New application of phenylethanol glycoside compound torenoside B
A technology of use and medicine, applied in the field of medicine, can solve the problems such as the efficacy of treating or preventing AD that have not been documented in the literature, and achieve the effect of huge potential value and broad application prospects.
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Embodiment 1
[0026] Example 1: Extraction and separation of Torenoside B.
[0027] (1) Extraction of medicinal materials:
[0028] 50kg of velvet antler medicinal material, cut into small pieces of 1~2cm, heated and refluxed with 15 times the amount of water to extract three times, each time for 2 hours. Combine the extracts, concentrate under reduced pressure at 70°C to 50L (relative density is 1.15~1.18), add 250L of 95% ethanol, stir while adding, let stand overnight, take the supernatant, and concentrate under reduced pressure at 70°C to 50L.
[0029] (2) AB-8 resin purification:
[0030] Resin dosage: medicinal material quality / resin quality=1:1;
[0031] Sample loading speed: 1mL / min.cm 2 ;
[0032] Elution rate: 2~3mL / min.cm 2 ;
[0033] The diameter-to-height ratio of the resin bed is 1:10;
[0034] Take 50 kg of the processed AB-8 resin, put it into a stainless steel chromatographic column with a diameter of 30 cm, wash it with water, and set it aside. The extract was load...
Embodiment 2
[0064] Example 2: The effect of Torenoside B on the survival rate of SH-SY5Y nerve cells.
[0065] Adjust the concentration of SH-SY5Y cells to 1×10 5 cells / ml, seed the cell suspension in a 96-well plate, 100 μl per well, at 37°C, 5% CO 2 After culturing in the incubator for 24 hours, the experiments were performed in random groups.
[0066] The experiment was divided into normal cell control group, L-glutamate damage group and different concentrations of Torenoside B groups (50μM, 100μM, 200μM). 24h after the test drug was added to the cells, the model was established with L-glutamic acid (10mM). After modeling for 24 hours, add 10 μl MTT (5 mg / ml) to each well and incubate for 4 hours, pour off the supernatant, add 100 μl DMSO, and shake for 10 minutes. The absorbance value (OD) at 570 nm was measured with an automatic microplate reader 570nm ), used to quantify cell viability. Experiments were repeated three times. The cell survival rate was calculated with reference...
Embodiment 3
[0068] Example 3. The effect of Torenoside B on the morphology of SH-SY5Y nerve cells.
[0069] Depend on image 3 It can be seen that the SH-SY5Y cells in the normal cell control group (A) are larger in size and have strong cell adhesion ability; in the glutamic acid (10mM) injury group (B), the growth of SH-SY5Y cells is obviously inhibited under the microscope , the refractive index decreased, and some cells were lysed into fragments; the cell morphology in the groups (C and D) given Torenoside B (100, 200 μM) was obviously restored, and the number of cells increased. The above results indicated that different concentrations of Torenoside B could effectively resist the morphological changes of SH-SY5Y cells induced by glutamate.
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