Abstract
Objective To evaluate the effect of isoflurane on the apoptosis of SH-SYSY cells transfected with APPsw gene and the role of inositol 1,4,5-triphosphate (IP3) recepters.Methods The SH-SYSY ceils transfected with APPsw gene were seeded in culture flasks with the density of 1.2 × 104/cm2.The cells were randomly divided into 4 groups (n =6 each):control group (group C),IP3 receptor antagonist group (group Ⅹ),isoflurane group (group Ⅰ) and isoflurane + IP3 receptor antagonist group (group Ⅰ + Ⅹ).After the cells were cultured for 24 h and attached to the wall,the cells were cultured routinely in group C,and Xestospongin C 100 nmol/L (IP3 receptor antagonist) was added to DMEM culture medium in groups X and Ⅰ + X,and 30 min later the cells were exposed to 1.2 ％ sevoflurane for 8 h in groups Ⅰ and Ⅰ + X.The cells were collected for examination of the ultrastructure and for determination of cell apoptosis,intracellular free calcium ion concentration [Ca2 +] i (by flow cytometry) and expression of IP3 receptor protein (by Western blot).The apoptosis rate was calculated.Results Compared with group C,there was no significant change in the apoptosis rate,[Ca2 +]i or IP3 receptor protein expression in group Ⅹ (P ＞ 0.05),while the cell apoptosis rate and [Ca2 +] i were significantly increased and IP3 receptor protein expression was up-regulated in groups I and Ⅰ + Ⅹ (P ＜ 0.05 or 0.01).Compared with group Ⅰ,cell apoptosis rate and [Ca2+]i were significantly decreased and IP3 receptor protein expression was down-regulated in group Ⅰ + Ⅹ (P ＜ 0.01).The pathological changes of the cells happened in groups Ⅰ and Ⅰ + Ⅹ,and the pathological changes were severer in group Ⅰ than in group Ⅰ + Ⅹ.Conclusion Isoflurane can induce apoptosis of SH-SY5Y cells transfected with APPsw gene through increasing [Ca2+]i and up-regulating IP3 receptor protein expression. Key words: Isoflurane; Apoptosis; Genes; Transfection; Inositol 1,4,5-trisphosphate receptors
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