Volume 11, Issue 2, Pages 121-128 (October 2004) Direct inhibition by a statin of TNFα-induced leukocyte recruitment in rat pial venules — in vivo confocal microscopic study  Ruriko Obama, Hideyuki Ishida, Shunya Takizawa, Chizuko Tsuji, Hiroe Nakazawa, Yukito Shinohara  Pathophysiology  Volume 11, Issue 2, Pages 121-128 (October 2004) DOI: 10.1016/j.pathophys.2004.07.001 Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 Number of rolling leukocytes in the six groups. Rolling leukocytes were counted and expressed as the number of cells/mm2. Both simvastatin and pravastatin treatments tended to decrease the number of rolling leukocytes, but there was no statistically significant difference between each statin and the corresponding vehicle group. An asterisk denotes P < 0.01 as compared with the control group. Pathophysiology 2004 11, 121-128DOI: (10.1016/j.pathophys.2004.07.001) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Representative images of venules with adhered leukocytes in the six groups: (A) control; (B) TNFα alone; (C) vehicle of simvastatin with TNFα; (D) simvastatin with TNFα; (E) vehicle of pravastatin with TNFα; (F) pravastatin with TNFα. In the control group, there were essentially no adhered leukocytes (A). Abundant adhered leukocytes were observed in all groups with TNFα treatment (B–F). The number of adhered leukocytes in the simvastatin group was scant compared with that in the vehicle group (D). The calibration bar represents 100μm. Pathophysiology 2004 11, 121-128DOI: (10.1016/j.pathophys.2004.07.001) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 The number of adhered leukocytes in the six groups. Adhered leukocytes were counted and expressed as the number of cells/mm2. All values are means ± S.E. A single asterisk denotes P < 0.01 as compared with the control group and double asterisks denotes P < 0.01 as compared with the corresponding vehicle group. The number of adhered leukocytes was significantly fewer in the simvastatin group than in the corresponding vehicle group. The number of adhered leukocytes appeared to be fewer in the pravastatin group than in the corresponding vehicle group, but without statistical significance. Pathophysiology 2004 11, 121-128DOI: (10.1016/j.pathophys.2004.07.001) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 Representative staining of P-selectin and ICAM-1 in rat brain: (A) double staining of P-selectin and endothelial cells; (B) double staining of ICAM-1 and endothelial cells. P-selectin was visualized as brown staining, while vessels were shown in blue by alkaline phosphatase staining, a marker of vessels. ICAM-1 and P-selectin were visualized as brown staining in the whole vessel wall, including endothelial cells. The calibration bar represents 50μm. Pathophysiology 2004 11, 121-128DOI: (10.1016/j.pathophys.2004.07.001) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 Ratio of P-selectin-positive or ICAM-1-positive vessels in the six groups: (A) ratio of P-selectin-positive vessels in the brain; (B) ratio of ICAM-1-positive vessels in the brain. In the brain hemisphere of each rat, five fields were observed and P-selectin-positive or ICAM-1-positive vessels were counted. Results were expressed as the ratio of numbers of positive vessels (PV)/total vessels (TV). P-selectin-positive vessels tended to be more frequent in the groups treated with TNFα, but there was no statistically significant difference among groups. Neither TNFα nor statin pretreatment significantly increased ICAM-1-positive vessels. Pathophysiology 2004 11, 121-128DOI: (10.1016/j.pathophys.2004.07.001) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions

Fig. 6 eNOS mRNA levels in the six groups. The eNOS mRNA levels were similar in all groups. Pathophysiology 2004 11, 121-128DOI: (10.1016/j.pathophys.2004.07.001) Copyright © 2004 Elsevier Ireland Ltd Terms and Conditions