A preliminary investigation of cell damage and possible mechanisms in SH-SY5Y cells exposed to a combination of lead,cadmium,mercury and arsenic
Objective To investigate the damaging effect and potential underlying mechanism of combination exposure to lead(Pb),cadmium(Cd),mercury(Hg),and arsenic(As)on SH-SY5Y cells.Methods A co-culture cell model of SH-SY5Y and HMC3 was established using the Transwell system,alongside a single cell culture of SH-SY5Y cells serving as a tissue-negative control.The cells were exposed to concentrations of 1,10,20,and 40 times the simulated mixture of Pb,Cd,Hg,and As(metal mixture,MM),with the highest concentration of 40 MM comprising 2.00 μmol/L Pb,0.01 μmol/L Cd,0.07 μmol/L Hgand 0.06 μmol/L As.A negative control for the mixture exposure was established using DMEM/F12 medium containing 10％fetal bovine serum and 1％penicillin.Cell viability was evaluated using the CCK8 assay,the expression levels of APP and MT1G transcripts were analyzed by Real-time PCR,and the levels of TNF-α,IL-6 and IFN-γ in the culture medium supernatant were measured by ELISA.To assess the level of oxidative stress in the cells after exposure,flow cytometry was used to detect the level of ROS after exposure of SH-SY5Y/HMC3 to 0,1,10,20,and 40 MM of metal mixtures,and single-cell culture of SH-SY5Y in DMEM/F12 complete medium was used as a tissue-negative control.Results After the combined exposure of Pb,Hg,Cd and As,both the co-culture and mono-culture models of SH-SY5Y cells exhibited inhibited growth(P＜0.05),and increased expression of MT1G mRNA.No significant changes were observed in APP mRNA levels.The supernatant showed an increasing trend in TNF-α content and a dose-dependent decreasing trend in IL-6 and IFN-γ.Compared with the result of the single SH-SY5Y cell injury indicators,the co-culture model showed less growth inhibition of SH-SY5Y cells,relatively lower APP mRNA,higher MT1G mRNA,smaller increase in TNF-α content,and more obvious decreasing trend of IL-6 and IFN-γ.Relative ROS levels increased dose-dependently with increasing exposure concentrations(F(4,10)=320.4,P＜0.05).Relative ROS levels were positively correlated with MT1G(r=0.98,P=0.001 5).Conclusion The neuroinflammatory response to mixed exposure to heavy metals was associated with ROS generation,co-cultured HMC3 cells attenuated the extent of SH-SY5Y cell injury in the co-culture model,and MT1G may provide a protective role in the process of SH-SY5Y cell injury.
Heavy metal combined exposureNeuronal cell modelOxidative stressInflammatory responseMT1G
万方数据
维普
