Immunopharmacology and InflammationSimultaneous proteosome inhibition and heat shock protein induction by bortezomib is beneficial in experimental pancreatitis
Introduction
Acute pancreatitis is an inflammatory disease caused by the premature activation of pancreatic exocrine enzymes which leads to the injury of the gland and other organs (Figarella et al., 1988). Heat shock protein (HSP) induction and nuclear factor kappa B (NF-κB) inhibition were identified in the past as two effective mechanisms reducing the severity of the disease.
NF-κB is a transcription factor regulating the inflammatory process of acute pancreatitis (Chen et al., 2002, Storz et al., 2004). NF-κB binding elements have been found in the promoter region of many genes encoding inflammatory cytokines and enzymes. In the pancreas, NF-κB exists as a homo or heterodimer of two subunits called p65 and p50 (Gukovsky et al., 1998). Under physiological conditions, NF-κB is located in the cytosol of the cell and is bound to the inhibitory protein I-kappa B (I-κB). During the inflammatory processes, I-κB is phosphorylated by I-κB kinase followed by ubiquitination. This complex is then degraded by the proteosome, a protein complex with multiple proteolytic sites. The unbound NF-κB can translocate to the nuclei. Inhibition of NF-κB activation has been shown to exert anti-inflammatory effects in acute pancreatitis (Letoha et al., 2005a, Letoha et al., 2005b, Rakonczay et al., 2003a). Nf-κB is believed to be a key factor in the development of acute pancreatitis as it regulates the synthesis of tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1) and 6 (IL-6), inducible nitric oxide synthase (iNOS), cyclooxigenase-2 (COX-2) and many other molecules.
HSPs were identified in the past as protective factors during the course of acute pancreatitis (Rakonczay et al., 2003b). These proteins are molecular chaperones responsible for the correct folding, transportation of newly synthesized proteins and for the reconstruction of the three dimensional structure of damaged molecules. HSP synthesis is regulated through transcription factors called heat shock factors (HSF). These factors are bound to HSP70 in the resting cells. If the amount of denaturized intracellular proteins increases, HSP70 dissociates from HSF which allows the transcription factor to bind to its specific responsive element on the DNA. Induction of HSP synthesis in acute pancreatitis reduces the amount of denaturized proteins in the cells which otherwise would lead to the induction of cell death (Garrido et al., 2001, Kubisch et al., 2004, Nollen et al., 1999, Otaka et al., 1994, Parcellier et al., 2003, Rakonczay et al., 2002, Strowski et al., 1997, Wagner et al., 1996).
Bortezomib is a proteosome inhibitor synthesized in 1995 and approved for the treatment of patients with myeloma multiplex or mantle cell lymphoma (Chauhan et al., 2005, Pei et al., 2003, Zhao et al., 2008). Proteosomes are responsible for the degradation of ubiquitinylated proteins and also abnormal or misfolded proteins. One target of proteosomes is I-κB, but proteosomes are also responsible for the degradation of anti- or pro-apoptotic proteins, increasing the sensitivity of immortal tumor cells to chemotherapeutic agents (Grisham et al., 1999). Based on our present knowledge about the molecular mechanism of acute pancreatitis, we hypothesize that bortezomib can have a beneficial effect on the inflammatory process of acute pancreatitis.
The most commonly used experimental model of acute edematous pancreatitis is the cholecystokinin-octapeptide (CCK-8)-induced pancreatitis model in rats. CCK-8 consists of the C-terminal 8 amino acids of CCK and has almost the same biological effects as CCK. In rats, repeated supramaximal doses of CCK-8 lead to the development of a mild edematous pancreatitis. The model is characterized by elevated serum amylase and lipase activities, changes of free-radical-scavenger activities, edema of the pancreas and increased amounts of inflammatory cytokines in the pancreas and serum (Czako et al., 1998, Grady et al., 1996, Laszik et al., 1989, Takacs et al., 1994, Takacs et al., 1996). As CCK-8-induced pancreatitis is a rapidly developing pancreatitis model, it is ideal for the investigation of early events of pancreatitis. These characteristics and the high reproducibility of CCK-8-induced pancreatitis make it the most widely used model for testing the effect of a new compound in acute pancreatitis.
