ABSTRACT:Alterations in transporter expression may represent a compensatory mechanism of damaged hepatocytes to reduce accumulation of potentially toxic compounds. The present study was conducted to investigate the expression of hepatobiliary efflux transporters in livers from patients after toxic acetaminophen (APAP) ingestion, with livers from patients with primary biliary cirrhosis (PBC) serving as positive controls. mRNA and protein expression of multidrug resistance-associated protein (MRP) 1-6, multidrug resistance protein (MDR) 1-3/P-glycoprotein (P-gp), and breast cancer resistance protein (BCRP) in normal (n ‫؍‬ 6), APAP overdose (n ‫؍‬ 5), and PBC (n ‫؍‬ 6) human liver samples were determined by branched DNA and Western blot analysis, respectively. Double immunohistochemical staining of P-gp and proliferating cell nuclear antigen (PCNA), a marker of proliferation, was performed on paraffin-embedded tissue sections. Compared with normal liver specimens, MRP1 and MRP4 mRNA levels were elevated after APAP overdose and in PBC. Up-regulation of MRP5, MDR1, and BCRP mRNA occurred in PBC livers. Protein levels of MRP4, MRP5, BCRP, and P-gp were increased in both disease states, with MRP1 and MRP3 protein also being induced in PBC. Increased P-gp protein was confirmed immunohistochemically and was found to localize to areas of PCNA-positive hepatocytes, which were detected in APAP overdose and PBC livers. The findings from this study demonstrate that hepatic efflux transporter expression is up-regulated in cases of APAP-induced liver failure and PBC. This adaptation may aid in reducing retention of byproducts of cellular injury and bile constituents within hepatocytes. The close proximity of P-gp and PCNApositive hepatocytes during liver injury suggests that along with cell regeneration, increased efflux transporter expression is a critical response to hepatic damage to protect the liver from additional insult.The incidence of acetaminophen (APAP) overdose, which frequently leads to severe liver injury at supratherapeutic doses, has been steadily increasing in the United States. The number of APAP-related cases of liver failure has increased from 39% (1998 -2001) to nearly 50% (2003) of all cases of acute liver failure, with well over 100,000 cases of APAP overdose reported annually in the United States (Lee, 2004). Although APAP poisoning is often intentional, an increased incidence of unintentional overdose cases is gaining attention. A recent study reported that 50% of APAP overdose cases were attributable to unintentional exposure, most commonly because of APAP overutilization in multiple over-the-counter and prescription preparations (Larson et al., 2005).Mouse models of APAP-induced hepatotoxicity are widely used to recapitulate human exposures because of the similarities in APAP bioactivation and patterns of liver injury between mice and humans. Similar doses of APAP cause hepatotoxicity in both species, and the liver injury is dose-dependant (Mitchell et al., 1973;Davis et al., 1974). Another striking...

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Reducing protein accumulation is essential for treating Alzheimer's disease (AD) by attenuating a pathogenic cascade that leads to synaptic decline. Z‐Phe‐Ala‐diazomethylketone (PADK) increases lysosomal enzymes 2‐ to 9‐fold in vitro and in vivo. This enhancement clears AD‐related proteins and restores synaptic integrity. Here, PADK produced a dose‐dependent increase in cathepsin D without adverse effects. In the first of two transgenic models of AD, APPSwInd mice of 10–11 months exhibited deficits in coordination and spatial memory. PADK at 23 mg/kg × 9 d recovered open‐field intersession habituation (p<0.03) and improved balance beam and rotarod scores in the mice (p<0.001). 6E10 anti‐Aβ staining also revealed a 39% reduction in hippocampal CA1 sp. In a second model, 18–20‐month old APPswe/PS1dE9 mice exhibited a deficit in episodic spontaneous alternation behavior (SAB). PADK at 20 mg/kg × 10 d improved their SAB to control‐level (p=0.01). Synaptic markers GluR1 and NCAM180 were decreased 23–34% in APPswe/PS1dE9 hippocampus compared to non‐tg mice. Corresponding with the SAB improvement, PADK increased GluR1 and NCAM to non‐tg levels (p=0.0001–0.002). PADK also reduced 6E10 staining of CA1 neurons as well as number and size of plaque structures. Lysosomal modulatory drugs that enhance clearance mechanisms thus have the potential to slow the synaptic decline and cognitive deficits associated with AD.