The Antioxidation And Anti-inflammaion Of Ursolic Acid In Combination With Viili Exopolysaccharides, Astragalus Polysacharin

Primary carcinoma of the liver is one of the most common human malignancies, secondly to gastric cancer, esophageal cancer. The number of new cases in China takes up 50%of the global incidents. It has been shown that cyclooxygenase (COX)-2 is over expressed in hepatoma carcinoma cells. Ursolic acid (UA) and viili exopolysaccharides (EPS), Astragalus polysaccharin (APS) make differences in inhibiting cell growth and inducing apoptosis in some tumors. However, it is unknown the action mechanism of antioxidation and anti-inflammation in HepG2 by UA, viili EPS and APS, and their combination.First of all, to detect the antioxidation force of UA, viili EPS, APS and their combination by the total reducing power, scavenging of hydroxyl free radical or DPPH free radicals. Secondly, HepG2 cells as experimental model, by MTT method to detect the cell proliferation of HepG2 cells; Scanning electronic microscopy (SEM) to observe changes in cell morphology and flow cytometry instrument (FCM) to detect the cell cycles; Using the superoxide dismutase (SOD) and malondialdehyde (MDA) kits to demonstrate SOD activity and MDA content in HepG2 cells; Reverse transcription-quantitive PCR (RT-qPCR) and Western blot to detect the expression of COX-2, ELISA kits to detect COX-2 and PGE2 concentrations.The results have shown that samples have potent antioxidant force, can clean up hydroxyl free radical or DPPH free radicals, and inhibite the growth of HepG2 cells, the inhibition was dose and time dependent. Typical apoptosis morphology was observed by SEM. Flow cytometry assay also showed the ratio of apoptosis in different phase of cell cycle, HepG2 cells were accumulated in Gl. SOD activity decreased significantly in HepG2 cells, especially with 5μg/mL UA,50 pg/mL viili EPS,50 μg/mL APS and their combination, but MDA was largely decreased except low concentration of viili EPS and APS. COX-2 gene and protein expression were down-regulated demonstrated by (RT-qPCR) and Western blot. COX-2 and PGE2 concentration were also decreased in HepG2 cells.Our results indicate that UA in combination with viili EPS and APS particularly affects COX-2 inhibition that may trigger internal antioxidative pathway, which may actually create an oxidative environment to reduce cell proliferation. We also conclude that UA, viili EPS and APS have anti-cancer properties via apoptosis.