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
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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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