Potential of 99mTc-MIBI SPECT imaging for evaluating non-alcoholic steatohepatitis induced by methionine-choline-deficient diet in mice
© Rokugawa et al.; licensee Springer. 2014
Received: 30 July 2014
Accepted: 28 September 2014
Published: 9 October 2014
Hepatic mitochondrial dysfunction has been implicated in pathological conditions leading to non-alcoholic steatohepatitis (NASH). Technetium-99 m-2-methoxyisobutyl-isonitrile (99mTc-MIBI), a lipophilic cationic myocardial perfusion agent, is retained in the mitochondria depending on membrane potential. The aim of this study was to investigate the feasibility of 99mTc-MIBI for evaluating the hepatic mitochondrial dysfunction induced by methionine-choline-deficient (MCD) diet in mice.
Male C57Black6J/jcl mice were fed a MCD diet for up to 4 weeks. SPECT scan (N =6) with 99mTc-MIBI was performed at 2 and 4 weeks after MCD diet. Mice were imaged with small-animal SPECT/CT under isoflurane anesthesia. Radioactivity concentrations of the liver were measured, and the time of maximum (Tmax) and the elimination half-life (T1/2) were evaluated. After SPECT scan, liver histopathology was analyzed to evaluate steatosis and inflammation. Non-alcoholic fatty liver disease (NAFLD) activity score was obtained from the histological score of hepatic steatosis and inflammation. Blood biochemistry and hepatic ATP content were also measured (N =5 to 6).
Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were significantly elevated at 2 and 4 weeks after MCD diet. A decrease in hepatic ATP content was also observed in MCD-fed mice. 99mTc-MIBI SPECT imaging clearly showed the decrease of hepatic 99mTc-MIBI retention in MCD-fed mice compared to control mice. T1/2 after 99mTc-MIBI injection was significantly decreased in the liver of MCD-fed mice (control, MCD 2 weeks, and MCD 4 weeks, T1/2 = 57.6, 37.6, and 19.8 min, respectively), although no change in Tmax was observed in MCD-fed mice. SPECT data and histological score showed that the negative correlation (r = -0.74, p <0.05) between T1/2 and NAFLD activity score was significant.
Hepatic 99mTc-MIBI elimination was increased with increase in NAFLD activity score (NAS) in mice fed MCD diet for 2 and 4 weeks. These results suggest that 99mTc-MIBI SPECT imaging might be useful for detecting hepatic mitochondrial dysfunction induced by steatosis and inflammation such as NAFLD or NASH.
Keywords99mTc-MIBI NASH Mice MCD diet
Non-alcoholic fatty liver disease (NAFLD) is one of the most common forms of chronic liver disease recognized as a hepatic manifestation of metabolic syndrome without a history of alcoholic abuse . NAFLD encompasses a wide spectrum of conditions ranging from simple steatosis to non-alcoholic steatohepatitis (NASH) with or without fibrosis, cirrhosis, and hepatocellular carcinoma . A two-hit theory has been proposed for NASH pathogenesis. The first hit refers to factors that promote hepatic steatosis, and the second hit refers to factors leading from hepatic steatosis to steatohepatitis ,. One of the second hit factors is the formation of reactive oxygen species (ROS) ,. ROS directly damages respiratory chain polypeptides and oxidizes the unsaturated lipid of cytoplasmic hepatic fat deposits to cause lipid peroxidation. Both ROS and lipid peroxidation products attack mitochondrial DNA . Oxidative mitochondrial DNA lesions and mitochondrial DNA depletion may impair the synthesis of respiratory chain polypeptides. These effects may further block the flow of electrons in the respiratory chain to further increase mitochondrial ROS formation and decrease the mitochondrial membrane potential ,. Thus, NASH is considered to be a mitochondrial disease because mitochondrial dysfunction in the liver would be involved in all successive steps in the induction of NASH . It has been reported that patients with NASH present ultrastructural mitochondrial alterations , impairment of hepatic ATP synthesis , and increased ROS production ,. Therefore, mitochondrial dysfunction is considered to have an important role in the liver during NASH progression since mitochondrial dysfunction causes overproduction of ROS inducing lipid peroxidation, inflammation, and cell death.
