The Experimental Study Of Cajanonic Acid A On HepG2 Cells In Vitro Metabolism Of Sugar And Insulin Resistance Of HepG2 Cells

Objective:To explore the influence of Cajanonic acid A on HepG2 cells in vitro metabolism of sugar and improve insulin resistance of HepG2 cells and related mechanism is discussed in this paper.Methods:The First Part Research the influence of Cajanonic acid A on HepG2 cells in vitro metabolism of sugar1.Cultivation of HepG2 cells in vitro,detection by different concentration(10-3mol/L,10-4 mol/L,10-5 mol/L and 10-6 mol/L)Cajanonic acid A role within 24 h after HepG2 cells to the change of glucose metabolism;2.ELISA method to detect the different concentration(10-3 mol/L,10-4 mol/L,10-5 mol/L and 10-6 mol/L)Cajanonic acid A role within 24 h after HepG2 cells sugar metabolism related enzyme the activity of hexokinase(HK) and pyruvate dehydrogenase(PDH).The Second Part Cajanonic acid A role on the improvement of the insulin resistance HepG2 cells 1.By different concentration(10-6 mol/L,10-7 mol/L,10-8 mol/L and 10-9 mol/L)of insulin action HepG2 cells,select the best resistance concentration;the best resistance concentration insulin action HepG2 cells after different time pionts(12 h,24 h,36 h and48 h),select the best resistance time;2. Detection by different concentration(10-3mol/L,10-4 mol/L and 10-5 mol/L) of Cajanonic acid A role within 24 h after IR-HepG2 cells to the change of glucose metabolism;3.ELISA method to detect the different concentration(10-3 mol/L,10-4 mol/L and 10-5 mol/L)Cajanonic acid A role within 24 h after IR-HepG2 cells sugar metabolism related enzyme the activity of hexokinase(HK) and pyruvate dehydrogenase(PDH);4.Western blot test by different concentration(10-3 mol/L,10-4 mol/L and 10-5 mol/L) of Cajanonic acid A role within24 h after IR-HepG2 cells AMPK and AKT protein expression level.Results:The First Part 10-3 mol/L,10-4 mol/L and 10-5 mol/L Cajanonic acid A effect after 24 h HepG2 cells glucose consumption respectively(8.650 ± 0.319) mmol/L,(8.231 ±0.182) mmol/L and(8.136±0.402) mmol/L,compared with negative control group(6.521±0.056) mmol/L,the difference was statistically significant(P < 0.05);10-3mol/L and 10-4 mol/L Cajanonic acid A effect after 24 h in HepG2 cells hexokinase content respectively(882.91±30.72) pg/mL and(718.25±20.67) pg/mL,compared with negative control group(616.65±17.24) pg/m L,the activity of hexokinase increased;10-3 mol/L Cajanonic acid A effect after 24 h in HepG2 cells pyruvate dehydrogenase content is(1526.69±66.74) pg/mL,compared with negative control group(1204.73±55.72) pg/m L,the activity of pyruvate dehydrogenase increased,the difference was statistically significant(P<0.05).The Second Part 10-7 mol/L insulin action HepG2 cells after 24 h,can establish a stable model of insulin resistance;10-3mol/L,10-4 mol/L Cajanonic acid A effect after 24 h IR-HepG2 cells glucose consumption respectively(7.934±0.013) mmol/L,(7.032±0.032) mmol/L,compared with negative control group(5.952±0.015) mmol/L,the difference was statistically significant(P < 0.05);10-3 mol/L,10-4 mol/L and 10-5 mol/L Cajanonic acid A effect after 24 h in IR-HepG2 cells hexokinase content respectively(568.87±39.77) pg/mL,(495.43±35.43) pg/mL and(489.63±27.30) pg/mL,compared with negative control group(444.42±33.52) pg/mL,the activity of hexokinase increased;10-3 mol/L,10-4mol/L Cajanonic acid A effect after 24 h in IR-HepG2 cells pyruvate dehydrogenase content respectively(1765.54±135.26) pg/mL,(1620.48±149.81) pg/mL,compared with negative control group(1247.92±152.47) pg/m L,the activity of pyruvate dehydrogenase increased,the difference was statistically significant(P < 0.05);10-3mol/L and 10-4 mol/L Cajanonic acid A effect after 24 h can raise IR-HepG2 cells AMPK protein expression,10-3 mol/L Cajanonic acid A effect after 24 h can raise IR-HepG2 cells AKT protein expression,the difference was statistically significant(P<0.05).Conclusion: Cajanonic acid A in vitro can promote HepG2 cells to glucose uptake,with hypoglycemic activity,its action may be associated with enhanced glucose metabolism enzyme hexokinase and pyruvate dehydrogenase activity.Cajanonic acid A might also can improve insulin resistance of HepG2 cells,its action may be associated with enhanced glucose metabolism enzyme hexokinase and pyruvate dehydrogenase activity and raise AMPK and AKT protein expression.