Rat LIPC sera upregulated the mRNA expression of antioxidases in HUVECs.

<p>CAT, SOD-1, SOD-2 and GSH-Px-1 mRNA levels in HUVECs were detected by real-time PCR. The mRAN expression of antioxidases was presented by normalizing the antioxidases expression with GAPDH and taking control as 100%. All data are expressed as mean ± SEM, n = 6, *<i>P <</i> 0.05, **<i>P <</i> 0.01 vs model.</p

Rat LIPC sera reduced MDA and increased the activities of antioxidases in H₂O₂-injured HUVECs.

<p>Medium MDA concentration was measured spectrophotometrically at 532 nm. The activities of CAT, GSH-Px and total SOD were analyzed spectrophotometrically at 240 nm, 340 nm and 550 nm respectively. All data are expressed as mean ± SEM, n = 6, *<i>P</i> < 0.05, **<i>P</i> < 0.01 vs model.</p

List of primers for qPCR analysis.

<p>List of primers for qPCR analysis.</p

Rat LIPC sera enhanced Nrf2 localization into nucleus inH₂O₂-injured HUVECs.

<p>Localization of Nrf2 was performed by immunofluorescence and confocal microscopy. Nrf2 was stained with an anti-Nrf2 antibody and visualized with a secondary antibody conjugated with FITC (green). The nuclei were counterstained with DAPI (4’,-diamidino-2-phenylindole) staining indicating the location of the nucleus (blue). The merged image showed the nuclear location of Nrf2 protein.</p

Rat LIPC sera decreased ROS in H₂O₂-injured HUVECs.

<p>ROS level was determined by measuring the intensity of dihydroethidium (DHE) fluorescence. The relative fluorescence intensities of DHE were analyzed by Image-Pro Plus software taking the fluorescence intensity of the control as 100%. A: original images of the cells preloaded with DHE. B: pooled data are expressed as mean ± SEM, n = 5, **<i>P</i> < 0.01 vs model.</p

Effects of rat LIPC sera on Nrf2 translocation and cellular viability were affected by neither PI3K/Akt inhibitor nor MEK/ERK inhibitor in H₂O₂-injured HUVECs.

<p>The cells were cultured in medium for 24 h, after which the inhibitors U0126 (U0, 26 μM) and LY294002 (LY, 25 μM) were added 1 h before the especial serum and H_<b>2</b>O_<b>2</b> treatments. A: nucleus localization of Nrf2 determined by immunofluorescence and confocal microscopy. B: cells’ viability detected using MTT method. The data (mean ± SEM) were obtained from at least three independent experiments, **<i>P</i> < 0.01 vs model.</p

Rat LIPC sera increased the cells’ viability of H₂O₂-injured HUVECs.

<p>HUVECs were pretreated with 5% different sera for 12 h and followed by 2 h incubation with 1 mMH_<b>2</b>O_<b>2</b>. Cells’ viability was measured by MTT assay and expressed as the ratio of optical density (OD) with OD value of the control as 100%. The data (mean ± SEM) were obtained from at least three independent experiments, **<i>P</i> < 0.01 vs model. Control: cultured with normal medium without any intervention throughout the experiment. Model: cultured with normal medium for 12 h and then incubated with 1mM H_<b>2</b>O_<b>2</b> for 2 h. NPS, EPS and DPS: cultured with normal medium containing either 5% NPS (serum derived from rats after sham LIPC), 5% EPS (serum derived from rats 20 min after LIPC) or 5% DPS (serum derived from rats 24 h after LIPC) respectively for 12 h and followed by 2 h incubation with 1mM H_<b>2</b>O_<b>2</b>. The figure legends are same in following figures unless illustrated elsewhere.</p