Polygala tenuifolia extract for parkinson's disease and preparation method and application of polygala tenuifolia extract
A Parkinson's disease and extract technology, which is applied in the field of anti-Parkinson's disease Polygala extract and its preparation, can solve problems such as unclear mechanism of action, inability to develop new drugs, and ineffective clinical efficacy of drugs, and achieve broad development and application prospects Effect
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Embodiment 1
[0018] Example 1 Preparation of Polygala extracts and parts
[0019] Weigh 2287g Polygala, cut it to 1-2cm, add 10 times the amount of 95% ethanol to soak overnight, heat and reflux for extraction 3 times, 2 hours each time, combine the extracts, filter, and recover the filtrate under reduced pressure until it has no alcohol smell, to obtain alcohol extract: add 10 times of water to soak the remaining medicinal residue after filtration overnight, heat and reflux for extraction 3 times, each time for 2 hours, combine the extracts, filter, concentrate under reduced pressure to liquid extract, and obtain water extract; The extract and water extract were mixed in equal volumes, 1.5 times the volume of petroleum ether with a boiling point of 60°C-90°C was added, liquid-liquid extraction was performed 3 times, the extracts were collected and combined, the solvent was recovered, concentrated to an extract, and vacuum-dried at 60°C. Pulverize to obtain part A; add 1.5 times the volume...
Embodiment 2
[0020] Example 2 Screening of hDDC Inhibitory Activity of Polygala Extracts and Parts
[0021] 1. Principle of hDDC and PEPC (phosphoenolpyruvate carboxylase) enzyme activity assay
[0022] In order to specifically detect the activity of hDDC, we used an ornithine decarboxylase (ODC) detection method to detect the activity of purified DDC. The principle is based on an enzyme-linked reaction ( figure 2 ), levodopa (L-Dopa) produces dopamine under the action of dopa decarboxylase and releases CO 2 , CO 2 with H 2 O forms HCO 3 — , HCO 3 — It reacts with phosphoenolpyruvate (PPPA) to generate oxaloacetate, and then under the action of malate dehydrogenase (MDH), the consumption of oxaloacetate and nicotinamide adenine dinucleotide (NADH) decreases, with NADH generates non-absorbed NAD as the reaction proceeds + , so the absorbance value at OD340nm decreases. CO produced by deacidification of hDDC 2 There is a 1:1 relationship between the amount of NADH and the amount ...
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