Tsuchiya S, Tsuji M, Morio Y, Oguchi K. Involvement of endoplasmic reticulum in glycochenodeoxycholic acid-induced apoptosis in rat hepatocytes. Toxicol Lett. 2006;

Department of Pharmacology, School of Medicine, Showa University, Hatanodai 1-5-8, Shinagawa-Ku, Tokyo 142-8555, Japan.
Toxicology Letters (Impact Factor: 3.26). 11/2006; 166(2):140-9. DOI: 10.1016/j.toxlet.2006.06.006
Source: PubMed


In chronic cholestatic liver diseases, accumulation of hydrophobic bile acids is thought to damage hepatocytes. The mechanism of how cells die has been an open debate, but apoptotic pathways are known to involve activation of death receptors and mitochondrial dysfunction. Recently apoptosis via an endoplasmic reticulum (ER) stress-mediated pathway was also found. In this study, we examined whether ER stress is induced in rat hepatocytes by treatment with glycochenodeoxycholic acid (GCDCA, 50-300microM for 1-24h), and if so, whether ER stress-mediated apoptosis occurs in this system. We determined mobility of intracellular calcium ion, activities of calpain and caspase-12, specific to ER stress-mediated apoptosis, and Bip and Chop mRNA expressions, biomarkers of ER stress. We found that GCDCA induces ER-related calcium release within about ten seconds. Significant increases in activities of calpain and caspase-12 were observed after 15h of GCDCA treatment. Bip and Chop mRNA expressions were increased with the treated GCDCA dose and incubation time. Cytochrome c release from mitochondria peaked in about 2h of incubation. These results suggest that ER stress is actually induced by GCDCA, though its role in hepatocellular apoptosis may be smaller than mitochondria-mediated pathway. The presence of ER stress might be important in pathogenesis of cholestatic liver diseases.

