| ObjectiveThe water transportation plays an important role in cell normal metabolism. Aquaporins(AQPs) are a family of membrance channels that plays an important role in human body. Among aquaporins AQP1 is the mostly investigated,which is involved in osmotic pressure driven water transport, mediating pharmacological effects, participating in edema formation and so on. AQP1 which has an complex regulatory mechanism are expressing in the kidney, lung, brain, and eyes. In this experiment, to investigate the impact of energy metabolism on the AQP1, we applied tubulin inhibitor Nocodazole, phosphocreatine kinase inhibitor iodoacetamide, and respiratory chain inhibitors to the Balb/c 3T3 fibroblast,and observed the changes in the expression of AQP1.MethodsThe Balb/c 3T3 fibroblast were cultured in vitro. tubulin inhibitor nocodazole, phosphocreatine kinase inhibitor iodoacetamide, and respiratory chain inhibitors such as nitric oxide, antimycin A, sodium azide were used in the cells. Then, detection the expression of AQP1 in these cells at Oh,4hr,6hr and Ohr,2hr,4hr,6hr,8hr by Western blotting.ResultsAfter incubated with nocodazole, iodoacetamide, nitric oxide, and antimycin A fibroblasts became round at 4 hrs, enlargement of nuclei and dead at 6hrs. When the Sodium azide was applied to fibroblasts, the cell morphology had no significant changes in the light microscope. Immunoblot demonstrated that AQP1 expression began to increase at 4hrs and was upregulated significantly at 6hrs,and was dose-dependence after incubated with nocodazole and iodoacetamide. After sodium azide were added into the cells, AQP1 expression commence to increase at 2hrs and was upregulated significantly at 4hrs.However,Nitric oxide and Antimycin A had no effect on AQP1 expresson.ConclusionTubulin inhibitor Nocodazole, phosphocreatine kinase inhibitor iodoacetamide, and respiratory chain inhibitors induced AQP1 expression in fibroblasts, which indicated that upregulated AQP1 accumulation was energy dependent.
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