![Figure 2. R-flurbiprofen decreased mechanical hyperalgesia and cold sensitivity in HbSS-BERK mice. Mechanical hyperalgesia (allodynia) was defined as a decrease in mechanical withdrawal threshold of the hind paw. Littermates expressing normal human hemoglobin (HbAA-BERK) were used as the control. (A) R-flurbiprofen (intraperitoneal) decreased mechanical hyperalgesia in HbSS-BERK mice in a dose-dependent manner with an 50% effective dose of 6.54 ± 3.8 mg/kg (Prism software; GraphPad Software, Inc). For perspective, the open square represents the mean mechanical threshold in naive HbAA-BERK mice from panel D. *Different from vehicle (V; ratio of DMSO to Tween 80 to saline, 27%:0.5%:72.5%). Data were transformed to log10 for parametric statistical analysis. (F[3, 31] = 15.393; P < .001, 1-way ANOVA, N = 6-9 mice per group). (B) R-flurbiprofen (9 mg/kg intraperitoneally) acutely increased the mechanical threshold in HbSS-BERK, but not HbAA-BERK, mice. In addition, the same dose of S-flurbiprofen had no effect in HbSS-BERK mice. *Different from baseline (BL) and vehicle within HbSS-BERK mice (ratio of DMSO to Tween 80 to saline, 13.5%:0.5%:86%). #Different from S-flurbiprofen within HbSS-BERK mice at P < .001 (F[12, 99] = 4.71; P < .001, 2-way ANOVA repeated measures, treatment × time, N = 6-9 mice per group). (C) R-flurbiprofen (9 mg/kg intraperitoneally) reduced cold avoidance at 60 minutes post–drug injection. Data (median and 25% and 75% range) represent the amount of time mice spent on the colder plate (23°C). *Different from HbAA-BERK mice within vehicle treatment or naive condition. #Different from vehicle within HbSS-BERK mice P < .05, Kruskal-Wallis 1-way ANOVA (H[5] = 35.04 with Dunn multiple comparisons test; number within each bar represents sample size). (D) Intraplantar injection of R-flurbiprofen (30 μg/10 μL) suppressed mechanical allodynia in HbSS-BERK mice (F[9, 69] = 2.761; P = .008, 2-way ANOVA repeated measures, treatment × time, N = 5-8 mice per group) whereas the same dose of S-flurbiprofen or vehicle (ratio of DMSO to Tween 80 to saline, 13.5%:0.5%:86%), had no effect. *Different from HbAA-BERK mice within baseline (BL) at P < .001. #Different from other HbSS-BERK groups at 60 minutes and from BL within the same treatment group; P < .001; Bonferroni t test.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/133/18/10.1182_blood-2018-11-884346/3/blood884346f2.png?Expires=1724239650&Signature=Npcstb3xFOyzGORO3gCaAuE7lmwxQQuEKVFHf0rOzrS2520Pbw7LQDbgzuSns5KjAPGB0IvfZnkQoXaUu2jMLtoTza9uRCNzFPYq2yRMw04x7tNoOWDm0Cf6MC4vCm4Bas-dlaM8wKO795rzn6AbkjCXFVv0U2MMqZt24iBz24k1m-wt0L28AlrfWAe1G~BeKzHcGAqEhbftP152eNChRMkukkgGMh-z2esWxdsgbg7hizq2b9XFAgAwLKHyI5c7kgamc7XhA0q-fiz66Gm5W8rMpQuFIQUZl0reYMvtxJcZYXPZSxxWZU-8fOHrcKB1Hhv5GkiVFSdJQN~FpNjFeg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
R-flurbiprofen decreased mechanical hyperalgesia and cold sensitivity in HbSS-BERK mice. Mechanical hyperalgesia (allodynia) was defined as a decrease in mechanical withdrawal threshold of the hind paw. Littermates expressing normal human hemoglobin (HbAA-BERK) were used as the control. (A) R-flurbiprofen (intraperitoneal) decreased mechanical hyperalgesia in HbSS-BERK mice in a dose-dependent manner with an 50% effective dose of 6.54 ± 3.8 mg/kg (Prism software; GraphPad Software, Inc). For perspective, the open square represents the mean mechanical threshold in naive HbAA-BERK mice from panel D. *Different from vehicle (V; ratio of DMSO to Tween 80 to saline, 27%:0.5%:72.5%). Data were transformed to log10 for parametric statistical analysis. (F[3, 31] = 15.393; P < .001, 1-way ANOVA, N = 6-9 mice per group). (B) R-flurbiprofen (9 mg/kg intraperitoneally) acutely increased the mechanical threshold in HbSS-BERK, but not HbAA-BERK, mice. In addition, the same dose of S-flurbiprofen had no effect in HbSS-BERK mice. *Different from baseline (BL) and vehicle within HbSS-BERK mice (ratio of DMSO to Tween 80 to saline, 13.5%:0.5%:86%). #Different from S-flurbiprofen within HbSS-BERK mice at P < .001 (F[12, 99] = 4.71; P < .001, 2-way ANOVA repeated measures, treatment × time, N = 6-9 mice per group). (C) R-flurbiprofen (9 mg/kg intraperitoneally) reduced cold avoidance at 60 minutes post–drug injection. Data (median and 25% and 75% range) represent the amount of time mice spent on the colder plate (23°C). *Different from HbAA-BERK mice within vehicle treatment or naive condition. #Different from vehicle within HbSS-BERK mice P < .05, Kruskal-Wallis 1-way ANOVA (H[5] = 35.04 with Dunn multiple comparisons test; number within each bar represents sample size). (D) Intraplantar injection of R-flurbiprofen (30 μg/10 μL) suppressed mechanical allodynia in HbSS-BERK mice (F[9, 69] = 2.761; P = .008, 2-way ANOVA repeated measures, treatment × time, N = 5-8 mice per group) whereas the same dose of S-flurbiprofen or vehicle (ratio of DMSO to Tween 80 to saline, 13.5%:0.5%:86%), had no effect. *Different from HbAA-BERK mice within baseline (BL) at P < .001. #Different from other HbSS-BERK groups at 60 minutes and from BL within the same treatment group; P < .001; Bonferroni t test.