Application of calycosin in preparation of ALDH2 agonist medicines
A technology of calycosin and agonist, which is applied in the field of medicine, can solve the problems of unreported ALDH2 relationship, etc., and achieve the effect of improving the viability of cardiomyocytes and lowering the preparation cost
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Embodiment 1
[0028] Example 1 Calycosin inhibits alcohol-induced decline in cardiomyocyte viability
[0029] First use the MTS method to explore the effect of different doses of calycosin on the activity of cardiomyocytes, in order to choose the appropriate dose.
[0030] Adjust cell density to 1.5 x 10 5 / mL, 100 μL per well, 5 replicate wells per well. After culturing for 24 hours and waiting for the cells to adhere to the wall, 10% FBS was replaced with DMED to continue culturing for 24 hours, and the cells were observed to be in good shape. Different concentrations of calycosin (2.5, 5, 10, 20 μM) were added to stimulate the cells for 12 hours. Then the culture medium was removed, and 100 μL of MTS-containing working solution (0% FBS in DMEM:MTS=100:20) was added to each well. The MTS working solution should be protected from light and should be prepared and used immediately. After incubating in a 37°C incubator in the dark for 2 hours, the absorbance value was measured at a wavelen...
Embodiment 2
[0037] Example 2 Calycosin increases the activity of ALDH2.
[0038] In order to investigate the effect of calycosin on the activity of ALDH2, the inventors conducted intracellular and extracellular activity experiments.
[0039] Intracellular experiment: Treat cardiomyocytes with different concentrations of calycosin (2.5 μM, 5 μM, 10 μM) for 12 hours, and set up a blank control group, and then use the tissue acetaldehyde dehydrogenase 2 activity photometric quantitative detection kit (GENMED ) to detect ALDH2 activity and use Western Blot to detect protein expression (cell inoculation and Western Blot methods are the same as Example 2).
[0040] The results of Western Blot showed ( figure 2 A), there was no statistically significant difference between the groups (P=0.9567) ( figure 2 B). The results of the activity experiment showed that there were significant differences in the activity of ALDH2 between different groups. Compared with the blank control group, the acti...
Embodiment 3
[0043] Example 3 Molecular dynamics simulation to explore the interaction of ALDH2_Calycosin
[0044] To explore the ALDH2_Calycosin interaction, we performed molecular dynamics simulations, obtained the ALDH2 protein structure from the website https: / / www.rcsb.org / , and obtained the molecular structure of Calycosin from the website https: / / tcmsp-e.com / , Virtual docking of ALDH2 and Calycosin was performed using YASARA with default options.
[0045] The surface visualization model of the ALDH2_Calycosin complex at 0ns and 60ns as image 3 As shown in A and B. It can be seen from the results that calycosin was stably combined with ALDH2 until the end of MD. image 3 C shows the RMS evolution of heavy atoms. The RMSD of ALDH2 rises at 0ns Gradually ramp up to 20ns after the stable Fluctuates nearby, and the RMSD of the ligand calycosin fluctuates between Nearby, it can be seen that the association of calycosin and ALDH2 is stable.
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