多环芳烃光动力降解产物的细胞毒性检验

Cytotoxicological test of photodynamic degradation products of polycyclic aromatic hydrocarbons

对多环芳烃的光动力降解产物进行细胞毒性评价,将8种多环芳烃(菲、蒽、萘、■、荧蒽、苯并[a]芘、苯并[k]荧蒽和苯并[a]蒽)水溶液分为对照组和光动力试验组,分别与小鼠成纤维细胞L929细胞系共培养一段时间,通过倒置显微镜观察细胞形态的变化,磺酰罗丹明B试验计算细胞存活率,根据细胞核染色和Annexin V-FITC/PI染色配合流式细胞仪分析细胞凋亡情况来综合评价多环芳烃的光动力降解产物的细胞毒性。结果表明:经倒置显微镜观察,光动力试验组的死细胞数量变少;在25.0 ng/mL、12.5 ng/mL、3.1 ng/mL和1.6 ng/mL下,细胞存活率较之对照组分别增高了21.35%、20.22%、20.22%和20.23%;经细胞核染色发现,在50.0 ng/mL和25.0 ng/mL下,光动力试验组的细胞凋亡率分别为40%和14%,较之对照组明显降低;经Annexin V-FITC/PI染色及流式细胞仪分析,在50.0 ng/mL下,光动力试验组的细胞凋亡率比对照组降低了32.67%。试验结果证实了多环芳烃的光动力降解产物较之未降解的多环芳烃的细胞毒性降低。

The article is engaged in a study of the cytotoxicity of photodynamic degradation products of polycyclic aromatic hydrocarbons(PAHs). 8 PAHs(phenanthrene, anthracene, naphthalene, chrysene, fluoranthene, benzo [a] pyrene, benzo [k] fluoranthene, benzo [a] anthracene) mixed aqueous solution were divided into control group and photodynamic group. The concentrations of the two groups were set as 50.0 ng/mL, 25.0 ng/mL, 12.5 ng/mL, 6.3 ng/mL, 3.1 ng/mL and 1.6 ng/mL respectively. The two groups of solutions were co-cultured with mouse fibroblast L929 cell line for a period of time, and then the changes of cell morphology were observed by an inverted microscope. In addition, cell viability was calculated by sulfonyl rhodamine B test, and the cytotoxicity of PAHs photodynamic degradation products was comprehensively evaluated by Hoechst 33258 nuclear staining combined with fluorescence microscope imaging and Annexin V-FITC/PI staining combined with flow cytometry analysis, which are both cell apoptosis detection methods. Results show that the number of dead cells in the photodynamic group is reduced through observing with an inverted microscope. After incubation for 72 h, cell viability in the photodynamic group is significantly higher than that in the control group by 21.35%, 20.22%, 20.22% and 20.23%, at the concentration of 25.0 ng/mL, 12.5 ng/mL, 3.1 ng/mL and 1.6 ng/mL, respectively. Moreover, the cell apoptosis rate of the photodynamic group is 40% and 14% at the concentration of 50.0 and 25.0 ng/mL, which is significantly lower than that of the control group after being detected by Hoechst 33258 nuclear staining combined with fluorescence microscope imaging. Annexin V-FITC/PI staining combined with flow cytometry analysis showed that the apoptosis rate of the photodynamic group was lower than that of the control group at all concentrations. In addition, the apoptosis rate of the photodynamic group was 32.67% lower than that of the control group at the concentration of 50.0 ng/mL. The results of our experiments and investigations demonstrate that the cytotoxicity of the photodynamic degradation products of PAHs is lower than that of the undegraded PAHs.