Influences Of Division Growth And Relationship Between Cytotoxicityn And Apoptosis On Aedes Albopictus Cell Line (C6/36) Produced By Piperine

Objective To observe cell proliferation,apoptosis,the expression of Fas protein,growth state and recovery of damaged cell produced by botanical insecticide piperine on C6/36 cell strain of Aedes albopictus. Toxicity of piperine was studied at cellular level in order to compare its action on integer and microcosmic cells then further bring to light the mechanism and mode of action of piperine.Methods The growth rate and recovery of C6/36 cell was evaluated. MTT,inverted microscope observation was using in this study to observe the effects of piperine on cell multiplication. Annexin-V/PI double staining and Fas protein evaluation were using to study apoptosis and the relationship between apoptosis and cell death. Through all these we could study mode action and cytotoxicity on C6/36 cell by piperine.Results The main results are given as follow:(1) The growth rate of C6/36 cell had no change at the concentration of 0.018mmol/ L while was influenced and depressed apparently by piperine at the concentration of 0.14mmol/ L. And the damage was grate at the concentration of 0.56mmol/ L because the cells recovered slowly after a long term of stop. And it can cause the change of morphous of cell above the concentration of 0.035 mmol/L . The cells represent pantomorphia under the inverted microscope. They showed swollen or shrinkage and the space between cells were widened. A large number of cells assembled to be conglobations, and then ablated, disaggregated and death. Treat the cell with high concentration of piperine (0.56 mmol/L ) for 24 hours and 48 hours respectively. During a period of growth retardation, the cells of 24-hour-group can recover laggingly while the 48-hour-group show inconvertibility. It suggests that there is time dependence of the effect on C6/36 by piperine . The cytotoxicity would be increased if the treating time prolonged.(2) It was found that the LD50 of C6/36 cell 24 hours after treated by piperine was 0.32 mmol/L . Besides the cytotoxicity was increasing following the raise of concentration of piperine. The result of Annexin-V/PI double staining showed that the apoptosis was apparent when treated by the concentration above 0.28 mmol/Lwhile the rate of apoptosis was 19.4% at the concentration of 0.56 mmol/L . The expression of Fas protein increased at the concentration above 0.28 mmol/L but not apparent and the total death rate was 72.7% which all suggest that apoptosis is not the main mechanism of the cytotoxicity caused by piperine.Conclusion There are strong toxic action and growth inhibiting on C6/36 cell strain of Aedes albopictus by piperine and piperine should be the important functional material to research neotype insecticide.