Use of composition in prevention or treatment of neurodegenerative diseases
A neurodegenerative and composition technology, applied in the field of prevention or treatment of neurodegenerative diseases, composition, can solve problems such as incurable, highly disabling, and incompletely clear pathogenesis
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Embodiment 1
[0100] Cell culture and PD cell model establishment
[0101] Experimental materials: Human neuroblastoma cells (SH-SY5Y) were purchased from the Kunming Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences.
[0102] Cell culture:
[0103] The medium was RPMI-1640 medium containing 10% fetal bovine serum, and the culture condition was 5% CO 2 , 37 ℃ constant temperature.
[0104] Modeling and administration methods:
[0105] SH-SY5Y cells were seeded in 96-well plates (1×10 4 per hole), placed in 5% CO 2 , and incubated in a 37°C incubator. After 4 hours of cell attachment, the medium was discarded and replaced with a serum-free medium containing (or not) 100 μmol / L lipoic acid (LA) or 20-50% w / w sacha inchi oil (SIO) in the incubator After incubation for 1 h, add with (or without) 2mmol / L MPP + Incubate in serum-free medium for 24 h.
[0106] MTT assay for cell viability:
[0107] SH-SY5Y cells were seeded in 96-well plates (1×10 4 pe...
Embodiment 2
[0119] Effects of different concentrations of lipoic acid and / or Sacha Inchi oil on cell viability:
[0120] SH-SY5Y cells were seeded in 96-well plates (1×10 4 per hole), placed in 5% CO 2 , and incubated in a 37°C incubator. After 4 hours of cell attachment, the medium was discarded, and (1) containing MPP at a concentration of 0, 1, 2, 5mmol / L was added + or (2) serum-free medium containing 0, 12.5, 25, 50, 100 μmol / L LA or 20-50% w / w SIO, incubated in an incubator 24h; or (3) serum-free medium containing (or not containing) 0, 12.5, 25, 50, 100 μmol / L LA or 20-50% w / w SIO, after incubation in the incubator for 1 hour, add containing (or not Including) 2mmol / L MPP + Incubate in serum-free medium for 24 h. Discard the old medium, add 100 μl of MTT solution containing 0.5 mg / mL to each well, and incubate in the incubator for more than 4 hours. Remove the culture medium, add 100 μl of DMSO to each well to dissolve the blue-purple crystals, place the 96-well plate at 37°C...
Embodiment 3
[0124] Effects of different concentrations of lipoic acid and / or Sacha Inchi oil on mitochondrial membrane potential (Δψ):
[0125] SH-SY5Y cells were seeded in 96-well plates (1×10 4 per hole), placed in 5% CO 2 , and incubated in a 37°C incubator. After 4 hours of cell attachment, the medium was discarded and replaced with a serum-free medium containing (or not containing) 100 μmol / L LA and 20% or 50% w / w SIO. After incubation in the incubator for 1 hour, adding containing (or Excluding) 2mmol / L MPP + Incubate in serum-free medium for 24 h. After the incubation, add JC-1 fluorescent staining solution with a concentration of 10 μg / mL, and incubate in the incubator for 1 h. Finally, the fluorescence value was detected in a full-wavelength multifunctional microplate reader (green light excitation wavelength 485nm, emission wavelength 538nm; red light excitation wavelength 485nm, emission wavelength 585nm), and the final mitochondrial membrane potential result was presented ...
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