Section snippets
Experimental protocol
This study was approved by the Ethical Committee on Animal Experiments of the University of Szeged. Male Wistar rats weighing 230–250 g were kept at a constant room temperature (25 °C) with light–dark cycles of 12 h and were allowed free access to water and standard laboratory chow (Biofarm, Zagyvaszántó, Hungary). After 1 week of acclimatization, the rats were divided into three groups (n = 8 in each) and starved overnight before the beginning of the experiment. Animals in group P were injected
Results
Serum amylase activity was significantly elevated in both group P and BP compared to group C. Bortezomib pre-treatment did not influence the serum amylase activity of rats in group BP versus group P (Fig. 1A).
Pancreatic weight /body weight ratio was calculated to evaluate the degree of tissue edema in the pancreatic gland. The ratio was significantly elevated in the animals treated with CCK-8, but the elevation of the quotient was significantly reduced by bortezomib pre-treatment (Fig. 1B).
Discussion
The development of acute pancreatitis in the group treated with CCK-8 was demonstrated by the elevation of serum amylase activity, pancreatic weight/body weight ratio and by the result of the histological investigation where edema, vacuolization, inflammation, hyperemia and the calculated total damage score were significantly increased as compared to the control group. Necrosis was not present. Western blot analysis of the pancreatic tissue showed the degradation of I-κB-β in the animals with
Conclusion
The beneficial effects of bortezomib were demonstrated for the first time in acute experimental pancreatitis. The proteosome inhibitor bortezomib reduced the histological damage caused by CCK. One of the mechanisms behind these effects is the inhibition of I-κB degradation which leads to the reduction of Nf-κB activation. The HSP72 inducing effect of bortezomib contributed to the histological protection.
References (35)
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Anal. Biochem.
(1976)- et al.
NF-kappaB activation in pancreas induces pancreatic and systemic inflammatory response
Gastroenterology
(2002) - et al.
Heat shock proteins: endogenous modulators of apoptotic cell death
Biochem. Biophys. Res. Commun.
(2001) - et al.
Inhibition of NF-kappa B activation in vitro and in vivo: role of 26S proteasome
Methods Enzymol.
(1999) - et al.
Overexpression of heat shock protein Hsp27 protects against cerulein-induced pancreatitis
Gastroenterology
(2004) - et al.
Ultrastructural localization of Hsp-72 examined with a new polyclonal antibody raised against the truncated variable domain of the heat shock protein
Cell Stress Chaperones
(1999) - et al.
The proteasome inhibitor MG132 protects against acute pancreatitis
Free Radic. Biol. Med.
(2005) - et al.
Heat shock proteins, cellular chaperones that modulate mitochondrial cell death pathways
Biochem. Biophys. Res. Commun.
(2003) - et al.
Nontoxic heat shock protein coinducer BRX-220 protects against acute pancreatitis in rats
Free Radic. Biol. Med.
(2002) - et al.
NF-kappaB activation is detrimental in arginine-induced acute pancreatitis
Free Radic. Biol. Med.
(2003)
Hyperthermia induces heat shock protein expression and protection against cerulein-induced pancreatitis in rats
Gastroenterology
Bortezomib induces caspase-dependent apoptosis in Hodgkin lymphoma cell lines and is associated with reduced c-FLIP expression: a gene expression profiling study with implications for potential combination therapies
Leuk. Res.
Heat shock protein 70 prevents secretagogue-induced cell injury in the pancreas by preventing intracellular trypsinogen activation
J. Clin. Invest.
Inflammatory mediators in acute pancreatitis
J. Pathol.
Proteasome inhibitor therapy in multiple myeloma
Mol. Cancer Ther.
Involvement of oxygen-derived free radicals in L-arginine-induced acute pancreatitis
Dig. Dis. Sci.
Possible lysosomal activation of both human trypsinogen by cathepsin B and spontaneous acid activation of human trypsinogen.
Biol. Chem. Hopp-Seyler
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