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    • "Besides kinase signaling pathways, hydrophobic bile salts, such as GCDCA, may also induce apoptosis by increasing ER stress and induction of the unfolded protein response via transcription factors like CHOP [30], [31]. Induction of CHOP is a sensitive marker for ER stress and CHOP-knockout mice are protected against cholestasis-induced hepatocyte apoptosis [32], [33]. "
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    ABSTRACT: Angiotensin II (AT-II) is a pro-fibrotic compound that acts via membrane-bound receptors (AT-1R/AT-2R) and thereby activates hepatic stellate cells (HSCs). AT-II receptor blockers (ARBs) are thus important candidates in the treatment of liver fibrosis. However, multiple case reports suggest that AT-1R blockers may induce hepatocyte injury. Therefore, we investigated the effect of AT-II and its receptor blockers on cytokine-, oxidative stress- and bile salt-induced cell death in hepatocytes. Primary rat hepatocytes were exposed to TNF-α/Actinomycin D, the ROS-generating agent menadione or the bile salts: glycochenodeoxycholic acid (GCDCA) and tauro-lithocholic acid-3 sulfate (TLCS), to induce apoptosis. AT-II (100 nmol/L) was added 10 minutes prior to the cell death-inducing agent. AT-1R antagonists (Sartans) and the AT-2R antagonist PD123319 were used at 1 µmol/L. Apoptosis (caspase-3 activity, acridine orange staining) and necrosis (Sytox green staining) were quantified. Expression of CHOP (marker for ER stress) and AT-II receptor mRNAs were quantified by Q-PCR. AT-II dose-dependently reduced GCDCA-induced apoptosis of hepatocytes (−50%, p<0.05) without inducing necrosis. In addition, AT-II reduced TLCS-induced apoptosis of hepatocytes (−50%, p<0.05). However, AT-II did not suppress TNF/Act-D and menadione-induced apoptosis. Only the AT-1R antagonists abolished the protective effect of AT-II against GCDCA-induced apoptosis. AT-II increased phosphorylation of ERK and a significant reversal of the protective effect of AT-II was observed when signaling kinases, including ERK, were inhibited. Moreover, AT-II prevented the GCDCA-induced expression of CHOP (the marker of the ER-mediated apoptosis). Conclusion Angiotensin II protects hepatocytes from bile salt-induced apoptosis through a combined activation of PI3-kinase, MAPKs, PKC pathways and inhibition of bile salt-induced ER stress. Our results suggest a mechanism for the observed hepatocyte-toxicity of Sartans (angiotensin receptor blockers, ARBs) in some patients with chronic liver injury.
    PLoS ONE 12/2012; 7(12):e52647. DOI:10.1371/journal.pone.0052647 · 3.23 Impact Factor
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    • "The various different mechanisms that have been suggested to contribute to bile acid-induced apoptosis include oxidative stress, mitochondrial damage, ER damage, or the activation of cell death receptors, Fas and TRAIL-R2 [8], [9], [10], [11], [12], [13], [14], [15]. Interestingly, one study in colon cancer cells showed that deoxycholic acid (DCA)-induced apoptosis is associated with altered cytoplasmic ion concentrations [16]. "
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    ABSTRACT: Apoptosis resistance is a hallmark of cancer cells. Typically, bile acids induce apoptosis. However during gastrointestinal (GI) tumorigenesis the cancer cells develop resistance to bile acid-induced cell death. To understand how bile acids induce apoptosis resistance we first need to identify the molecular pathways that initiate apoptosis in response to bile acid exposure. In this study we examined the mechanism of deoxycholic acid (DCA)-induced apoptosis, specifically the role of Na(+)/H(+) exchanger (NHE) and Na(+) influx in esophageal cells. In vitro studies revealed that the exposure of esophageal cells (JH-EsoAd1, CP-A) to DCA (0.2 mM-0.5 mM) caused lysosomal membrane perturbation and transient cytoplasmic acidification. Fluorescence microscopy in conjunction with atomic absorption spectrophotometry demonstrated that this effect on lysosomes correlated with influx of Na(+), subsequent loss of intracellular K(+), an increase of Ca(2+) and apoptosis. However, ethylisopropyl-amiloride (EIPA), a selective inhibitor of NHE, prevented Na(+), K(+) and Ca(2+) changes and caspase 3/7 activation induced by DCA. Ouabain and amphotericin B, two drugs that increase intracellular Na(+) levels, induced similar changes as DCA (ion imbalance, caspase3/7 activation). On the contrary, DCA-induced cell death was inhibited by medium with low a Na(+) concentrations. In the same experiments, we exposed rat ileum ex-vivo to DCA with or without EIPA. Severe tissue damage and caspase-3 activation was observed after DCA treatment, but EIPA almost fully prevented this response. In summary, NHE-mediated Na(+) influx is a critical step leading to DCA-induced apoptosis. Cells tolerate acidification but evade DCA-induced apoptosis if NHE is inhibited. Our data suggests that suppression of NHE by endogenous or exogenous inhibitors may lead to apoptosis resistance during GI tumorigenesis.
    PLoS ONE 08/2011; 6(8):e23835. DOI:10.1371/journal.pone.0023835 · 3.23 Impact Factor
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    • "Intrahepatic accumulation of bile acids induces hepatocellular injury. Glycochenodeoxycholic acid depletes ER Ca 2þ and induces a UPR and apoptosis (Tsuchiya et al. 2006). It is unclear to what extent the decreasing [Ca 2þ ] ER directly contributes to the pathology. "
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    ABSTRACT: The endoplasmic reticulum (ER) as an intracellular Ca(2+) store not only sets up cytosolic Ca(2+) signals, but, among other functions, also assembles and folds newly synthesized proteins. Alterations in ER homeostasis, including severe Ca(2+) depletion, are an upstream event in the pathophysiology of many diseases. On the one hand, insufficient release of activator Ca(2+) may no longer sustain essential cell functions. On the other hand, loss of luminal Ca(2+) causes ER stress and activates an unfolded protein response, which, depending on the duration and severity of the stress, can reestablish normal ER function or lead to cell death. We will review these various diseases by mainly focusing on the mechanisms that cause ER Ca(2+) depletion.
    Cold Spring Harbor perspectives in biology 03/2011; 3(6). DOI:10.1101/cshperspect.a004317 · 8.68 Impact Factor
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