Application of homoplantaginin and derivatives of homoplantaginin as Nrf-2 activators
A technology of homopsyllogenin and derivatives, which is applied in the preparation of sugar derivatives, sugar derivatives, sugar derivatives, etc., to achieve the effect of anti-oxidation and protection of endothelial cells and endothelial cell damage
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Embodiment 1
[0050] Example 1: Preparation of Homopsyllogen and Its Derivatives
[0051] The specific operation is as follows:
[0052] 1) Pulverize the raw material of lychee grass, add 10 times the amount of 95% ethanol solution, heat and reflux for extraction twice, each time for 4 hours, combine the extracts, recover ethanol under reduced pressure, and obtain a concentrated solution;
[0053] 2) Extraction and column separation: use ethyl acetate / n-butanol-water as a two-phase solvent system, take the upper layer as the acetyl acetate / n-butanol phase, and the lower layer as the water phase, take the upper layer acetyl acetate / n-butanol phase and concentrate it into For the extract, dilute and filter the above-mentioned concentrated solution with an appropriate amount of water, put it on a macroporous resin column, elute with 30%, 50%, 85%, and 95% ethanol aqueous solution, monitor by thin layer, collect high psyllogenin fractions, concentrate and wash Dehydration;
[0054] 3) Impurit...
Embodiment 2
[0056] Example 2: High psyllogen activates Nrf2
[0057] 1. Experimental method
[0058] Inoculate the endothelial cells in a 96-well culture dish, and when the cell tiled area reaches 80%, set up the control group: culture under normal conditions without serum; experimental group: add concentrations of 0.1 μM and 1 μM under serum-free conditions, respectively. and 10 μM homopsyllogenin S3 and its derivative S1, cultured for 24 hours, fixed with 10% TCA for one hour, rinsed with deionized water for 5 times, and dried naturally, added 50 μL of SRB dye to each well, and shaken to mix After homogenizing for 10 min, rinse with deionized water for 5 times, and after drying naturally, add 100 μL Trisbase to each well to dissolve, and measure the absorbance at 540 nm with a microplate reader. Luci-hela cells (purchased and ATCC) were planted in 96-well plates, and after the cell tiled area reached 80%, set up the control group: culture under normal conditions without serum; experime...
Embodiment 3
[0062] Example 3: Homopsyllogen and its derivatives can activate Nrf2 downstream antioxidant protein HO-1
[0063] 1. Experimental method
[0064] Endothelial cells were inoculated in a culture dish with a diameter of 6 cm. Normal group: under normal conditions without serum, low-density lipoprotein (LDL) was added for culture; control group: under normal conditions without serum, oxidized low-density lipoprotein (ox -LDL) culture; experimental group: cultured with S1 and S3 at concentrations of 0.1 μM, 1 μM and 10 μM respectively under the condition of removing serum. 37°C, CO 2 After culturing in the incubator for 6 hours, the cells were lysed, the cell lysate was collected, centrifuged at 12,000 g at 4°C for 15 min, the supernatant was taken, and the expression level of HO-1 in endothelial cells was detected by western blot.
[0065] 2. Experimental results
[0066] Experimental results such as image 3 As shown, the results showed that: compared with the ox-LDL group o...
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