Technetium-99 m-2-methoxy-isobutyl-isonitrile (99mTc-MIBI) is a SPECT imaging probe which is used for myocardial perfusion imaging . This lipophilic cationic imaging probe predominantly accumulates in the mitochondria, where it is retained in response to the electrical potential generated across the membrane bilayer -. Therefore, 99mTc-MIBI retention in the mitochondria might be related to mitochondrial function ,. Masuda et al. have recently reported that 99mTc-MIBI scintigraphy is useful for discriminating NASH from simple steatosis in clinical studies . These findings suggest the possibility of 99mTc-MIBI imaging for the evaluation of hepatic mitochondrial function in the liver disease state such as NASH and NAFLD. The present study was undertaken to investigate the feasibility of 99mTc-MIBI imaging for evaluating hepatic dysfunction using mice fed methionine-choline-deficient (MCD) diet, one of the most commonly used NASH animal models ,.
Animals and MCD diet
Male C57Black6J/jcl mice, aged 8 weeks, were purchased from CLEA Japan (Shizuoka, Japan). Mice were studied at 2 and 4 weeks after MCD diet (Dyets, Bethlehem, PA, USA) feeding or normal diet feeding (control group). They were allowed free access to chow and tap water and housed in a temperature-controlled room maintained on a 12-h light/dark cycle with lights on at 8:00 am. The experimental protocols were reviewed and approved by the Institutional Animal Care and Use Committee of Shionogi Research Laboratories, Osaka University Graduate School of Medicine, and Animal Care Committee of Chiba University.
All SPECT data were reconstructed by a 3D-ordered subset expectation maximization (3D-OSEM) algorithm method with two subsets and five iterations in FLEX-RECON software. Imaging data were analyzed using AMIDE 0.9.2 software. 3D region of interest (ROI) was put on the liver tissue except for the portal area and estimated liver time-activity curves (TACs). TACs were normalized to liver activity peak. The half-life (T1/2), peak time (Tmax), and area under the curve (AUC) for each individual animal were calculated from the data of TAC using WinNonlin. All the parameters were then averaged for all the control mice and the mice fed the MCD diet for 2 and 4 weeks. Because the hepatic TACs of 99mTc-MIBI did not reach a constant level within 45 min, we used the first-order intermediate equation to fit the kinetic data and generate the best fitted liver curves .
Hepatic ATP content
The content of hepatic ATP was measured by HPLC according to the method of Dai et al. . Briefly, tissue samples were homogenized in homogenized buffer (0.3 M HClO4 and 1 mM EDTA-Na) and centrifuged at 14,000 rpm for 2 min. A 20-μL portion of the supernatant was injected into an HPLC system for ATP content determination. HPLC analysis was performed with Chemcosorb 5-I-C18 (4.6 mm × 300 mm, Chemco Scientific Co., Ltd., Osaka, Japan) at 1 mL/min. The composition of the mobile phase was 0.2 M NH4H2PO4 adjusted to pH =4.1 with 1 N HCl. The detection wavelength was 254 nm and the retention time of ATP was 16.3 min. ATP was calculated using an internal standard method.
Histology and blood biochemistry
After SPECT scans, mice were sacrificed by exsanguination under isoflurane anesthesia. Plasma was collected and assayed for the content of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (TC), and high-density lipoprotein cholesterol (HDLC). The right hepatic lobes were fixed in 10% formalin and sectioned, and the sections were stained with hematoxylin and eosin (H&E). The liver histopathology was scored as follows: steatosis (0 to 4), inflammation (0 to 3), and ballooning (0 to 2). NAFLD activity score (NAS) was calculated by using the sum of each histological score.
Quantitative data were expressed as means ± standard error of the mean (SEM). Means were compared using Dunnett's test. p values <0.05 were considered statistically significant. The Pearson product-moment correlation coefficient was used to evaluate the relationship between T1/2 of 99mTc-MIBI in the liver and the liver histological score or plasma.
