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Application of mesenchymal stem cell exosome in preparation of preparations for promoting mitophagy

A technology of mitophagy and mesenchymal stem cells, applied in the biological field, can solve problems such as cell damage, damaged mitochondria aggregation, malignant transformation, etc., and achieve the effects of promoting mitophagy, correcting mitochondrial dysfunction, and improving the ability to promote mitophagy

Active Publication Date: 2019-06-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PINK1 and PARKIN are closely related to maintaining the integrity of the mitochondrial network and the normal function of cells. Mutations in their coding genes can lead to the accumulation of damaged mitochondria, which in turn leads to cell damage or malignant transformation

Method used

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  • Application of mesenchymal stem cell exosome in preparation of preparations for promoting mitophagy
  • Application of mesenchymal stem cell exosome in preparation of preparations for promoting mitophagy
  • Application of mesenchymal stem cell exosome in preparation of preparations for promoting mitophagy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 AMSC-Exo promotes mitophagy in liver cells

[0043] The human liver cell line HL-7702 was treated with gradient concentrations of hAMSC-Exo (5 μg / mL, 20 μg / mL, 80 μg / mL) for 24 hours, and then the cells were collected for Western Blot detection of mitophagy-related molecules. Or add gradient concentrations of mAMSC-Exo (5 μg / mL, 20 μg / mL, 80 μg / mL) to primary mouse hepatocytes, collect cells after 24 hours of treatment, and perform Western Blot detection of mitophagy-related molecules. The WB bands were all scanned by ChemiScope Western BlotImaging System (Clinx Science Instruments Co., Ltd), and then analyzed by Image J software (Rawak Software, Inc. Germany) for gray ratio analysis. Such as figure 1 It was shown that AMSC-Exo can significantly promote mitophagy in hepatic cell lines and primary hepatocytes, specifically manifested as a dose-dependent up-regulation of the levels of mitophagy-related molecules PARKIN and PINK1 in hepatocytes.

Embodiment 2

[0044] Example 2 AMSC-Exo promotes macrophage mitophagy

[0045] Macrophages RAW264.7 were treated with gradient concentrations of mAMSC-Exo (5 μg / mL, 20 μg / mL, 80 μg / mL) for 24 hours, and then the cells were collected for Western Blot detection of mitophagy-related molecules. WB bands After scanning with ChemiScope Western Blot Imaging System (Clinx Science Instruments Co., Ltd), image J software (Rawak Software, Inc. Germany) was used for gray ratio analysis. Such as figure 2 It was shown that mAMSC-Exo can significantly promote the mitophagy of macrophages, which is manifested in the dose-dependent up-regulation of the levels of mitophagy-related molecules PARKIN and PINK1.

[0046] Adding hAMSC-Exo to THP-1 or U937-derived macrophages induced by PMA also had the above effects.

Embodiment 3

[0047] Example 3 AMSC-Exo can relieve drug-induced mitochondrial damage in hepatocytes

[0048] HL-7702 cells were cultured with gradient concentrations of AMSC-Exo (5 μg / mL, 20 μg / mL, 80 μg / mL) for 12 hours, then treated with 5 mM APAP for 12 hours to induce mitochondrial damage, and then analyzed by MitoSOX Red staining and flow cytometry Mitochondrial ROS (mtROS) level; JC-1 probe staining combined with flow cytometry was used to detect mitochondrial membrane potential; Western Blot was used to detect TOM20 protein level to reflect mitochondria clearance.

[0049] Such as image 3 It was shown that AMSC-Exo can effectively reduce APAP-induced hepatocyte mtROS secretion; correct APAP-induced abnormal mitochondrial membrane potential in hepatocytes; and clear damaged mitochondria in hepatocytes caused by APAP.

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Abstract

The invention provides an application of a mesenchymal stem cell exosome in preparation of preparations for promoting mitophagy. By preparing an exosome derived from the adipose mesenchymal stem cells, Western Blot and flow experiments at the cell level confirm that the regulation and control effect of AMSC-Exo on the mitophagy of parenchyma cells and immune cells; at the same time, in-vivo experiments confirm the effect of the AMSC-Exo on mitophagy pathway in liver tissue; and biological preparations or medicaments for promoting mitophagy are further prepared. Meanwhile, by modifying the AMSC-Exo, the effect for promoting mitophagy of the AMSC-Exo is further improved. The exosome can promote mitophagy and correct mitochondrial dysfunction; has mitophagy promoting effect on both parenchymacells and immune cells; can be used for preparing preparations or medicines for treating diseases related to mitochondrial dysfunction; and can further improve ability for promoting the mitophagy after specific modification.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a new application of exosomes derived from mesenchymal stem cells, in particular to an application of exosomes from adipose-derived mesenchymal stem cells (AMSC) in preparing preparations for promoting mitophagy. Background technique [0002] Mitochondria are the energy factories of the cell, and healthy mitochondria are like a power plant that converts energy through the tricarboxylic acid cycle and oxidative phosphorylation. The function of mitochondria can determine the life and death of cells. On the one hand, it generates ATP through oxidative phosphorylation to provide the energy demand of cells, and transports calcium ions in the cytoplasm to mitochondria for storage through calcium ion channels, so that cells can maintain normal functions; on the other hand Abnormal accumulation of reactive oxygen species (ROS) in mitochondria and the production of apoptosis-related proteins, s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P43/00C12N5/0775
Inventor 刘艳宁楼国华郑敏
Owner ZHEJIANG UNIV
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