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Application of FG-4592 or salt thereof in aspect of proliferation of neural stem cells

A technology of FG-4592, 1. FG-4592 is applied in the application field of FG-4592 or its salt in the expansion of neural stem cells to achieve the effect of promoting proliferation

Inactive Publication Date: 2020-07-28
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, so far, whether FG-4592 can activate the HIF signaling pathway and have the effect of neural stem cell expansion has not been reported

Method used

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  • Application of FG-4592 or salt thereof in aspect of proliferation of neural stem cells
  • Application of FG-4592 or salt thereof in aspect of proliferation of neural stem cells
  • Application of FG-4592 or salt thereof in aspect of proliferation of neural stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1F

[0020] Example 1 Effect of FG-4592 on Neural Stem Cell Expansion in Vitro

[0021] 1. FG-4592 drug

[0022] (1) Treatment of FG-4592: FG-4592 was purchased from Selleck, CAS number: 808118-40-3, molecular weight 352.34. Take 100mg of FG-4592 powder, add 4ml of dimethyl sulfoxide (DMSO), oscillate to make it fully dissolved, add DMSO to a final volume of 5.676ml, mix well, and the final concentration of this stock solution is 50mM. Store at -20°C after aliquoting.

[0023] (2) Treatment of neural stem cells: The experiment used isolated and cultured rat embryonic neural stem cells. The third-generation neural stem cells were used to adjust the density to 1×10 6 / ml, 6 hours after inoculation, add the prepared FG-4592 to the culture medium to make the final concentration 10μM, mix gently, and place at 37°C, 5% CO 2 Continue culturing for 24 h in the cell culture incubator.

[0024] (3) Control group: the control group was divided into negative control group and positive con...

Embodiment 2HI

[0031] Example 2 HIF-1α mediates autophagy and proliferation of neural stem cells

[0032] In order to prove that hypoxia does regulate the autophagy and proliferation of neural stem cells through the mediation of HIF-1α, we knocked down the expression of HIF-1α by RNA interference technology (si-HIF-1α), and then detected the neural stem cells by Western Blot. Altered autophagy and proliferative activity. The results showed that the autophagy and proliferation of neural stem cells were weakened after knocking down the expression of HIF-1α ( figure 2 ). The above results indicate that HIF-1α mediates the proliferation of neural stem cells, and may regulate proliferation by affecting autophagy activity.

Embodiment 3

[0033] Example 3 Effect of Autophagy Inhibitor 3-MA Treatment on Neural Stem Cell Proliferation

[0034] Next, we treated cells with autophagy inhibitor 3-MA, and observed the growth of neural stem cells at different time points in normoxia and hypoxia ( image 3). Figure A shows the growth state of neural stem cells photographed under different oxygen conditions and at different times, and Figure B is the statistical result of the diameter of the neurospheres of the photographed cells. The results showed that the diameter of neurospheres was significantly reduced after inhibiting autophagy, suggesting that increased autophagy activity contributes to the proliferation of neural stem cells.

[0035] From the above results, we conclude that FG-4592 can induce neural stem cell autophagy by increasing the expression of HIF-1α, thereby stimulating the proliferation of neural stem cells.

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Abstract

The invention discloses an application of FG-4592 or a salt thereof in the aspect of proliferation of neural stem cells. The FG-4592 or the salt thereof is added to an isolated neural stem cell culture medium, so that proliferation of the neural stem cells can be promoted, and HIF-1alpha overexpression and autophagy are induced at the same time. Under constant oxygen conditions, the neural stem cells are treated in vitro with the FG-4592 for 24 hours, so that proliferation of the neural stem cells can be significantly promoted, HIF-1alpha overexpression and autophagy are induced at the same time, and a proliferation promoting effect of low oxygen on the neural stem cells and the effects of inducing HIF-1alpha overexpression and enhancing autophagy are completely simulated.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of FG-4592 or its salt in neural stem cell expansion. Background technique [0002] The decrease in the number of neural stem cells is the main reason why neurodegenerative diseases or nerve damage are difficult to repair themselves. The number of neural stem cells in adults is limited, and it is difficult to be activated and expanded from a resting state to perform functions. Transplantation of neural stem cells after in vitro expansion provides a feasible candidate for patients. In addition, if the proliferation of neural stem cells can be effectively promoted in vivo, it will bring bright prospects for the treatment of such diseases. The maintenance of self-renewal, proliferation and multi-directional differentiation potential of neural stem cells requires a certain microenvironment, which is called "stem cell niche" in the field of stem cell research. Thi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797A61K31/472A61P25/00
CPCA61K31/472A61P25/00C12N5/0623C12N2500/32
Inventor 吴丽颖朱玲玲巩生辉何云凌赵名成祥韩莹赵彤范明
Owner ACADEMY OF MILITARY MEDICAL SCI
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