Physiological characteristics and hepatic pathology
Plasma parameters in mice fed control and MCD diet for 2 and 4 weeks
MCD 2 weeks
MCD 4 weeks
90.9 ± 2.55
57.7 ± 1.67**
32.9 ± 2.84**
110 ± 18.4
9.80 ± 1.77**
14.7 ± 1.81**
63.8 ± 21.3
312 ± 84.5**
324 ± 65.8**
26.7 ± 8.02
191 ± 41.7**
239 ± 31.4**
50.9 ± 1.72
30.8 ± 1.58**
14.4 ± 1.63**
Hepatic ATP content
Hepatic ATP content in mice fed control and MCD diet for 2 and 4 weeks
MCD 2 weeks
MCD 4 weeks
3.04 ± 0.15
2.57 ± 0.03*
2.47 ± 0.02**
Hepatic 99m Tc-MIBI kinetics in mice fed control and MCD diet for 2 and 4 weeks
MCD 2 weeks
MCD 4 weeks
57.6 ± 7.73
37.6 ± 5.80*
19.8 ± 2.05**
3.59 ± 0.35
2.71 ± 0.28
2.63 ± 0.17
99mTc-MIBI heart scintigraphy is used to evaluate cardiac mitochondrial dysfunction in patients with cardiomyopathy ,. In addition, 99mTc-MIBI leg scintigraphy has been used to detect mitochondrial dysfunction in progressive supranuclear palsy patient skeletal muscle . 99mTc-MIBI, a lipophilic cationic myocardial perfusion agent, is considered to be retained in the mitochondria by the higher membrane potential, and the loss of mitochondrial membrane potential results in rapid washout of 99mTc-MIBI from the myocyte . In the present study, we showed that rapid washout of 99mTc-MIBI indicating mitochondrial dysfunction was observed in the liver of MCD-fed mice. The MCD-fed rodent model is widely used as a NASH model because of its similarity with human NASH pathology. Rodents fed MCD diet showed mitochondrial dysfunction such as reduction of the activity of mitochondrial respiratory chain and hepatic ATP depletion . These findings suggest that the cause of rapid washout of 99mTc-MIBI is the loss of mitochondrial membrane potential since 99mTc-MIBI is retained to the mitochondrial membrane in response to membrane potential. Rogers et al. showed that prolonged exposure of preadipocytes to fatty acid led to mitochondrial dysfunction such as decrease of ATP content and reduction of mitochondrial inner membrane potential . We also confirmed the decrease of ATP content in the liver of MCD-fed mice. Therefore, our and previous findings indicate that the rapid clearance of hepatic MIBI might be due to mitochondrial dysfunction including reduction of ATP content in MCD-fed mice. Regarding magnetic resonance imaging (MRI), it has been reported that Tmax and T1/2 after injection of gadolinium-ethoxybenzyl-diethylenetriamine penta-acetic acid (Gd-EOB-DTPA) were significantly prolonged in a NASH rat model . Furthermore, Tmax and T1/2 after Gd-EOB-DTPA injection significantly correlated with the fibrosis rate . It is well known that organic anion-transporting polypeptide (oatp) 1 mediates the uptake of Gd-EOB-DTPA and multidrug resistance-associated protein 2 (mrp2) mediates biliary excretion of Gd-EOB-DTPA in rats ,. These reports suggest that the elimination of 99mTc-MIBI also might be influenced by transporter expression in NASH pathology since 99mTc-MIBI passively diffuses into hepatocytes and the biliary excretion is mediated by P-glycoprotein (P-gp) . However, Canet et al. recently reported that the liver protein expression of P-gp did not alter in a MCD-fed mouse . This report suggests that the role of efflux transporter via P-gp is considered to be small for 99mTc-MIBI washout in this MCD-fed mouse.
In the present study, the most important finding is to show the negative correlation between 99mTc-MIBI clearance (T1/2) and NAFLD activity score including steatosis and inflammation score. Steatosis and inflammation were histologically observed at 2 and 4 weeks after MCD diet in this study. The MCD diet also seemed to aggravate the inflammation at 4 weeks compared with that at 2 weeks after feeding, although the severity of steatosis was not different between 2 and 4 weeks. These data suggest that the 99mTc-MIBI washout is likely to correlate with pathology including both steatosis and inflammation in MCD mice. Masuda et al.  have reported that hepatic 99mTc-MIBI uptake is correlated with the NAFLD activity score in a clinical study. These non-clinical and clinical findings suggest that hepatic 99mTc-MIBI SPECT imaging might be useful for evaluating NASH progression pathology such as mitochondrial dysfunction. However, further study will be needed to clarify the relation between mitochondrial membrane potential and 99mTc-MIBI binding activity.
Hepatic retention of 99mTc-MIBI was decreased with increase in NAFLD activity score in MCD-fed mice. This study indicates that 99mTc-MIBI SPECT imaging might be useful for evaluating hepatic mitochondrial dysfunction such as NAFLD or NASH.
