Application of chondroitin sulfate nano selenium in preparing medicine for treating nervous system diseases related to cognitive function decline
A technology for nervous system diseases and chondroitin sulfate, which is applied in the field of medicine, can solve the problems of high toxicity and poisoning of inorganic selenium, and achieve good protective effects, inhibition of aggregation, and low cytotoxicity
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Embodiment 1
[0022] The preparation of embodiment 1 chondroitin sulfate nano selenium
[0023] Take 100mmol Na 2 SeO 3 , 200mmol shark CS (Na 2 SeO 3 : CS molar ratio=1:2) was dissolved in 80ml deionized water, and 50mM L-cysteine (L-cys) solution (Na 2 SeO 3 :L-cys molar ratio=1:2), magnetic stirring at 30°C for 1.5h. After the MWCO 1000 dialysis bag was dialyzed against water for 48 hours (change the water every 2 hours), it was freeze-dried to obtain pink chondroitin sulfate nano-selenium.
Embodiment 2
[0024] Example 2 Effect of CS@Se on the structure of Aβ fibrils
[0025] Take 1mg Aβ 1-42 Dissolve in 2.216mL sterile water and filter through a 0.22μm microporous membrane to obtain a 100μM Aβ solution for later use. Take the Aβ solution and add the final concentration of CS@Se (25 μg / mL and 50 μg / mL), CS (48.55 μg / mL), nano-Se (1.45 μg / mL) solution, and the blank control group is 100 μM Aβ 1-42 . Then incubate the sample at 37°C for 3 days, take the sample for electron microscope observation experiment, drop the sample onto a 200-mesh Formvar-coated copper grid, dry at 25°C for 1min, and negatively stain with 2% phosphotungstic acid, Aβ 1～42 Fibril morphology JEM-1400 transmission electron microscope was used to observe the effect of CS@Se on the formation of Aβ fibril structure. The result is as figure 1 As shown, the inhibitory effect of CS@Se on the ultrastructure of Aβ fibers was observed by transmission electron microscope, Aβ 1-42 Samples incubated without CS@Se f...
Embodiment 3
[0026] Example 3 CS@Se to Aβ 1-42 inhibition of aggregation
[0027] To characterize the effect of CS@Se on Aβ 1-42 Inhibition of Aggregation Using a ThT Fluorescent Dye with Aβ 1-42 Fibrillar structure-specific binding characteristics examined for Aβ 1-42 polymerization kinetics. Aβ 1-42 The solution (100 μM) was incubated with CS@Se (25 μg / mL and 50 μg / mL), CS (48.55 μg / mL), nano-Se (1.45 μg / mL) at 37 °C in the dark for 72 h, at different time points ( 0h, 10h, 24h, 48h, 72h, 100h) take out 30μL Aβ 1-42 The solution was mixed with 400 uL ThT (15 μM) solution, incubated at room temperature in the dark for 30 min, and then the fluorescence intensity was measured by a fluorescence spectrophotometer. The excitation wavelength is 440nm, the emission wavelength is 480nm, and the slit width is 20nm. The result is as figure 2As shown, when Aβ 1-42 After incubation for 72h, Aβ 1-42 The kinetic growth curves of β-β were semi-S-shaped, which was consistent with the research ...
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