Preparation method of active substance capable of resisting neurodegenerative disease
A neurodegenerative, active substance technology, applied in neurological diseases, microorganism-based methods, biochemical equipment and methods, etc., can solve problems such as irreversible disease, adverse reactions to acetylcholine, etc.
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Embodiment 2
[0031] Example 2 The preparation method of the anti-neurodegenerative disease active substance of Phomopsis cryptifera according to the present invention
[0032] (1) The fermented liquid obtained in Example 1 was vacuum-filtered with 9 layers of gauze to collect the bacteria;
[0033] (2) The bacteria were dried under reduced pressure and low temperature in a constant temperature vacuum drying oven at 40°C for 24 hours;
[0034] (3) After drying, the cells were soaked in 3L methanol, ultrasonicated for 15 minutes, soaked for 3 days, repeated 3 times, filtered to combine the extracts, and distilled methanol under reduced pressure at 45°C to obtain an aqueous solution of the cell extracts;
[0035] (4) Add twice the volume of pure water to the aqueous solution of the bacterial cell extract, then extract with 4 / 3 of the total volume of n-butanol, repeat 4 times, wash the n-butanol part with water once, and combine the water phases, 45 Concentrate under reduced pressure at ℃ to ...
Embodiment 3
[0037] Example 3 Screening research on in vitro Aβ42 aggregation model for hiding the anti-neurodegenerative disease activity of Phomopsis sp. according to the present invention:
[0038]ThT analysis uses synthetic Aβ42 polypeptide, 37 ° C for 2 hours, add each sample with a final concentration of 100 μg / mL, use a fluorescent microplate reader to test the fluorescence intensity of the reaction system, the excitation wavelength is 444nm, and the emission wavelength is 485nm. Using DMSO as a negative control and EGCG as a positive control, the inhibitory activity of the components on A-beta aggregation can be judged according to the difference in the fluorescence intensity of the system. For experimental results, see figure 1 . The experimental results show that the relative fluorescence intensity of many samples is low, and they have a significant inhibitory effect on A-beta42 aggregation, and the inhibitory effect of some samples is stronger than that of the positive control ...
Embodiment 4
[0039] Example 4 The in vitro Poly Q aggregation model screening research on the hidden anti-neurodegenerative disease activity of Phomopsis sp. according to the present invention:
[0040] Purified Poly Q protein was used for ThT analysis, let stand at 37°C for 2 hours, add each sample at a final concentration of 100 μg / mL, and test the fluorescence intensity of the reaction system with a fluorescent microplate reader, the excitation wavelength is 444nm, and the emission wavelength is 485nm. Using DMSO as the negative control and EGCG as the positive control, the inhibitory activity of the components on Poly Q aggregation can be judged according to the difference in the fluorescence intensity of the system. For experimental results, see figure 2 . The experimental results show that the relative fluorescence intensity of many samples is low, which has a significant inhibitory effect on Poly Q aggregation, and the inhibitory effect of some samples is close to that of the posi...
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