Endoplasmic Reticulum Stress Accounts For Sevoflurane-induced Parthanatos In Developing Brain

Objective:To explor the effects of endoplasmic reticulum stress on sevoflurane-induced Parthanatos in order to clarify the possible mechanism of sevoflurane-induced neurotoxicity in developing brain.Method:Selecting Human SH-SY5 Y neuroblastoma cells and rat primary hippocampal neurons as the research objects.The cells were grouped as follows: blank control groups(C groups),3-Aminobenzamide groups(3AB groups),N-acetylcysteine groups(NAC groups),4-Phenylbutyric acid groups(4-PBA groups),sevoflurane groups(Sevo groups),sevoflurane+3-Aminobenzamide groups(Sevo+3AB groups),sevoflurane+N-acetylcysteine groups(Sevo+NAC groups)and sevoflurane+4-Phenylbutyric acid groups(Sevo+4-PBA groups).C groups were not treated with drugs.3AB groups,NAC groups and 4-PBA groups were treated with 3AB(500 (?)mol/L),NAC(5 mmol/L)and 4-PBA(3 mmol/L)for 13 h respectively.Sevo groups were treated with different concentrations of sevoflurane(2%,4%,8%)for 6 h,12 h and 24 h.Sevo+3AB groups,Sevo+NAC groups and Sevo+4-PBA groups were pretreated with 3AB(500 (?)mol/L),NAC(5 mmol/L)and 4-PBA(3 mmol/L)for 1 h respectively,and then treated with different concentrations of sevoflurane(4%,8%)for 12 h.Cell viability assay(MTT)was used to detect the viability of Human SH-SY5 Y neuroblastoma cells and rat primary hippocampal neurons.LDH release assay was used to detect cells mortalities.Western blot was used to detect the expression of Parthanatos-related proteins and endoplasmic reticulum stress-related proteins in Human SH-SY5 Y neuroblastoma cells and rat primary hippocampal neurons.DCFH-DA was used to detect the levels of reactive oxygen species(ROS)in Human SH-SY5 Y neuroblastoma cells and rat primary hippocampal neurons.Results:Sevoflurane induced cells death of Human SH-SY5 Y neuroblastoma cells and rat primary hippocampal neurons in a time and dose-dependent manner(P<0.01),decreased cells activity(P<0.01),promoted the expression of Parthanatos-related proteins(PARP-1,PAR,AIF)in cells(P<0.01).3AB significantly reduced cells mortalities(P<0.01),reduced the inhibition of sevoflurane on cells(P<0.01),decreased the expression of Parthanatos-related proteins(PARP-1,PAR,AIF)in cells(P<0.01).Sevoflurane induced overproduction of ROS in SH-SY5 Y cells and rat primary hippocampal neurons(P<0.01).NAC can significantly inhibited the overproduction of ROS induced by sevoflurane.NAC also reduced cells mortalities(P<0.01),reduced the inhibition of sevoflurane on cells(P<0.01),decreased the expression of Parthanatos-related proteins(PARP-1,PAR,AIF)in cells(P<0.01).Sevoflurane promoted the expression of endoplasmic reticulum stress-related proteins(GRP78、ATF-6、p-IRE-1、p-PERK、p-e IF-2α、ATF-4)in SH-SY5 Y cells and rat primary hippocampal neurons(P<0.01).4-PBA significantly decreased the expression of endoplasmic reticulum stress-related proteins(GRP78、ATF-6、p-IRE-1、p-PERK、p-e IF-2α、ATF-4)in cells,and also inhibited the overproduction of ROS induced by sevoflurane,reduced cells mortalities(P<0.01),reduced the inhibition of sevoflurane on cells(P<0.01),decreased the expression of Parthanatos-related proteins(PARP-1,PAR,AIF)in cells(P<0.01).Conclusion:Sevoflurane exposure induces neuronal cell Parthanatos mediated by endoplasmic reticulum stress via improvement of intracellular reactive oxygen species in SH-SY5 Y cells and rat primary hippocampal neurons.