Application of Sangsin N in the preparation of products for preventing and treating diseases related to ferroptosis
A technology of Sangxinsu and products, which is applied in the application field of products to achieve the effect of increasing the content level
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Embodiment 1
[0046] Example 1: Inhibitory Effect of Moranginin N on Neuronal Cell Death Induced by Ferroptosis Inducing Agents
[0047] 1. Using Erastin as a ferroptosis inducer
[0048] Erastin (CAS No: 571203-78-6) is one of the common inducers of ferroptosis, which directly inhibits
[0049] -Inhibits the activity of the cysteine / glutamate antiporter system (Xc), activates endoplasmic reticulum stress, thereby reducing glutathione content, thereby inducing cell ferroptosis.
[0050] The details of the test method are as follows: the samples are Sangshin N and Quercetin.
[0051] The HT22 cell line is a mouse hippocampal neuron cell line. HT22 cells were cultured in Dulbecco's Modified Eagle's medium (DMEM), which was added with 10% fetal bovine serum, 50 units / mL penicillin and 50 μg / mL streptomycin. cells at 37°C in carbon dioxide (5% CO 2 ) for cultivation. After three passages, ferroptosis induction experiments were performed. Cells were cultured in a 96-well plate with a cell ...
Embodiment 2
[0066] Example 2: Inhibition of Sangsin N on Neuronal Cell Death Induced by Glutamic Acid
[0067] HT22 cells were seeded in 96-well cell culture plates at a density of 3000 cells / well. After culturing at 37°C for 24 h, glutamic acid with a final concentration of 6 mM and different concentrations of the test compound (sanginin N or quercetin, with final concentrations of 0.25, 0.5, 1, 2, 5, and 10 μM) were added respectively. culture medium for incubation. After 12 hours, discard the old medium, add 110 μL of DMEM medium containing CKK8 solution (CKK8 solution: DMEM medium = 1:10), incubate at 37°C in the dark for 2 hours, and measure the absorbance at 450 nm with a microplate reader. Cell viability was calculated according to Equation 1. Wherein A control means the cells treated with only 0.5% DMSO medium; A sample means the cells treated with Sangsin N or quercetin or 0.5% DMSO and glutamic acid. The experiment was repeated three times, and the statistical results are sho...
Embodiment 3
[0071] Example 3: Effects of Sangsin N on GSH Levels in Nerve Cells
[0072] HT22 cells were treated with 4×10 4 Cells / well were inoculated in 12-well cell culture plates, and after 24 hours of attachment, different concentrations of Sangsin N (0, 0.25, 0.5, 1 μM) and ferroptosis inducers (0.4 μM erastin or 0.15 μM RSL3) or 6mM glutamate medium for treatment. After 12 hours of drug treatment, discard the drug-containing medium, rinse twice with phosphate buffered saline (PBS), add serum-free DMEM medium containing Monobromobimane (final concentration: 10 μM), incubate at 37°C for 25 minutes, collect cells, and wash with PBS After 3 passes, cells were resuspended in Hank's balanced salt solution and seeded into black 96-well plates. Fluorescent changes were detected by a multifunctional microplate reader (excitation wavelength was 380 nm, emission wavelength was 450 nm). The fluorescence levels of the control group were normalized and calculated as a percentage.
[0073] Th...
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