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Molecular marker for early warning of atherosclerosis and application of molecular marker

A technology of atherosclerosis and molecular markers, applied in the medical field, can solve the problem of unclear effect of Hcy on endothelial cell apoptosis, and achieve the effect of helping clinical intervention in time and delaying the development of the disease

Inactive Publication Date: 2021-01-01
NINGXIA MEDICAL UNIV +1
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Problems solved by technology

lncRNA DINO (DNA damage-induced non-coding RNA), as a member of lncRNA (long non-coding RNA), plays an important role in cancer cell migration and apoptosis, but its role in Hcy-induced endothelial cell apoptosis is not known yet. clear

Method used

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  • Molecular marker for early warning of atherosclerosis and application of molecular marker
  • Molecular marker for early warning of atherosclerosis and application of molecular marker
  • Molecular marker for early warning of atherosclerosis and application of molecular marker

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Embodiment Construction

[0036] The technical solution of the present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0037] A molecular marker for early warning of atherosclerosis, the specific embodiment is:

[0038] 1 Material preparation

[0039] 1.1 Materials and reagents Human umbilical vein endothelial cells (HUVECs) (China, Mingjin Biotechnology Co., Ltd.); homocysteine ​​(US, Sigma); fetal bovine serum, DMEM medium (Australia, Gibco); Streptomycin, trypsin digestion solution, NP-40 (ethylphenyl polyethylene glycol) (China, Beyentian Biotechnology Research Institute); Cell Viability Fluorescence Assay Kit (Switzerland, Roche Company); PMSF (China, Beijing Suolaibao Technology Co., Ltd.); Bax (Bcl-2-related X protein) antibody, Bcl-2 (B lymphocytoma-2) antibody (UK, Abcam), β-actin antibody, horseradish peroxidase Labeled secondary antibody (China, Beijing Jinqiao Co., Ltd.); skimmed milk powder (USA, BD company); c...

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Abstract

The invention discloses a molecular marker for early warning of atherosclerosis. A method comprises the following steps that (1) the influence of Hcy (Homocysteine) with different concentrations on endothelial cell proliferation is detected by an MTT colorimetric method, and a proper concentration is screened out; (2) after cells is intervened with the Hcy with optimal concentration, cell viability staining detection is carried out; (3) after the cells is intervened with the Hcy with optimal concentration, the apoptosis of endothelial cells is detected by flow cytometry; (4) after the cells isintervened with the Hcy with optimal concentration, the expression of LncRNA (long noncoding RNA) DINO (Damage Induced Noncoding) is detected through fluorescent quantitative PCR in real time; (5) transfecting of the LncRNA DINO siRNA is carried out, after the LncRNA DINO is interfered, and then cell viability staining detection is carried out; (6) LncRNA DINO siRNA is constructed, after the LncRNA DINO is interfered, and the apoptosis of endothelial cells is detected by flow cytometry; and (7) LncRNA DINO siRNA is constructed, after the LncRNA DINO is interfered, and then the expression of apoptosis-related protein is detected by protein immunoblotting. A novel molecular target for early diagnosis and early warning of atherosclerosis is provided, and a new direction is provided for treatment.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a molecular marker for early warning of atherosclerosis and its application. Background technique [0002] Atherosclerosis (Atherosclerosis, AS) is a chronic progressive disease that is characterized by endothelial cell apoptosis, smooth muscle cell proliferation, and foam cell formation, and is prone to occur in large and medium-sized arteries. Major cause of high morbidity and mortality. In recent years, with the rapid development of society and economy, people's living standards have been continuously improved. Affected by factors such as changes in diet structure and increased work pressure, the incidence of AS (atherosclerosis) has shown an increasing trend year by year and is gradually becoming younger. It can be seen that early detection, prevention and delay of the process of atherosclerosis are the top priority in the field of clinical science today. There...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/113C12N15/11A61K45/00A61K31/713A61P9/10
CPCC12Q1/6883A61K45/00A61K31/713A61P9/10C12Q2600/158C12Q2600/178
Inventor 姜怡邓张慧萍张辉刘圆马芳杨安宁马胜超
Owner NINGXIA MEDICAL UNIV
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