Medicine for preventing and treating alzheimer's disease and preparative method thereof
A technology for Alzheimer's disease and medicine, which is applied in the field of medicines for preventing and treating Alzheimer's disease and their preparation, and can solve the problem of increasing coronary flow, anti-virus, separation and identification of no active components, and reports of pharmacological effects. and other problems, to achieve the effect of inhibiting Aβ aggregation and toxicity, reducing memory damage, and easy access
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Embodiment 1
[0022] Example 1: Isolation and identification of RNFG
[0023] Panax notoginseng was collected in Wenshan, Yunnan Province, and dried naturally. After the crude drug identification, the sample certificate was preserved in the Department of Biology, Hong Kong University of Science and Technology. American ginseng and Korean ginseng were purchased from the Hong Kong market.
[0024] 1 kg of the original medicinal material Radix Notoginseng was pulverized, passed through a 200-mesh sieve, and extracted with 70% ethanol (twice, 3 liters each). The ethanol solution was concentrated in vacuo, the concentrate was heated and dissolved with 500 ml of water, centrifuged for 15 min, the supernatant was taken, passed through a polyamide column, and eluted with water, 30% ethanol and 50% ethanol in sequence, and the 30% ethanol eluted part was dissolved in 100 mL of methanol, as the crude flavonoid fraction. Crude flavonoids were further subjected to Sephadex LH-20 column chromatography...
experiment example 2
[0032] Experimental Example 2: Verification of estrogenic activity of RNFG
[0033] 1. Cell Culture Conditions
[0034] MCF-7 (containing ERE gene) or MCF-7 cells were cultured in MEM medium containing 10% fetal calf serum, penicillin 100U / ml, gentamicin 100ug / ml, 1mM amino acid and 0.1mM sodium pyruvate, Incubator temperature 37°C, containing 5% CO 2 air and saturation humidity. Three days before the experiment, the medium was changed to phenol red-free MEMα medium containing 2% activated carbon / dextran-treated fetal bovine serum, and other conditions were the same as above.
[0035] 2. Determination of estrogen level
[0036] (1) Determination of Estrogen Response Element-Luciferase Activity (ERE-luciferase)
[0037] MCF-7 cells (containing ERE gene) were planted on 24-well plates, and the cell density per well was about 5x10 4 Let the cells adhere to the wall overnight, and add RNFG (1-100 μ M) from the next day, every 24 hours, and replace the culture medium before ea...
experiment example 3
[0040] Experimental example 3: neuroprotective effect of RNFG
[0041] 1. Cell Culture Conditions
[0042] PC12 cells were cultured in DMEM medium, containing 6% fetal bovine serum, containing 6% horse serum, penicillin 100U / ml, gentamicin 100ug / ml, incubator temperature 37°C, containing 7.5% CO 2 air and a certain humidity. The primary cultured cortical nerve cells were obtained by dissecting the cerebral cortex of rat embryos (18 days). The cerebral cortex cells were cultured in neural basal medium containing B27 and 0.5mM GlutaMAX for more than two weeks. The temperature of the incubator was 37°C, containing 5 %CO 2 air and saturation humidity.
[0043] 2. Determination of reactive oxygen species (ROS)
[0044] PC-12 cells were planted on a 96-well plate with a cell density of about 5x10 per well 4 Let the cells adhere to the wall overnight, start adding the drug RNFG (1-100μM) the next day, and replace the culture medium before adding the drug. 24 hours after adding ...
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