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Application of astragaloside in preparation of drug for promoting neural stem cell regeneration

A technology of neural stem cells and astragalus saponins, applied in the field of medicine, can solve problems such as low proliferation and differentiation

Inactive Publication Date: 2018-11-02
陈曦
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Unfortunately, these activated endogenous neural stem cells are in a state of low-level proliferation and differentiation, and the vast majority of neural stem cells stop proliferation and differentiation due to apoptosis. Therefore, it is very urgent to obtain endogenous neural stem cells that can expand Proliferation of Stem Cells and Drugs to Promote Differentiation of Neural Stem Cells

Method used

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  • Application of astragaloside in preparation of drug for promoting neural stem cell regeneration
  • Application of astragaloside in preparation of drug for promoting neural stem cell regeneration
  • Application of astragaloside in preparation of drug for promoting neural stem cell regeneration

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Experimental program
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Effect test

Embodiment 1

[0085] The astragaloside VI monomer is extracted from the traditional Chinese medicine Astragalus membranaceus, and a white flocculent powder is obtained. Further spectral verification was performed on the extracted components. Among them, the spectrum of ESI / MS (electrospray ionization mass spectrometry, electrospray ionization-mass spectrometry) analysis is as follows figure 1 As shown, the C spectrum is as figure 2 As shown, the H spectrum is as image 3 shown. The chromatogram of HPLC-LESD (high performance liquid chromatography) analysis is as Figure 4 as shown, Figure 4 The results are shown in Table 1 below.

[0086] Table 1: HPLC-LESD analysis results

[0087]

[0088] The above results show that, after verification, the extracted component is indeed astragaloside VI. The chemical molecular formula of this astragaloside VI is C 47 h 78 o 19 , the molecular weight is 946, and the structural formula is shown in formula (6).

Embodiment 2

[0090] Primary culture of neural stem cells C17.2

[0091] Female rats with 14 days to 15 days of pregnancy were taken, sacrificed under anesthesia, opened the cranial cavity under aseptic conditions, and took out the brain, placed in a petri dish of ice-cold PBS solution, and washed several times with PBS solution. The blood vessels and meninges were carefully peeled off under the microscope, the lateral walls of the subventricular zone on both sides were cut off, the tissues were cut into pieces, and centrifuged for 5 minutes. Discard the supernatant, digest the precipitate and enzyme mixture in a water bath at 37°C for 10 minutes, add an equal amount of trypsin inhibitor, gently pipette with a 1mL pipette to avoid the formation of air bubbles, and then centrifuge at 110g for 7 minutes, discard the supernatant, and add the precipitate to DMEM / F12 Mix well after blowing, slowly settle and centrifuge, then slowly roll over a 100-mesh sieve, collect the sieved cell suspension, ...

Embodiment 3

[0093] Astragaloside VI promotes the proliferation of neural stem cells C17.2 cultured in vitro

[0094] Firstly, neurosphere assay (Neurosphere assay) was used to measure the self-renewal ability of neural stem cells: as follows, the neurospheres within 2 to 5 passages were blown into a single cell suspension, and seeded in a 96-well plate at a density of 500 cells per well. In the medium, use the proliferation medium that adds growth factor to culture, add the astragaloside VI (HQ22) of 5nmol / L, 10nmol / L, 20nmol / L, 100nmol / L respectively, the control group (ctrl) does not add. The BrdU (cell proliferation marker) incorporation method was used to determine the proliferation ability of neural stem cells. After 7 days of treatment, the number of neurospheres formed under different treatments and the average diameter of the neurospheres were counted under a microscope. The results of different dosing concentrations by BrdU fluorescent staining are as follows: Figure 5 shown. ...

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Abstract

The invention relates to application of astragaloside in preparation of drug for promoting neural stem cell regeneration. Research shows that astragaloside is capable of effectively promoting self-renewing and multiplication of neural stem cells, and promoting neural stem cell in mouse brain subjected to ischemia reperfusion injury to produce new neuron cells, astrocyte cells or synaptic glial cells, and the like. Therefore, the regeneration capacity of the neural stem cell can be effectively recovered.

Description

technical field [0001] The invention relates to the field of medicines, in particular to the application of astragaloside in the preparation of medicines for promoting the regeneration of neural stem cells. Background technique [0002] Stroke is the general term for acute cerebrovascular diseases in traditional Chinese medicine. Because of the sudden onset, multiple symptoms, and rapid changes in the condition, it is similar to the characteristics of wind's good deeds, so it is called stroke and apoplexy. Stroke is a type of cerebral blood circulation disorder disease, often accompanied by sudden fainting, unconsciousness, accompanied by crooked mouth corners, poor speech and other main symptoms such as hemiplegia. Due to the characteristics of high morbidity, high mortality, high disability rate, high recurrence rate and many complications of stroke, the medical profession listed it as one of the three major diseases threatening human health along with coronary heart disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61K38/17A61P25/00
CPCA61K31/7048A61K38/179A61K2300/00
Inventor 陈曦沈剑刚
Owner 陈曦
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