Application of neohesperidin in preparing of diabetes preventive treatment medicines
A technology of neohesperidin and diabetes, which is applied in the field of medicine, can solve the problems such as no reports on the effect of neohesperidin, achieve good clinical application prospects, reduce adverse reactions, and improve the effect of glucose and lipid metabolism
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Embodiment 1
[0022] One of the sources of neohesperidin in the present invention is obtained from the white cortex of the citrus plant Huyou, and its preparation method and results are as follows:
[0023] Preparation of the crude extract powder of the macroporous resin of the white cortex of Huyou: 16 g of the dried white cortex of Huyou was extracted twice with 80% ethanol (solid-to-liquid ratio 1:25) ultrasonically (40°C), centrifuged at 4000 rpm for 20 min, Combine the two supernatants and evaporate to dryness on a rotary evaporator to obtain an extract, which is dissolved in ddH 2 In O, about 400 mL of D101 macroporous resin was used, the column bed volume (BV) was 20 mL, and the sample loading flow rate was 2 mL / min. The D101 macroporous resin after sample loading was first washed with water for 4 BV to remove impurities, and then naringin and Neohesperidin, wash 4 BV for each gradient, the elution flow rate is 2 mL / min, combine 10%-60% eluent, and evaporate to dryness on a rotary e...
Embodiment 2
[0028] The hypoglycemic activity of human hepatocytes was studied on the natural source neohesperidin extracted in Example 1, and it was found that neohesperidin can reduce the glucose concentration in the cell culture medium, and its glucose consumption is significantly higher than that of the control group. The experimental methods and results are as follows:
[0029] Human HepG2 hepatocytes were cultured in high-glucose DMEM containing 10% fetal bovine serum at 37°C and 5% CO 2 Incubate in a cell incubator, replace with fresh culture medium every other day, and passage once every 2 to 3 days. During the experiment, HepG2 cells were seeded in 96-well plates, and blank control wells without cells were set. After the cells grow to 80-90% confluence, discard the original medium, wash twice with PBS buffer, replace with serum-free 1640 culture medium containing 0.2% BSA, and add test substances in groups.
[0030]Set up DMSO solvent control group, metformin positive control gr...
Embodiment 3
[0034] The present invention finds that neohesperidin can obviously up-regulate the phosphorylation level of glucolipid metabolism-related signal proteins after acting on liver cells. The experimental methods and results are as follows:
[0035] Human liver HepG2 cells were inoculated in culture dishes, respectively set up DMSO solvent control group, metformin control group (final concentration 1 mmol / L), neohesperidin and naringin 1 μg / ml treatment groups, and neohesperidin and naringin at 0.5 μg / ml each, the cells were collected after 24 hours of treatment, and the phosphorylated protein levels of the target proteins AMPK and ACC were detected by Western blotting. The same experiment was repeated 3 times.
[0036] Protein extraction and concentration detection: The collected cell suspension was transferred to a centrifuge tube and centrifuged at 1000 rpm for 5 min. Remove the culture medium, wash the cells twice with cold PBS buffer, store at -80°C or directly lyse. Whe...
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