Synergistic Effects Of Calycosin And Isoferulic Acid On Antioxidant And Inhibiting Proliferation Of HepG2 Cells

Some researchers have demonstrated that the complex interaction of the natual plant active components lead to synthetic effects.Calycosin and isoferulic acid are widely distributed in plant food.They have various pharmacological activities such as cardiovascular protection effect,anti-tumor,decreasing blood pressure effect,lowering blood lipids,antioxidant and other biological characteristics.On the basis of previous reserach,the synergistic effects of calycosin and isoferulic acid were analyzed on antioxidant and inhibiting proliferation of HepG2 cells.The research on synergistic mechanism may provide some theoretical basis for the purported synergistic efficiency of natural active components as functional foods.The major results of my study were as below:1.The DPPH and ABTS radical scavenging assay were applied to evaluate the antioxidant capacity of calycosin and isoferulic acid.The combination of calycosin and isoferulic acid showed synergistic effects in scavenging DPPH radicals.While,as for ABTS radical scavenging assay,the combination of isoferulic acid and calycosin showed antagonistic effect.There were some differences in the vitro antioxidant assay.Only a single method can not give a comprehensive and accurate evaluation of antioxidant efficacy.2.It was found that calycosin and isoferulic acid have shown protective effects on the H₂O₂-induced oxidative damage of HepG2 cells at low concentrations,and the protective effects of both were of a dose-dependent manner effect relationship.The combination of calycosin and isoferulic acid have shown a synergistically antioxidant activity when used together under the condition of certain concentrations.3.It was found that calycosin and isoferulic acid significantly inhibitied the growth of HepG2,the inhibitory effects was associated with the concentration of them.The IC₅₀ value of calycosin was less then it of isoferulic acid.They have shown a synergistically anticancer activity when used together under the condition of certain concentrations.4.To analysis of cell cycle of treated Hep G2 with calycosin employing the flow cytometric,the proportion in G1 phase were increased obviously in a dose-effect relationship comparing to negative control group.In test concentration isoferulic acid have no effect in arresting the G1 phase.But the combination of calycosin and isoferulic acid have shown a synergistically blocking in the G1 phase.5.Detecting the protein level of cyclinD1,CDK6 and p21 in HepG2 by Western Blot method,the experimental results indicates that the combination of isoferulic acid and calycosin obviously suppress the expression of cyclinD1 and CDK6,meanwhile increase the expression of p21.The study confirmed that the combination of calycosin and isoferulic acid suppress the proliferation of HepG2 cells by p21-mediated cell cycle regulation and down-regulating cyclinD1 and CDK6.