Application of plantamajoside in preparing medicines for treating Parkinson's disease
A technology of psyllium and Parkinson's disease, applied in the field of medical use
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experiment example 1
[0039] In vitro inhibitory activity of psyllogen on monoamine oxidase B (MAO-B):
[0040] 1. Experimental principle:
[0041] MAO-B catalyzes the substrate kynuramine to generate 4-hydroxyquinoline under suitable reaction conditions, and 4-hydroxyquinoline produces 400nm emission light under the excitation light of 310nm. By detecting the change of the fluorescence intensity at 310 / 400nm, the change of the production amount of the product was detected, and then the inhibitory effect of the inhibitor on MAO-B was calculated.
[0042] 2. Experimental materials:
[0043] Test article: great psyllidin;
[0044] Positive control: safinamide;
[0045] Enzyme source: purchased commercial MAO-B, stored at -20°C. Dilute it to the working solution concentration (9U / mL) with 12.5mM phosphate buffer solution (pH 7.4) immediately before use.
[0046] Test article and positive control treatment: dissolve in 12.5mM phosphate buffer solution (pH 7.4) containing 10% dimethyl sulfoxide, th...
experiment example 2
[0063] In vitro agonist activity of psyllogen on PINK1 protein kinase:
[0064] 1. Experimental principle
[0065] Ubiquitin is an effective substrate of PINK1. Under the activation of PINK1, ubiquitin will use ATP to phosphorylate and generate ADP. After ADP is derivatized with fluorescence, the amount of ADP generated can be detected by chemiluminescence, representing PINK1 For the activation of ubiquitin, according to the enzyme reaction rate method, the ADP generation rate can reflect the activity of PINK1.
[0066] 2. Experimental materials
[0067] Test article: great psyllidin;
[0068] Buffer: 40mM Tris-HCl (pH 7.5), 20mM MgCl 2 , 0.1mg / mL BSA;
[0069] Enzyme and ATP treatment: purchase, store at -80°C. Dilute to 0.25 μM before use.
[0070] Substrate: Construct a plasmid according to the ubiquitin nucleotide sequence queried on Genbank, transfect it into Escherichia coli BL21 for expression, purify it with Ni-IDA, store it at -80°C, and dilute it to 0.25 μM bef...
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