Application of Alpha-mangostin in preparing medicines for treating Alzheimer's disease
A technology of Alzheimer's disease and mangostin, applied in the field of pharmacology and chemical biology, to achieve the effect of improving pathological symptoms, enhancing normal physiological functions, and improving intervention
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Embodiment 1
[0029] Example 1 Computer Simulation of the Binding Model of α-Mangostin and Aβ
[0030] Import Aβ from the protein structure database PDB 1-40The nuclear magnetic resonance NMR structure (PDB: 1BA4) of the molecular simulation software MOE. After removing water molecules and other heteromolecules added to obtain nuclear magnetic resonance experiments, and filling in the missing hydrogen atoms in nuclear magnetic resonance experiments, the Aβ is established by using the charge balance program and energy optimization program in the MOE software 1-40 3D simulation structure.
[0031] The two-dimensional simulation structure of α-mangostin was established in the molecular simulation software MOE, and the three-dimensional simulation structure of the molecule was established by using the charge balance program and energy optimization program in the MOE software.
[0032] remove Aβ 1-40 There is no specific conformational region (positions 1 to 13) in the three-dimensional simul...
Embodiment 2
[0034] Example 2 Determination of α-Mangostin Inhibition of Aβ Aggregation and Deposition by Fluorescence Kinetic Method
[0035] Dissolve 1 mg of Aβ in 500 microliters of hexafluoroisopropanol, place it at room temperature for 120 minutes, shake intermittently, dry the hexafluoroisopropanol gently with high-purity nitrogen, and then add 100 microliters of dimethyl Sulfoxide was prepared into a 2.3 mmol / L Aβ stock solution and stored at -20°C. After diluting the Aβ stock solution with dimethyl sulfoxide, take 2 microliters and add it to 16 microliters of 0.215 mol / liter sodium phosphate buffer solution with a pH value of 8.0, so that the concentration of Aβ in the final 100 microliters system is 25 μmol / L, add 2 μl of 25 μmol / L α-mangostin or other inhibitors or corresponding blank solvent, incubate for 30 minutes, add 80 μl of 50 mmol / L Thioflavin T Glycine-sodium hydroxide solution at pH 8.5 per liter was transferred to a fluorescence detection microwell plate, and placed o...
Embodiment 3
[0037] Example 3 ELISA Determination of α-Mangostin Depleting Aβ Oligomers
[0038] Dissolve 1 mg of Aβ in 500 μl of hexafluoroisopropanol and let stand at room temperature. Transfer 100 microliters of the solution to a clean 1.5 milliliter centrifuge tube, add 900 microliters of sterilized deionized water, and let stand at room temperature. After centrifugation, transfer the supernatant to another clean 1.5 ml centrifuge tube, and gently dry the hexafluoroisopropanol with high-purity nitrogen. Then add a micro-stirring bar to the solution, stir and incubate at 22 degrees Celsius and 500 rpm for 48 hours, and obtain a certain volume of Aβ oligomers diluted to 1 micromol / liter for the obtained product, wherein the final concentration of 5 micromol / liter is added. mol / L, 2 micromol / L, 1 micromol / L, 0.5 micromol / L and 0.2 micromol / L of α-mangostin, continue to shake and incubate at 22 degrees Celsius and 500 rpm for 12 hours. Or add α-mangostin at final concentrations of 5 micr...
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