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Application of POU2F1 gene expression inhibitor in preparation of drugs treating tissue fibrosis diseases

A POU2F1, 1. POU2F1 technology is applied to the POU2F1 gene expression inhibitor, the application field in the preparation and treatment of tissue fibrosis diseases, and can solve the problems of unsatisfactory application and the like

Active Publication Date: 2020-04-14
PEKING UNIV THIRD HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there have been many studies on cardiac fibrosis, but the clinical application of the components found is not ideal. Therefore, it is still necessary to find new members that can regulate many cardiac fibrosis signaling pathways to prevent the occurrence of heart failure. an important research direction

Method used

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  • Application of POU2F1 gene expression inhibitor in preparation of drugs treating tissue fibrosis diseases
  • Application of POU2F1 gene expression inhibitor in preparation of drugs treating tissue fibrosis diseases
  • Application of POU2F1 gene expression inhibitor in preparation of drugs treating tissue fibrosis diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. In vitro experiments, using different matrix stiffnesses to simulate the effects of changes in extracellular matrix stiffness in different pathological conditions on the expression levels of transcription factor POU2F1 and myofibroblast markers fibronectin and αSMA in cardiac fibroblasts.

[0027] Isolation and culture of cardiac fibroblasts: Cardiac fibroblasts were obtained from 8- to 10-week-old male C57BL / 6 mice. The heart tissue from the atrial appendage was cut into pieces and then digested and separated using 0.1% type II collagenase (300U, 17101-015, Gibco, Carlsbad, CA, USA) in a water bath at 37°C. Then centrifuge at 1000 rpm at room temperature for 5 minutes, discard the supernatant, and wash the cell pellet with DMEM (12800-058, Gibco, Carlsbad, CA) containing 10% fetal bovine serum (15140-122, Gibco, Carlsbad, CA, USA). , USA) resuspended, then transferred to a 10cm petri dish, and cultured at 37°C and 5% carbon dioxide. This is the primary card...

Embodiment 2

[0037] Example 2. Detection of POU2F1 protein levels in the infarcted area and remote area at different time points during cardiac tissue surgery in a mouse model of myocardial infarction.

[0038] 10-week-old male C57BL / 6 mice were used to establish the myocardial infarction model. The mice were randomly divided into myocardial infarction group and random 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 experiment, the infarcted area and remote area of ​​the heart tissue of the mice were collected at 1 day, 4 days, and 7 days after the operation.

[0039] We detected the protein level of the endogenous transcription factor POU2F1 in the heart tissue of the infarcted area and remote area of ​​the mouse model of myocardial infarction by Wester...

Embodiment 3

[0040] Example 3. Using chromatin immunoprecipitation experiments to determine that the transcription factor POU2F1 has the ability to bind to the promoters of its possible downstream genes.

[0041] Chromatin immunoprecipitation experiments: 1% formaldehyde was used to cross-link proteins and their associated DNA in live cardiac fibroblasts. Cell lysates were disrupted by sonication after cell fixation, and the size of DNA fragments was determined to be 500-600 bases by agarose gel electrophoresis. The antibody of POU2F1 (ab178869, abcam, Cambridge, MA, USA) and the control rabbit monoclonal antibody IgG (ab172730, abcam, Cambridge, MA, USA) were used for immunobinding, and then A / G beads were added for immunoprecipitation. After the precipitate was centrifuged, it was washed with washing solutions of different salt concentrations, and then eluted and cross-linked, and then real-time quantitative PCR was used to detect the levels of IL1R2, CD69, and TGIF2 promoters.

[0042]...

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Abstract

The invention provides a new application of POU2F1 (POU domain class 2transcription factor 1) as a new important target for treating fibrosis. Specifically, the invention provides a treatment method,the expression of the transcription factor POU2F1 is interfered through small interfering RNA so as to block the inhibiting effect of the transcription factor POU2F1 on downstream genes, so that multiple fibrosis inhibiting factors on the downstream can play a role, thus inhibiting cardiac fibrosis.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of POU2F1 gene expression inhibitors in the preparation and treatment of tissue fibrosis diseases. Background technique [0002] Cardiac fibrosis is a common pathological process, which is very common in heart diseases such as myocardial infarction, aortic stenosis, dilated cardiomyopathy, and diabetic heart disease. The main pathological changes of cardiac fibrosis are increased extracellular matrix in the intercellular matrix of cardiac tissue, increased hardness of cardiac tissue, and decreased parenchymal cells. Continuous development will lead to structural damage and functional decline of the heart, and eventually lead to heart failure, which seriously threatens human health. health and life. Sustained increases in extracellular matrix and cardiac tissue stiffness are not only the result of cardiac fibrosis, but also further aggravate cardiac fibrosis. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/713A61P9/00A61P11/00C12Q1/6883G01N33/68
CPCA61K45/00A61K31/713A61P9/00A61P11/00C12Q1/6883G01N33/6893C12Q2600/158G01N2800/323G01N2800/324G01N2800/325
Inventor 肖晗张幼怡李明喆
Owner PEKING UNIV THIRD HOSPITAL
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