Pterostilbene induces accumulation of autophagic vacuoles followed by cell death in HL60 human leukemia cells.

Pterostilbene, a naturally occurring structural analog of resveratrol, has been reported to exert antiproliferative and proapoptotic effects in various cancer types. Recently, it has been demonstrated to induce both autophagy and apoptosis in human bladder and breast cancer cell lines. The aim of this study was to evaluate the effects of pterostilbene on HL60 human leukemia cells. Cell morphology was examined using confocal and electron microscopy. Cell viability was determined by MTT, neutral red uptake and trypan blue exclusion assays. LC3 processing was studied based on Western blotting and immunofluorescence analyses. Flow cytometry was used to study cell cycle distribution, phosphatidylserine externalization, caspase activation, disruption of mitochondrial membrane potential and intracellular production of reactive oxygen species. DNA degradation was examined by gel electrophoresis. We found that treatment of HL60 cells with pterostilbene at the IC90 concentration resulted in the G0/G1 cell cycle arrest. Pterostilbene induced conversion of cytosolic LC3-I to membrane-bound LC3-II and accumulation of large LC3-positive vacuolar structures. Pterostilbene also led to phosphatidylserine externalization, internucleosomal DNA fragmentation, caspase activation and disruption of mitochondrial membrane potential. Moreover, it did not induce oxidative stress. Our results suggest that pterostilbene induces accumulation of autophagic vacuoles followed by cell death in HL60 cells.

Ultrastructural response of arcuate nucleus neurons to fasting in aged rats.

The arcuate nucleus of the hypothalamus (ARH) is involved in the control of energy homeostasis. Leptin - an adipocyte derived hormone - is known to act on the hypothalamic nuclei and thus to control body weight by food intake reduction. Oxidative stress is believed to be implicated in leptin signalling. However, its relevance for leptin-induced signal transduction within ARH remains unclear. The goal of the study was to investigate the effect of fasting on morphological alterations of the neuronal endoplasmic reticulum/Golgi network as well as on the expression of leptin receptors in the arcuate nucleus of aged rats. Male Wistar rats, aged 24 months, were fasted for 96 hours. The control animals were fed ad libitum. Membranous whorls in the ARH neurons were visualized using the electron microscopy technique. Leptin receptors in the membranes of ARH neurons were determined immunohistochemically (IHC), and soluble leptin receptors in the plasma as well as plasma isoprostanes were quantified immunochemically (ELISA). An intense formation of membranous whorls was observed, directly associated with the cisternae of the rough endoplasmic reticulum, as well as lamellar bodies. Interestingly, the whorls were often localized near a well-developed Golgi complex. Moreover, some Golgi complexes displayed an early stage of whorl formation. Groups of residual lipofuscin granules were found in the immediate proximity of the whorls. An increased immunoreactivity with neuronal leptin receptors suggests that hypersensitive neurons may still effectively respond to the fasting serum levels of leptin, mediating ultrastructural transformation of ARH neurons during short-term fasting. Having observed a significant accumulation of lipofuscin granules and a marked increase of total 8-isoprostane serum level in the fasting rats, we hypothesize that signal transduction within the neurons of ARH is dependent on oxidative stress phenomena.