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The role of circRNA MNCR in regulating cardiomyocyte necrosis and myocardial ischemia injury

A cardiomyocyte, myocardial ischemia technology, applied in DNA/RNA fragments, cardiovascular system diseases, recombinant DNA technology, etc., to achieve the effects of inhibiting the degree of myocardial ischemia damage, easy to use, and friendly to the treatment environment

Active Publication Date: 2020-09-18
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the signaling pathway of cardiomyocyte necrosis is still far from elucidated, and the pathway of circRNA in regulating the process of cardiomyocyte necrosis needs further study. Therefore, it is necessary to search for circRNA specifically expressed in the heart and participate in cardiomyocyte necrosis, and to clarify its mechanism of action. The diagnosis and treatment of heart disease with circRNA as a strategy has extremely important significance and application prospects

Method used

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  • The role of circRNA MNCR in regulating cardiomyocyte necrosis and myocardial ischemia injury
  • The role of circRNA MNCR in regulating cardiomyocyte necrosis and myocardial ischemia injury
  • The role of circRNA MNCR in regulating cardiomyocyte necrosis and myocardial ischemia injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: H 2 o 2 circRNA MNCR expression during induced cardiomyocyte necrosis

[0022] This embodiment uses the experimental model of myocardial cell necrosis, and adopts the method established in this laboratory to cultivate the primary cardiomyocytes of rat suckling mice (rat suckling mice were purchased from Beijing Medical University, the rat strain is Wistar rats, rat suckling mice The preparation of primary cardiomyocytes can be found in the following literature: Tan WQ, Wang K, Lv DY, Li PF. Foxo3a inhibits cardiomyocyte hypertrophy through transactivating catalase. The Journal of biological chemistry. 2008; 283:29730-29739), 500uM H 2 o 2 Treat the primary cardiomyocytes of rat neonatal rats, and at the same time set without using 500uM H 2 o 2 In the control experiment (Control group) of primary cardiomyocytes of rat suckling rats, RNA was extracted after the experiment, and the expression level of circRNA MNCR was detected by real-time fluorescent quant...

Embodiment 2

[0023] Example 2: MNCR-siRNA targeted silencing of circRNA MNCR

[0024] In this example, primary cultured cardiomyocytes were used as a model. Primary cardiomyocytes were transfected with MNCR-siRNA adenovirus. At the same time, a negative control group transfected with MNCR-sc adenovirus and a blank group without transfection were set. After 24 hours, The RNA of primary cardiomyocytes in the three groups of experiments was extracted, and the expression level of circRNA MNCR was detected by real-time fluorescent quantitative PCR technology, as shown in figure 2 The shown MNCR-siRNA can target and silence circRNA MNCR, that is, MNCR-siRNA can effectively reduce the expression level of circRNA MNCR in cardiomyocytes.

Embodiment 3

[0025] Example 3: Inhibition of circRNA MNCR expression can promote cardiomyocyte necrosis

[0026] In this example, primary cultured cardiomyocytes were used as a model. Primary cardiomyocytes were transfected with MNCR-siRNA, and at the same time transfected with MNCR-sc random sequence as a control group, and a blank group without transfection. After 24 hours, the three groups experiment while using H 2 o 2 The cells were induced for 12 hours, and then propidium iodide (PI) staining was used to mark necrotic cardiomyocytes, and DAPI staining was used to mark the nuclei of cardiomyocytes, and the percentage of necrotic cells to the number of cardiomyocytes was calculated to detect cardiomyocyte necrosis. image 3 a is the percentage of the number of necrotic cells in the number of cardiomyocytes in the three groups of experiments. From the figure, it can be concluded that after circRNA MNCR is inhibited, H 2 o 2 Induced increased cardiomyocyte necrosis. image 3 bUsing m...

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Abstract

The invention discloses circRNA MNCR nucleotide, a circRNA MNCR nucleotide containing pharmaceutical composition and application thereof. The pharmaceutical composition comprises the circRNA MNCR nucleotide, auxiliary materials, a virus vector or an embedding carrier, wherein the embedding carrier is cholesterol, nanoparticles or liposome and is preferably liposome; the virus vector is one or more of an adenovirus vector, a lentiviral vector and a retroviral vector and is an adenovirus vector preferably; the auxiliary material is one or more of mannitol, phosphate buffer and normal saline and is phosphate buffer preferably; infection titer of the circRNA MNCR virus vector is 10<16>PFU, and a mass ratio of the circRNA MNCR nucleotide to the liposome is 1:1.25; a mass ratio of the virus vector or the embedding vector to the auxiliary material is 1:200. The circRNA MNCR nucleotide and the circRNA MNCR nucleotide containing pharmaceutical composition can be used for prevention and treatment of myocardial hypertrophy, myocardial fibrosis, coronary heart disease and heart failure and prevention of myocardial fibrosis and myocardial remodeling. A preparation process of the pharmaceutical composition is simple, and evident efficacy, convenience in use and environment friendliness in treatment are realized.

Description

Technical field: [0001] The invention belongs to the technical field of biopharmaceuticals, and relates to a circRNA MNCR nucleotide expressed in the process of myocardial necrosis, and a pharmaceutical composition containing the nucleotide, and the corresponding siRNA (circRNA MNCR) of the pharmaceutical composition can target silencing circRNA MNCR, through oral or injection, is used to prevent and treat heart disease and improve heart function, which can prevent myocardial fibrosis and myocardial remodeling. Background technique: [0002] Cardiovascular disease is the number one killer of human health, and more than 17 million people die of cardiovascular disease every year worldwide; with the improvement of people's living standards and changes in diet structure, the mortality rate of cardiovascular disease has increased significantly Cancer has surpassed cancer as the leading cause of death. According to the latest statistical results released by the Ministry of Health...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A61K31/7105A61K48/00A61P9/00A61P9/10A61P9/04
CPCA61K31/7105C12N15/113C12N2310/532
Inventor 王昆李培峰周露玙王胤刘翠云
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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