The Toxic Effects Of Triethylamine In HBE Cells

Objective:Study the toxic effects of triethylamine on human bronchial epithelial cells, provide the basis for a comprehensive evaluation of the toxicity of triethylamine.Methods:(1) The HBE cells were exposed with different concentration of triethylamine(0, 5, 7.5, 10, 15, 20 mmol/L) of different times(6, 12, 24 and 48 h);The cell viability were detected by living cells count kit(CCK- 8);(2) The HBE cells were exposed with different concentration of triethylamine(0, 5, 7.5, 10, 15, 20 mmol/L) of different times(6, 12, 24 and 48 h), the reactive oxygen species(ROS) level of HBE cells were measured by flow cytometry instrument;The malondialdehyde(MDA)content were detected by intracellular malondialdehyde detection kits;(3) The HBE cells were exposed with different concentration of triethylamine(0, 5, 7.5, 10, 15, 20mmol/L) of different times(6, 12, 24 and 48 h);Single cell gel electrophoresis was applied for quantitative analysis of DNAdamage in HBE cells induced by triethylamine.With different concentration of triethylamine(0, 5, 7.5, 10, 15 mmol/L)infected HBE cells for 24 hours, flow cytometry instrument detected HBE cells changes in cell cycle.Results:(1)Statistics showed that: in the 6 hours and 12 hours exposure period,exposed to the concentration of 15 mmol/L,compared with the control group, the difference of HBE cells viability is statistically significant(P < 0.05);In the 24 hours and 48 hours exposed period, exposed to the concentration of 10 mmol/L, compared with the control group, the difference of HBE cells viability is statistically significant(P < 0.05); When the concentration of the triethylamine is less than or equal to 10mmol/L, there was no statistically significant difference between each group;When the concentration of triethylamine was 15 mmol/L,compared with the 6 hours group,the results were significantly decreased(P < 0.05);The experimental results show that the viability of the HBE cells decreased with the increase of the exposed concentration and times, and there was a significant dose- response relationship;(2) Statistics showed that:The level of ROS and the content of MDA in HBE cells was significantly increased compare to the control(P<0.05).With the increase of exposure time andconcentration;The level of ROS and the content of MDA in HBE cells increased obviously, and presents a certain dose- response relationship;(3)The results of SCGE showed that in the terms of the tail length,the tail DNA% and the Olive tail moment,for the same exposure period,the difference between the exposure group and the control group was significant(P < 0.05);(4) Under the function of triethylamine,there are significant changes in cell cycle(P < 0.05).The G1 phase cells were decreased,S phase and G2 phase cells were increased significantly.The difference was significant between the exposed groups and the control(P < 0.05).Conclusion:(1) Triethylamine can obviously inhibit HBE cells proliferation, and survival rate of HBE cells decreased with the increase of concentration and exposure time of triethylamine significantly, and there was an obvious dose- response relationship;(2)The level of ROS and the content of MDA in HBE cells were increased sinificantly with the exposed of triethylamine, and there was an obvious dose- response relationship,which indicated that triethylamine caused severe oxidative damage in HBE cells;(3) Triethylamine caused HBE cells DNA damage and the cell cycle of HBE cells may also be affected.