Pharmaceutical composition containing miRNA-324 and application of pharmaceutical composition

A technology of miRNA-324 and a composition, which is applied in the field of biopharmaceuticals, can solve the problems that the prevention, diagnosis and treatment of heart disease cannot achieve satisfactory results, the pathogenesis of heart disease is not completely clear, etc., and can resist myocardial ischemia and regeneration. Perfusion injury, reducing myocardial infarction size, and improving the effect of injury

Inactive Publication Date: 2015-04-08
QINGDAO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Pharmaceutical composition containing miRNA-324 and application of pharmaceutical composition
  • Pharmaceutical composition containing miRNA-324 and application of pharmaceutical composition
  • Pharmaceutical composition containing miRNA-324 and application of pharmaceutical composition

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Detection of miRNA-324 expression during myocardial ischemia and cardiomyocyte hypoxia-induced cardiomyocyte apoptosis. In this example, for the cardiomyocyte apoptosis experimental model, the method established in this laboratory was used to cultivate the primary myocardium of rat neonatal rats. Cells (rat suckling mice were purchased from Beijing Medical University, the rat strain is Wistar rats, the preparation of primary cardiomyocytes of rat suckling mice can be found in the following literature: W.-Q.Tan, et al, Foxo3a Inhibits Cardiomyocyte Hypertrophy through Transactivating Catalase J Biol Chem.2008October 31; 283(44):29730-29739), cultured in a hypoxic incubator (oxygen concentration lower than 1%) for different time, extracted RNA, real-time fluorescent quantitative PCR technology to detect the expression level of miRNA-324, Such as figure 1 As shown in A, the expression level of miRNA-324 was significantly down-regulated within 10 min to 180 min of hypoxia t...

Embodiment 2

[0021]In the experiment of miRNA-324 inhibiting cardiomyocyte apoptosis, the miRNA-324 adenovirus involved in this example was constructed using the pSilencer Adeno 1.0-CMV system of Ambion Company. Construct the corresponding vector and adenovirus according to the company's instructions. The specific method is as follows: extract the genome of the rat heart, use it as a template, and PCR amplify miRNA-324 and its upstream and downstream sequences of nearly 200 bp each (PCR system 50 μL, The PCR conditions are as follows: 95°C for 3min for one cycle; 95°C for 30s, 56°C for 30s, 72°C for 1min, a total of 28 cycles; and finally 72°C for 5min). The adenoviral vector pSilencer Adeno CMV1.0 shuttle vector (pSilencer Adeno CMV1.0shuttle vector vector) (purchased from Ambion Company) was double digested and ligated with the digested fragment, that is, the fragment was cloned into the adenoviral vector, PacⅠ The vector and the adenovirus LacZ Backbone were digested and linearized, and...

Embodiment 3

[0024] The overexpression of miRNA-324 inhibits myocardial ischemia injury experimental verification. The mice designed in this example were injected with miRNA-324 mimics through coronary arteries, which can significantly inhibit myocardial ischemia-reperfusion injury. C57 mice were used as experimental objects, and the following methods were used: The miRNA-324 mimic (miRNA-324mimic) and the miRNA-324 mimic negative control (miRNA-324mimic NC) were injected, and sham-operated mice were used as the control group, and C57 mice were used as the wild type. The mice were weighed and anesthetized, fixed in the supine position, sheared and disinfected with iodine; the trachea was cut midline in the neck and intubated, and the animal ventilator was used for positive pressure ventilation. Cut the skin about 3 cm, bluntly separate the subcutaneous tissue and muscle layer by layer, open the chest, carefully lift the pericardium and cut it to fully expose the heart; dilute 2×10 11 pfu (...

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Abstract

The invention relates to a pharmaceutical composition containing miRNA-324 and application of the pharmaceutical composition. The pharmaceutical composition comprises an effective dose of miRNA-324 and pharmaceutically acceptable vectors, viruses or accessories, wherein the vectors are cholesterol, cholesterin, lipidosome or nano-particles; the sequence of miRNA-324 antisense nucleotide is 5'-CGCAUCCCCUAGGGCAUUGGUGU-3'. The pharmaceutical composition is used for treating the heart disease such as myocardial infarction, myocardial ischemial injury, myocardial hypertrophy or myocardial fibrosis in an oral taking or injection manner. The pharmaceutical composition has the beneficial effects that the preparation process is simple, the pharmaceutical effect is obvious, the use is convenient, and the treatment is environmentally friendly.

Description

Technical field: [0001] The invention belongs to the technical field of biopharmaceuticals, and relates to a new drug of endogenous non-coding small RNAs and its application, especially a pharmaceutical composition containing miRNA-324 and its application for preventing or treating heart disease. Background technique: [0002] Cardiovascular disease is a major threat to human health and life, and is the number one killer of human health. Worldwide, more than 17 million people die from cardiovascular disease every year, 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, surpassing cancer as the number one cause of death. According to the latest statistical results recently released by the Ministry of Health, the prevalence of hypertension among residents aged 18 and over in my country is 18.8%, and it is estim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P9/10A61P9/00
Inventor 王昆李培峰
Owner QINGDAO UNIV
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