Application of D-erythro-Sphingosine in central nervous system

A sphingosine and myelin technology, which can be used in nervous system cells, nervous system diseases, vertebrate cells, etc., and can solve problems such as unclear downstream signal transduction pathways.

Inactive Publication Date: 2014-08-20
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But its downstream signal tra

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  • Application of D-erythro-Sphingosine in central nervous system
  • Application of D-erythro-Sphingosine in central nervous system
  • Application of D-erythro-Sphingosine in central nervous system

Examples

Experimental program
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Embodiment 1

[0011] Example 1: Culture of SD rat oligodendrocytes

[0012] Take the brains of SD newborn rats and separate the bilateral cerebral cortex, D. Rinse with Hanks solution, pipette to make cell suspension, filter with 200 mesh cell screen, centrifuge at 180×g for 5 min, discard the supernatant, add 10% fetal bovine serum in high glucose DMEM medium to resuspend the cells to (1.0 -2.0) *106 density planted in 0.025% polylysine pretreated 75c㎡ culture flask 5% CO2 culture for 8-11 days, after the cells are fused and form obvious stratification, start OPCs shaking separation and purification (the culture flask Place in a constant temperature air bath vibrating shaker, 37 degrees, 150r / min, shake for 2h to remove microglia, etc. D. Hanks solution rinse, change the solution; 5%CO2 incubator equilibrate 2h; 230r / min, 16-18h , 280r / min, 1-2h; Collect the cell suspension, centrifuge at 400×g for 5 min, resuspend the cells in high glucose DMEM medium containing 10% fetal bovine serum, and ...

Embodiment 2

[0013] Example 2: See figure 1 , 2 We found that culturing primary cultured rat OPC cells with myelin fragments as a matrix inhibits the differentiation and maturation of OPC cells in its differentiation medium. Adding sphingosine reverses the inhibition of myelin fragments to the differentiation and maturation of OPC cells. effect. figure 1 It shows that the primary cultured rat OPC cells cultured with myelin fragments as the matrix were stained with MBP after 5 days of differentiation in the differentiation medium. The poor differentiation of OPC cells in the control group was manifested by shorter neurite outgrowth and a significant decrease in the total area. The OPC cells differentiated well after adding sphingosine, and the total area of ​​longer branches increased significantly. figure 2 :Statistical analysis of the total area of ​​the control group (DMSO) and sphingosine treatment (DES 100 nM concentration) OPC cells grown on myelin debris for 5 days, the area of ​​the c...

Embodiment 3

[0015] Example 3: Preparation of EAE rat model and evaluation of exercise capacity after sphingosine treatment.

[0016] Strain: 20 Wistar rats, 6 weeks to 8 weeks old, weight (200 ± 20) g, female; (Shanghai Experimental Animal Center, Chinese Academy of Sciences).

[0017] Modeling method: According to the literature and the basis of previous work, the synthesis of MOG and Freund’s adjuvant (the ratio is 1:1 by volume, each rat uses 50ul complete Freund’s adjuvant plus 50ul total 50ug normal saline Rat MOG (35-55 small peptide) was injected subcutaneously into multiple points. After 3 days of immunization, 20 female Wistar rats were randomly divided into 2 groups: EAE + DMSO group (control group), EAE + intraperitoneal injection of sphingosine group [0.5 mg / (kg·d) ]. Observe the rats' exercise performance scores at different time points after treatment, and the results are shown in Table 1:

[0018] EAE rat score detection. EAE rat score: 0.5 distal tail paralysis, 1 complete tai...

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Abstract

The invention relates to an application of a compound D-erythro-Sphingosine (DES) in a central nervous system. During the multiple sclerosis disease course, many inhibitory factors are generated in disintegrated axon myelin. These inhibitory factors inhibit differentiation and maturation and remyelination of newborn oligodendrocytes. It proves through results of compound screening that DES can relieve inhibition of myelin debris on differentiation and maturation of newborn oligodendrocytes and promote recovery of EAE demyelination mouse athletic ability. Thus, it prompts that the compound can be used for treating demyelination disease of the central nervous system.

Description

technical field [0001] The invention relates to the application of a compound sphingosine (D-erythro-Sphingosine, DES) in the central nervous system. Background technique [0002] In the pathogenesis of central nervous system demyelinating diseases such as multiple sclerosis, it is accompanied by the differentiation, maturation and remyelination of OPC cells. At present, the research on the molecular mechanism of how to promote the process of remyelination in patients with multiple sclerosis is a hot spot both at home and abroad. During the pathogenesis of multiple sclerosis, many inhibitory factors are produced in the disintegrated axon myelin sheath. These inhibitors inhibit maturation and remyelination of nascent oligodendrocytes. The use of antibodies antagonizing these inhibitors was found to induce partial recovery of function in experimentally demyelinated rats. But its downstream signal transduction pathway is unknown. Contents of the invention [0003] One of ...

Claims

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Application Information

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IPC IPC(8): C12N5/079A61K31/133A61P25/28
Inventor 徐晓辉栗娇娇王庆金
Owner SHANGHAI UNIV
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