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Application of siRNA-golph3 in the preparation of drugs for preventing or treating cardiomyocyte hypertrophy

A cardiomyocyte and hypertrophy technology, which is used in drug combinations, pharmaceutical formulations, cardiovascular system diseases, etc.

Active Publication Date: 2020-10-23
NANHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether apelin can regulate the function of Golgi apparatus through GOLPH3-dependent Golgi autophagy and induce cardiomyocyte hypertrophy has not been reported.

Method used

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  • Application of siRNA-golph3 in the preparation of drugs for preventing or treating cardiomyocyte hypertrophy
  • Application of siRNA-golph3 in the preparation of drugs for preventing or treating cardiomyocyte hypertrophy
  • Application of siRNA-golph3 in the preparation of drugs for preventing or treating cardiomyocyte hypertrophy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Determination of the impact of Apelin-13 on Golgi autophagy

[0026] After apelin stimulation and APJ receptor antagonist blocking, Golgi apparatus markers (cis membrane vesicle marker GM130-RFP and trans membrane vesicle marker TGN46-RFP) and autophagy marker proteins were observed in rat cardiomyocytes LC3B-GFP is the result of fluorescent co-localization. To detect the effect of Apelin-13 on the expression of Golgi apparatus stress marker protein GOLPH3 and the protein interaction with LC3B in H9c2 cardiomyocytes. The results of transmission electron microscopy were used to determine whether Apelin-13 could induce Golgi autophagy in rat cardiomyocytes.

[0027] 1. Determination of the effect of Apelin-13 on the fluorescence co-localization of Golgi markers and autophagy marker protein LC3B-GFP

[0028] Select cells with a growth density of 50%-70% in the culture plate for plasmid transfection. First dilute Lipofectamine with optimized medium TM 3000, P...

Embodiment 2

[0033] Example 2 Measuring the effect of interfering with GOLPH3 expression on Apelin-13-induced cardiomyocyte hypertrophy

[0034] After interfering with GOLPH3 stimulation of apelin, Golgi markers (cis membrane vesicle marker GM130-RFP and trans membrane vesicle marker TGN46-RFP) and autophagy marker protein LC3B-GFP were fluorescently co-localized in rat cardiomyocytes result. The effect of interfering with GOLPH3 on Apelin-13-induced Golgi autophagy in rat cardiomyocytes was determined by transmission electron microscopy. Effect of interference with GOLPH3 on the upregulation of diameter and volume of H9c2 cardiomyocytes by Apelin-13.

[0035] 1. Determination of the effect of si-GOLPH3 on the fluorescence colocalization of Apelin-13-induced Golgi markers and autophagy marker protein LC3B-GFP

[0036] After GM130-RFP, TGN46-RFP, and LC3B-GFP were all transferred into the cells for 24 hours, rat cardiomyocytes were treated with Apelin-13, Apelin-13+si-GOLPH3 respectively,...

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Abstract

The invention discloses an application of Golgi apparatus outer membrane protein GOLPH3 as a biomarker in predicting cardiomyocyte hypertrophy, and an application of siRNA-GOLPH3 in preparing a medicine for preventing or treating cardiomyocyte hypertrophy. The Golgi apparatus outer membrane protein GOLPH3 can promote Golgiphagy, the novel selective autophagy can mediate Apelin-13 to promote cardiomyocyte hypertrophy, and the expression of interference GOLPH3 can inhibit Golgiphagy in cardiomyocytes and the effect of increasing diameter and volume of cardiomyocytes by Apelin-13.

Description

technical field [0001] The present invention relates to Golgi body autophagy mediated by GOLPH3, which itself can participate in cardiomyocyte hypertrophy induced by apelin. GOLPH3 inhibitor siRNA-GOLPH3 itself can inhibit apelin-induced cardiomyocyte hypertrophy and cardiac hypertrophy by inhibiting Golgi autophagy. Background technique [0002] APJ is a G protein-coupled receptor first discovered by O'Dowd et al. in 1993, and in 1998, Tatemoto et al. extracted the endogenous ligand apelin of APJ for the first time in bovine stomach. The precursor apelin has 77 amino acids, which can be hydrolyzed into some amino acid fragments of different lengths by endopeptidase, including apelin-12, apelin-13, apelin-17, apelin-36. Among these active fragments of apelin, Apelin-13 has the strongest biological activity. The Apelin / APJ system has very rich biological functions in the cardiovascular system. Apelin / APJ system can promote angiogenesis, maintain fluid homeostasis, induce p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/713A61P9/00
Inventor 陈临溪陆丽群李兰芳罗旭灵戚芷豪唐名珠张开
Owner NANHUA UNIV
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