Thiopental protects human neuroblastoma cells from apoptotic cell death - Potential role of heat shock protein 70.

Abstract

Abstract Aims Thiopental, a general anesthetic, is recommended for the treatment of severe brain injury in the presence of intracranial hypertension and during surgical procedures where adequate perfusion of the brain may be at risk leading to neuronal programmed cell death, also known as apoptosis. The exact cellular mechanisms of the neuroprotective effects associated with thiopental in this context have not been identified yet. As we have previously shown that thiopental is able to protect human t lymphocytes from apoptosis by inducing heat shock protein-70, we hypothesized that the same mechanism may be responsible in a neuronal cell model. Main methods Human SK-N-SH neuroblastoma cells were preincubated with thiopental (100–500 μg/ml) before the induction of apoptosis (staurosporine, 2 μM). DNA-binding of heat shock factor (HSF)-1 was analyzed by electrophoretic mobility shift assay, mRNA-expression of heat shock protein (hsp)70 by Northern Blot, activity of caspase-3 by fluorogenic caspase activity assay and cleavage of pro-caspase-3 by Western blot. Apoptosis was assessed by flow cytometry after annexin V-fluorescein isothiocyanate staining. Induction of Hsp70 was inhibited using N-formyl-3,4-methylenedioxy-benzylidene-gamma-butyrolaetam. Data are given as mean ± standard deviation or median and 25/75% percentile. Key findings Dobutamine induced DNA-binding of HSF-1 and mRNA expression of hsp70. Pre-incubation with thiopental inhibited staurosporine-induced LDH release (p Significance Thiopental protects human neuroblastoma cells from apoptotic cell death possibly by the induction of Hsp70.