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Application of a long non-coding RNA and its interfering RNA in the treatment of atherosclerosis

An atherosclerosis, long-chain non-coding technology, applied in the field of biomedicine, can solve problems such as unclear function of lncRNA, and achieve the effect of avoiding the occurrence of clinical time, delaying or even reversing the disease

Active Publication Date: 2021-07-23
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, research on lncRNAs has progressed rapidly, but the functions of most lncRNAs are still unclear

Method used

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  • Application of a long non-coding RNA and its interfering RNA in the treatment of atherosclerosis
  • Application of a long non-coding RNA and its interfering RNA in the treatment of atherosclerosis
  • Application of a long non-coding RNA and its interfering RNA in the treatment of atherosclerosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Pathological staining after establishment of atherosclerosis mouse model at different stages

[0027] 1. Materials

[0028] 1.1 Animals

[0029] The Apoe mice of the present invention are purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., and the experimental animals are randomly divided into groups, respectively, at the age of 12 weeks (starting high-fat feeding for 4 weeks at the age of 8 weeks, EAR group) and at the age of 24 weeks (8 weeks). At the age of 16 weeks, high-fat feeding was started for 16 weeks, and the aorta was obtained from the ADV group). The use of experimental animals was approved by the Ethics Committee of Harbin Medical University.

[0030] 1.2 Reagents

[0031] High-fat feed (10% lard, 4% milk powder, 2% cholesterol, 0.5% sodium cholate) was purchased from Nanjing Senbao Biological Products Co., Ltd., and the improved oil red O staining solution was purchased from Beijing Suolaibao Co., Ltd.

[0032] 2....

Embodiment 2

[0041]Example 2: Expression of LncRNA-AFIAR in arterial tissue of mice with atherosclerosis at different stages

[0042] 1. Materials

[0043] 1.1 Organization

[0044] Apoe of the present invention - / - The mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. The experimental animals were randomly divided into groups, and the main materials were collected at the age of 8 weeks (NOR group) and 24 weeks of age (high-fat feeding for 16 weeks at the age of 8 weeks, ADV group). artery. The use of experimental animals was approved by the Ethics Committee of Harbin Medical University.

[0045] 1.2 Reagents

[0046] TRIzol Reagent was purchased from Invitrogen, USA. The reverse transcription kit (04897030001) was purchased from Roche, Germany. The SYBRGreen Master (ROX) kit used for real-time quantitative PCR (polymerase chain reaction) was purchased from Roche, Germany. Real-Time PCR specific primers were designed and synthesized by Ruibo Xin...

Embodiment 3

[0063] Example 3: Construction of the silencing model of lncRNA-AFIAR in RAW264.7 cells

[0064] 1. Materials

[0065] 1.1 Cells and siRNA

[0066] The RAW264.7 cells used in the present invention are purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences, and the cell culture application contains 10% inactivated fetal bovine serum (Science Cell, USA), penicillin (100 U / mL), streptomycin (100 μg / mL) In DMEM medium, the culture conditions were 37°C, 5% CO2.

[0067] Design related silencing lncRNA-AFIAR (siRNA) sequence, sense strand (Sence): GCAGAUAACUCGUUAGCAAUU (shown in SEQ ID NO.4), antisense strand (Antisense): UUGCUAACGAGUUAUCUGCUU (shown in SEQ ID NO.5). The design and synthesis of small interfering RNA (siRNA) were completed by Suzhou Gemma Gene Co., Ltd.

[0068] 1.2 Reagents

[0069] X-tremeGENE siRNA Transfection Reagent used for small interfering RNA (siRNA) transfection was purchased from Roche Company in Germany; DMEM medium was purchased fr...

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Abstract

The invention discloses the application of a long-chain non-coding RNA and its interference RNA in treating atherosclerosis. In the present invention, through high-throughput sequencing of the aortic tissue of the mouse model of atherosclerosis, a long non-coding RNA (lncRNA) molecule was found to be significantly expressed in the aortic tissue of normal and advanced atherosclerosis mice At the same time, the lncRNA molecule was highly expressed in atherosclerotic tissue, and the lncRNA molecule was named lncRNA‑AFIAR. Experiments have proved that lncRNA-AFIAR has the functions of enhancing macrophage proliferation and inhibiting macrophage apoptosis, and is involved in the process of promoting the progression of atherosclerosis. Inhibition of lncRNA-AFIAR can inhibit the proliferation of macrophages, reduce plaque formation, and thus inhibit the progression of AS. Therefore, the proposal of the present invention provides new molecular markers and intervention targets for the diagnosis and treatment of atherosclerosis, and also provides new technical means for the treatment of atherosclerosis.

Description

technical field [0001] The invention relates to the application of a long-chain non-coding RNA and its interference RNA in the treatment of atherosclerosis. The invention belongs to the technical field of biomedicine. Background technique [0002] In January 2018, the National Center for Cardiovascular Diseases released the "China Cardiovascular Disease Report 2017". He pointed out that in 2017, the number of patients with coronary atherosclerotic heart disease in my country reached 11 million. Overall, the prevalence and mortality of cardiovascular diseases in China are still on the rise, and the number of patients with cardiovascular diseases will remain unchanged in the next 10 years. will grow rapidly. As a high incidence of cardiovascular disease, coronary heart disease is one of the diseases with the highest mortality rate in the world. According to the report of the World Health Organization in 2011, the death toll of coronary heart disease in China ranks second in t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12Q1/6883A61K45/00A61P9/10
CPCA61K45/00A61P9/10C12N15/113C12N2310/10C12N2310/14C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 于波田进伟孙长斌袭祥文顾霞刘新新
Owner HARBIN MEDICAL UNIVERSITY
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