Effect of paclitaxel on mechanical hyperalgesia in mice.

<p>Effect of a single administration of paclitaxel (2, 5 mg/kg, i.p.) on mechanical hyperalgesia in mice. The nociceptive threshold was determined by the von Frey filament test. Each point represents the mean ± S.E.M of 8 mice. **P < 0.01, ***P < 0.001 vs. respective 10% DMSO group.</p

Effect of pretreatment with RKT on paclitaxel (single injection)-induced mechanical hyperalgesia in mice.

<p>(A) Time course of the effect of RKT (0.1, 0.3, 1 g/kg p.o.) on the mean mechanical threshold in paclitaxel-treated mice. (B) Effect of RKT (0.1, 0.3, 1 g/kg p.o.) on the mean mechanical threshold in paclitaxel-treated mice on day 3. RKT was administered 24 hours before the injection of paclitaxel (5 mg/kg, i.p.). RKT was administered 1 hour before each nociceptive test. The nociceptive threshold was determined by the von Frey filament test. Each point represents the mean ± S.E.M of 8–9 mice. *P < 0.05, ***P < 0.001 vs. 10%DMSO-DW group. ^###P < 0.001 vs. paclitaxel-DW group.</p

Effect of paclitaxel on the protein levels of spinal NFκB and pNFκB in mice.

<p>Immunoblots of (A) NFκB and (B) pNFκB were normalized by GAPDH. The NFκB column represents the mean ± S.E.M of 5–7 mice. The pNFκB column represents the mean ± S.E.M of 3–5 mice. ** P < 0.01 vs. 10%DMSO group.</p

Effect of the NFκB inhibitor, PDTC, on paclitaxel-induced mechanical hyperalgesia in mice.

<p>(A) Time course of the effect of PDTC (100 ng/5μl, i.t.) on the mean mechanical threshold in paclitaxel-treated mice. (B) Effect of PDTC (100 ng/5μl, i.t.) on the mean mechanical threshold in paclitaxel-treated mice on day 3. PDTC was administered 30 minutes before the injection of paclitaxel (5 mg/kg, i.p.). The nociceptive threshold was determined by the von Frey filament test. Each point represents the mean ± S.E.M of 8–10 mice. ***P < 0.001 vs. saline-10%DMSO group. ^###P < 0.001 vs. saline-paclitaxel group.</p

Effect of RKT on the protein levels of spinal NFκB and pNFκB in mice treated with paclitaxel.

<p>Immunoblots of (A) NFκB and (B) pNFκB were normalized by GAPDH. The NFκB column represents the mean ± S.E.M of 5–7 mice. The pNFκB column represents the mean ± S.E.M of 3–5 mice. **P < 0.01 vs. 10%DMSO-DW group. ^#P < 0.05 vs. paclitaxel-DW group.</p

Effect of RKT on paclitaxel (repeated injections)-induced mechanical hyperalgesia in mice.

<p>RKT (1g/kg, p.o.) was administered before the injection of paclitaxel (2 mg/kg, i.p.) on days 1, 3, 5 and 7. RKT was administered 1 hour before each nociceptive test. The nociceptive threshold was determined by the von Frey filament test. Each point represents the mean ± S.E.M of 7–8 mice. *** P < 0.001 vs. 10%DMSO-DW group. ^##P < 0.01 vs. paclitaxel-DW group.</p

Liver fibrotic area was not observed in <i>Nrd1^−/−</i> mice fed the CDAA diet.

<p>A. Liver fibrosis was determined by Sirius red staining (red) in <i>Nrd1^+/+</i> and <i>Nrd1^−/−</i> mice at 4 (upper), 12 (middle), and 20 (lower) weeks in the livers of <i>Nrd1^+/+</i> and <i>Nrd1^−/−</i> mice fed the CSAA or CDAA diet. Fibrotic changes were not observed in <i>Nrd1^+/+</i> or <i>Nrd1^−/−</i> mice fed the CSAA diet (left). Fibrotic changes were prominent in <i>Nrd1^+/+</i> mice, but not in <i>Nrd1^−/−</i> mice fed the CDAA diet (right). Bars indicate 100 µm. B. Quantification of fibrotic areas. Fibrotic areas was observed and increased in a time-dependent manner only in the livers of <i>Nrd1^+/+</i> mice fed the CDAA diet. n = 5–8, each. *<i>P</i><0.05.</p

Liver fibrogenesis was not observed in <i>Nrd1^−/−</i> mice fed the HFD.

<p>A. Steatosis was observed in both <i>Nrd1^+/+</i> and <i>Nrd1^–/–</i> mice after 20-week HFD administration (right), but not in those fed a normal control diet (left). Bars indicate 100 µm. B. Quantification of triglyceride in the liver. Triglyceride was elevated in the livers of both <i>Nrd1^+/+</i> and <i>Nrd1^–/–</i> mice after 20-week HFD administration, although it was significantly higher in <i>Nrd1^+/+</i> mice. n = 4, each. *<i>P</i><0.05. C. Serum ALT levels were significantly elevated in <i>Nrd1^+/+</i> mice upon administration of the HFD, but were not elevated in other mouse groups. *<i>P</i><0.05. D. Fibrotic area was less prominent in <i>Nrd1^−/−</i> mice than in <i>Nrd1^+/+</i> mice (right). Bars indicate 100 µm. E. Fibrotic area was observed only in the livers of <i>Nrd1^+/+</i> mice fed the HFD (right). n = 5, each. *<i>P</i><0.05.</p

TNF-α was expressed in <i>Nrd1^+/+</i> and <i>Nrd1^−/−</i> mice fed the CDAA diet.

<p>A. Immunohistochemistry showed that TNF-α protein (red, arrowheads) was expressed in F4/80-positive Kupffer cells or macrophages (green, arrowheads) in the livers of both <i>Nrd1^+/+</i> and <i>Nrd1^−/−</i> mouse fed the CDAA diet for 20 weeks (right), but not in mice fed the CSAA diet for 20 weeks (left). A blue color indicates DAPI-positive nuclei. Bars indicate 50 µm. B. The number of F4/80-positive cells/×100 high-power field (HPF) in livers slightly increased (approximately 1.2 times) only in <i>Nrd1^+/+</i> mice fed the CDAA diet (right). C. Relative expression of mRNA are shown as relative values compared to those at 0 w. The mRNA expression level of CCR2 was increased in <i>Nrd1^+/+</i> mice fed the CDAA diet, and the levels were significantly higher than respective values in <i>Nrd1^−/−</i> mice fed the CDAA diet. *<i>P</i><0.05.</p

CDAA diet caused hepatic steatosis in <i>Nrd1^+/+</i> and <i>Nrd1^−/−</i> mice.

<p>A. Histology of the livers of <i>Nrd1^+/+</i> and <i>Nrd1^−/−</i> mice fed the CSAA (left) or CDAA (right) diets. Representative changes of the liver with regard to fat deposition at 4 (upper), 12 (middle), and 20 (lower) weeks during the experiments are depicted. Fat deposits were confirmed by oil red O staining shows (orange, inset). CV indicates central vein, and PT marks portal triad. Bars indicate 100 µm. B. Quantification of triglyceride in the liver. Triglyceride in the liver was increased in the livers of both <i>Nrd1^+/+</i> and <i>Nrd1^−/−</i> mice during administration of the CDAA or CSAA diets, although it was significantly more prominent in <i>Nrd1^+/+</i> mice. n = 4–5, each. *<i>P</i><0.05. C. There was no significant difference in the liver/body weight ratio between <i>Nrd1^+/+</i> and <i>Nrd1^−/−</i> mice during the experiments.</p