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Application of pax4 gene expression inhibitor in the preparation of drugs for inhibiting fibrosis

A technology of gene expression and fibrosis, applied in the field of genes, can solve the problem that the research on the function of PAX4 is still blank

Active Publication Date: 2022-01-18
PEKING UNIV THIRD HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on the function of PAX4 in the heart is still blank

Method used

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  • Application of pax4 gene expression inhibitor in the preparation of drugs for inhibiting fibrosis
  • Application of pax4 gene expression inhibitor in the preparation of drugs for inhibiting fibrosis
  • Application of pax4 gene expression inhibitor in the preparation of drugs for inhibiting fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1, animal pathological model experiment, constructing a mouse cardiac fibrosis model, taking heart tissue, using the method of immunohistochemistry and Western blot experiment, detecting the position and content of PAX4 protein expression, as well as the myofibroblast markers fibronectin and αSMA, the content of Col I in the extracellular matrix.

[0032] Preparation of angiotensin II-induced cardiac fibrosis model in mice: 10-week-old male C57BL / 6 mice were randomly divided into two groups, the operation group and the sham operation group, and the mice were treated with angiotensin (3 mg·kg-1· day-1) A micro-osmotic pump buried pump (Alzet MODEL 1007D, DURECT, Cupertino, CA) was used to construct the fibrosis model for 7 days. Preparation of the micro-osmotic pressure pump: 1 day before the operation, inject angiotensin II (dissolved in sterile PBS buffer) into the micro-osmotic pump with a 1mL syringe, soak the micro-osmotic pressure pump in sterile PBS buffer...

Embodiment 2

[0040] Example 2. Detecting the PAX4 protein level in the infarct area and remote area at different time points during heart tissue surgery in a mouse model of myocardial infarction.

[0041] 10-week-old male C57BL / 6 mice were used to establish the myocardial infarction model. Mice were randomly divided into myocardial infarction group and sham operation control group. In the myocardial infarction group, left coronary artery stenosis was used to induce myocardial infarction. The control group underwent sham surgery. The operation was performed under gas anesthesia, and mice were gas anesthetized by inhalation of 2% isoflurane. In the experimental group, the infarction area, border area and remote area of ​​the heart tissue of the mice were collected at 1 day, 4 days, and 7 days after operation to collect heart tissue.

[0042] The protein level of the endogenous transcription factor PAX4 in the heart tissue of the infarct area, border area and remote area of ​​the mouse myo...

Embodiment 3

[0043] Example 3. Using cardiac fibroblasts, the cells were treated with 1 μM angiotensin for three days, and the protein levels of PAX4, fibrosis markers fibronectin, αSMA and Col I were detected by immunofluorescence and Western blot experiments.

[0044] Isolation and culture of adult mouse cardiac fibroblasts: 8-week-old male C57 / BL6 mice were killed by neck dislocation, quickly soaked in 75% alcohol for about half a minute, and the heart was immediately removed from the chest in an ultra-clean workbench. Wash twice in PBS buffer at 4 degrees Celsius, cut off the blood vessels at the atrium and the bottom of the heart, then cut the ventricle into small pieces, and wash it once with PBS to wash away some residual blood. Add 0.1% type II collagenase (330U, Worthington, Columbia, NJ, USA / Sigma, St.Louis, MO, USA) prepared in PBS balanced salt solution for digestion. The entire digestion process was carried out under the condition of constant temperature stirring at 36-37 degr...

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Abstract

The invention discloses the application of PAX4 gene expression inhibitor in the preparation of medicine for inhibiting fibrosis. Experiments of the present invention have confirmed that the expression of small interfering RNA interference transcription factor PAX4 can block its inhibitory effect on downstream genes, allowing multiple downstream fibrosis inhibitors to function; the expression of PAX4 reduced by small interfering RNA also reduces fiber The expression level of pro-fibrotic factors can be inhibited, and the effect of pro-fibrosis can be inhibited, thereby inhibiting the occurrence of cardiac fibrosis. Therefore, PAX4 in the heart is a potential new important target for the treatment of cardiac fibrosis and the prevention of heart failure.

Description

technical field [0001] The invention relates to the field of genes, in particular to the application of PAX4 gene expression inhibitors in the preparation of drugs for inhibiting fibrosis. Background technique [0002] Cardiac fibrosis is an important component of most cardiac pathological conditions. The manifestation of cardiac fibrosis is the excessive deposition of extracellular matrix in cardiac tissue, which leads to the destruction of physiological cardiac tissue structure, eventually leading to heart failure, which seriously threatens human health and life. Fibroblast-to-myofibroblast transdifferentiation is key to the initiation and maintenance of cardiac fibrotic responses. Myofibroblasts have important contractile and secretory functions, and are characterized by the expression of α-smooth actin (α-smoothactin, αSMA), fibronectin and type I collagen (Collagen I, Col I). Angiotensin II is a polypeptide preparation recognized in current research to regulate vasoco...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K31/7088A61P9/00A61P11/00A61P1/18
CPCA61K45/00A61K31/7088A61P9/00A61P11/00A61P1/18
Inventor 肖晗张幼怡李明喆胡国民
Owner PEKING UNIV THIRD HOSPITAL
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