Fisetin reduces biochemical changes in Akita mouse kidneys.

(A) The diabetes-dependent increase in TBARS is prevented by fisetin. Kidneys from wild type, wild type + fisetin, Akita and Akita + fisetin mice were homogenized in PBS containing phosphatase and protease inhibitors and equal amounts of extract were assayed for TBARS. Kidney samples from 6–7 mice were analyzed and the amount of TBARS normalized to protein. (B) The diabetes-dependent up-regulation of osteopontin (OPN) is prevented by fisetin. Kidneys from wild type, wild type+fisetin, Akita and Akita + fisetin mice were homogenized in PBS containing phosphatase and protease inhibitors and equal amounts of protein were analyzed by SDS-PAGE and Western blotting with an antibody to OPN and actin as a loading control. The expression of GCL (C) and the RAGE receptor (D) in kidneys were analyzed by Western blotting as described in (B). Kidney samples from 6–7 mice per group were analyzed and the average intensity of the GCL, RAGE, and OPN bands normalized to actin is shown in the graphs. A representative blot is shown for each. ***(p<0.001), **(p<0.01) and *(p<0.05) indicate significantly different from wild type or Akita alone as determined by ANOVA followed by Tukey's post test.