Volume 41, Issue 5, Pages 837-844 (November 2004) Regional and transient ischemia/reperfusion injury in the liver improves therapeutic efficacy of allogeneic intraportal hepatocyte transplantation in low-density lipoprotein receptor deficient Watanabe rabbits  Masoumeh Attaran, Andrea Schneider, Christiane Grote, Caroline Zwiens, Peer Flemming, Klaus F. Gratz, Andrea Jochheim, Matthias J. Bahr, Michael P. Manns, Michael Ott  Journal of Hepatology  Volume 41, Issue 5, Pages 837-844 (November 2004) DOI: 10.1016/j.jhep.2004.07.014 Copyright © 2004 European Association for the Study of the Liver Terms and Conditions

Fig. 1 Histological analysis of rt-IRP 24h after rt-IRP (90min). (A) Almost complete necrosis of the liver tissue in a representative histological section. (B) Regeneration of the liver tissue after 16 days. Specimen were stained with hematoxylin and eosin solution (original magnification 100×). [This figure appears in colour on the web.] Journal of Hepatology 2004 41, 837-844DOI: (10.1016/j.jhep.2004.07.014) Copyright © 2004 European Association for the Study of the Liver Terms and Conditions

Fig. 2 Histological grading of liver tissue specimen after rt-IRP. The extend of tissue necrosis as judged by an experienced liver pathologist showed a gradual decline over a period of 16 days. Necrotic cell structures (i.e., hepatocytes with lost or condensed nuclei) were counted and expressed as percent of the total number of cells in a microscopic field. Journal of Hepatology 2004 41, 837-844DOI: (10.1016/j.jhep.2004.07.014) Copyright © 2004 European Association for the Study of the Liver Terms and Conditions

Fig. 3 BrdU incorporation and hepatobiliary sequence scintigraphy after rt-IRP injury. In sections derived from the rt-IRP treated liver lobe BrdU positive cell nuclei were numerous at day 4 of the regeneration phase (A) but not seen in the untreated liver lobes (C) of the same animal. Cell nuclei were counterstained by DAPI (B and D) (original magnification 400×). Before rt-IRP treatment hepatobiliary sequence scintigraphy showed homogenous tracer uptake of the liver (E). One day after rt-IRP of the left liver lobe the Technetium-99m-IDA tracer signal was reduced indicating selective necrosis of the liver tissue. In addition to the liver the figure shows excretion of the radioactive material into the intestine (F). [This figure appears in colour on the web.] Journal of Hepatology 2004 41, 837-844DOI: (10.1016/j.jhep.2004.07.014) Copyright © 2004 European Association for the Study of the Liver Terms and Conditions

Fig. 4 Analysis of rabbit serum samples. Total serum protein and serum bilirubin levels did not change throughout the observation period of 28 days after rt-IRP. AST and ALT enzyme activities increased one day after rt-IRP and declined to baseline levels within 28 days after rt-IRP. Journal of Hepatology 2004 41, 837-844DOI: (10.1016/j.jhep.2004.07.014) Copyright © 2004 European Association for the Study of the Liver Terms and Conditions

Fig. 5 Repeated HcTx in WHHL rabbits. In the sham treated group I (rt-IRP, w/o HcTx,) LDL cholesterol levels slightly increased during the observation time of 119 days. In the experimental group II repeated HcTx and rt-IRP lowered blood LDL cholesterol levels from 731±32mg/dl to 416±15mg/dl (P<0.001). In group III (w/o rt-IRP, with HcTx) LDL cholesterol levels were decreased from mean pretreatment values of 660±63 to 587±49mg/dl after 119 days (P<0.001). Analysis of variance showed that LDL levels of all groups were significantly different with a P value of <0.05 from day 21 on and with a P<0.001 from day 63 on (A). Differences of LDL cholesterol levels at the end of the four months period in %±SEM of the corresponding pre-treatment values of all three groups (B). [This figure appears in colour on the web.] Journal of Hepatology 2004 41, 837-844DOI: (10.1016/j.jhep.2004.07.014) Copyright © 2004 European Association for the Study of the Liver Terms and Conditions

Fig. 6 Detection of LDL receptor mRNA in the liver of rabbits. The LDL receptor mRNA fragment was not detected in liver tissue from WHHL-rabbits, but was present in NZR liver tissue and in samples from transiently ischemic lobes of four representative hepatocyte transplanted WHHL rabbits. PCR fragments were sequenced to confirm identity. Journal of Hepatology 2004 41, 837-844DOI: (10.1016/j.jhep.2004.07.014) Copyright © 2004 European Association for the Study of the Liver Terms and Conditions

Fig. 7 Western blot of LDL receptor protein in liver tissue of New Zealand, WHHL and transplanted WHHL rabbits. New Zealand rabbits showed a major immunoreactive band at 130,000D representing the mature receptor and a minor band at 90,000D (precursor protein). WHHL rabbits had a minor band at 120,000D, but large amounts of the 90,000D precursor protein. Transplanted WHHL rabbits show a major band at 130,000D and a substantial decrease of the precursor protein at 90,000D. Journal of Hepatology 2004 41, 837-844DOI: (10.1016/j.jhep.2004.07.014) Copyright © 2004 European Association for the Study of the Liver Terms and Conditions