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Application of natural product small molecular alpha-arbutin (Arb) to preparation of medicine for preventing and controlling Parkinson's disease

A technology of natural products and small molecules, which is applied in the direction of medical preparations containing active ingredients, drug combinations, nervous system diseases, etc., to achieve the effect of reducing the level of active oxygen in cells and increasing the amount of active oxygen

Inactive Publication Date: 2018-08-03
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antioxidant small molecule (Arb), as a natural tyrosinase inhibitor, is widely added in cosmetics, but so far there is no report of its application in the treatment of Parkinson's disease

Method used

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  • Application of natural product small molecular alpha-arbutin (Arb) to preparation of medicine for preventing and controlling Parkinson's disease
  • Application of natural product small molecular alpha-arbutin (Arb) to preparation of medicine for preventing and controlling Parkinson's disease
  • Application of natural product small molecular alpha-arbutin (Arb) to preparation of medicine for preventing and controlling Parkinson's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Rotenone stimulated cells to establish a PD model at the cellular level and α-arbutin activity screening SH-SY5Y cells were seeded in a 96-well plate, and when the cells grew to about 70%, different concentrations of rotenone (0, 0.125, 0.25, 0.5, 1, and 2μM) to treat the cells for 12h and 24h. Then add XTT, put it in a 37°C incubator and incubate for 2h, measure the absorbance value at λmax=450nm with a microplate reader, and then calculate the cell survival rate.

[0021] After confirming the PD model, the cells were pretreated with different concentrations of α-arbutin (1, 10, 100μM) for 6h, and then stimulated with 0.25uM rotenone for 24h, and then added XTT to measure the absorbance value to calculate the cell survival rate.

Embodiment 2

[0023] α-Arbutin protects SH-SY5Y cells by interfering with intracellular biological indicators related to oxidative stress (reactive oxygen species ROS, mitochondrial membrane potential MMP, ATP, glutathione GSH)

[0024] 1. Determination of ROS level

[0025] Place SH-SY5Y cells in a 10mm glass-bottom culture dish. When the cells grow to about 70%, treat the cells with α-arbutin or rotenone. Then wash the cells with PBS three times, and change to the culture medium containing 10uM DCFH-DA probe. The medium was incubated in an incubator for 30 min, and then the cells were washed three times with PBS and imaged under a confocal microscope.

[0026] 2. MMP determination

[0027] Connect SH-SY5Y cells in a 6-well plate, and when the cells grow to about 70%, treat the cells with α-arbutin or rotenone. Then change the medium containing 10 μM DJ-1 probe into the incubator and incubate for 30 minutes, and then use Cells were washed three times with PBS and imaged on a multi-functi...

Embodiment 3

[0032] Example 3 Protein co-immunoprecipitation screening signaling pathway (AMPK and p62 expression)

[0033] The cells were placed in a 10 cm culture dish, and when the cells grew to about 70%, the cells were treated as above, washed with PBS three times, and collected by centrifugation. Then add RIPA lysate to lyse the cells on ice for 10min, centrifuge at 13000g, 4°C for 15min, and take the supernatant for protein content determination. The supernatant was added to the sample buffer and boiled for 10 min, and the sample was loaded with a controlled protein content of 40ug per well. After electrophoresis, membrane transfer, and blocking, primary antibodies (AMPK, P-AMPK, P62, and β-actin) were incubated overnight at 4°C, washed three times with TBST, 10 min each time, then incubated with secondary antibodies at room temperature for 1 hour, washed three times with TBST, each time After 10 minutes, the images were imaged on a gel chemiluminescence instrument, and grayscale a...

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Abstract

The invention provides application of a natural product small molecular alpha-arbutin (Arb) to preparation of a medicine for preventing and controlling Parkinson's disease. The research result shows that the compound can be used for reducing the oxidative stress level in a nerve cell system and protecting neuronal cells. Furthermore, influences, caused by the Arb, on AMPK (Adenosine MonophosphateActivated Protein Kinase) and downstream protein p62 thereof are inspected through protein detection, and finds out that the Arb protects the neuronal cells from being injured by oxidative stress through inhibiting AMPK activation and p62 expression for the first time, so that the effect of preventing and controlling the Parkinson's disease is realized. The Arb is a novel autophagy activating agent derived from a natural product and has a good market application prospect.

Description

technical field [0001] The invention relates to the medical application of Arb, an active ingredient of natural products, in particular to the application of Arb in preventing and treating Parkinson's disease, and belongs to the field of traditional Chinese medicine. Background technique [0002] Parkinson's disease (PD) is a degenerative disease of the central nervous system that commonly occurs in middle-aged and elderly people. At present, PD has become one of the main diseases that lead to the decline of the living standards of the elderly, and it is also a problem that needs to be overcome in the field of social medical and health care. The early intervention and treatment of PD has become a hot spot of social concern and is of great practical significance. It is also in line with the current national conditions of my country's aging population. [0003] The main pathological feature of PD is the progressive loss of dopamine (DA) neurons in the substantia nigra compact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7034A61P25/16
CPCA61P25/16A61K31/7034
Inventor 李林丁亚琦张承武辛陈琦倪赟张航白磊赵燕菲于海东刘金华王延宾黄维
Owner NANJING UNIV OF TECH
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