Increase of heat shock protein gene expression by melatonin in AR42J cells.
Heat shock proteins (HSPs) have been reported to protect the pancreatic cells from the acute damage produced by caerulein overstimulation. However the effects of caerulein, melatonin or hyperthermia preconditioning on mRNA signal for HSP60 in the pancreatic acinar cells has not been examined yet. The aims of this study were: 1. To investigate the gene expression for HSP60 in the pancreatic AR42J cells stimulated by melatonin, caerulein or combination of both these substances. 2. To compare above changes with mRNA signal for HSP60 in pancreatic AR42J cells subjected to hyperthermia preconditioning. AR42J cells were incubated in standard medium at 37 degrees C for: 0, 1, 3, 5, 12 or 24 h, under basal conditions. Above cells were then subjected to heat shock (42 degrees C) for 0, 1 or 3 h. In the next part of the study AR42J cells were incubated in presence of caerulein (10(-11), 10(-9) or 10( -7) M), melatonin (10(-8) or 10(-6) M), or combination of above under basal conditions or following heat shock pretreatment. Gene expression for HSP60 was determined by RT-PCR. The mRNA signal for HSP60 has been observed in AR42J cells under basal conditions, and this signal was markedly and time-dependently increased in these cells subjected to hyperthermia preconditioning. Incubation of AR42J cells in presence of melatonin (10(-8) or 10(-6) M) resulted in the significant and dose-dependent increase of gene expression for HSP60 in both groups of AR42J cells: preconditioned and in those, which were not subjected to hyperthermia. Caerulein stimulation reduced mRNA signal for HSP60. The strongest signal has been observed after the exposition of AR42J cells to hyperthermia preconditioning, combined with melatonin and caerulein. We conclude that: 1. Gene expression for HSP60 has been detected in pancreatic AR42J cells under basal conditions. 2. Hyperthermia preconditioning resulted in a significant and time-dependent increase of HSP60 signal in pancreatic AR42J cells. 3. HSP60 gene expression was significantly increased in pancreatic AR42J cells stimulated by melatonin whereas caerulein reduced this signal. 4. The strongest gene expression for HSP60 has been found in the cells subjected to the combination of hyperthermia preconditioning, caerulein and melatonin.
Bonior
J
J
Department of Medical Physiology Health Care Faculty, School of Medicine, Jagiellonian University, Cracow, Poland.
Jaworek
J
J
Konturek
S J
SJ
Pawlik
W W
WW
D000199:Actins / Q000096:biosynthesis; D000818:Animals; D002460:Cell Line; D002108:Ceruletide / Q000494:pharmacology; D018834:Chaperonin 60 / Q000096:biosynthesis / Q000235:genetics; D005334:Fever / Q000503:physiopathology; D016166:Free Radical Scavengers / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D008550:Melatonin / Q000494:pharmacology*; D010179:Pancreas / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D013268:Stimulation, Chemical
D000199:Actins; D018834:Chaperonin 60; D016166:Free Radical Scavengers; D006360:Heat-Shock Proteins; D012333:RNA, Messenger; D002108:Ceruletide; D008550:Melatonin