1. Volume 117, Issue 5, Pages 1198-1204 (November 1999)
Rapamycin inhibits hepatic stellate cell proliferation in vitro and limits fibrogenesis in an in vivo model of liver fibrosis 
Jianliang Zhu, Jian Wu, Edward Frizell, Shu-Ling Liu, Reza Bashey, Raphael Rubin, Pamela Norton, Mark A. Zern 
Gastroenterology 
Volume 117, Issue 5, Pages (November 1999)
DOI: /S (99)
Copyright © 1999 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Trichrome preparations of representative liver samples from the (A) group treated with CCI4 and vehicle and (B) group treated with both CCl4 and rapamycin.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

3. Fig. 1
Trichrome preparations of representative liver samples from the (A) group treated with CCI4 and vehicle and (B) group treated with both CCl4 and rapamycin.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

4. Fig. 2
Total collagen quantitation performed in defatted liver samples from the 3 experimental groups: control and vehicle, CCl4 and vehicle, and CCl4 and rapamycin. Liver samples were defatted, hydrolyzed, and assayed for hydroxyproline as described in Materials and Methods. *P < 0.05 compared with CCl4 + vehicle.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

5. Fig. 3
Representative transcripts of 28S, collagen type I, and TGF-β1 by Northern blot analysis. (A) Autoradiograms of Northern blots performed with RNA extracted from the livers of animals from the 3 experimental groups. Lanes 1 and 2, control and vehicle; lanes 3 and 4, CCl4 and vehicle; lanes 5 and 6, CCl4 plus rapamycin. Total RNA (20 μg) was loaded into each well, electrophoresed, transferred, and hybridized with different cDNA probes as described in Materials and Methods. The first row shows 28S ribosomal RNA; the second row shows collagen type I (α2), and the third row shows TGF-β1. (B) Results of densitometry scanning of the transcripts. All transcripts from each group were scanned, and the density of tracing is expressed as means ± SEM. **P < 0.01 compared with CCl4 + vehicle.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

6. Fig. 3
Representative transcripts of 28S, collagen type I, and TGF-β1 by Northern blot analysis. (A) Autoradiograms of Northern blots performed with RNA extracted from the livers of animals from the 3 experimental groups. Lanes 1 and 2, control and vehicle; lanes 3 and 4, CCl4 and vehicle; lanes 5 and 6, CCl4 plus rapamycin. Total RNA (20 μg) was loaded into each well, electrophoresed, transferred, and hybridized with different cDNA probes as described in Materials and Methods. The first row shows 28S ribosomal RNA; the second row shows collagen type I (α2), and the third row shows TGF-β1. (B) Results of densitometry scanning of the transcripts. All transcripts from each group were scanned, and the density of tracing is expressed as means ± SEM. **P < 0.01 compared with CCl4 + vehicle.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

7. Fig. 4
tTG activity in the livers of rats from the 3 experimental groups. tTG activity was measured using the filter paper assay of Lorand et al.22 Levels in the CCl4 + vehicle group were greater than those of the control group (P < 0.01), and levels in the CCl4 + rapamycin group were less than those in the CCl4 + vehicle group (*P < 0.01).
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

8. Fig. 5
(A) Effect of rapamycin on the proliferation of hepatic stellate cells stimulated by PDGF. Hepatic stellate cells were cultured for 2 days and treated with PDGF alone or plus rapamycin. DMSO was used for dissolving rapamycin and was also included as DMSO control at the concentration for dissolving the highest concentration of rapamycin (10−9 mol/L). [3H]Thymidine incorporation into hepatic stellate cells was used as an indication of the cell proliferation as described in Materials and Methods. The data are summarized from 3 independent experiments. ΔP < 0.05 and ΔΔP < 0.01 compared with controls; **P < 0.01 compared with PDGF alone. (B) Effect of rapamycin or FK506 on the proliferation of hepatic stellate cells stimulated by PDGF. Stellate cells were cultured and treated with PDGF alone or with the immunosuppressant agent as indicated. The data represent n = 6 in each group and are summarized from 2 independent experiments. **P < 0.01 compared with PDGF alone.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions

9. Fig. 5
(A) Effect of rapamycin on the proliferation of hepatic stellate cells stimulated by PDGF. Hepatic stellate cells were cultured for 2 days and treated with PDGF alone or plus rapamycin. DMSO was used for dissolving rapamycin and was also included as DMSO control at the concentration for dissolving the highest concentration of rapamycin (10−9 mol/L). [3H]Thymidine incorporation into hepatic stellate cells was used as an indication of the cell proliferation as described in Materials and Methods. The data are summarized from 3 independent experiments. ΔP < 0.05 and ΔΔP < 0.01 compared with controls; **P < 0.01 compared with PDGF alone. (B) Effect of rapamycin or FK506 on the proliferation of hepatic stellate cells stimulated by PDGF. Stellate cells were cultured and treated with PDGF alone or with the immunosuppressant agent as indicated. The data represent n = 6 in each group and are summarized from 2 independent experiments. **P < 0.01 compared with PDGF alone.
Gastroenterology  , DOI: ( /S (99) )
Copyright © 1999 American Gastroenterological Association Terms and Conditions