CircRNA CHIF nucleotide and pharmaceutical composition comprising such nucleotide and application thereof

A composition and nucleotide technology, applied in the field of medicine and biology, can solve problems such as the incomplete understanding of the pathogenesis of heart disease, the unsatisfactory effect of heart disease prevention, diagnosis and treatment, etc., and achieve pharmacological reliability and environmental protection. The effect of friendliness and heart function improvement

Active Publication Date: 2016-12-14
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the pathogenesis of heart disease is not completely clear, and the prevention, diagnosis and treatment of heart disease cannot achieve satisfactory results. It is urgent to develop new technologies and methods for the diagnosis, prevention and treatment of heart disease.

Method used

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  • CircRNA CHIF nucleotide and pharmaceutical composition comprising such nucleotide and application thereof
  • CircRNA CHIF nucleotide and pharmaceutical composition comprising such nucleotide and application thereof
  • CircRNA CHIF nucleotide and pharmaceutical composition comprising such nucleotide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Changes in the expression level of circRNA CHIF in cell and animal hypertrophy models

[0020] In this example, the established method was used to cultivate the primary cardiomyocytes of rat suckling rats, and the cardiomyocyte hypertrophy model was induced by 10uM ISO to detect the expression level of circRNA CHIF, and it was found that the expression level of circRNA CHIF in the cell model decreased under hypertrophy stimulation ( figure 1 A). Using the TAC-induced cardiac hypertrophy animal model, it was found that the expression level of circRNA CHIF in the animal model decreased under hypertrophy stimulation ( figure 1 B). The results suggested that the expression level of circRNA CHIF decreased during cardiomyocyte hypertrophy.

[0021] Among them, the animal model of cardiac hypertrophy was prepared as follows: mice were anesthetized by intraperitoneal injection of a mixture of ketamine (80 mg kg), xylazine (20 mg kg) and atropine (0.6 mg kg). 20G v...

Embodiment 2

[0023] Example 2: circRNA CHIF adenoviral vector increases the expression level of CHIF

[0024] The process of the circRNA CHIF adenovirus vector in this embodiment is as follows: PCR amplification obtains the circRNA CHIF sequence, subclones and connects it into the pSilencer Adeno 1.0-CMV system of Ambion Company, and constructs the circRNA CHIF overexpression adenovirus, and the circRNA sequence SEQ ID NO: 1, the PCR primer sequence is: upstream primer: 5'-TGAGCTGACCAGGACCATTTTACCAG-3', downstream primer: 5'-CAATTTCAGCAGGCAGCTGCTTGATT-3'.

[0025] ISO-induced cardiomyocyte hypertrophy model: After the primary cardiomyocytes were infected with circRNA CHIF adenovirus, the cardiomyocytes were treated with 10uM ISO, and after culturing for 60 minutes, the total RNA of the cells was extracted, and the expression level of circRNA was detected by real-time fluorescent quantitative PCR. Such as figure 2 As shown in A, the expression level of circRNA CHIF in rat primary cardiomy...

Embodiment 3

[0027] Example 3: Detection of overexpression of circRNA CHIF inhibiting cardiac hypertrophy in ISO-induced cardiomyocyte hypertrophy model

[0028] The primary cardiomyocytes were treated with ISO as described in Example 2, and various cardiomyocyte hypertrophy indexes were detected. Such as image 3 As shown in A, the surface area of ​​cardiomyocytes increases after ISO treatment, and overexpression of circRNA CHIF can effectively reduce the increase of ISO-induced cardiomyocyte surface area. Such as image 3 As shown in B, the ratio of cardiomyocyte protein / DNA increased after ISO treatment, and overexpression of circRNACHIF could effectively reduce the increase of ISO-induced cardiomyocyte protein / DNA ratio. The above results indicate that overexpression of circRNA CHIF can inhibit ISO-induced cardiomyocyte hypertrophy at the cellular level.

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Abstract

The invention discloses circRNA CHIF nucleotide and a pharmaceutical composition comprising the circRNA CHIF nucleotide. The pharmaceutical composition comprises the circRNA CHIF nucleotide, an auxiliary, and a viral vector or an embedding vector, the embedding vector is cholesterol, nanoparticles or lipidosome, preferably lipidosome, the viral vector is one or more of adenovirus vector, lentiviral vector and retroviral vector, preferably adenovirus vector, and the auxiliary is one or more of mannitol, phosphate buffer solution and normal saline, preferably phosphate buffer solution, the circRNA MNCR viral vector is 1016 PFU in infectivity titer, and a mass ratio of the circRNA CHIF nucleotide to the lipidosome is 1:1.25; a mass ratio of the viral vector or embedding vector to the auxiliaries is 1:200. The pharmaceutical composition is used for preventing and treating cardiac hypertrophy, cardiac fibrosis, coronary heart disease and heart failure, has reasonable formula, is simple to manufacture, has significant therapeutic effect, is widely applicable and is friendly to service environments.

Description

Technical field: [0001] The invention belongs to the field of medical biotechnology, and relates to a circRNA CHIF nucleotide for inhibiting myocardial hypertrophy, and a pharmaceutical composition containing the nucleotide, which is used to inhibit the problem of myocardial hypertrophy and realize the protection of cardiomyocytes. Background technique: [0002] Cardiovascular disease mainly includes hypertension, coronary heart disease and congestive heart failure, etc. Cardiovascular disease is the number one killer of human health, and its mortality rate is close to the sum of all cancer mortality rates. With the improvement of people's living standards and the change of diet structure, the mortality rate of cardiovascular diseases is on the rise. According to the latest statistical results released recently by the "2014 China Cardiovascular Disease Report", at present, cardiovascular disease death accounts for the first cause of death among urban and rural residents, wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K31/7088A61P9/00A61P9/10A61P9/04
CPCA61K31/7088C12N15/113C12N2310/10
Inventor 王昆李培峰周露玙刘翠云
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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