Application of koumine to preparation of drugs for treating hepatocyte damage
A technology of hepatocyte injury and glucosinolates, which is applied in the field of biomedicine and can solve the problems that there are no literature reports on the prevention and treatment effects of glucosinolates, no new use descriptions, etc.
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Embodiment 1
[0036] Example 1: Protective effect of kinesin on human liver cell L02 cells exposed to aflatoxin
[0037] Aflatoxin (AFB1)-exposed cell model was made using normal human liver cells L02 cells:
[0038] (1) Use RPMI-1640 medium (90%) + FBS (10%) + double antibody (1%) to culture L02 cells, passage once every 2 to 3 days, and the culture environment is 37°C, containing 5% CO 2 Incubator, when the density reaches more than 80%, subculture is carried out, the subculture ratio is 1:2, and the cells between the 10th generation and the 40th generation are selected for the experiment;
[0039] (2) 5 mg AFB1 was fully dissolved in 250 μL dimethyl sulfoxide to a 20 mg / ml stock solution, and then diluted to a concentration of 10 μM with RPMI-1640 medium;
[0040] (3) Fully dissolve 2 mg of kelkinin with 100 μL dimethyl sulfoxide to a 20 mg / mL stock solution, and then dilute it to a concentration of 20 μM or 5 μM with RPMI-1640 medium;
[0041] (4) L02 cells were treated with 1×10 5 T...
Embodiment 2
[0045] Example 2 Kiskinin increases the expression level of growth-related genes in human liver L02
[0046] The method in Example 1 was used to make aflatoxin AFB1-exposed cell model. After 33 hours, the blank control group, the hooked kiss treatment group and the negative control group all discarded the culture medium, washed twice with 100 μL / well of PBS, removed the residual medium, added 50 μL TRIzol to each well, repeatedly blown and beat the cells to fully lyse, and collected them in In EP tube.
[0047] 1. RNA extraction:
[0048] A. Take an appropriate amount of cell samples, add 600 μL Lysis / Binding Buffer, and homogenate; add 30 μL miRNAHomogenateAdditive, mix well and ice-bath for 10 minutes; B. Add an equal volume of phenol or chloroform, centrifuge at the maximum speed for 5 minutes at room temperature, and take the supernatant; C. Add 1. 25 times the volume of 100% ethanol and add the mixture to the spin column (up to 700 μL), room temperature, 13000 rpm, 30 s...
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