| ObjectivesTo study Simvastatin effects expression ofα-sunbunit of epithelial sodium channel on A549 cell membrane in vitro.Methods1. A549 cells were regularlly subcultured in vitro and were divided into two group: different concentrations of simvastatin group(A group)and simvastatin for different time group(B group).A goup divided intoA1 group( blank group) , A2 group (simvastatin 2.5uM group) , A3 group(simvastatin 5uM group) ,A4 group(simvastatin 10uM group) ,cultured for 24 hours.And B goup divided into B1 group( blank group), B2 group(simvastatin 10uM 12h group),B3 group(simvastatin10uM 24h group), B4 group(simvastatin 10uM 48h group) .2. The protein level ofα-ENaC in A549 cell was analyzed by Western blotting.The expression ofα-ENaC in A549 cell mRNA was detected by RT-PCR.Results1. The result of Western Blot indicated that in A group the expression of protein ofα-ENaC in the membrane of A549 cell was increased in A3 group (P<0.05),and was significant increased in A4 group(P<0.01);At the same time , the result of RT-PCR indicated that the mRNA ofα-ENaC was increased in A3 group(P<0.05),and that was significant increased in A4 group(P<0.01) .2. The result of Western Blot indicated that in B group the protein ofα-ENaC in the membrane of A549 cell in B3 group and B4 group were higher than those in B1 and B2 group (P<0.05).The RT-PCR indicated the same rerult .But there were no significant difference in the former two and the latter two in either protein expression or mRNA expression(P>0.05).Conclusions1. Simvastatin can up-regulated the expression the protein and mRNA ofα-ENaC in the membrane of A549 cell with dose-dependent. high-dose Simvastatin could significant improve the expression levels ofα-ENaC protein andα-ENaCmRNA in the membrane of A549 cell.2. The effect of simvastatin increase the expression of the protein and mRNA ofα-ENaC in the membrane of A549 cell after simvastatin was administrated 24hs ,but not in 12hs . |
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