Buspirone partially protects an astrocytic cell line from the ethanol

ROSSI AR, EVRARD SG, RAMOS AJ, BRUSCO A

Buzios, Brasil

Congreso; 1er Congreso IBRO/LARC de Neurociencias de América Latina, Caribe y Península Ibérica (NeuroLatAm); 2008

SAN

The human astrocytoma cell line 1321N1 expresses 5HT1A receptors. Our objectives were to study i) the response of this cell line to ethanol (E) ;ii) if buspirone (B, a 5HT1A partial agonist) has a protective effect. The 1321N1 cells were cultured in slides with the following culture medium: DMEM, high glucose, 1% w/v glutamine, 10% v/v fetal bovine serum and 1% penicillin-streptomycin at 37° C, 5% CO2. When confluence reached 80% the culture medium was replaced by OPTIMEM for 3 hours before the experimental treatment for three days with: a) OPTIMEM only (control, C); b) OPTIMEM + 400nM B; c) OPTIMEM + 25mM E; d) OPTIMEM + 25mM E and 400nM B; e) OPTIMEM + 75mM E; f) OPTIMEM + 75mM E and 400nM B. Cells were fixed with 4% p/v paraformaldehyde in phosphate buffer 0.1M pH 7.4 for 15 min at room temperature. We assessed nuclear morphology in Hoechst 33342 stained cells. The expression of 5HT1A receptors, GFAP and S100B protein were assessed by immunoflurescence. The results were: 1) a tendency to an increase in E25 in the number of abnormal nuclei/field with an increase in E75 (C: 9.06 ± 4.15; E25: 20.18 ± 7.98; E75: 39.53 ± 22.74, P<0.01) reversed when B was added (E25+B: 14.27 ± 6.97; E75+B: 19.85 ± 10.51); 2) a maximal decrease of total cell number/field in the 75mM E treatment (C: 78.6 ± 56.49; E25: 40.08 ± 29.54; E75: 15.6 ± 7.8) partially reversed when B was added (E75+B: 39.92 ± 19.45); 3) more than 80% of the cells coexpressed S100B, GFAP and 5HT1A receptors. E altered nuclear morphology and the number of processes in each cell. These results show that buspirone partially protects the astroglioma cell line from the detrimental effects of the ethanol. Since this cell line has 5HT1A receptors, they are likely to be involved in the protective effects.