The Effects Of Perfluorocarbon On Sodium Channels In A549 Cells

Objective:To explore the impaction of perfluorocarbon and new perfluorocarbon emulsion on A549 cells; To investigate the protection and mechanism of PFC and NPE on TNF-αattacked A549 cells.Methods:1. The exploration of PFC: A549 cells were divided into four groups: Control group untreated as blank control; TNF-αgroup was incubated with 100ng/ml TNF-αand PFC group with PFC at a 1/10 volume; we used the same dose of TNF-αand PFC in treat group. After treatment for 2,4,6 hours,the activity of cells were determined by MTT assay. The mRNA levels of ENaC-α,β,γsubunits and protein levels of ENaC-αwere detected by real-time PCR and western-blot, respectively.2. The exploration of NPE: A549 cells were divided into four groups: Control group untreated as blank control; TNF-αgroup was incubated with 100ng/ml TNF-αand NPE group with NPE at a 1/10 volume; we used the same dose of TNF-αand NPE in treat group. After treatment for 2,4,6 hours,the activity of cells were determined by MTT assay. The mRNA levels of ENaC-α,β,γsubunits and protein levels of ENaC-αwere detected by real-time PCR and western-blot, respectively. Results:1. Compared with control group, the cell morphology of other groups had no significant change after treated for 2 hours; however, when it came to 4 and 6 hours, the cells in TNF-αgroup grew alas, and many cells died and float. Meanwhile, the cells in PFC group and treat group had no difference with control group (P﹥0.05).2. The impaction of cell activity: after treatment for 2 hours, the optical densities of cells had no significant change among each group, compared with control group (P﹥0.05). As treated for 4 and 6 hours, the optical densities of cells in TNF-αgroup and treat group decreased, however, treat group has higher densities than TNF-αgroup.(P﹤0.05).3.The mRNA levels of ENaC-α,β,γsubunits: after treated for 2 hours,the mRNA levels of any group had no difference(P﹥0.05). Afeter treated for 4 and 6 hours,the mRNA levels of TNF-αgroup and treat group significant decreased compared with control group (P﹤0.05), and there was no difference between TNF-αgroup and treat group(P﹥0.05). There was no significant change in PFC group compared with control group after treated for 4 and 6 hours( P﹥0.05).4. The protein levels of ENaC-α: the protein levels of ENaC-αhad no significant change among each groups after treated for 2 hours. When treated for 4 and 6 hours, the protein levels of ENaC-αin TNF-αgroup and treat group decreased compared with control group, there was no difference between treat group and TNF-αgroup(P﹥0.05). The mRNA levels of PFC group had no difference with group C(P﹥0.05).5. The cell morphology of each group had no significant difference after treated for 2 hours; however, when incubated for 4 and 6 hours, the cells in TNF-αgroup grew bad, and many cells died and float. Meanwhile, the cells in NPE group and treat group had no difference with control group (P﹥0.05).6. The impaction of cell activity: after treatment for 2 hours, the optical densities of cells had no significant change among each group, compared with control group (P﹥0.05). As treated for 4 and 6 hours, the optical densities of cells in TNF-αgroup and treat group decreased, however, treat group has higher densities than TNF-αgroup.(P﹤0.05).7. The mRNA levels of ENaC-α,β,γsubunits: after treated for 2 hours,the mRNA levels of any group had no difference(P﹥0.05). Afeter treated for 4 and 6 hours,the mRNA levels of TNF-αgroup and treat group significant decreased compared with control group (P﹤0.05), and the treat group had a higer level than TNF-αgroup (P﹤0.05). There was no significant change in NPE group compared with control group after treated for 4 and 6 hours( P﹥0.05).8. The protein levels of ENaC-α: the protein levels of ENaC-αhad no significant change among each groups after treated for 2 hours. When treated for 4 and 6 hours, the protein levels of ENaC-αin TNF-αgroup and treat group decreased compared with control group, and the level of treat group was higer than TNF-αgroup (P﹤0.05). But the NPE group has the same level as control group (P﹥0.05).Conclusion:1. Incubated with TNF-αimpacted the morphology and cell activity of A549 cell.When the incubation last to 4 and 6 hours, the mRNA expression of ENaC-α,β,γsubunits and protein level of ENaC-αwere decreased.2. PFC protected A549 cells attacted with TNF-αby physical barriers, but has no ameliorations in the mRNA and protein expression.3. NPE released the depression of TNF-α, and aliveiated the downregulation of TNF-αto the mRNA and protein expression of ENaC in A549 cells.