] After 24 hours of drug treatment, add 100 μl to each well Reagent ( 3 / 7 Assaypromega company, article number: G8090), gently mix the contents of each well for about 30 se...

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Abstract

The invention discloses polypeptide GPR5 and application thereof to promoting of hepatocyte proliferation and inhibiting of hepatocyte apoptosis. The invention discloses polypeptide and application thereof to promoting of hepatocyte proliferation and / or inhibiting of hepatocyte apoptosis. According to the polypeptide capable of promoting hepatocyte proliferation and / or inhibiting hepatocyte apoptosis, compared with an amino acid sequence shown in SEQ ID NO:1, the amino acid sequence of the polypeptide has 3 / 5 or above sequence consistency, the polypeptide length is 5 amino acids, the polypeptide has the advantages of being small in molecular weight, easy to synthesize, low in immunogenicity and the like, and therefore the polypeptide has broad application prospects in treatment of hepatopathy.

Description

technical field [0001] The invention relates to the technical field of cell proliferation and apoptosis, in particular to a polypeptide GPR5 and its use for promoting liver cell proliferation and / or inhibiting liver cell apoptosis. 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...

Claims

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Application Information

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IPC IPC(8): C07K14/75A61K38/36A61P1/16
CPCC07K14/75A61P1/16A61K38/00Y02A50/30
Inventor 宋燕孟鹤卢圆黄玉影
Owner LINK HEALTH GRP
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