Use of coniferaldehyde or coniferaldehyde and vanadium compound composition for preventing and treating neurodegenerative diseases
A neurodegenerative disease, vanadium compound technology, applied in the field of medicine, can solve problems such as drug failure, achieve the effect of protecting nerve cells, restoring cell mitochondrial function, and inhibiting hyperphosphorylation of Tau protein
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Embodiment 1
[0048] Preparation of coniferaldehyde (4H3M) and coniferaldehyde (4H3M) combined with vanadyl acetylacetonate (VAC) compound (4H3M+VAC) composition preparation
[0049] Weigh 17.18mg (0.1mmol) coniferaldehyde (4H3M), dissolve in 10mL DMSO, and configure coniferaldehyde (4H3M) mother solution with a concentration of 10mM; take 100 μL coniferaldehyde (4H3M) mother solution (10mM) and dilute it in 9.9mL of DMEM (containing 10% serum) prepared 200 μM coniferaldehyde (4H3M) solution; then diluted 200 μM coniferaldehyde (4H3M) solution to 100 μM, 50 μM, 10 μM, 5 μM, 1 μM, 0.5 μM, 0.1 μM to obtain different concentrations Coniferaldehyde (4H3M) solution.
[0050] Weigh 26.51mg (0.1mmol) vanadyl acetylacetonate (VAC), take 10mL of distilled water to dissolve, configure the concentration of 10mM vanadyl acetylacetonate (VAC) mother solution; take 10 μL of vanadyl acetylacetonate (VAC) mother solution (10mM) Diluted in 9.99mL of DMEM (containing 10% serum) to prepare 10 μM vanadyl acet...
Embodiment 2
[0052] MTS assay to detect cell viability
[0053] Cell pre-cultivation and passage: cells were placed at 37°C, 5% CO 2 cultured under the conditions of DMEM (containing 10% fetal bovine serum), 100 μM G418, 100 U / ml penicillin and 100 μg / ml streptomycin. (1) Preheat medium, trypsin and PBS in a 37°C water bath; (2) Discard the old medium in the culture bottle, take 2ml PBS and add 25cm 3 (3) Add 1 mL of 0.25% trypsin to digest the cells, and observe under the microscope that the cell shape changes slightly, then pour out the trypsin in a clean bench Take about 100 μL, and place the cells in a 37°C cell culture incubator and incubate for 3 minutes until the digestion is complete; (4) Stop the digestion with 2 mL of cell culture medium, blow the wall of the culture bottle, transfer the cells into a centrifuge tube, and centrifuge at 1000 rpm for 3 minutes; ( 5) Discard the supernatant after centrifugation, add 1mL of fresh DMEM (containing 10% fetal bovine serum) to the cultu...
Embodiment 3
[0058] MitoTracker Mitochondrial Selection Probe Method to Detect Mitochondrial Morphology
[0059] Red CMXRos is a cell-permeable X-rosamine derivative containing a weakly sulfhydryl-reactive chloromethyl functional group that labels mitochondria. It is an oxidized red fluorescent dye (Ex=579nm, Em=599nm), which can be passively transported through the cell membrane and directly aggregated on the active mitochondria after simply incubating cells, and can be used to observe the shape of mitochondria. (1) According to Example 2, SH-SY5Y Neo, APPwt and APPswe cells were respectively spread on 35mm 2(2) After the cells are about 40-50% confluent, add a solution of 100 μM coniferaldehyde (4H3M) for 36 hours; (3) Remove the old medium and add serum-free medium containing 50 nM MitoTracker Red CMXRos based on 37 ° C Incubate in an incubator for 10 minutes; (3) After 10 minutes, remove the original medium, add serum-free medium and shake gently, and wash three times; (4) Confocal ...
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