PLC ameliorates blood flow recovery.

<p>(A) Representative diagram of rat skin flap. On the left, the proximal (P), medial (M), and distal (D) segments. Center, representative laser doppler images of blood flow before (basal) and just after flap elevation (T0). Low or no blood perfusion is displayed in dark blue, and the highest perfusion level is displayed in red. Right, it was reported the means ± SEM of blood perfusion relative units (RU) in proximal, medial and distal portions of the flap before (baseline) and just after flap creation (T0). (B, C) Representative laser Doppler images of blood flow (B) and time course (C) of in skin flap in vehicle- (closed symbols) and PLC-treated (open symbols) rats. PLC was administered daily, dissolved in tap water at the dose of 100mg/kg. Points are mean ± SEM of 15 rats, in relative units of blood perfusion at 2, 4, 6 and 8 days after flap elevation. (C) Left, blood perfusion in distal portion of flap (repeated measures ANOVA: * indicates p< 0.05, PLC-treated vs vehicle-treated rats). Center, blood perfusion in medial portion of flap (ANOVA:** indicates p< 0.001,PLC-treated vs vehicle-treated rats). Right, blood perfusion in proximal portion of flap (ANOVA:* and ** indicate p< 0.05 and p< 0.001, respectively, PLC-treated vs vehicle-treated rats). (D,E) Representative images of flap skin and necrotic area measurement after 8 days. Necrotic area (black) is clearly demarcated from survival regions. (F) Values represent mean ± SEM of 15 rats. Repeated measures ANOVA: treatment, p< 0.0001; Dunnett’s test: * and ** indicate p< 0.001 and p< 0.0001, respectively. (F) Bar graph showing ROS levels, inversely proportional to fluorescence intensity (F.I), in rat skin flap. Values represent mean ± SEM of 15 rats. t-Student: * indicates p< 0.05. (G) iNOS protein expression and NO level (expressed in optical density, OD), in rat skin flap. Values represent mean ± SEM of 15 rats. t-Student: * indicates p< 0.05. Abbreviation: α-tub, α-tubulin.