Use of ginsenoside M1 for treating huntington's disease
A technology of ginsenoside and chorea, applied in the field of treating Huntington's disease
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Embodiment 1
[0050] Example 1: Ginsenoside M1 significantly reduces cytotoxicity in mHtt expressing cells
[0051] Conditionally immortalized wild-type STHdh Q7 Striatal neuronal precursors expressing endogenous normal htt (termed wild-type striatal cells), homozygous STHdh from knock-in mice Q109 The homozygous mutant STHdh Q109 Striatal neuronal precursor cells expressing mutant htt with 109 glutamic acids (termed mutant striatal cells). wxya Q7 and STHdh Q109 Cells were a generous donation of Dr. Elena Cattaneo and Marta Valenza (Department of Pharmacology and Center for Stem Cell Research, University of Milan, Italy). These cells were maintained in a culture room at 33°C in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS).
[0052] STHdh were treated with 30 μM ginsenoside M1 or vehicle (DMSO) Q7 and STHdh Q109 Cells for 24 hours. Quantification of STHdh by MTT reduction assay Q7 and STHdh Q109 cell death. For the MTT reduction assay, 3-...
Embodiment 2
[0053] Example 2: Ginsenoside M1 significantly reduces STHdh Q109 Reactive Oxygen Species (ROS) Production in Cells
[0054] Oxidative stress is what triggers STHdh Q109 One of the important events of cell death in cells. To investigate whether ginsenoside M1 inhibits STHdh by reducing oxidative stress Q109 Cell death of cells, intracellular ROS indicator fluorescent probe 2,7-dichlorodihydrofluorescein diacetate (H 2 In the presence of DCFDA; Molecular Probes, Inc., Eugene, Oregon, USA), STHdh was treated with 1, 10 and 30 μM ginsenoside M1 or vehicle (DMSO) Q109 Cells for 1 hr. Cellular ROS content was measured by detecting fluorescence intensity (excitation / emission: 488nm / 510nm) using a fluorescent plate reader (Fluoroskan Ascent; Thermo Electron Corporation, Woburn, MA, USA). Such as figure 2 As shown, 10 and 30 μM ginsenoside M1 significantly reduced ROS production. Results are expressed as mean ± SEM of triplicate samples. Groups will be analyzed using one-way ...
Embodiment 3
[0055] Example 3: Ginsenoside M1 significantly reduces STHdh Q109 Phosphorylation of AMPK in cells
[0056] Activation of AMPK contributes to the pathogenesis of HD [37]. To investigate whether ginsenoside M1 inhibits AMPK activation, STHdh were treated with 10 and 30 μM ginsenoside M1 or vehicle (DMSO) Q7 and STHdh Q109 Cells for 24 hours. The degree of phosphorylation of AMPK was measured by Western blotting. Such as image 3 As shown, compared with untreated (vehicle-treated) STHdh Q7 Cells compared to untreated (vehicle-treated) STHdh Q109 The degree of phosphorylation of AMPK in cells was significantly increased. Notably, compared with untreated (vehicle-treated) STHdh Q109 10 and 30 μM ginsenoside M1 significantly decreased STHdh compared to cells Q109 Phosphorylation of AMPK in cells. Western blot results represent three different experiments and histograms show quantification expressed as the mean ± SEM of these three experiments. Groups will be analyzed usi...
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