Inhibits LDH release from cells treated with ox-LDLA 24 h exposure to 100 mg/L 4-Deoxyuridine structure ox-LDL resulted in a marked and significant facilitation of LDH release from the cells, compared with the vehicle control group (555.15 ?9.67 U/L vs. 269.12 ?97.80 U/L, p < 0.01). In contrast, the LDH activity in the supernatant was significantly decreased in the samples pretreated with 0.1 mg/L EOFAZ compared with the samples treated with ox-LDL alone (328.68 ?61.29 U/L vs. 555.15?9.67 U/L, p < 0.01; Figure 2).Table 1 Protective effects of EOFAZ on HUVECs' injury induced by ox-LDLGroups Control ox-LDL ox-LDL+EOFAZ Dose (mg/L) -- 100 100+0.1 100+0.01 ox-LDL+PRA# #OD570 1.85?.11 0.98?.12##Inhibition of cell damage ( )Data were expressed as mean ?S.E.M (standard error of the mean) of at least three independent experiments. The differences in mean values among the experimental groups were measured using two-tailed analysis of variance (ANOVA) followed by Dunnett's test. Differences were considered statistically significant at p < 0.05.1.25?.20** 1.09?.25 1.19?.21*31.0 12.6 24.100+10 mol/Lp < 0.01, compared with the control group; *p < 0.05 and ** p < 0.01, compared with the ox-LDL group.Shen et al. BMC Complementary and Alternative Medicine 2012, 12:174 http://www.biomedcentral.com/1472-6882/12/Page 5 ofFigure 1 Effects of EOFAZ on trypan blue exclusion staining in HUVECs' injury induced by ox-LDL. HUVECs were pretreated PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/27362935 for 30 min with EOFAZ or PRA, and then exposed to 100 mg/L ox-LDL for 24 h. A cell suspension was mixed with a 0.4 trypan blue dye for 5 min at 25 , and the unstained (viable) and stained (nonviable) cells were counted in a hemacytometer. The values shown are mean ?S.E.M. of three experiments (four or five cultures per experiment). ##p < 0.01, vs. control group; *p < 0.05, **p < 0.01, vs. ox-LDL group.EOFAZ reduces the MDA contents and increases the GSH contents in cells treated with ox-LDLIn the present study, incubation of HUVECs with 100 mg/L ox-LDL for 24 h resulted in a significant increase in the MDA contents, which was significantly attenuated by EOFAZ or PRA pretreatment (Figure 3). However, the exposure of HUVECs to ox-LDL 100 mg/L for 24 h resulted in a significant decrease in GSH contents, which was ameliorated by preincubation with EOFAZ or PRA (Figure 4).EOFAZ upregulates the activities of antioxidant enzymesEOFAZ significantly ameliorated the activities of SOD, CAT and GSH-Px in the cell lysates. Preincubation with 0.01 mg/L EOFAZ enhanced the activities of these enzymes, however, these results were not statistically significant. PRA (10 mol/L) alleviated the decreased activities of SOD and CAT (**p < 0.01, compared with the ox-LDL treatment), but did not improve the GSH-Px activity.The main scavengers responsible for inactivation and termination of free oxygen radicals are SOD, CAT, and the glutathione system. Our results indicated that the activities of the anti-oxidative enzymes were significantly decreased after HUVECs were exposed to 100 mg/L oxLDL, and that EOFAZ and PRA improved these activities (Figures 5, 6 and 7). Preincubation with 0.1 mg/LDiscussion To the best of our knowledge, the present study demonstrates for the first time that EOFAZ (the composition was significantly different from previous reports and from the essential oil extract from the fruit of A. zerumbet) protects against endothelial cell injury induced by ox-LDL via ameliorating oxidative stress, which may be the main underlying mechanism. Human umbilic.