Dataset Information

Caffeine protects against experimental acute pancreatitis by inhibition of inositol 1,4,5-trisphosphate receptor-mediated Ca2+ release.

ABSTRACT:

Objective

Caffeine reduces toxic Ca²⁺ signals in pancreatic acinar cells via inhibition of inositol 1,4,5-trisphosphate receptor (IP₃R)-mediated signalling, but effects of other xanthines have not been evaluated, nor effects of xanthines on experimental acute pancreatitis (AP). We have determined effects of caffeine and its xanthine metabolites on pancreatic acinar IP₃R-mediated Ca²⁺ signalling and experimental AP.

Design

Isolated pancreatic acinar cells were exposed to secretagogues, uncaged IP₃ or toxins that induce AP and effects of xanthines, non-xanthine phosphodiesterase (PDE) inhibitors and cyclic adenosine monophosphate and cyclic guanosine monophosphate (cAMP/cGMP) determined. The intracellular cytosolic calcium concentration ([Ca²⁺]_C), mitochondrial depolarisation and necrosis were assessed by confocal microscopy. Effects of xanthines were evaluated in caerulein-induced AP (CER-AP), taurolithocholic acid 3-sulfate-induced AP (TLCS-AP) or palmitoleic acid plus ethanol-induced AP (fatty acid ethyl ester AP (FAEE-AP)). Serum xanthines were measured by liquid chromatography-mass spectrometry.

Results

Caffeine, dimethylxanthines and non-xanthine PDE inhibitors blocked IP₃-mediated Ca²⁺ oscillations, while monomethylxanthines had little effect. Caffeine and dimethylxanthines inhibited uncaged IP₃-induced Ca²⁺ rises, toxin-induced Ca²⁺ release, mitochondrial depolarisation and necrotic cell death pathway activation; cAMP/cGMP did not inhibit toxin-induced Ca²⁺ rises. Caffeine significantly ameliorated CER-AP with most effect at 25?mg/kg (seven injections hourly); paraxanthine or theophylline did not. Caffeine at 25?mg/kg significantly ameliorated TLCS-AP and FAEE-AP. Mean total serum levels of dimethylxanthines and trimethylxanthines peaked at >2?mM with 25?mg/kg caffeine but at <100?µM with 25?mg/kg paraxanthine or theophylline.

Conclusions

Caffeine and its dimethylxanthine metabolites reduced pathological IP₃R-mediated pancreatic acinar Ca²⁺ signals but only caffeine ameliorated experimental AP. Caffeine is a suitable starting point for medicinal chemistry.

SUBMITTER: Huang W

PROVIDER: S-EPMC5284483 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

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Publications

Caffeine protects against experimental acute pancreatitis by inhibition of inositol 1,4,5-trisphosphate receptor-mediated Ca2+ release.

Huang Wei W Cane Matthew C MC Mukherjee Rajarshi R Szatmary Peter P Zhang Xiaoying X Elliott Victoria V Ouyang Yulin Y Chvanov Michael M Latawiec Diane D Wen Li L Booth David M DM Haynes Andrea C AC Petersen Ole H OH Tepikin Alexei V AV Criddle David N DN Sutton Robert R

Gut 20151207 2

<h4>Objective</h4>Caffeine reduces toxic Ca<sup>2+</sup> signals in pancreatic acinar cells via inhibition of inositol 1,4,5-trisphosphate receptor (IP<sub>3</sub>R)-mediated signalling, but effects of other xanthines have not been evaluated, nor effects of xanthines on experimental acute pancreatitis (AP). We have determined effects of caffeine and its xanthine metabolites on pancreatic acinar IP<sub>3</sub>R-mediated Ca<sup>2+</sup> signalling and experimental AP.<h4>Design</h4>Isolated pancre ...[more]

PMID: 26642860

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Project description:AIMS:Phospholamban (PLB) is the key regulator of the cardiac Ca2+ pump (SERCA2a)-mediated sarcoplasmic reticulum Ca2+ stores. We recently reported that PLB is highly concentrated in the nuclear envelope (NE) from where it can modulate perinuclear Ca2+ handling of the cardiomyocytes (CMs). Since inositol 1,4,5-trisphosphate (IP3) receptor (IP3R) mediates nuclear Ca2+ release, we examined whether the nuclear pool of PLB regulates IP3-induced nuclear Ca2+ handling. METHODS AND RESULTS:Fluo-4 based confocal Ca2+ imaging was performed to measure Ca2+ dynamics across both nucleus and cytosol in saponin-permeabilized CMs isolated from wild-type (WT) or PLB-knockout (PLB-KO) mice. At diastolic intracellular Ca2+ ([Ca2+]i?=?100?nM), the Fab fragment of the monoclonal PLB antibody (anti-PLB Fab) facilitated the formation and increased the length of spontaneous Ca2+ waves (SCWs) originating from the nuclear region in CMs from WT but not from PLB-KO mice. We next examined nuclear Ca2+ activities at basal condition and after sequential addition of IP3, anti-PLB Fab, and the IP3R inhibitor 2-aminoethoxydiphenyl borate (2-APB) at a series of [Ca2+]i. In WT mice, at 10?nM [Ca2+]i where ryanodine receptor (RyR2) based spontaneous Ca2+ sparks rarely occurred, IP3 increased fluorescence amplitude (F/F0) of overall nuclear region to 1.19?±?0.02. Subsequent addition of anti-PLB Fab significantly decreased F/F0 to 1.09?±?0.02. At 50?nM [Ca2+]i, anti-PLB Fab not only decreased the overall nuclear F/F0 previously elevated by IP3, but also increased the amplitude and duration of spark-like nuclear Ca2+ release events. These nuclear Ca2+ releases were blocked by 2-APB. At 100?nM [Ca2+]i, IP3 induced short SCWs originating from nucleus. Anti-PLB Fab transformed those short waves into long SCWs with propagation from the nucleus into the cytosol. In contrast, neither nuclear nor cytosolic Ca2+ dynamics was affected by anti-PLB Fab in CMs from PLB-KO mice in all these conditions. Furthermore, in WT CMs pretreated with RyR2 blocker tetracaine, IP3 and anti-PLB Fab still increased the magnitude of nuclear Ca2+ release but failed to regenerate SCWs. Finally, anti-PLB Fab increased low Ca2+ affinity mag-fluo 4 fluorescence intensity in the lumen of NE of nuclei isolated from WT but not in PLB-KO mice. CONCLUSION:PLB regulates nuclear Ca2+ handling. By increasing Ca2+ uptake into lumen of the NE and perhaps other perinuclear membranes, the acute reversal of PLB inhibition decreases global Ca2+ concentration at rest in the nucleoplasm, and increases Ca2+ release into the nucleus, through mechanisms involving IP3R and RyR2 in the vicinity.

Dataset Information

Caffeine protects against experimental acute pancreatitis by inhibition of inositol 1,4,5-trisphosphate receptor-mediated Ca2+ release.

Objective

Design

Results

Conclusions

Publications

Caffeine protects against experimental acute pancreatitis by inhibition of inositol 1,4,5-trisphosphate receptor-mediated Ca2+ release.

Similar Datasets

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