Article Figures & Data
Figures
- Fig. 1.
Dosing A/J mice with ibuprofen and ISDN in the chow. Data are displayed in color to indicate each of the studies we performed. Red, cohort 1; blue, cohort 2; orange, cohort 3; black, cohort 4. Please consult Table 1 for the concentrations of drugs used in each. (A) Food consumed per mouse per day (mean ± S.D.). (B) Body weights of mice by diet type, plotted for individual mice. Data are expressed as mean ± S.D. (C) Scattergram of drug concentrations measured in the serum of treated and control A/J mice. The ibuprofen concentrations in mice that were dosed simultaneously with ISDN, and the ISDN concentrations in mice dosed simultaneously with ibuprofen, are given separately. Control values are negligible. The y-axes in (A and B) are curtailed. See Materials and Methods for details.
- Fig. 2.
Torque measurements in dosed and control mice before and after LSI. Contractile torque was determined from control (n = 13), ibuprofen-treated (n = 11), and ISDN-treated (n = 12) mice, as well as from mice treated with both drugs (Ibu + ISDN, n = 13). (A) Initial torque levels are not affected by drug dosing. (B) Torque immediately following injury on day 0 shows no effect of the drugs. (C) Torque measured on day 3 indicates no beneficial effect of the drugs in these grouped data. (D) Torque levels from day 0 postinjury and day 3 are directly compared. None of the differences are significant at P < 0.05. Data are means ± S.D.
- Fig. 3.
Torque measurements as a function of serum levels of drugs. Graphs show the relationship between percentage of recovery from LSI on day 3 versus serum concentration of ibuprofen and ISDN, in micrograms per milliliter. (A) Higher ibuprofen levels lead to an inhibited recovery (r2 = 0.68; P = 0.012). (B) ISDN at increasing concentrations does not inhibit recovery significantly. (C and D) Ibuprofen serum levels (C) and ISDN serum levels (D) in ibuprofen + ISDN-treated animals give similar results. Even in the presence of ISDN, higher levels of ibuprofen significantly inhibit the ability of the muscle to recover (r2 = 0.83; P < 0.0001), but higher levels of ISDN do not. Lines are best fits by linear regression. Dashed curves represent the 95% confidence interval of the best-fit line.
- Fig. 4.
Histologic analysis of necrotic fibers and macrophages in treated muscles. Morphologic studies used H&E to identify and quantitate necrotic fibers, and anti-CD68 antibodies and immunofluorescence to identify and quantitate macrophages. (A) Stained (a) and immunofluorescent (b) images showing regions of muscles on day 3 after LSI under control conditions (1), with ibuprofen (2), with ISDN (3), and with both drugs (4). (B) Necrotic fibers decrease by a small but significant amount at day 3 following injury in mice treated with ibuprofen + ISND, compared with control and ibuprofen alone. **P < 0.01; *P < 0.05. (C) The extent of macrophage infiltration does not differ significantly among groups. Values shown are mean ± S.E.M.
- Fig. 5.
Necrotic fibers and macrophages quantitated as a function of drug levels in serum. As in Fig. 3, but for the values for necrotic fibers and macrophages measured as in Fig. 4. (A–D) Data for necrotic fibers show that increasing concentrations of ibuprofen are associated with a trend to increased necrosis (A). (E–H) Ibuprofen tends to increase macrophage infiltration to a small extent at higher concentrations with or without ISDN. None of the changes shown reach significance, however. Lines are best fits by linear regression. Dashed curves represent the 95% confidence interval of the best-fit line.
- Fig. 6.
Long-term dosing of BlAJ mice with ibuprofen: effects on body weight, creatine kinase, rotarod performance, and treadmill running. (A) Food consumption by BlAJ mice fed with chow containing ibuprofen (green), BlAJ mice fed control chow (red), and C57BL/6J mice fed control chow (blue). The y-axis in this panel is curtailed. The results show that ibuprofen stimulates food consumption in BlAJ mice. (B) Body weights of mice at 14 months of age. Chow containing ibuprofen has no significant effect on body weight of BlAJ mice (dark gray) compared with BlAJ (light gray) and C57BL/6J (white) mice on the control diet. The same shading is used in (C–E). (C) Plasma levels of creatine kinase (CK) are significantly elevated in BlAJ mice compared with C57BL/6J, with or without ibuprofen (*P < 0.05), but are not significantly altered by ibuprofen. (D) Rotarod performance is not significantly different between BlAJ and C57BL/6J mice, nor is it significantly affected by ibuprofen. (E) BlAJ and C57BL/6J mice run equally well on a treadmill, but the performance of BlAJ mice is significantly impaired by ibuprofen in the chow (*P < 0.05).
- Fig. 7.
Volumes and composition of hind limb muscles after long-term treatment with ibuprofen. Volumes and fat and water contents were measured by MRI when mice were 12 months of age. (A and B) Volumes of the gluteus (A) and psoas (B) muscles were significantly reduced in the BlAJ mice compared with C57BL/6J controls, but were not affected by ibuprofen. (C and D) Fat (C) and water (D) contents of gluteus muscles elevated (fat) or diminished (water) in BlAJ mice compared with C57BL/6J controls, but were not affected by ibuprofen. ***Indicate P < 0.001 for differences between C57BL/6 and BlAJ, with or without ibuprofen treatment.
- Fig. 8.
Morphology of muscles after long-term treatment with ibuprofen. Frozen sections of muscles were collected from mice at 14 months of age and stained with H&E to quantitate centrally nucleated fibers (CNFs) (A), muscle fiber size (B), and the distribution of muscle sizes from small to large (C). Compared with control C57BL/6J mice, treated and untreated BlAJ mice contained significantly more CNFs (**P < 0.002), but did not differ in fiber size (P > 0.35). Necrotic fibers (D) and macrophages (E) were quantitated as in Fig. 5. Ibuprofen had no significant effects on these properties of BlAJ muscle, which were, however, significantly different from those in C57BL/6J controls (*P < 0.01; ††P < 0.02; †P < 0.05).
Tables
Short-Term Cohort No. Ibuprofen (mg/g chow) ISDN (mg/g chow) Intended Ibuprofen Dosage (mg/kg per day) Estimated Ibuprofen Dosage (mg/kg per day) Intended ISDN Dosage (mg/kg per day) Estimated ISDN Dosage (mg/kg per day) 1 0.289 0.177 50 31.1 30.6 17.5 2 0.576 2.95 50 25.9 30.6 19.0 3 0.576 2.95 50 29.5 30.6 19.8 4 2.13 1.82 200 191.3 20.0 9.5
