Figure 3.

Inhibition of lymphangiogenesis augments intrarenal inflammation in cisplatin (Cp)-induced acute kidney injury. Total RNA was isolated from whole kidney tissue at day 3 and analyzed for expression of colony stimulating factor-1 (Csf1; A), chemokine (C-C motif) ligand 2 (Ccl2; B), tumor necrosis factor-α (Tnfα; C), and interleukin-6 (Il6; D) by real-time PCR. Data were normalized to Gapdh and expressed as fold changes compared with vehicle controls. Data are expressed as means ± SE; n = 6–19 animals per group. Protein lysates from whole kidney tissue were analyzed for expression of phosphorylated (p)-p65 (NF-κB), p-Akt (RAC-α serine-threonine-protein kinase), p-JNK, and IL-6 (E) and heme oxygenase-1 (HO-1; F). Anti-β-actin was used as a loading control. Densitometric values were calculated and expressed in arbitrary units (AU). Data are expressed as means ± SE; n = 6–19 animals per group. Data were analyzed using one-way ANOVA followed by a Tukey’s multiple comparisons test. G: real-time PCR of kidneys at day 3 for expression of the HO-1 gene (Hmox1). Data were normalized to Gapdh and expressed as fold changes compared with vehicle controls. Data are expressed as means ± SE; n = 5–9 animals per group. *P < 0.05 vs. vehicle control; #P < 0.05 vs. Cp + DMSO using one-way ANOVA followed by a Tukey’s multiple comparisons test.