Application of micromolecule RNA hsa-miR-451a in preparation of medicine for treating cerebral arterial thrombosis

A technology for ischemic stroke and ischemic stroke, applied in DNA/RNA fragments, gene therapy, drug combination, etc., can solve problems such as reducing the expression of pro-inflammatory proteins

Pending Publication Date: 2021-06-08
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Intraventricular injection of miR-3473b inhibitor may reduce expression of pro-inflammatory proteins by targeting SOCS3 during ischemic stroke

Method used

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  • Application of micromolecule RNA hsa-miR-451a in preparation of medicine for treating cerebral arterial thrombosis
  • Application of micromolecule RNA hsa-miR-451a in preparation of medicine for treating cerebral arterial thrombosis
  • Application of micromolecule RNA hsa-miR-451a in preparation of medicine for treating cerebral arterial thrombosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Detection of hsa-miR-451a expression level in ischemic stroke brain tissue

[0048] Total RNA was extracted from ischemic stroke (MCAO) brain tissue by Trizol method. Wash the cells twice with PBS, add 1mL Trizol to each bottle, pipette to mix, and incubate at room temperature for 5min; transfer the cell solution into a DEPC-treated 1.5mL centrifuge tube, add 0.2mL chloroform to each tube, shake vigorously for 15s to mix, Stand on ice for 3-5 minutes to separate the layers; centrifuge at 12,000 rpm for 15 minutes at 4°C; carefully absorb the upper layer liquid, transfer it to another 1.5mL centrifuge tube treated with DEPC, add an equal volume of isopropyl alcohol that has been pre-cooled at 4°C alcohol, upside down and gently mix for 15-30s, let stand on ice (room temperature) for 5-10min; , centrifuge at 7500rpm for 10min, discard the supernatant, invert the centrifuge tube to dry, and add 30μL of DEPC water to dissolve the RNA. The concentration and puri...

Embodiment 2

[0052] Example 2: hsa-miR-451a reduces the infarct size of MACO mice

[0053] Prepare an in vivo transfection solution by mixing 3.5 μL of hsa-miR-451a mimic or hsa-miR-451a inhibitor with 3.5 μL of siRNA-Mate transfection reagent. The transfection solution was injected into the ventricles of the affected side (the side of MCAO) of C57BL / 6 mice at the positions of bregma: −2.5 mm, ventral: 1 mm, lateral opening: 1.5 mm). One day after the intracerebroventricular injection, MCAO was performed. After 48 hours, the mouse brain tissue was frozen and sectioned. The brain sections were immersed in Nissl staining solution for 15 minutes, washed with distilled water, and incubated with 95% ethanol for 5 seconds. Nissl-stained images of the brain were loaded into Adobe PhotoShop for infarct volume measurement ( figure 2 ).

Embodiment 3

[0054] Example 3: hsa-miR-451a reduces apoptosis in MCAO mice

[0055] hsa-miR-451a mimics and hsa-miR-451a inhibitors are transfected into the affected side of C57BL / 6 mice in vitro, the method is as in Example 2, and the frozen sections of the mouse brain tissue are treated with 4% paraformaldehyde Fix for 20 minutes, soak the cells in the osmotic solution on ice for 2 minutes, carefully absorb the excess liquid around the tissue on the slide with filter paper, immediately add 20 μL Tunnel reaction solution to the slice, put it in a wet box and react at 37°C for 1 hour, and observe it under a fluorescence microscope. Observation of cell staining showed that there were more apoptotic cells in the brain tissue of mice transfected with hsa-miR-451a inhibitor, and the number of apoptotic cells in the brain tissue of mice transfected with hsa-miR-451a mimic decreased ( image 3 ).

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Abstract

The invention relates to application of micromolecule RNAhsa-miR-451a in preparation of a medicine for treating cerebral arterial thrombosis, and belongs to the field of biotechnology research. The invention discloses the application of micromolecule RNAhsa-miR-451a in preparation of the medicine for treating the cerebral arterial thrombosis. The application provided by the invention can effectively protect cells from ischemia-induced death, reduce the infarct area and effectively improve the treatment effect of preparing the medicine for treating the cerebral arterial thrombosis.

Description

technical field [0001] The invention belongs to the field of biotechnology research, and specifically relates to the application of a small molecule RNA hsa-miR-451a in the preparation of medicines for treating ischemic stroke. Background technique [0002] Stroke affects millions of people worldwide every year and is one of the leading causes of death [1-2]. Ischemic stroke is the most common type of stroke, accounting for 80% of all stroke cases [3]. The pathogenesis of ischemic stroke stems from insufficient blood supply to the affected brain tissue, whereby the brain tissue receives less oxygen and glucose than it needs. As a result, brain damage occurs, which can lead to permanent and irreversible cell death. For many years, there have been few therapeutic breakthroughs in the treatment of ischemic stroke. Accordingly, extensive research has focused on identifying effective therapies to rescue affected cells from death. [0003] In the study of ischemic stroke drugs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K31/7088A61P9/10
CPCC12N15/113A61K31/7088A61P9/10C12N2310/141C12N2320/30
Inventor 郎明非白鹰王蒙蒙孙晶康秀芝
Owner DALIAN UNIV
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