LECT2/Tie1/Src signaling regulates the oxidative stress response in endothelial cells and liver ischemia-reperfusion injury

Figure 2 Tie1 knockdown or Tie1-Ig3 treatment reduces leukocyte-derived chemotaxin-2-induced hypoxia-reoxygenation injury in EA. hy926 cells. A: MRNA expression of the Tie1 decreased after transfection with siTie1 RNA into EA.hy926 cells. Student’s t-test; B: Protein expression of the Tie1 decreased after transfection with Tie1-small interfering RNA (siRNA) into EA.hy926 cells; C: A cell counting kit-8 assay was used to measure cell proliferation capacity in the siTie1 group and Tie1-siRNA + recombinant leukocyte-derived chemotaxin-2 (rLECT2) group at different time points during reoxygenation after 8 hours of hypoxia. Two-way ANOVA; D: Superoxide dismutase activity was measured in the siTie1 group and Tie1-siRNA + rLECT2 group at different time points during reoxygenation after 8 hours of hypoxia to determine the level of cellular oxidative stress. Two-way ANOVA; E: Lactate dehydrogenase release was measured in the siTie1 group and Tie1-siRNA + rLECT2 group at different time points during reoxygenation after 8 hours of hypoxia to assess cellular cytotoxicity. Two-way ANOVA; F: A cell counting kit 8 assay was used to assess cell proliferation capacity at different time points during reoxygenation after 8 hours of hypoxia. Two-way ANOVA; G: Superoxide dismutase activity was measured at different time points during reoxygenation after 8 hours of hypoxia to assess the level of cellular oxidative stress. Two-way ANOVA; H: Lactate dehydrogenase levels were measured at different time points during reoxygenation after 8 hours of hypoxia to assess cellular cytotoxicity. Two-way ANOVA. Data are representative of three independent experiments. All data are presented as the mean ± SEM. ^aP < 0.05, ^bP < 0.01, and ^cP < 0.001. rLECT2: Recombinant leukocyte-derived chemotaxin-2; siRNA: Small interfering RNA; H/R: Hypoxia/reoxygenation.