Caffeine activates preferentially α1-isoform of 5'AMP-activated protein kinase in rat skeletal muscle.

[Aim]: Caffeine activates 5′AMP-activated protein kinase (AMPK), a signalling intermediary implicated in the regulation of glucose, lipid and energy metabolism in skeletal muscle. Skeletal muscle expresses two catalytic α subunits of AMPK, α1 and α2, but the isoform specificity of caffeine-induced AMPK activation is unclear. The aim of this study was to determine which α isoform is preferentially activated by caffeine in vitroand in vivo using rat skeletal muscle. [Methods]: Rat epitrochlearis muscle was isolated and incubated in vitro in the absence or presence of caffeine. In another experiment, the muscle was dissected after intravenous injection of caffeine. Isoform-specific AMPK activity, the phosphorylation status of AMPKα Thr172 and acetyl-CoA carboxylase (ACC) Ser79, the concentrations of ATP, phosphocreatine (PCr) and glycogen, and 3-O-methyl-D-glucose (3MG) transport activity were estimated. [Results]: Incubation of isolated epitrochlearis muscle with 1 mM of caffeine for 15 min increased AMPKα1 activity, but not AMPKα2 activity; concentrations of ATP, PCr and glycogen were not affected. Incubation with 3 mM of caffeine activated AMPKα2 and reduced PCr and glycogen concentrations. Incubation with 1 mM of caffeine increased the phosphorylation of AMPK and ACC and enhanced 3MG transport. Intravenous injection of caffeine (5 mg kg−1) predominantly activated AMPKα1 and increased 3MG transport without affecting energy status. [Conclusion]: Our results suggest that of the two α isoforms of AMPK, AMPKα1 is predominantly activated by caffeine via an energy-independent mechanism and that the activation of AMPKα1 increases glucose transport and ACC phosphorylation in skeletal muscle

The role of peritoneal immunity and the tumour-bearing state on the development of wound and peritoneal metastases after laparoscopy

BackgroundThe effect of the tumour-bearing state and alterations in peritoneal immune function on the incidence of port-site and peritoneal metastases was investigated after laparoscopy with and without CO2 pneumoperitoneum.MethodsA suspension of viable adenocarcinoma cells was introduced into the left upper quadrant of the peritoneal cavity of syngeneic tumour-bearing rats at laparotomy, laparoscopy with CO2, and gasless laparoscopy. Control rats did not have pre-existing tumours. A group of non-tumour-bearing rats were also injected intraperitoneally with endotoxin 4 h before intraperitoneal tumour cell injection. Six days later the peritoneal cavity and surgical wounds were examined for macroscopic evidence of implanted tumour. Peritoneal macrophages were obtained from tumour-bearing rats subjected to different laparoscopic procedures and the activation state measured following exposure to lipopolysaccharide in vitro.ResultsIn the control rats, tumour implantation in the surgical wounds and peritoneum was significantly greater in the rats that had undergone laparoscopy with CO2. The presence of a pre-existing tumour was associated with increased tumour spread in all treatment groups and at most sites. Injection of endotoxin also resulted in increased tumour spread. Peritoneal macrophages from control and tumour-bearing rats who underwent laparoscopy with CO2 produced significantly less TNF-alpha in vitro, compared to gasless laparoscopy or laparotomy.ConclusionsCarbon dioxide insufflation enhances tumour spread and implantation. The underlying immune or metabolic status of the host, as influenced by the tumour-bearing state or modification of the peritoneal environment, also has a marked independent effect on tumour spread and implantation. The immune and metabolic status of the peritoneum including the extent of macrophage activation is implicated in this effect