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PK (Pyruvate Kinase) M2 mutant and application thereof to cardiovascular diseases

A pyruvate kinase and mutant technology, applied in the field of molecular biology, can solve the problems of unclear myocardial fibrosis and slow progress in the development of anti-myocardial fibrosis drugs, and achieve the effect of inhibiting myocardial fibrosis and heart failure

Active Publication Date: 2020-04-03
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the mechanism of myocardial fibrosis is not very clear, which also leads to slow progress in the development of anti-myocardial fibrosis drugs

Method used

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  • PK (Pyruvate Kinase) M2 mutant and application thereof to cardiovascular diseases
  • PK (Pyruvate Kinase) M2 mutant and application thereof to cardiovascular diseases
  • PK (Pyruvate Kinase) M2 mutant and application thereof to cardiovascular diseases

Examples

Experimental program
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Effect test

Embodiment 1

[0025] 1. RT-PCR

[0026] (1) Remove the primary milk rat cell culture medium, and wash the cells with PBS 3 times.

[0027] (2) Add 1mL Trizol, blow down the cells, pipette into an RNAase-free centrifuge tube (cell sample), and lyse for 5 minutes. .

[0028] (3) Add 200 μL of chloroform to each EP tube, vibrate vigorously, and after mediation for a few seconds, place it on ice for 10 minutes for lysis.

[0029] (4) Centrifuge at 4°C, 12000rpm, 15min, carefully draw the supernatant (500μL) and transfer it to a new EP tube.

[0030] (5) Add an equal volume of isopropanol, mix upside down, and let stand at 4°C for 10 minutes.

[0031] (6) Centrifuge at 4° C., 12000 rpm for 15 min, discard the supernatant, and wash with 75% ethanol.

[0032] (7) Centrifuge at 4°C, 12000rpm, 5min, discard the supernatant, air-dry, add DEPC water (20-50μL), and detect the RNA concentration.

[0033] (8) The reverse transcription reaction system procedure of RNA is as follows:

[0034]

[0...

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Abstract

The invention relates to a PK (Pyruvate Kinase) M2 mutant and application thereof to cardiovascular diseases. The nucleotide sequence of the PKM2 mutant is shown as SEQ ID NO. 2. The activity of the PKM2 in cells is recovered by using a specific PKM2 agonist or transfecting and modifying site mutation plasmids into cardiac muscle cells, so that the occurrence of myocardial fibrosis and congestiveheart failure can be effectively inhibited.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to a mutant of pyruvate kinase M2 and its application in cardiovascular diseases. Background technique [0002] Cardiovascular disease is the number one cause of death in the world. According to statistics, from 1990 to 2010, the death rate of cardiovascular disease increased by 1 / 3. Myocardial fibrosis is the common pathological basis of various heart diseases. The process of myocardial fibrosis is the proliferation of cardiac fibroblasts and the conversion to a myofibroblast phenotype, including increased collagen synthesis and abnormal deposition of collagen in the extracellular matrix (ECM), including smooth muscle actin (a-SMA ), fibronectin (Fibronectin) and type I collagen (Collagen I) secretion increased. Extensive, multifocal fibrosis with atrophy and hypertrophy of adjacent myocardial fibers is common under light microscopy, and sarcoplasmic vacuoliz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/12C12N15/54A61K38/45A61P9/00G01N33/573
CPCC12N9/1205A61P9/00G01N33/573C12Y207/0104A61K38/00G01N2800/32
Inventor 季勇罗姗姗周苗田佳馨周雪纯林喆
Owner NANJING MEDICAL UNIV
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