Embolus at the origin of the occluded MCA. (A) Schematic representation of a ventral view of a rat brain is shown. The enlarged boxed area from panel A shows the intracranial segment of the ICA and the MCA. A large fragment of Evans blue dye (B, red) was detected within the origin of the MCA and intracranial segment of the ICA, which blocked plasma perfusion (panel B, green) from a representative rat treated with tPA alone. The combination treatment with atorvastatin at 4 h and tPA at 6 h resulted in only a small fragment of Evans blue dye (C, red) at the origin of the MCA and the intracranial segment of the ICA, which were perfused by FITC-dextran (panel C, green). (D) shows the quantitative data of an embolus (n=4 per group). Bar in panels B and C=400 μm.

Infarct volume. Bar graph shows the effects of monotherapy with tPA and atorvastatin and the combination therapy on infarct volume assessed 7 days after MCA occlusion. Values are mean ± s.e. *P < 0.05 as compared with that of the saline-treated group. ^†P < 0.05 as compared with that of the tPA alone group. ^‡P < 0.05 as compared with that of the atorvastatin alone group.

Thrombocyte, fibrin/fibrinogen, MPO, ICAM-1, MMP9, and collagen type IV immunoreactive cerebral vessels and FITC-dextran perfusion. Double immunofluorescent staining (A–F) shows thrombocyte (panels A and B, green), fibrin/fibrinogen (panels C and D, green), and MPO (panels E and F, green) in cerebral blood vessels (Eba, panels A–F, red) of representative rats treated with tPA alone (panels A, C, and E) and with a combination of atorvastatin at 4 h and tPA at 6 h (panels B, D, and F). (G–L) The cerebral microvessels perfused with FITC-dextran in a normal rat (panel H), and in representative rats treated with saline (panel I), tPA (panel J), atorvastatin (panel K), and a combination of tPA at 4 h and atorvastatin at 6 h (panel L) 30 h after MCA occlusion (n=4 per group) is shown. (M–P) Represents the quantitative data of the number of thrombocyte (panel M), fibrin/fibrinogen (panel N), MPO immunoreactive cerebral vessels (panel O), and microvessels perfused with FITC-dextran (panel P). Double immunostaining shows that cerebral blood vessels with diffused collagen type IV immunoreactivity (Q, R, T, U; red) were ICAM-1 (panels Q and R, green) or MMP9 positive (panels T and U, green) in representative rats treated with tPA alone (panels Q and T), and with a combination of atorvastatin at 4 h and tPA at 6 h (panels R and U). Panel S and (V) show intensive collagen type IV immunoreactivity (red) in contralateral homologous areas wherein ICAM-1 (panel C, green) and MMP9 (panel F, green) were absent from a representative rat treated with tPA alone. Double immunostaining (W–Y) shows that ICAM-1 (panels W and Y, green; arrows) and MMP9 (panels X and Y, brown; arrowheads) immunoreactivities were in different vessels of a representative rat treated with tPA alone. Panel Y is a merged image. Ctx, cortex; Str, striatum; CC, corpus callosum. *P < 0.05 as compared with that of saline-treated group. ^†P<0.05 as compared with that of the tPA alone group. ^‡P<0.05 as compared with that of the atorvastatin alone group. ^§P<0.05 for treatment interaction (synergistic effect). Bars for panels A–F=40 μm; panels H–L=1 mm; panels Q–Y=40 μm.