TR participated in the design of the study, performed all the experiments, and drafted the manuscript. TU and YH performed the SPECT experiments. SM performed the histological study. AO drafted the manuscript. KA and YA participated in the study coordination and the design of the study and drafted the manuscript. All authors read and approved the final manuscript.
The authors thank Mr. Momosaki Sotaro, Department of Drug Metabolism and Pharmacokinetics, Research Laboratory for Development, Shionogi & Co., Ltd., for preparing and feeding the animals.
- Bhala N, Angulo P, van der Poorten D, Lee E, Hui JM, Saracco G, Adams LA, Charatcharoenwitthaya P, Topping JH, Bugianesi E, Day CP, George J: The natural history of nonalcoholic fatty liver disease with advanced fibrosis or cirrhosis: an international collaborative study. Hepatology 2011, 54: 1208–1216. 10.1002/hep.24491PubMed CentralView ArticlePubMedGoogle Scholar
- Angulo P: Nonalcoholic fatty liver disease. N Engl J Med 2002, 346: 1221–1231. 10.1056/NEJMra011775View ArticlePubMedGoogle Scholar
- Day CP, James OF: Hepatic steatosis: innocent bystander or guilty party? Hepatology 1998, 27: 1463–1466. 10.1002/hep.510270601View ArticlePubMedGoogle Scholar
- Fukunishi S, Nishio H, Fukuda A, Takeshita A, Hanafusa T, Higuchi K, Suzuki K: Development of fibrosis in nonalcoholic steatosis through combination of a synthetic diet rich in disaccharide and low-dose lipopolysaccharides in the livers of Zucker (fa/fa) rats. J Clin Biochem Nutr 2009, 45: 322–328. 10.3164/jcbn.09-50PubMed CentralView ArticlePubMedGoogle Scholar
- Pessayre D, Fromenty B: NASH: a mitochondrial disease. J Hepatol 2005, 42: 928–940. 10.1016/j.jhep.2005.03.004View ArticlePubMedGoogle Scholar
- Pessayre D, Fromenty B, Mansouri A: Mitochondrial injury in steatohepatitis. Eur J Gastroenterol Hepatol 2004, 16: 1095–1105. 10.1097/00042737-200411000-00003View ArticlePubMedGoogle Scholar
- Gao D, Wei C, Chen L, Huang J, Yang S, Diehl AM: Oxidative DNA damage and DNA repair enzyme expression are inversely related in murine models of fatty liver disease. Am J Physiol Gastrointest Liver Physiol 2004, 287: G1070-G1077. 10.1152/ajpgi.00228.2004View ArticlePubMedGoogle Scholar
- Yang S, Zhu H, Li Y, Lin H, Gabrielson K, Trush MA, Diehl AM: Mitochondrial adaptations to obesity-related oxidant stress. Arch Biochem Biophys 2000, 378: 259–268. 10.1006/abbi.2000.1829View ArticlePubMedGoogle Scholar
- Serviddio G, Sastre J, Bellanti F, Vina J, Vendemiale G, Altomare E: Mitochondrial involvement in non-alcoholic steatohepatitis. Mol Aspects Med 2008, 29: 22–35. 10.1016/j.mam.2007.09.014View ArticlePubMedGoogle Scholar
- Caldwell SH, Swerdlow RH, Khan EM, Iezzoni JC, Hespenheide EE, Parks JK, Parker WD Jr: Mitochondrial abnormalities in non-alcoholic steatohepatitis. J Hepatol 1999, 31: 430–434. 10.1016/S0168-8278(99)80033-6View ArticlePubMedGoogle Scholar
- Cortez-Pinto H, Chatham J, Chacko VP, Arnold C, Rashid A, Diehl AM: Alterations in liver ATP homeostasis in human nonalcoholic steatohepatitis: a pilot study. Jama 1999, 282: 1659–1664. 10.1001/jama.282.17.1659View ArticlePubMedGoogle Scholar
- Hensley K, Kotake Y, Sang H, Pye QN, Wallis GL, Kolker LM, Tabatabaie T, Stewart CA, Konishi Y, Nakae D, Floyd RA: Dietary choline restriction causes complex I dysfunction and increased H(2)O(2) generation in liver mitochondria. Carcinogenesis 2000, 21: 983–989. 10.1093/carcin/21.5.983View ArticlePubMedGoogle Scholar
- Perez-Carreras M, Del Hoyo P, Martin MA, Rubio JC, Martin A, Castellano G, Colina F, Arenas J, Solis-Herruzo JA: Defective hepatic mitochondrial respiratory chain in patients with nonalcoholic steatohepatitis. Hepatology 2003, 38: 999–1007. 10.1053/jhep.2003.50398View ArticlePubMedGoogle Scholar
- Jones AG, Abrams MJ, Davison A, Brodack JW, Toothaker AK, Adelstein SJ, Kassis AI: Biological studies of a new class of technetium complexes: the hexakis(alkylisonitrile)technetium(I) cations. Int J Nucl Med Biol 1984, 11: 225–234. 10.1016/0047-0740(84)90004-4View ArticlePubMedGoogle Scholar
- Chiu ML, Kronauge JF, Piwnica-Worms D: Effect of mitochondrial and plasma membrane potentials on accumulation of hexakis (2-methoxyisobutylisonitrile) technetium(I) in cultured mouse fibroblasts. J Nucl Med 1990, 31: 1646–1653.PubMedGoogle Scholar
- Crane P, Laliberte R, Heminway S, Thoolen M, Orlandi C: Effect of mitochondrial viability and metabolism on technetium-99 -sestamibi myocardial retention. Eur J Nucl Med 1993, 20: 20–25. 10.1007/BF02261241View ArticlePubMedGoogle Scholar
- Ikawa M, Kawai Y, Arakawa K, Tsuchida T, Miyamori I, Kuriyama M, Tanaka M, Yoneda M: Evaluation of respiratory chain failure in mitochondrial cardiomyopathy by assessments of 99mTc-MIBI washout and 123I-BMIPP/99mTc-MIBI mismatch. Mitochondrion 2007, 7: 164–170. 10.1016/j.mito.2006.11.008View ArticlePubMedGoogle Scholar
- Strauss HW: MIBI and mitochondria. J Am Coll Cardiol 2013, 61: 2018–2019. 10.1016/j.jacc.2013.02.026View ArticlePubMedGoogle Scholar
- Masuda K, Ono M, Fukumoto M, Hirose A, Munekage K, Ochi T, Okamoto N, Akagi N, Ogawa Y, Saibara T: Usefulness of technetium-99 m-2-methoxy-isobutyl-isonitrile liver scintigraphy for evaluating disease activity of non-alcoholic fatty liver disease. Hepatol Res 2012, 42: 273–279. 10.1111/j.1872-034X.2011.00923.xView ArticlePubMedGoogle Scholar
- Romestaing C, Piquet MA, Letexier D, Rey B, Mourier A, Servais S, Belouze M, Rouleau V, Dautresme M, Ollivier I, Favier R, Rigoulet M, Duchamp C, Sibille B: Mitochondrial adaptations to steatohepatitis induced by a methionine- and choline-deficient diet. Am J Physiol Endocrinol Metab 2008, 294: E110-E119. 10.1152/ajpendo.00407.2007View ArticlePubMedGoogle Scholar
- Vetelainen RL, Bennink RJ, de Bruin K, van Vliet A, van Gulik TM: Hepatobiliary function assessed by 99mTc-mebrofenin cholescintigraphy in the evaluation of severity of steatosis in a rat model. Eur J Nucl Med Mol Imaging 2006, 33: 1107–1114. 10.1007/s00259-006-0125-3View ArticlePubMedGoogle Scholar
- Liu Z, Chen L, Liu S, Barber C, Stevenson GD, Furenlid LR, Barrett HH, Woolfenden JM: Kinetic characterization of a novel cationic (99 m)Tc(I)-tricarbonyl complex, (99 m)Tc-15C5-PNP, for myocardial perfusion imaging. J Nucl Cardiol 2010, 17: 858–867. 10.1007/s12350-010-9262-yPubMed CentralView ArticlePubMedGoogle Scholar
- Dai DZ, Zhu Y, Tian Y, Chen SH, Wang H: Comparison of protection by propranolol, bepridil, verapamil, and captopril on depleting ATP, ADP, and AMP in heart, brain, and liver by anoxia plus isoprenaline. Drug Dev Res 1996, 39: 125–130. 10.1002/(SICI)1098-2299(199610)39:2<125::AID-DDR2>3.0.CO;2-RView ArticleGoogle Scholar
- Matsuo S, Nakae I, Tsutamoto T, Okamoto N, Horie M: A novel clinical indicator using Tc-99 m sestamibi for evaluating cardiac mitochondrial function in patients with cardiomyopathies. J Nucl Cardiol 2007, 14: 215–220. 10.1016/j.nuclcard.2006.10.022View ArticlePubMedGoogle Scholar
- Unno K, Isobe S, Izawa H, Cheng XW, Kobayashi M, Hirashiki A, Yamada T, Harada K, Ohshima S, Noda A, Nagata K, Kato K, Yokota M, Murohara T: Relation of functional and morphological changes in mitochondria to myocardial contractile and relaxation reserves in asymptomatic to mildly symptomatic patients with hypertrophic cardiomyopathy. Eur Heart J 2009, 30: 1853–1862. 10.1093/eurheartj/ehp184View ArticlePubMedGoogle Scholar
- Chang YY, Lee CH, Lan MY, Wu HS, Chang CC, Liu JS: A new noninvasive test to detect mitochondrial dysfunction of skeletal muscles in progressive supranuclear palsy. Ann N Y Acad Sci 2005, 1042: 76–81. 10.1196/annals.1338.046View ArticlePubMedGoogle Scholar
- Serviddio G, Bellanti F, Tamborra R, Rollo T, Romano AD, Giudetti AM, Capitanio N, Petrella A, Vendemiale G, Altomare E: Alterations of hepatic ATP homeostasis and respiratory chain during development of non-alcoholic steatohepatitis in a rodent model. Eur J Clin Invest 2008, 38(4):245–252. 10.1111/j.1365-2362.2008.01936.xView ArticlePubMedGoogle Scholar
- Rogers C, Davis B, Neufer PD, Murphy MP, Anderson EJ, Robidoux J: A transient increase in lipid peroxidation primes preadipocytes for delayed mitochondrial inner membrane permeabilization and ATP depletion during prolonged exposure to fatty acids. Free Radic Biol Med 2014, 67: 330–341. 10.1016/j.freeradbiomed.2013.11.012PubMed CentralView ArticlePubMedGoogle Scholar
- Tsuda N, Okada M, Murakami T: Potential of gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) for differential diagnosis of nonalcoholic steatohepatitis and fatty liver in rats using magnetic resonance imaging. Invest Radiol 2007, 42: 242–247. 10.1097/01.rli.0000258058.44876.a5View ArticlePubMedGoogle Scholar
- Tsuda N, Okada M, Murakami T: New proposal for the staging of nonalcoholic steatohepatitis: evaluation of liver fibrosis on Gd-EOB-DTPA-enhanced MRI. Eur J Radiol 2010, 73: 137–142. 10.1016/j.ejrad.2008.09.036View ArticlePubMedGoogle Scholar
- van Montfoort JE, Stieger B, Meijer DK, Weinmann HJ, Meier PJ, Fattinger KE: Hepatic uptake of the magnetic resonance imaging contrast agent gadoxetate by the organic anion transporting polypeptide Oatp1. J Pharmacol Exp Ther 1999, 290: 153–157.PubMedGoogle Scholar
- Pascolo L, Cupelli F, Anelli PL, Lorusso V, Visigalli M, Uggeri F, Tiribelli C: Molecular mechanisms for the hepatic uptake of magnetic resonance imaging contrast agents. Biochem Biophys Res Commun 1999, 257: 746–752. 10.1006/bbrc.1999.0454View ArticlePubMedGoogle Scholar
- Swift B, Yue W, Brouwer KL: Evaluation of (99 m)technetium-mebrofenin and (99 m)technetium-sestamibi as specific probes for hepatic transport protein function in rat and human hepatocytes. Pharm Res 2010, 27: 1987–1998. 10.1007/s11095-010-0203-xPubMed CentralView ArticlePubMedGoogle Scholar
- Canet MJ, Hardwick RN, Lake AD, Dzierlenga AL, Clarke JD, Cherrington NJ: Modeling human nonalcoholic steatohepatitis-associated changes in drug transporter expression using experimental rodent models. Drug Metab Dispos 2014, 42: 586–595. 10.1124/dmd.113.055996PubMed CentralView ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.