Abstract

BENEFICIAL EFFECTS OF PYRIDOXAL PHOSPHATE ON HYDROGEN PEROXIDE INDUCED STRESS ON ISOLATED PANCREATIC ACINAR CELLS

Rajanna Ajumeera and Vijayalakshmi Venkatesan*

Abstract

Present study addressed the cytoprotective and anti oxidant effects of pyrodoxal phospahe (PLP) along with increased expression levels of important transcription factors of pancreas acinar cells from 3 month WNIN rat pancreas in vitro. The experiments were carried out with acinar cells were primed with or without PLP (5mM) against H2O2 (100μM) induced stress in cell culture conditions. These cells were divided in to four groups as untreated group 1 (without any treatment), group 2 (30 minutes H2O2 exposed), group 3 (24 hours PLP primed) and group 4 (PLP exposure for 23.5 hours followed by 30 minutes H2O2 exposure. The cell viability was estimated with MTT assay, group 1 considered as 100%, while in group 2 it was decreased to 82% due to H2O2 induced tress; in group3 it was 99.8%; fallowed 99.5% in group 4 indicated PLP has protected from H2O2 impact. Increased levels of reactive species (ROS) was induced with H2O2 in group 2 was estimated 1.47 folds more compared to group 1, while PLP presence minimised to 1.02 fold in group 4. Increased expressions of pancreatic important proteins fold change were observed as amylase -2.01, neurogenin-3-9.51, PDX-1- 23.6 and insulin-13.5 in group 3 compared to group 1. The more or less similar changes were observed with group 4. The specific PLP’s response was confirmed by amino oxy acetic acid (AOAA), which is a PLP inhibitor. The increased amylase protein localization with PLP was confirmed by confocal microscopy. Insulin secretion efficiency of acinar cells was observed to be 6.13 folds higher at basal and 24.63 fold higher at stimulated levels in group3 compared to group1. Taken together PLP proved as potential molecule to protect acinar cells from the stress.

Keywords: acinar cells; PLP; amylase; insulin secretion; FACS; viability; PDX-1.