miRNA markers related to auxiliary diagnosis of myocardial fibrosis diseases and application of miRNA markers

Abstract

The invention discloses miRNA markers related to auxiliary diagnosis of myocardial fibrosis diseases and application of the miRNA markers. Hsa-miR-6802-3p and hsa-miR-1229-3p which are differentially expressed in patients suffering from myocardial fibrosis diseases are screened out by a high-throughput sequencing analysis technology, further cell experiments verify that the miRNA biomarkers hsa-miR-6802-3p and hsa-miR-1229-3p can be used in clinical auxiliary diagnosis of myocardial fibrosis, have important significance in early diagnosis of myocardial fibrosis, provide drug targets for clinical preparation of drugs for myocardial fibrosis, and are helpful for realizing targeted therapy of myocardial fibrosis.

Description

technical field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to miRNA markers related to the auxiliary diagnosis of myocardial fibrosis diseases and applications thereof. Background technique

[0002] Cardiovascular disease is the number one deadly disease in the world. According to the "China Cardiovascular Health and Disease Report 2019" released by the National Center for Cardiovascular Diseases, cardiovascular disease death is the number one cause of death in China. There are many types of diseases , the etiology is complex. Among them, the continuous and / or repeated aggravated ischemia and hypoxia of myocardial fibers caused by moderate and severe coronary atherosclerotic stenosis will lead to myocardial fibrosis (Myocardial fibrosis, MF) disease, which is characterized by myocardial fibroblasts Excessive proliferation, activation of cardiac fibroblasts into myofibroblasts, and secretion of type I collagen, resulting in ...

Examples

Embodiment 1

[0073] Example 1 Screening biomarkers related to myocardial fibrosis

[0074] 1. Research object

[0075] Three cases of patients with myocardial fibrosis provided by the cardiology department of the hospital were collected, and their blood samples were collected respectively. At the same time, three cases of healthy people were collected as healthy controls, and their blood samples were collected respectively. All patients were excluded from malignant tumors, acute infection, trauma and severe liver and kidney diseases, cerebral embolism, pulmonary embolism, lower extremity venous thrombosis, DIC and acute renal insufficiency and other diseases.

[0076] All research subjects were informed about this study and signed the informed consent form, which was approved by the organizational ethics committee of the hospital.

[0077] 2. Sequencing experiments

[0078] The sequencing experiments used the Illumina TruseqTM RNA sample prep Kit method for strand-specific library construc...

Embodiment 2

[0117] Example 2 Detection of the effects of miRNA on the proliferation and migration of cardiac fibroblasts

[0118] 1. Cell culture

[0119] The human primary cardiac fibroblasts (HCF) preserved in liquid nitrogen were taken out and resuscitated, inoculated in DMEM medium containing 10% fetal bovine serum, placed at 37°C, 95% humidity and 5% CO 2 Cells in the logarithmic growth phase were subcultured in a cell incubator for experiments.

[0120] 2. Digestion and passage of primary cardiac fibroblasts

[0121] When the fusion rate of the cells reaches about 80%-90% of the bottom area of ​​the full culture dish and the culture medium turns yellow, suck up the liquid in the culture dish, add 4mL trypsin, put it in the incubator for digestion for 2min, take it out and put it in Observe under a microscope, after the cells retract and the intercellular space widens, quickly absorb the trypsin and add 2 mL of culture medium containing 10% fetal bovine serum to stop the digestion;...

technical field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to miRNA markers related to the auxiliary diagnosis of myocardial fibrosis diseases and applications thereof. Background technique

[0002] Cardiovascular disease is the number one deadly disease in the world. According to the "China Cardiovascular Health and Disease Report 2019" released by the National Center for Cardiovascular Diseases, cardiovascular disease death is the number one cause of death in China. There are many types of diseases , the etiology is complex. Among them, the continuous and / or repeated aggravated ischemia and hypoxia of myocardial fibers caused by moderate and severe coronary atherosclerotic stenosis will lead to myocardial fibrosis (Myocardial fibrosis, MF) disease, which is characterized by myocardial fibroblasts Excessive proliferation, activation of cardiac fibroblasts into myofibroblasts, and secretion of type I collagen, resulting in ...

Examples

Embodiment 1

[0073] Example 1 Screening biomarkers related to myocardial fibrosis

[0074] 1. Research object

[0075] Three cases of patients with myocardial fibrosis provided by the cardiology department of the hospital were collected, and their blood samples were collected respectively. At the same time, three cases of healthy people were collected as healthy controls, and their blood samples were collected respectively. All patients were excluded from malignant tumors, acute infection, trauma and severe liver and kidney diseases, cerebral embolism, pulmonary embolism, lower extremity venous thrombosis, DIC and acute renal insufficiency and other diseases.

[0076] All research subjects were informed about this study and signed the informed consent form, which was approved by the organizational ethics committee of the hospital.

[0077] 2. Sequencing experiments

[0078] The sequencing experiments used the Illumina TruseqTM RNA sample prep Kit method for strand-specific library construc...

Embodiment 2

[0117] Example 2 Detection of the effects of miRNA on the proliferation and migration of cardiac fibroblasts

[0118] 1. Cell culture

[0119] The human primary cardiac fibroblasts (HCF) preserved in liquid nitrogen were taken out and resuscitated, inoculated in DMEM medium containing 10% fetal bovine serum, placed at 37°C, 95% humidity and 5% CO 2 Cells in the logarithmic growth phase were subcultured in a cell incubator for experiments.

[0120] 2. Digestion and passage of primary cardiac fibroblasts

[0121] When the fusion rate of the cells reaches about 80%-90% of the bottom area of ​​the full culture dish and the culture medium turns yellow, suck up the liquid in the culture dish, add 4mL trypsin, put it in the incubator for digestion for 2min, take it out and put it in Observe under a microscope, after the cells retract and the intercellular space widens, quickly absorb the trypsin and add 2 mL of culture medium containing 10% fetal bovine serum to stop the digestion;...

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