Application of 3,4-methyl dihydroxybenzoate in preparation of drugs for inducing directional differentiation of neural stem cells or neural precursor cells

A technology of methyl dihydroxybenzoate and neural precursor cells, applied in the field of medicine, can solve problems such as no reports on the directional differentiation of neural stem cells/neural precursor cells, avoid false positive results, and have good cell purity and activity. , to ensure the effect of purity and activity

Active Publication Date: 2017-11-21
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, it is reported that it has antioxidant activity and promotes the growth of neurons, but there is no report that it induces the directed differentiation of neural stem cells / neural precursor cells

Method used

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  • Application of 3,4-methyl dihydroxybenzoate in preparation of drugs for inducing directional differentiation of neural stem cells or neural precursor cells
  • Application of 3,4-methyl dihydroxybenzoate in preparation of drugs for inducing directional differentiation of neural stem cells or neural precursor cells
  • Application of 3,4-methyl dihydroxybenzoate in preparation of drugs for inducing directional differentiation of neural stem cells or neural precursor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 The effect of MDHB on the morphological differentiation of neural stem / precursor cells

[0031] 1. Culture of rat hippocampal neural stem / precursor cells:

[0032] Take the required number of P0-2 newborn SD rats (Guangdong Animal Center), soak them in 75% (v / v) ethanol solution for 30s, cut the skin and skull at the head, take out the rat brain, and place the brain In the HBSS buffer solution, remove the rat hippocampus, remove the hippocampus meninges, move the hippocampus into a penicillin bottle containing 3ml HBSS solution, cut it to 1mm with ophthalmic scissors, and add 2ml 0.25%(w / v) to the bottle Trypsin + EDTA, gently pipet to mix, and put it in a cell culture incubator (37°C, 5% carbon dioxide, constant humidity) for 15 min. Use 10% (v / v) FBS+DMEM / F12 to terminate the digestion, gently pipette several times, then add 900U DNaseI (deoxyribonuclease I) to remove DNA, gently pipette several times to make the solution fully and DNaseI (deoxyribonuclease I) ...

Embodiment 2

[0037] Example 2 MDHB improves the differentiation of neural stem / precursor cells into neuron-specific antibody Tuj1 (axon-specific protein) and reduces the expression of glial cell-specific antibody GFAP (glial fibrillary acidic protein)

[0038] The 12mm cell slide (neural stem / precursor cell slide) was plated on a 24-well plate the night before, and 500 microliters of 0.0125g / ml PLL was added to the plate overnight. The next day, the PLL was aspirated and washed with 1×PBS for 2 minutes. Then, add 100 microliters of 10μg / ml lamnin (laminin) to each slide for 2h, inoculate the digested neural stem / precursor cells on the slide, inoculate 30,000 cells per well, use DMEM / F12+ Suspend the cells in 10% (v / v) FBS, then place them in a cell culture incubator and incubate for 60 minutes. The cells adhere to the wall, discard the complete medium, add differentiation medium, and add it in proportion (0μM, 8μM, 16μM and 32μM) MDHB, change the medium half the next day, aspirate the medium ...

Embodiment 3

[0039] Example 3 MDHB improves immunofluorescence staining of neuron-specific antibody MAP2 (microtubule-associated protein 2) during neural stem / precursor cell differentiation.

[0040] The 12mm cell slide was plated on a 24-well plate the night before, and 500 microliters of 0.0125g / ml PLL was added to the plate overnight. The next morning, the PLL was aspirated and washed twice with 1×PBS for 2 minutes each time. 100 microliters were added to the slide. Incubate 10μg / ml lamnin in an incubator for 2h, then inoculate the digested neural stem / precursor cells on the slide, inoculate 30,000 cells per well, and suspend the cells in DMEM / F12+10% FBS and put them into the cells Incubate in an incubator for 60 minutes, the cells adhere to the wall, discard the complete medium, add differentiation medium, and add MDHB in proportions (0 μM, 8 μM, 16 μM, and 32 μM), change the medium half the next day, aspirate the medium after 7 days, and add Wash 2 times with pre-cooled 1×PBS, 3 min eac...

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Abstract

The invention discloses application of 3,4-methyl dihydroxybenzoate (MDHB) in preparation of drugs for inducing directional differentiation of neural stem cells or neural precursor cells. The invention discovers that MDHB has the effect of directionally inducing differentiation of neural stem cells or neural precursor cells to cholinergic neurons, can be used for developing drugs for curing related diseases (SCI, AD and the like) of cholinergic neuron damage and loss, can be used for cell transplantation therapy of related diseases (SCI, AD and the like) of cholinergic neuron damage and loss, and can be used for developing directional inducers and the like of neural stem cells or neural precursor cells. The new pharmacological activity of MDHB is determined, the illumination of a pesticide effect mechanism provides reference for developing new drugs with proprietary intellectual property rights, MDHB is applied to basic experiments in the field of neurosciences, and a novel method is provided for searching models of cholinergic neurons.

Description

Technical field [0001] The invention belongs to the field of medicine, and particularly relates to the application of methyl 3,4-dihydroxybenzoate in the preparation of drugs for inducing the directional differentiation of neural stem cells / neural precursor cells. Background technique [0002] Cholinergic neurons are widely distributed in the central nervous system, such as spinal anterior horn motor neurons, including the collateral branches that burst out to runshau cells, and specific sensory projection neurons on the posterior ventral side of the thalamus are all cholinergic Cholinergic neurons, various links of the ascending activation system of the brainstem network structure, striatum, forebrain basal nucleus, the piriform area of ​​the limbic system, amygdala, hippocampus and other parts all contain cholinergic neurons. The injury and death of cholinergic neurons in different parts leads to a variety of important neurological diseases, such as spinal cord injury (SCI), Al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0793A61K35/30A61P25/00A61P25/28
CPCA61K35/30C12N5/0619C12N2501/999
Inventor 罗焕敏潘君平信怡榕胡阳米象男王嘉惠高勤
Owner JINAN UNIVERSITY
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