Application of teprenone in resisting neurotoxicity caused by methamphetamine
A technology of methamphetamine and teprenone, which is applied in the field of medicine to achieve the effect of high safety, significant curative effect and low cost
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Embodiment 1
[0020] Example 1: Determination of the protective effect of teprenone on methamphetamine-stimulated PC12 cells by tetrathiazolium blue (MTT) method
[0021] 1. Culture of PC12 cell line
[0022] PC12 cells were cultured in RPMI1640 medium containing 10% horse serum, 5% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin at 37°C, 5% CO 2 and 95% humidity for adherent culture. Replace the medium every 1-2 days, and when the cells reach about 70%-80% confluence, passage or use for experiments.
[0023] 2. Arrangement of PC12 cells
[0024] The adherent PC12 cells were digested with 0.25% trypsin and centrifuged at 1500rpm for 5min to remove trypsin. Cells were evenly arranged in a 96-well plate with a density of 1×10 4 Cell / well, in RPMI1640 medium containing 10% horse serum, 5% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin, at 37°C, 5% CO 2 and 95% humidity for adherent culture.
[0025] 3. Stimulate PC12 cells in groups
[0026] The ce...
Embodiment 2
[0034] Example 2: Western Blot detection teprenone restores methamphetamine-induced reduction of pro-caspase3 protein
[0035] 1. Cell line culture and drug stimulation
[0036] PC12 cells were cultured in 1640 medium containing 10% horse serum, 5% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin at 37°C, 5% CO 2 Adhesive culture is carried out at 95% humidity, and the culture medium is replaced every 2-3 days. When the cells reach about 70%-80% confluency, they are passaged or used for experiments.
[0037] The cells were seeded in six-well plates, and after overnight culture, the cells were treated with drugs. The groups were treated as follows: the control group did not receive drugs, and the experimental group received: (1) 2 mM methamphetamine; (2) 10 μM teprenone pre-stimulated 4 hours in advance, and then added 2 mM methamphetamine; ( 3) 10 μM teprenone alone was pre-stimulated 4 hours in advance. After the above treatments, place at 37 °C and 5% CO...
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