Figure 1
Scavenging ROS by apocynin (Apo) reduced Nox expression, cell injury and inflammatory response in cisplatin (Cis)-treated HK2 cells. (a) DCFH-DA assay in Cis-treated HK2 cells. Treatment of Apo decreased the production of intracellular reactive oxygen species (ROS) in Cis-treated HK2 cells. (b) Western blotting analysis of Noxs in HK2 cells. Treatment of Apo inhibited Cis-induced upregulation of kidney-related NADPH oxidases, including Nox2 and NADPH oxidase 4 (Nox4). (c, d) Western blotting analysis and real-time PCR of kidney injury molecule-1 (KIM-1) in HK2 cells. Treatment of Apo significantly reduced the protein and mRNA levels of KIM-1. (e) Lactate dehydrogenase (LDH) assay. Apo suppressed LDH release in Cis-treated HK2 cells. (f) Immunofluorescence and quantitative analysis of KIM-1. Treatment of Apo decreased KIM-1 protein in Cis-treated HK2 cells. (g) Immunofluorescence and quantitative analysis of TNF-α. Apo decreased the percentage of TNF-α-positive cells in response to Cis. (h) Real-time PCR of inflammatory indexes. Apo reduced mRNA levels of inflammation cytokines, including TNF-α, IL-1β and IL-6, in Cis-treated HK2 cells. Data represent the mean±s.e.m. for 3–4 independent experiments in vitro. *P<0.05, **P<0.01, ***P<0.001 versus normal; ##P<0.01, ###P<0.001 versus Cis-treated group.
