Experimental Study on the Apoptosis of MDEC-07114Cell Induced by Methomyl
|School||Zunyi Medical College,|
|Keywords||Methomyl Musca Domestica Embryonic Cell apoptosis mitochondrial|
Objective:To explore the apoptosis and mechanism of Musca domestica embryonic cell line (MDEC-07114) induced by Methomyl.Methods:1.Experimental groups:the different concentrations of Methomyl groups (1mg/ml group,2mg/ml group,4mg/ml group,8mg/ml group)、normal group、1%acetone group;2. The morphology of apoptosis cells was observed by hematoxylin-eosin stainings,and the apoptosis index(AI) was measured.3.The apoptosis of MDEC-07114cell was detected by TUNEL.4.The ultra-structure changes of MDEC-07114was observed by transmission electron microscopy.5. Rhodamine-labeled MDEC-07114cells,effect of Methomyl on mitochondrial membrane potential were detected by flow cytometry.Result:1.From the2mg/ml groups of Methomyl, the typical apoptotic cell began to appear. Moreover, with the increase of the concentration,the apoptosis index(AI) was gradually increased.2. TUNEL assay:at the2mg/ml groups, MDEC-07114cells appeared the visible green fluorescence. The green fluorescence in cells was enhanced at the4mg/ml groups. It was the strongest at the8mg/ml groups.3.Transmission electron microscopy observations:The cell was observed nuclear chromatin condensation and margination at1mg/ml group.From the2mg/ml group, apoptotic cells increased with the increase of the concentration. Vacuoles in the cytoplasm, karyopycnosis, mitochondrial injury and apoptotic bodies were observed.4.Flow cytometry assay:The Rh-123fluorescentintensity in MDEC-07114cells was reduced after treatment for lh,and fluorescence peak shift left, the mitochondrial membrane potential was degressed.The declining rate was typical in time-dependent manner.Conclusion:1.Methomyl can induce apoptosis of MDEC-07114cells;2. The cell apoptosis of Methomyl induced may be involved in mitochondrial pathway.