Application of atractyloside in preparation of medicine for treating fatty liver
A technology of fatty liver and atractylodes glycosides, which is applied in the direction of drug combinations, pharmaceutical formulas, medical preparations containing active ingredients, etc., can solve the problems of lack of specificity, achieve low liver and kidney toxicity, broad social significance, and promote fat degradation Effect
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Embodiment 1
[0057] Example 1: Effect of atractyloside on the proliferation of cultured hepatocytes and the ratio of ADP / ATP
[0058] The liver cancer cell line HepG2 was selected and cultured at 37°C, 5% CO 2 , 95% humidity in a cell culture incubator, when the cell density reaches 80%, spread in a 6-well plate (the density is 1×10 6 each / hole), and different concentrations of ATR were added to the culture medium for treatment, and the cell proliferation rate and ADP / ATP ratio were measured after the treatment; the results were as follows: figure 1 with figure 2 As shown, atractyloside has no significant effect on the proliferation of HepG2 cells at the concentration of 2.5 and 7.5 μmol / L, but can significantly increase the ADP / ATP ratio in the cultured cells (1.2-1.5 times higher than that of the control group). After 24 hours of drug action The ratio reaches a maximum.
[0059] This example illustrates that under the premise that low dose atractyloside has no effect on the prolifera...
Embodiment 2
[0060] Example 2: Effect of atractyloside on fatty degeneration of HepG2 cells cultured in vitro
[0061] HepG2 cells were selected and plated in two 6-well plates (at a density of 1×10 6 cells / well), after using free fatty acid FFA to treat cells for 24h to induce fatty degeneration of liver cells, give low doses of ATR to intervene for 24h, one plate is used to collect cells to measure triglyceride (TG) level in cells, and one plate is used for oil Red O staining detection; the results are as follows image 3 with Figure 4 As shown, ATR of 5, 7.5 and 10 μmol / L significantly reduced the content of triglyceride and oil red O in HepG2 cells, and the ATR dose of 7.5 μmol / L showed that the content of triglyceride in HepG2 cells could be reduced by 50%. , The content of oil red O is 20%.
[0062] This example shows that low-dose ATR can significantly reduce the lipid content of fatty liver cells and improve the degree of fatty degeneration of liver cells.
Embodiment 3
[0063] Example 3: Atractyloside’s effect on AMP-dependent protein kinase (Adenosine5′-monophosphate-activated protein kinase, AMPK for short) and mammalian target of rapamycin (mTOR for short) in in vitro induced steatosis HepG2 cells Expression Effect Experiment
[0064] HepG2 cells were selected and plated in a 6-well plate (at a density of 1×10 6 cells / well), treated the cells with free fatty acid FFA for 24 hours to induce fatty degeneration of hepatocytes, gave low-dose ATR intervention for 24 hours, collected cells to extract total protein, and detected the expression levels of AMPK and mTOR total protein and their phosphorylated protein by Western blot; results Such as Figure 5 with Image 6 As shown, ATR increased the ratio of AMPK-α, P-AMPK-α and P-AMPK-α / AMPK-α in steatotic liver cells, and decreased the ratio of mTOR, P-mTOR and P-mTOR / mTOR.
[0065] This example suggests that ATR activates the AMPK-mTOR signaling pathway in HepG2 cells with steatosis.
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