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Application of miR-674-3p in preparation of drugs for preventing or treating stress hypertension

A high blood pressure and stress technology, which can be used in drug combinations, pharmaceutical formulations, organic active ingredients, etc., and can solve problems such as limited

Active Publication Date: 2021-09-14
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 miR-674-3p in preparation of drugs for preventing or treating stress hypertension
  • Application of miR-674-3p in preparation of drugs for preventing or treating stress hypertension
  • Application of miR-674-3p in preparation of drugs for preventing or treating stress hypertension

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The establishment of embodiment 1 stress hypertensive rat model

[0037] The Animal Experiment Center provided 50 healthy and qualified 7-week-old adult male SD (Sprague-Dawley) rats, weighing 230-260 g. Raise in SPF grade animal room, drink water and eat freely, keep room temperature at 23±1°C, relative humidity 50%, standard 12:12 hours day and night cycle. The rats were randomly divided into two groups, the stress-induced hypertension group and the normal control group, 25 rats in each group, and the model was established after 3 days of adaptive feeding.

[0038] The rats in the stress hypertensive rat group were placed in a modeling cage (22cm×22cm×28cm), and the soles of their feet were subjected to intermittent electric shock stimulation (voltage 50V, duration 50ms each time, interval 2s), and accompanied by There is continuous noise stimulation (produced by buzzer, 90dB). The whole process was controlled by a computer program, and they were stressed once a day...

Embodiment 2

[0042] Example 2 miRNA-seq reveals that miR-674-3p is significantly highly expressed in the rostral ventrolateral region of the medulla in stress-induced hypertensive rats

[0043] Three stress-induced hypertensive rats and three normal control rats were randomly selected, and the total RNA of the rostral ventrolateral region of the medulla was extracted from six rats by Trizol (Invitrogen) method. Using agarose gel electrophoresis, Nanodrop ND-2000 ultra-micro spectrophotometer (Thermo), The quality of total RNA was assessed by a Fluorometer (Invitrogen) and an Agilent 2100 Bioanalyzer (Agilent). It is required that three bands of 28S, 18S and 5S can be seen clearly after agarose gel electrophoresis, and the brightness of 28S is twice that of 18S, and 5S is the darkest; the OD260 / OD280 value is between 1.8-2.0; the RNA integrity value (RIN) is greater than 8.0.

[0044] The Tru Seq Small RNA Sample Prep Kits (Illumina) kit was used for miRNA-seq library construction, includ...

Embodiment 3

[0048] Example 3, qPCR verification of miR-674-3p expression

[0049] Total RNA in the rostral ventrolateral region of the medulla was extracted from stress-induced hypertensive rats and normal control rats using Trizol (Invitrogen). use Ⅱ1st Strand cDNA Synthesis Kit (Yeasen) kit was used for reverse transcription to synthesize cDNA. 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 instrument is Bio-Rad CFX96touch qPCR system (Bio-Rad). The reaction procedure is as follows: the first step is 95° C., 5 min. The second step is 40 cycles, including 95°C, 10s; 60°C, 20s; 72°C, 20s. U6 was used as an internal reference gene. see results image 3 , the expression of miR-674-3p in the rostral ventrolateral medulla of stress-induced hypertensive rats was significantly higher than that of normal control rats (*P<0.05), consistent with miRNA-seq results.

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Abstract

The invention discloses an application of miR-674-3p in preparation of drugs for preventing or treating stress hypertension. The application of the miR-674-3p in preparation of the drugs for preventing or treating the stress hypertension reveals that the heart rate, renal sympathetic nerve activity, mean arterial pressure, systolic pressure and diastolic pressure of a rat with the stress hypertension can be effectively reduced by inhibiting the expression of the miR-674-3p. The miR-674-3p can be used as a new molecular target for treating the stress hypertension and is used for guiding the screening, research and development of related drugs.

Description

[0001] (1) Technical field [0002] The invention relates to the application of miR-674-3p in the preparation of drugs for preventing or treating stress hypertension. [0003] (2) Background technology [0004] With the rapid development of social economy, the pace of life is accelerating, the pressure of work is increasing, and all kinds of fierce competition make people's spirits and emotions continue to be tense and stress intensified, leading to high blood pressure. 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 hypertension. Stress leads to changes in the expression and regulation of a series of genes in the rostral ventrolateral medulla, causing sympathetic nerve excitation and increasing blood pressure. Therefore, it is of great ...

Claims

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Application Information

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Patent Type & Authority Applications(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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