ABSTRACT
[ ABSTRACT] AIM:To prove the purinergic signaling mechanism of the neuroprotective action of hydrogen sulfide by observing the effects of sodium hydrosulfide ( NaHS) , a donor of hydrogen sulfide, on the cell viability, intracellular Ca2+concentration ( [ Ca2+] i ) and the change of membrane permeability in the PC12 cells injured by adenosine triphos-phate ( ATP) .METHODS: PC12 cells in logarithmic growth phase were randomly divided into 4 groups.In control group, the cells were cultured without ATP treatment.In ATP group, the cells were treated with ATP after cultured for 24 h.In NaHS+ATP group, the cells were incubated with NaHS for 30 min before treated with ATP, and NaHS always exis-ted in the reaction system.In KN-62+ATP group, the cells were pretreated with KN-62 for 30 min, and the other treat-ments were as the same as those in NaHS+ATP group.The cell viability was assessed by MTT assay.The [ Ca2+] i was detected by Fura-2/AM staining.The membrane permeability was observed by staining with fluorescent dye YO-PRO-1. RESULTS:ATP at concentration of 0.3 mmol/L showed no injury effect on the cells.However, the cell viability was dropped gradually in a dose-dependent manner as the ATP at doses of 1, 3, 5 and 10 mmol/L.The decline of cell viability by ATP was obviously reversed by 200 μmol/L of NaHS in the PC12 cells (P<0.05), but exasperated by 800μmol/L of NaHS (P<0.05).At the same time, ATP evoked the increase in [Ca2+]i in a dose-dependent manner, which was inhib-ited by NaHS ( P<0.05) .Furthermore, the YO-PRO-1 uptake induced by ATP in a dose-dependent and time-dependent manner was also reduced by NaHS ( P<0.05) .CONCLUSION:Hydrogen sulfide has protective effect on the PC12 cells injured by ATP.The mechanism may be related to the reverse of the increased [ Ca2+] i and YO-PRO-1 uptake.