Volume 125, Issue 3, Pages 882-890 (September 2003) Sinusoidal obstruction syndrome (veno-occlusive disease) in the rat is prevented by matrix metalloproteinase inhibition 1   Laurie D. DeLeve, Xiangdong Wang, Jeffrey Tsai, Gary Kanel, Steven Strasberg, Zoltan A. Tokes  Gastroenterology  Volume 125, Issue 3, Pages 882-890 (September 2003) DOI: 10.1016/S0016-5085(03)01056-4

Figure 1 Gelatin zymography for the detection of matrix metalloproteinases. (A) Lanes 1 and 6 are purified MMP-2 and -9 standards identifying the pro (68- and 92-kilodalton) and active (62- and 84-kilodalton, respectively) forms of the enzymes. Representative liver samples of untreated rats, and rats 12, 24, and 48 hours after monocrotaline treatment are illustrated in lanes 2–5. (B) Graphic illustration of increased combined availability of pro and active MMP-2 (open bars) and -9 (black bars) after monocrotaline (Mct) treatment. Combined availability was determined by measuring the total diminished absorbance of Coomassie blue-stained gelatin gels at the band locations of the pro and active forms of MMPs. P < 0.05 for MMP-9, control vs. 12, 24, or 48 hours; P < 0.05 for MMP-2, control vs. 48 hours (n = 3); unt, untreated. Gastroenterology 2003 125, 882-890DOI: (10.1016/S0016-5085(03)01056-4)

Figure 2 MMP activity. SEC were plated on DQ gelatin, fluorescein conjugate. Cells were exposed to monocrotaline (Mct; 0, 1, 2 or 4 mmol/L) for 4 hours (n = 4); in each panel, columns from left to right indicate monocrotaline concentrations ranging from 0 to 4 mmol/L. (A) Response of SEC exposed to monocrotaline 0–4 mmol/L for 4 hours. (B) SEC were treated with monocrotaline 0–4 mmol/L plus 100 μmol/L of the MMP-2/MMP-9 inhibitor 2-[(4-biphenylsulfonyl)amino]-3-phenyl-propionic acid. (C) Cells were pretreated with 5 μmol/L of the F-actin stabilizer jasplakinolide, followed by 4 hours of monocrotaline 0–4 mmol/L. Two-factor analysis of variance with replication: P < 0.05. Least-significant difference (LSD) for comparison with control: ¶P < 0.001. LSD for comparison with monocrotaline alone: ∗P < 0.025; ∗∗P < 0.005; ∗∗∗P < 0.001. Gastroenterology 2003 125, 882-890DOI: (10.1016/S0016-5085(03)01056-4)

Figure 3 Monocrotaline effect on F-actin. Confocal microscopy image of SEC stained with rhodamine/phalloidin to visualize F-actin (polymerized form). Control SEC were plated for 4 hours (A), and treated SEC were exposed for 4 hours to 4 mmol/L of monocrotaline (B). The graph (C) depicts viability and F-actin polymerization in SEC exposed to 2 mmol/L of monocrotaline for 16 hours with (hatched columns) or without (solid columns) pretreatment with jasplakinolide to stabilize F-actin. Cell viability and F-actin fluorescence are expressed as a percentage of control SEC (i.e., not treated with monocrotaline). Analysis of variance was significant for both 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) and actin fluorescence; by least-significant difference, with vs. without jasplakinolide: ∗P < 0.02 (n = 3); ∗∗P < 0.001 (n = 4). Gastroenterology 2003 125, 882-890DOI: (10.1016/S0016-5085(03)01056-4)

Figure 4 Effect of MMP inhibitors on the histology of monocrotaline-treated rats. Rats were treated with monocrotaline 160 μg/kg IG and killed on day 4. The effect of MMP inhibition by doxycycline (A) can be contrasted with the effect of isochlorotetracycline (B), a chemically modified tetracycline with a weak MMP inhibitory effect. (B) Full-blown early SOS with loss of sinusoidal and venular endothelial cells and centrilobular necrosis, hemorrhage, and congestion; of note, the nucleated cells adherent within the central venule are monocytes. (C) The effect of the MMP-2/MMP-9 inhibitor 2-[(4-biphenylsulfonyl)amino]-3-phenyl-propionic acid on monocrotaline-induced SOS. Gastroenterology 2003 125, 882-890DOI: (10.1016/S0016-5085(03)01056-4)

Figure 5 Proposed mechanism of sinusoidal obstruction syndrome. RBC, red blood cell. Gastroenterology 2003 125, 882-890DOI: (10.1016/S0016-5085(03)01056-4)