Application of silymarin in preparation of medicine for preventing and/or treating Parkinson's disease
A technology for silymarin and Parkinson's disease, which is applied to the application field of silymarin in the preparation of medicines for preventing and/or treating Parkinson's disease, and can solve the problem that the prevention and control effect has not yet been reported.
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Embodiment 1
[0023] Example 1 Silymarin inhibits the killing effect of rotenone on human SH-SY5Y cells
[0024] Milk thistle (purchased from Chengdu Ruifensi Co., Ltd.), when used, is prepared into a mother solution with DMSO, and administered after dilution.
[0025] Human SH-SY5Y cells (purchased from China Jikai Biotechnology Co., Ltd.), and rotenone (purchased from Sigma Company) were used to make a cell model of "neuron damage caused by Parkinson's disease".
[0026] Human SH-SY5Y cells were inoculated in DMEM medium in 96-well plates, incubated overnight in a carbon dioxide incubator, and incubated with different concentrations of silymarin (2.0mmol / L and 4.0mmol / L) 1 hour in advance, and then After adding rotenone (30 μmol / L), 8 hours later, the cell death rate was detected by LDH (lactate dehydrogenase) release method.
[0027] Experimental results such as figure 1 shown. The results show that: silymarin alone has no killing effect on human SH-SY5Y cells, and rotenone alone has ...
Embodiment 2
[0033] Example 2 Silymarin inhibits DNA double-strand breaks in human SH-SY5Y cells caused by rotenone
[0034] DNA is the carrier of genetic information, and DNA double-strand breaks are the basis for DNA dissolution and cell death. In order to illustrate the protective effect of silymarin on cells, this experiment detected the effect of silymarin on DNA double-strand breaks induced by rotenone.
[0035] (1) Inoculate human SH-SY5Y cells in DMEM medium in a 6cm-diameter petri dish, in a carbon dioxide incubator overnight, and incubate with silymarin (2.0mmol / L and 4.0mmol / L) 1 hour in advance, then add Rotenone (30 μmol / L), the cells were collected 8 hours later. The collected cells were homogenized, differentially centrifuged, cytoplasmic components, mitochondrial components and nuclear components were separated, the protein concentration of each component was determined by BCA method, and Western blotting analysis was performed. The primary antibody used is the marker pro...
Embodiment 3
[0041] Example 3 Silymarin inhibits the endoplasmic reticulum stress response caused by rotenone
[0042] Endoplasmic reticulum stress is one of the important pathological bases leading to Parkinson's disease. Excessive endoplasmic reticulum stress can induce nuclear translocation of CHOP, leading to cell death.
[0043] In order to illustrate the protective effect of silymarin on cells, this experiment detected the effect of silymarin on the endoplasmic reticulum stress response induced by rotenone by Western blot analysis. Human SH-SY5Y cells were inoculated in DMEM medium in a 6cm-diameter petri dish, incubated overnight in a carbon dioxide incubator, and incubated with different concentrations of silymarin (2.0mmol / L and 4.0mmol / L) 1 hour in advance, and then Rotenone (30 μmol / L) was added, and the cells were collected 8 hours later. The collected cells were homogenized, centrifuged at a differential speed, and the cytoplasmic fraction and the nuclear fraction were separa...
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