The Role Of Mapksignaling Pathway In Death Induced By Aluminum In SH-SY5Y Cells

Objective To explore the role of MAPK signaling pathway in apoptosis and necroptosis inducedby aluminum in SH-SY5Y cells.Methods To imitate neural cell death induced by aluminium, we used AlCl36H2O(4mmol/L) totreat SH-SY5Y cells. Nec-1, the specific inhibitor for necroptosis, and zVAD-fmk, the specificinhibitor for apoptosis, were added into cultures for inhibiting the occurrence of necroptosis andapoptosis.And MAPK signaling pathways inhibitors were added into cultures for inhibiting thep-MAPK.Cell viability was performed to measure cell death, flow cytometry was used todistinguish apoptosis and necrosis，and western-blot were used to detect MAPK protein.Results1.Compared with blank control group, solvent control group, Nec-1control group andzVAD-fmk control group, cell viability of Al3+exposed group, Al3+plus Nec-1group and Al3+plus zVAD-fmk group showed statistically significant difference（P<0.01）. Compared with Al3+exposed group, Al3+plus Nec-1group and Al3+plus zVAD-fmk group showed statisticallysignificant difference（P<0.05）.Necroptotic rates and apoptotic rates in Al3+exposed group, Al3+plus Nec-1group and Al3+plus zVAD-fmk group obviously increased compared with the controlgroups, and the differences between those three groups were statistically significant（P<0.01）.Compared with control groups, expression of p-p38in Al3+exposed group, Al3+plus Nec-1group and Al3+plus zVAD-fmk group increased obviously, and expression of p-ERK decreasedsignificantly. Compared with Al3+exposed group, expression of p-p38decreased, but p-ERKincreased in Al3+plus Nec-1group;and expression of p-ERK increased in Al3+plus zVAD-fmkgroup.Compared with the control groups, expression of p-JNK in Al3+exposed group, Al3+plus Nec-1group and Al3+plus zVAD-fmk group have no statistical significance.2.(1)1）Compared with blank control group, solvent control group, SB203580control group,cellviability of Al3+exposed group,Al3+plus SB203580group，Al3+plus SP600125group and Al3+plus U0126group showed statistically significant difference（P<0.01）. And compared with Al3+exposed group, cell viability of Al3+plus SB203580group increased, Al3+plus U0126groupdecreased（P<0.01）.（2）Necroptotic rates and apoptotic rates in Al3+exposed group, Al3+plusSB203580group, Al3+plus SP600125group and Al3+plus U0126group obviously increasedcompared with the control groups（P<0.05）, and compared with Al3+exposed group, necroptotic rates and apoptotic rates of Al3+plus SB203580group decreased（P<0.01）, and Al3+plus U0126group increased（P<0.01）.（3）1) Compared with blank control group, solvent control group, expression of p-p38in p38inhibitor SB203580control group obviously decreased（P<0.01）, Al3+exposed group and Al3+plus SB203580group increased（P<0.01）; compared with Al3+exposed group, expression ofp-p38in Al3+plus SB203580group increased.2)Compared with blank control group, solventcontrol group, expression of p-JNK in SP600125control group,Al3+exposed group and Al3+plus SP600125group have no statistical significance（P>0.05）.3) Compared with blank controlgroup, solvent control group, expression of p-ERK in U0126control group, Al3+exposed groupand Al3+plus U0126group decreased（P<0.01）,and the difference between Al3+exposed groupand Al3+plus U0126group showed statistically significant difference（P<0.01）.Conclusion1.The dominant type of aluminum-induced SH-SY5Y cells death is necroptosis.2.The ERK and p38MAPK signaling pathways are involved in aluminum-induced SH-SY5Y cellsnecroptosis, and ERK is involved in apoptosis,JNK is not involved in neither aluminum-induced apoptosis nor necroptosis in this experiment.