Application of a kind of mir-674-3p in the preparation of drugs for preventing or treating stress-induced hypertension

A technique for irritability and high blood pressure, applied in the direction of drug combinations, pharmaceutical formulations, organic active ingredients, etc., can solve limited and other problems

Active Publication Date: 2022-08-02
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the relationship between miRNA regulation and stress hypertension is very limited, and there is no report on the role of miR-674-3p in the ventrolateral region of the rostral medulla in regulating stress hypertension.

Method used

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  • Application of a kind of mir-674-3p in the preparation of drugs for preventing or treating stress-induced hypertension
  • Application of a kind of mir-674-3p in the preparation of drugs for preventing or treating stress-induced hypertension
  • Application of a kind of mir-674-3p in the preparation of drugs for preventing or treating stress-induced hypertension

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Establishment of stress-induced hypertension rat model

[0032] The animal experiment center provided healthy and qualified 7-week-old adult male SD (Sprague-Dawley) rats, 50, weighing 230-260 g. The animals were reared in SPF animal room with free access to water and diet, room temperature maintained at 23±1°C, relative humidity 50%, and a standard 12:12-hour day-night cycle. Rats were randomly divided into two groups, the stress-induced hypertension group and the normal control group, with 25 rats in each group. The model was established after 3 days of adaptive feeding.

[0033] The rats in the stress hypertensive rat group were placed in a modeling cage (22cm×22cm×28cm), and their soles were stimulated by intermittent electric shocks (voltage 50V, each time duration 50ms, interval 2s), accompanied by There is continuous noise stimulation (buzzer generation, 90dB). The whole process was controlled by a computer program and was stressed once in the morning...

Embodiment 2

[0037] Example 2 miRNA-seq revealed that miR-674-3p was significantly highly expressed in the rostral ventrolateral region of the medulla oblongata in stress-induced hypertensive rats

[0038] Three stress-induced hypertensive rats and three normal control rats were randomly selected, and total RNA was extracted from the rostral ventrolateral region of the medulla oblongata from six rats by Trizol (Invitrogen). Using agarose gel electrophoresis, Nanodrop ND-2000 ultra-micro spectrophotometer (Thermo), The quality of total RNA was assessed with a Fluorometer Fluorometer (Invitrogen) and an Agilent 2100 Bioanalyzer (Agilent). It is required that three bands of 28S, 18S and 5S are obvious after agarose gel electrophoresis, and the brightness of 28S is twice that of 18S, and 5S is the darkest; OD260 / OD280 value is between 1.8-2.0; RNA integrity value (RIN) is greater than 8.0.

[0039] The Tru Seq Small RNA Sample Prep Kits (Illumina) kit was used for miRNA-seq library construct...

Embodiment 3

[0043] Example 3. qPCR verification of miR-674-3p expression

[0044] Trizol (Invitrogen) was used to extract total RNA from the rostral ventrolateral region of the medulla oblongata in stress-induced hypertensive rats and normal control rats. use II 1st Strand cDNA Synthesis Kit (Yeasen) was used to synthesize cDNA by reverse transcription. 20ul qPCR system includes: 7.2μL ultrapure water, 10μL qPCR SYBR Green Master Mix (Yeasen), 0.4 μL upstream primer, 0.4 μL downstream primer, and 2 μL cDNA. The reaction apparatus was Bio-Rad CFX96touch qPCR system (Bio-Rad). The reaction procedure is as follows: the first step is 95°C, 5min. The second step was 40 cycles, including 95°C, 10s; 60°C, 20s; 72°C, 20s. U6 was used as the internal reference gene. see the results image 3 , the expression of miR-674-3p in the rostral ventrolateral region of the medulla oblongata in stress-induced hypertensive rats was significantly higher than that in normal control rats (*P<0.05), whic...

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Abstract

The invention discloses an application of miR-674-3p in the preparation of a drug for preventing or treating stress-induced hypertension. The invention discloses that inhibiting the expression of miR-674-3p can effectively reduce the heart rate and kidney function of stress-induced hypertensive rats. Sympathetic nerve activity, mean arterial pressure, systolic blood pressure, and diastolic blood pressure. miR‑674‑3p can be used as a new molecular target for the treatment of stress-induced hypertension and guide the screening and development of related drugs.

Description

(1) Technical field [0001] The invention relates to the application of miR-674-3p in the preparation of drugs for preventing or treating stress-induced hypertension. (2) Background technology [0002] With the rapid development of social economy, the rhythm of life is accelerating, the pressure of work is increasing, and various fierce competitions make people's mental and emotional tension and stress intensify, leading to the occurrence of hypertension, which is called high stress. blood pressure. Stress-induced hypertension is an important risk factor for stroke, coronary heart disease and other diseases. The rostral ventrolateral medulla is one of the key central nuclei for the regulation of cardiovascular activity and plays an important regulatory role in stress-induced hypertension. Stress leads to a series of gene expression and regulation changes in the rostral ventrolateral medulla oblongata, which causes sympathetic nerve excitation and increases blood pressure. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/7088A61P9/12
CPCA61K31/7088A61P9/12
Inventor 张帅陈高俊邢孟愉杜东书杜羽
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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