1. Volume 39, Issue 4, Pages 538-546 (October 2003)
Oxidative stress, KLF6 and transforming growth factor-β up-regulation differentiate non- alcoholic steatohepatitis progressing to fibrosis from uncomplicated steatosis in rats 
Peter Stärkel, Christine Sempoux, Isabelle Leclercq, Michel Herin, C Deby, Jean-Pierre Desager, Yves Horsmans 
Journal of Hepatology 
Volume 39, Issue 4, Pages (October 2003)
DOI: /S (03)00360-X

2. Fig. 1
Histology in animals fed an orotic acid (OA) and methionine-choline deficient (MCD) diet for 6 weeks. After 6 weeks of OA treatment, (A) a diffuse microvesicular steatosis was observed in the hepatic lobule, in absence of any inflammation. (hematoxylin–eosin, ×185); In the MCD group (B), a severe and diffuse macrovesicular steatosis was present associated with foci of inflammation (arrow) (hematoxylin–eosin, ×185).
Journal of Hepatology  , DOI: ( /S (03)00360-X)

3. Fig. 2
α-Smooth muscle actin immunohistochemistry in animals fed an orotic acid (OA) and methionine-choline deficient (MCD). After 6 weeks of OA treatment, (A) α-smooth muscle actin-positive cells were only observed in vascular smooth muscle layers and around the central veins, whereas in the MCD group (B), α-smooth muscle actin positive hepatic stellate cells appeared in the sinusoids (arrows) in addition to vessels. After 16weeks of MCD diet (C), activated hepatic stellate cells strongly positive for α-smooth muscle actin (arrows) were detected adjacent to fibrotic and inflammatory areas (immunoperoxidase, ×450).
Journal of Hepatology  , DOI: ( /S (03)00360-X)

4. Fig. 3
Long-term histological outcome of rats fed the orotic acid (OA) and methionine-choline deficient (MCD) diet. Sirius red staining on liver sections to assess development of liver fibrosis. (A) In the OA model, no fibrosis was observed after 18 weeks. (B) In the MCD diet model, a perivenular fibrosis with septa extending into the liver parenchyma developed after 16 weeks. Note the regression of steatosis, only located in the centrolobular area (Sirius red, ×200).
Journal of Hepatology  , DOI: ( /S (03)00360-X)

5. Fig. 4
Overall liver lipid peroxidation assessed by quantification of thiobarbituric acid reactive substances (TBARs) in control animals and rats treated with orotic acid (OA) or a methionine-choline deficient (MCD) diet for 2, 6, and 16 weeks. Results in the different treatment groups are expressed as TBARs formation in nmol per g of liver. Mean values±SEM (n=4 per group) are shown. P values in treatment groups compared to controls are indicated at the top of each column. A dramatic increase in lipid peroxidation was observed in MCD animals from the second week of treatment onwards whereas only minor changes occurred in the OA animals.
Journal of Hepatology  , DOI: ( /S (03)00360-X)

6. Fig. 5
Quantitative PCR of TGFβ1 mRNA expression in controls and different treatment groups (OA, orotic acid; MCD, methionine-choline deficient). Mean values±SEM are represented. Fourteen (n=14) animals were used in the control group and, at least, seven (n=7) in the treatment groups at each time point. P values in treatment groups compared to controls are indicated at the top of each column. TGFβ1 expression did not change significantly in the OA group. In contrast, TGFβ1 levels started to rise in the MCD group from the sixth week of treatment onwards.
Journal of Hepatology  , DOI: ( /S (03)00360-X)

7. Fig. 6
Quantitative PCR of KLF6 mRNA expression in controls and different treatment groups (OA, orotic acid; MCD, methionine-choline deficient). Mean values±SEM are represented. Fourteen (n=14) animals were used in the control group and, at least, seven (n=7) in the treatment groups at each time point. P values in treatment groups compared to controls are indicated at the top of each column. After an initial and early increase in KLF6 mRNA in the OA group, KLF6 levels rapidly returned to control values at 6 weeks of treatment. In the MCD group, however, KLF6 mRNA expression increased very significantly at the sixth week of treatment and remained elevated at 16 weeks.
Journal of Hepatology  , DOI: ( /S (03)00360-X)