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Application of circTMEM165 in preparation of products for diagnosing and/or treating cardiovascular diseases

A cardiovascular and disease technology, applied in the field of cardiovascular disease drugs, can solve problems that do not involve detailed research on circRNA

Inactive Publication Date: 2021-09-17
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, there is no detailed research involving circRNA in the detection and treatment of atherosclerosis

Method used

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  • Application of circTMEM165 in preparation of products for diagnosing and/or treating cardiovascular diseases
  • Application of circTMEM165 in preparation of products for diagnosing and/or treating cardiovascular diseases
  • Application of circTMEM165 in preparation of products for diagnosing and/or treating cardiovascular diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0133] This example is a study on the expression downregulation of circTMEM165 in clinical samples and animal models of atherosclerosis. The research results are as follows figure 1 shown;

[0134] The expression of circTMEM165 was detected by qRT-PCR in the ascending aorta tissue of 8 pairs of healthy people and atherosclerosis patients and 20 pairs of human serum (Pfigure 1 C). It was preliminarily concluded that circTMEM165 may have a protective effect on atherosclerosis.

[0135] The inventors used the red fluorescent probe of circTMEM165 labeled with cy5 to label healthy SD rats and rats with carotid balloon injury, and found that circTMEM165 was significantly down-regulated in the injured rats (Figure 1D). The RNA of each cell line was collected, and the expression level of circTMEM165 in various human cell lines was detected by qRT-PCR (Pfigure 1 E, where VSMC: human vascular smooth muscle cell line; HUVEC: human endothelial cell line; THP-1: human monocyte cell line)....

Embodiment 2

[0137] This embodiment is the structural verification of the ring-shaped TMEM165, and the verification results are as follows figure 2 shown;

[0138] In order to further determine the head-to-tail circular structure of circTMEM165, Sanger sequencing was first performed, and it is not difficult to find from the sequence that circTMEM165 is indeed a head-to-tail circular structure ( figure 2 A-B). Subsequently, the gDNA and cDNA of HUVEC were extracted respectively, and the expression of circular TMEM165 and linear TMEM165 was detected by qRT-PCR. No obvious linear TMEM165 was found in the gDNA ( figure 2 C). The total RNA of HUVEC was treated with RNA exonuclease and actinomycin B respectively, and the expression of circular TMEM165 and linear TMEM165 was re-extracted and detected. It was found that the circular RNA was not degraded because of its stability, while the linear Significant degradation of RNA occurs ( figure 2 D-E).

[0139] In order to better observe the...

Embodiment 3

[0141] This example is a study of circTMEM165 inhibiting the inflammation and adhesion of HUVEC. The research results are as follows image 3 shown;

[0142] The inventors designed the overexpression vector of circTMEM165 ( image 3 A) qRT-PCR was used to detect the knockdown and overexpression efficiency of circTMEM165. It was found that the expression of circTMEM165 was significantly reduced in HUVEC transfected with si-circTMEM165, while the expression of circTMEM165 was significantly increased in HUVEC transfected with circTMEM165 overexpression vector High, (P image 3 B). Subsequently, the interaction between THP-1 cells and HUVEC was stained with CFSE, and the adhesion effect of HUVEC was observed by fluorescence microscope. It was found that the knockdown of circTMEM165 could promote the adhesion of endothelial cells, while circTMEM165 inhibited the adhesion effect of endothelial cells. Quantitative analysis ( image 3 C). HUVEC transfected with NC / si-circTMEM165 / ve...

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Abstract

The invention discloses an application of circTMEM165 in preparation of a product for diagnosing and / or treating cardiovascular diseases. According to the invention, circTMEM165 can play a binding role with a downstream target gene miR-192-3p, and the circTMEM165 can play a good role in repairing cell damage caused by the miR-192-3p on downstream target protein SCP2 of the miR-192-3p; the circTMEM165 is enabled to regulate and control inflammation adhesion, mitochondrial division and endothelial cell apoptosis effects of the endothelial cells through the circTMEM165 / miR-192-3p / SCP2 axis; therefore, a new target spot and a new idea are provided for prevention, treatment and diagnosis of atherosclerosis.

Description

technical field [0001] The present invention relates to the technical field of cardiovascular disease drugs, in particular to the application of circTMEM165 in the preparation of products for diagnosing and / or treating cardiovascular diseases. Background technique [0002] Cardiovascular disease (Cardiovascular diseases, CVD) is one of the most serious diseases threatening human beings in the world today, and it is the only killer of the world's population. According to the "China Cardiovascular Disease Report 2019", at present, the death rate of cardiovascular disease in my country is the first cause of death among urban and rural residents, accounting for 45.5% in rural areas and 43.16% in urban areas. That is, 2 out of every 5 deaths are due to cardiovascular disease. The mortality rate of coronary heart disease among urban and rural residents in China has continued to rise since 2012, and the mortality rate of coronary heart disease in rural areas has an obvious upward ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/53A61K31/7105A61P9/10
CPCC12Q1/6883G01N33/5308A61K31/7105A61P9/10C12Q2600/178C12Q2600/158
Inventor 于涛杨艳艳
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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