Use of natural compound as anti-diabetes drug
A technology of diabetes drugs and compounds, which is applied in the application field of 4-hydroxymelphenytoin in the preparation of drugs for treating diabetes, and can solve the problems that there are no research reports on the anti-diabetic effect of 4-hydroxymelphenytoin
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Embodiment 1
[0037] Embodiment 1 regulates the effect of phase secretion of insulin
[0038] Scheme and implementation: With reference to the detection of the effect of 4Q on the survival rate of this kind of cells, the low and high doses of 4Q were selected as 0.1 and 30 μM, respectively.
[0039] Rin-5f islet β cells were inoculated in 100mL culture dish (10 7 cells / well) after 24 hours of culture, the cells were cultured with high glucose 1640 medium (glucose content of 16.4mM). According to different treatments, they were divided into high glucose model group and 4Q low, middle and high dose groups. Under high glucose environment, 4Q acted for 15min, 60min and 120min respectively. Afterwards, 100 μL of the culture supernatant was taken, and the insulin content in the culture supernatant was detected, and the measurement unit of insulin was represented by International Viability Unit (IU, 1IU=45.4 μg).
[0040] Experimental results: In a high-glucose environment (medium with a glucos...
Embodiment 2
[0041] Example 2 Improving the Effect of Glucose Consumption in the Insulin Resistance Model
[0042] Scheme and implementation: With reference to the detection of the effect of 4Q on the survival rate of this kind of cells, the low, medium and high doses of 4Q were selected as 0.1, 1 and 10 μM respectively.
[0043] Establishment and testing of insulin resistance model: set up 2 groups, normal glucose and normal insulin group, high glucose and high insulin group, 6 samples in each group. Adipocytes were cultured in low-sugar DMEM medium (containing 5.5 mM glucose) containing 1% BSA for 12 hours, and then intervened in groups. The normoglucose normoinsulin group contained 1%BSA, 5.5mM glucose, 10 -9 The DMEM medium of M insulin was cultivated; the high glucose and high insulin group contained 1% BSA, 25mM glucose, 10 -6 M insulin was cultured in DMEM medium. After 24 hours of intervening culture, the culture medium was discarded, washed with PBS for 3 times, and the glucose...
Embodiment 3
[0046] Example 3 Detection of the toxicity of Rin-5f to pancreatic islet β cells and 3T3-L1 adipocytes
[0047] Protocol and implementation: Rin-5f islet β cells and 3T3-L1 adipocytes were seeded in 96-well plates (10 4 cells / well), after culturing for 24 hours, add different concentrations of 4Q (final concentrations are 1, 3, 5, 10 μM), set up three parallel wells for each concentration, control group without drug treatment, put them in the incubator for incubation 24h. Then add 10 μL of freshly prepared MTT (final concentration 5 mg / mL) solution, incubate for 4 h, pour off the supernatant, add 200 μL of DMSO (dimethyl sulfoxide) to each well, and measure the optical density at 570 nm (OD ).
[0048] Experimental results: if Figure 3A and 3B Shown are the toxicity test results of different concentrations of 4-hydroxymephenytoin on Rin-5f pancreatic beta cells and 3T3-L1 adipocytes in this example. The toxicity of -5f islet β cells and 3T3-L1 adipocytes is small, and th...
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