1. Volume 44, Issue 3, Pages 568-575 (March 2006)
Salvage effect of the vascular endothelial growth factor on chemically induced acute severe liver injury in rats 
Tadashi Namisaki, Hitoshi Yoshiji, Hideyuki Kojima, Junichi Yoshii, Yasuhide Ikenaka, Ryuichi Noguchi, Shinya Sakurai, Koji Yanase, Mitsuteru Kitade, Masaharu Yamazaki, Kiyoshi Asada, Masahito Uemura, Mitsutoshi Nakamura, Hiroshi Fukui 
Journal of Hepatology 
Volume 44, Issue 3, Pages (March 2006)
DOI: /j.jhep
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

2. Fig. 1
The therapeutic effect of the recombinant rat VEGF (rVEGF) on the survival rate of rats with Gal-N+LPS-induced acute severe liver injury (ALI). The rats were injected with Gal-N (500mg/kg) and LPS (50μg/kg) intraperitoneally, and randomly divided into two groups. In one group, rVEGF (50ng/kg) was injected into the tail vein at 0 and 12h after Gal-N+LPS intoxication, while the other group received the same amount of phosphate buffer saline (PBS), and was designated as a control group. Fifteen rats from each experimental group were used for monitoring the survival rate chronologically. Without rVEGF treatment, more than half of the rats died within 24h after Gal-N+LPS intoxication. The survival rate decreased to 27% at 48h in this group. On the other hand, all the rats survived in the rVEGF-treated group even at 48h. No rats died from 48h thereafter in both groups. Dash and solid lines represent rVEGF-treated group and untreated group, respectively.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

3. Fig. 2
The effect of rVEGF on the serum ALT level in the liver following Gal-N+LPS intoxication. Injection of Gal-N+LPS resulted in a marked increase of the serum ALT level between 12 and 72h following the intoxication, and reached a peak level at 24h. The serum ALT level in the rVEGF-treated group was significantly lower than in the untreated group between 12 and 48h following intoxication. Dashed and solid lines represent rVEGF-treated group and untreated group, respectively. Each point represents the mean±SD (n=5). *, ** Statistically significant differences between the experimental groups (P<0.01 and <0.05, respectively).
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

4. Fig. 3
The chronological effect of rVEGF on the VEGF expression in the liver following Gal-N+LPS intoxication. The hepatic VEGF expression increased 24h after Gal-N+LPS intoxication, and thereafter became gradually prominent until 168h later. The rVEGF administration resulted in a more than two-fold increase of the VEGF expression at 24h, and returned to a level similar to that of the untreated liver at 48h. Dashed and solid lines represent rVEGF-treated group and untreated group, respectively. Each point represents the mean±SD (n=5). * Statistically significant differences between the experimental groups (P<0.01).
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

5. Fig. 4
The chronological proliferation of the hepatocytes and sinusoidal endothelial cells (SEC) in the liver following Gal-N+LPS intoxication. (A) The number of hepatocytes showing DNA synthesis started to increase 24h after intoxication, with a peak at 36h, and decreased gradually thereafter. The proliferation rate of the rVEGF-treated group was significantly higher than that of the untreated group from 36 to 96h. (B) The number of labeled SEC exhibited a delayed and slower regenerative response in comparison to the hepatocytes, reaching a peak at 96h. The proliferation rate of SEC in the rVEGF-treated group was higher from 24 to 96h as compared with the untreated group. The black and white bars represent with or without rVEGF-treated group, respectively. The data represent the mean±SD (n=5). *, ** Statistically significant differences between the experimental groups (P<0.01 and <0.05, respectively).
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

6. Fig. 5
Representative features of the effect of rVEGF on the maintenance of SEC structure. SEC were visualized by immunostaining of SE-1, which is a specific marker for SEC. As compared with before intoxication (A), destruction of the SEC structure in the untreated group (B) already started to appear even at 6h after intoxication. At 24h, the initial SEC construction in the untreated group was mostly obliterated (C). The rVEGF treatment significantly prevented the destruction of SEC architecture at 6 and 24h after Gal-N+LPS intoxication (D and E, respectively). Original magnification ×40.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

7. Fig. 6
Semiquantitative analysis of the SE-1-immunopositive vessels in the liver following Gal-N+LPS intoxication. The microvessel density already significantly decreased even at 6h after intoxication. The destruction of SEC progressed thereafter. The rVEGF treatment significantly prevented the destruction of SEC architecture in the liver following Gal-N+LPS intoxication even at 6h after intoxication, and the preventive effect persisted until 48h. The black and white bars represent with or without rVEGF-treated group, respectively. The data represent the mean±SD (n=5). *, ** Statistically significant differences between the indicated groups (P<0.01 and <0.05, respectively). (−) and (+) represent rVEGF-untreated group and -treated group, respectively. MV, microvessel.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

8. Fig. 7
Effect of rVEGF on the in vitro cytotoxicity (A) and apoptosis (B) of EC induced by Gal-N+LPS intoxication. The cell cytotoxicity and apoptosis were measured by the MTT assay and ELISA system, respectively, as described in Section 2. (A) The viable cells were already significantly lower at 6h after Gal-N+LPS intoxication as compared with the untreated group, and progressed until 24h. The rVEGF treatment significantly improved the cell survival at all indicated points. (B) The Gal-N+LPS intoxication significantly induced the apoptosis of EC even at 6h. The treatment of rVEGF markedly attenuated the apoptosis of EC following Gal-N+LPS intoxication at all indicated points. The black and white bars represent with or without rVEGF-treated group, respectively. The data represents the mean±SD (n=6). *, ** Statistically significant differences between the indicated groups (P<0.01 and <0.05, respectively).
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions