Pulmonary, Gastrointestinal and Urogenital PharmacologyExperimental non-alcoholic fatty liver disease results in decreased hepatic uptake transporter expression and function in rats
Introduction
The liver plays a crucial role in the elimination of many clinically relevant drugs from the body. While drug-metabolizing enzymes are critical in hepatic drug clearance, often, hepatic transport is required to initiate and complete this process. Several basolateral transmembrane proteins are capable of transporting organic ions from plasma into the hepatocyte, including the organic anion transporting polypeptides (OATP), organic anion transporters (OAT); organic cation transporters (OCT); and the sodium-dependent bile salt uptake transporter Na+-taurocholate co-transporting polypeptide (NTCP). Coordinately, the overlapping affinities of these transporters provide an efficient means of extracting ionic xenobiotics from the sinusoidal blood (Kullak-Ublick and Meier, 2000).
Variations in patient response to drug therapy are a significant safety issue. Severe adverse drug effects among hospitalized patients in the United States may number as high as two million per year and as many as 100,000 of them prove fatal (Lazarou et al., 1998). Remarkably, the majority of these adverse reactions are in patients given the correct drug and proper dosage. It is estimated that less than 20% of these cases are due to genetic polymorphisms that result in altered metabolism: the vast majority of the adverse reactions are due to individual host or environmental factors such as age, nutrition, or disease state (Ingelman-Sundberg and Rodriguez-Antona, 2005). Both pre-clinical and clinical studies have indicated that hepatic disease may result in alterations in the pharmacokinetics and elimination of a drug (Williams and Mamelok, 1980, Westphal and Brogard, 1997). Alterations in drug disposition have been well documented for liver conditions such as sepsis (Pea et al., 2005, Kim et al., 2000) and primary biliary cirrhosis (Jorquera et al., 2001). In addition, altered expression of cytochrome P450 enzymes and hepatic transporters observed in viral and alcoholic hepatitis patients have been associated with variability in drug response, resulting in either drug toxicity or decreased therapeutic effectiveness (Morgan, 2001, Heinrich et al., 1990, Renton, 2005). In contrast, relatively little data has been reported with respect to the effect of non-alcoholic fatty liver disease (NAFLD) on hepatic drug transporter.
NAFLD encompasses a spectrum of symptoms ranging from steatosis (simple fatty liver, SFL), to steatohepatitis (fatty liver with liver cell damage, inflammation and progressive fibrosis) (Reynaert et al., 2005). The prevalence of NAFLD is estimated to be between 14% and 24% in the world population, and though once thought an adult disease, it is now known to afflict children as well (Browning and Horton, 2004). Simple steatosis is characterized by micro and macrovesicular steatosis and predisposes the liver to the more severe non-alcoholic steatohepatitis (NASH) (Koteish and Mae, 2002). NASH is histologically characterized by micro and macrovesicular steatosis, lobular inflammation, and hepatocellular damage, including accumulation of Mallory's hyaline, ballooning degeneration, necrosis, and fibrosis (zone 1 and zone 3) (Peters et al., 1975). NASH occurs in 2–3% of all patients with excess fat accumulation in the liver, and accounts for approximately 10% of all newly diagnosed cases of chronic liver disease.
The classic rodent model of NASH involves inducing steatohepatitis through a methionine-choline-deficient (MC−) diet. This model of “fibrosing steatohepatitis” is characterized by the development of fibrotic strands that surround hepatocytes in a fashion identical to those in human disorders of lipid-associated hepatic fibrosis. Additionally, steatosis, chronic hepatocyte injury, and hepatic inflammation precede activation of stellate cells and fibrosis by several weeks, a sequence of events that is analogous to that which occurs in NASH (George et al., 2003).
Previous studies have demonstrated that this model causes significant changes in hepatic efflux transporters resulting in a significant shift in the disposition of acetaminophen metabolites from the bile to the plasma (Lickteig et al., 2007a). In the current study, high-fat (SFL model) and MC− (NASH model) diets were used as models to determine the effect of SFL and NASH on hepatic uptake transporter expression and whether the resulting change in expression causes a functional alteration of xenobiotic pharmacokinetics.
Section snippets
Chemicals
Urethane, bromosulfophthalien (BSP), and NaOH were purchased from Sigma (St. Louis, MO). Heparin and sterile 0.9% sodium chloride solution (w/v) were obtained from Baxter Healthcare Corporation (Deerfield, IL). PE-10 and PE-50 polyethylene tubing were purchased from Braintree Scientific Inc. (Braintree, MA).
Animals
Male Sprague Dawley rats (8–9 weeks) were obtained from Harlan (Indianapolis, IN). All animals were acclimated in 12 h light and 12 h dark cycles in a University of Arizona AAALAC-certified
Liver histology
Control, low-fat isocaloric, and MC+ livers displayed no signs of steatosis or fibrosis (Fig. 1). High-fat livers showed primarily microvesicular steatosis with mild lobular inflammation, while MC− livers showed marked diffuse macrovesicular hepatic steatosis with mild lobular inflammation as well as early signs of bridging fibrosis. NAFLD Activity Scoring (NAS) for normal, low-fat isocaloric, high-fat, MC+ and MC− rat livers are summarized in Table 1. The histology and NAS scoring observed in
Discussion
Simple fatty liver (early stage NAFLD) is typically associated with obesity. In 1994, 22.5% of Americans were determined to be clinically obese, a statistic which is projected to reach almost 40% by 2025 (Kopelman, 2000). Due to this increase in obesity, information pertaining to liver function during the various stages of NAFLD is of increasing importance. In 2005, Geier et al. suggested that alterations in hepatic transporters may render fatty livers more vulnerable to various xenobiotics.
Acknowledgments
This work was supported by the National Institutes of Health grants ES007091 (C.D.F), ES011646 and DK068039 (to N.J.C). This work has been presented in part at the International Society for the Study of Xenobiotics meeting, Maui, Hawaii 2005.
References (43)
- et al.
Molecular mechanisms of reduced glutathione transport: role of the MRP/CFTR/ABCC and OATP/SLC21A families of membrane proteins
Toxicol. Appl. Pharmacol.
(2005) - et al.
Characterization of organic anion transporter regulation, glutathione metabolism and bile formation in the obese Zucker rat
J. Hepatol.
(2005) - et al.
Principles of hepatic organic anion transporter regulation during cholestasis, inflammation and liver regeneration
Biochim. Biophys. Acta
(2007) - et al.
Lipid peroxidation, stellate cell activation and hepatic fibrogenesis in a rat model of chronic steatohepatitis
J. Hepatol.
(2003) - et al.
Regulation of hepatocyte bile salt transporters by endotoxin and inflammatory cytokines in rodents
Gastroenterology
(1996) - et al.
Animal models of steatohepatitis
Best. Pract. Res. Clin. Gastroenterol.
(2002) - et al.
Mechanisms of cholestasis
Clin. Liver Dis.
(2000) - et al.
Alterations in transporter expression in liver, kidney, and duodenum after targeted disruption of the transcription factor HNF1alpha
Biochem. Pharmacol.
(2006) - et al.
Substrate specificity of sinusoidal bile acid and organic anion uptake systems in rat and human liver
Hepatology
(1997) - et al.
Glutathione metabolism and antioxidant enzymes in patients affected by nonalcoholic steatohepatitis
Clin. Chim. Acta
(2005)
Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues
Anal. Biochem.
A pilot trial of pentoxifylline in nonalcoholic steatohepatitis
Am. J. Gastroenterol.
Coordinated expression of multidrug resistance-associated proteins (Mrps) in mouse liver during toxicant-induced injury
Toxicol. Sci.
Xenobiotic and endobiotic transporter mRNA expression in the blood–testis barrier
Drug Metab. Dispos.
Molecular mediators of hepatic steatosis and liver injury
J. Clin. Invest
Localization of organic anion transporting polypeptide 4 (Oatp4) in rat liver and comparison of its substrate specificity with Oatp1, Oatp2 and Oatp3
Pflugers Arch.
Lipopolysaccharide-mediated regulation of hepatic transporter mRNA levels in rats
Drug Metab. Dispos.
Inflammatory cytokines, but not bile acids, regulate expression of murine hepatic anion transporters in endotoxemia
J. Pharmacol. Exp. Ther.
Substrate specificities of rat oatp1 and ntcp: implications for hepatic organic anion uptake
Am. J. Physiol. Gastrointest. Liver Physiol.
Interleukin-6 and the acute phase response
Biochem. J.
Simvastatin inhibits leptin-induced hypertrophy in cultured neonatal rat cardiomyocytes
Acta Pharmacol. Sin.
Cited by (103)
Impact of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) on the expression and function of hepatobiliary transporters: A comprehensive mechanistic review
2024, Biochimica et Biophysica Acta - Molecular Basis of DiseaseOAT3 mediates methotrexate resistance in the treatment of rheumatoid arthritis
2022, Biomedicine and PharmacotherapyPharmacokinetic herb-disease-drug interactions: Effect of ginkgo biloba extract on the pharmacokinetics of pitavastatin, a substrate of Oatp1b2, in rats with non-alcoholic fatty liver disease
2021, Journal of EthnopharmacologyCitation Excerpt :Despite the high prevalence of NAFLD, little is known about NAFLD-related changes in PK profiles and possible alterations in therapeutic effects and adverse reactions. Existing evidence indicated that changes in the drug transporter mRNA and protein expressions of NAFLD may disturb drug disposition (Fisher et al., 2009b; Canet et al., 2014). NAFLD at different stages can cause varying changes in liver organic anion transporting polypeptide (OATP), which leads to alterations in OATP-mediated drug transport from the blood into hepatocytes (Fisher et al., 2009b).
Probiotic-prebiotic-synbiotic modulation of (YAP1, LATS1 and NF2 mRNAs/miR-1205/lncRNA SRD5A3-AS1) panel in NASH animal model
2021, Biomedicine and PharmacotherapyIncreased penetration of diphenhydramine in brain via proton-coupled organic cation antiporter in rats with lipopolysaccharide-induced inflammation
2021, Brain, Behavior, and Immunity - HealthPost-transcriptional regulation of OATP2B1 transporter by a microRNA, miR-24
2020, Drug Metabolism and Pharmacokinetics