Composition for activating cardiac regeneration and application of composition

A technology of heart regeneration and composition, applied in the field of compositions for activating heart regeneration, can solve problems such as poor heart regeneration ability, and achieve the effect of effective treatment and reduction of mortality

Inactive Publication Date: 2015-11-18
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problem of poor regenerative ability of the adult heart, the prese

Method used

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Examples

Experimental program
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Effect test

experiment example 1

[0027] TL wild-type zebrafish were anesthetized in 4 mg / mL 3-aminobenzoic acid ethyl ester-methanesulfonate (purchased from Sigma, catalog number E10521) solution, and the thorax was opened with ophthalmic forceps and the pericardium was removed to expose the ventricle , using microdissecting scissors to remove the apex of the ventricle (about 20% of the entire ventricle), and then put it back into normal feeding water. The postoperative zebrafish were divided into 5 groups, and DMSO (control group), 10 μDPI, 10 μDPI+10 μMBCI, 100 μMapocynin, 100 μMapocynin+10 μMBCI were administered daily during 7-14 dpa (7-14 days after heart resection) by pleural injection, At the same time, 2.5mg / mL BrdU was injected to label the dividing cardiomyocytes. The heart was taken at 14dpa, fixed with 4% paraformaldehyde, and routinely paraffin-embedded. BrdU+ / Mef2C+ cardiomyocytes were labeled with conventional immunohistochemical methods (BrdU antibody was purchased from Sigma, catalog number ...

experiment example 2

[0031] Zebrafish apexectomy and paraffin section preparation were performed as described in Experimental Example 1. Zebrafish duox gene sequence was used as a template, T7 RNA polymerase (purchased from Promega, catalog number P2075) and digoxin-labeled single nucleotide were used. The mixture (purchased from Roche, catalog number 11277073910) was used to synthesize duoxRNA probes. The probe was used to perform in situ hybridization experiments on paraffin sections of zebrafish hearts at different stages after injury. After the sections were dewaxed to water, digested with proteinase K, and blocked with triethanolamine-acetic anhydride, they hybridized with the probe at 55°C overnight. After steps such as gradient SSC washing and blocking, it was incubated with alkaline phosphatase-labeled digoxin antibody (purchased from Roche, catalog number 11094273910) for 2 hours at 37°C. NBT / BClP substrate (purchased from Roche Company, catalog number 11697471001) was used for color dev...

experiment example 3

[0033] The Tol2-mediated transgenic system (for vector information and methods, see reference 1) was used to construct transgenic fish that specifically express Hyper (for the working principle, see reference 2) regulated by the promoter of the cardiomyocyte marker gene myl7. Using this transgenic fish for heart surgery and H 2 o 2 concentration determination. See Experimental Example 1 for apexectomy in zebrafish. The heart was taken out at the scheduled time, soaked in Tyrode'sBuffer (Tyrode'sBuffer formula: 150mMNaCl, 5.4mMKCl, 1.5mMMgSO4, 0.4mMNaH2PO4, 2mMCaCl2, 10mMglucose, 10mMHEPES, pH7.4. All reagents were purchased from sigma company.), remove the atrium Stop the heart. Use a ZeissLSM700 laser confocal microscope for real-time shooting, use 405nm and 488nm for excitation, and calculate the ratio of F488 / F405 to get H 2 o 2 real-time concentration. The Hyper-overexpressed hearts after surgery were soaked in different concentrations (5, 10, 20, 50, 100 and 200 μM)...

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Abstract

The invention provides a composition for activating cardiac regeneration. The composition contains an Erk1/2 phosphorylation level positive regulator. The invention further relates to application of Mek1/2 protein, Erk1/2 dephosphorylated inhibitor, hydrogen peroxide or NOX family proteins in preparing a cardiac regeneration activating medicine. Upon experiments, the invention discovers a signal pathway which is capable of activating cardiac regeneration capacity, and by regulating the signal pathway from various links, the cardiac regeneration capacity is activated. By operating a heart injury surface and activating the cardiac regeneration capacity, the composition can be used for effectively treating heart injury and reducing death rate.

Description

technical field [0001] The present application relates to a composition for activating heart regeneration and its application. Background technique [0002] Due to the extremely limited regenerative ability of the adult heart, heart injury poses a great threat to human life. Finding a drug that can activate the regenerative capacity of the adult heart for the treatment of adult heart injury has become a major task for the scientific community. Contents of the invention [0003] In order to solve the problem of poor regeneration ability of the adult heart, the present invention provides a composition capable of activating the regeneration ability of the heart. [0004] As one aspect of the present invention, it relates to a composition for activating heart regeneration, which contains a positive regulator of Erk1 / 2 phosphorylation level. The positive regulator of phosphorylation level in the present invention can be any substance that can directly or indirectly increase t...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K38/45A61K38/44A61K33/40A61P9/00
Inventor 熊敬维周小海
Owner PEKING UNIV
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