Assessment of the mRNA level and the release of IL-1β and TNF-α in RAW264.7 cell culture treated with 30 mM high glucose over time.

<p>(A-B) RAW264.7 cells were exposed to 30 mM D-glucose and cultured for indicated time periods of 0, 4, 8, 24, 36 and 48h. The mRNA levels of two inflammatory cytokines, IL-1β (A, <i>df</i> = 5, <i>f</i> = 62.812, <i>p</i> < 0.001) and TNF-α (B, <i>df</i> = 5, <i>f</i> = 5.492, <i>p</i> = 0.03), were assessed by real-time PCR. Each data point is the mean ± SEM of <i>n</i> = 3 and normalized against corresponding <i>GAPDH</i> mRNA level with the value at 0h arbitrarily set as 1. (C-D) The release of IL-1β (C, <i>df</i> = 5, <i>f</i> = 27.324, <i>p</i> < 0.001) and TNF-α (D, <i>df</i> = 5, <i>f</i> = 48.74, <i>p</i> < 0.001) in culture medium was assessed by ELISA. Each data point is the mean ± SEM of <i>n</i> = 3. *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001.