论文部分内容阅读
目的观察三氯乙烯(TCE)诱导离体培养的人角质形成细胞(KC)Caspase-3活力变化及细胞凋亡情况,探讨TCE诱导KC凋亡的可能信号通路。方法以不同浓度(0.125、0.250、0.500、1.000、2.000 mmol/L)TCE对离体分离培养的KC分别染毒至4、8、12、24 h;Caspase-3抑制剂(Z-DEVD-FMK)预处理组,先用100μmol/L Z-DEVD-FMK预处理细胞1 h,然后再用2.000 mmol/L TCE染毒12 h。用分光光度法检测细胞Caspase-3活力变化,借助Annexin-V/PI双染和流式细胞仪检测细胞凋亡情况。结果与空白对照相比,TCE染毒4 h,各TCE剂量组Caspase-3活力无明显变化(P>0.05);染毒8 h,1.000 mmol/L TCE组Caspase-3活力和2.000 mmol/L TCE组Caspase-3活力,与对照组相比差异有显著性(P<0.05);TCE染毒12 h和24 h,各TCE剂量组Caspase-3活力明显升高,与对照组相比差异有显著性(P<0.05),且呈剂量-效应关系。各TCE剂量组,在TCE浓度达到0.250 mmol/L以后,细胞凋亡率与对照组相比差异有显著性(P<0.05),且明显呈剂量-效应关系。Z-DEVD-FMK预处理组,与2.000 mmol/L TCE组比较,Caspase-3活力和细胞凋亡率明显得到抑制(P<0.01),而与空白对照相比差异无显著性(P>0.01)。结论在TCE诱导离体培养的KC凋亡中,Caspase-3的活化可能发挥了重要的作用。
Objective To observe the changes of activity and apoptosis of human keratinocyte (KC) Caspase-3 induced by trichlorethylene (TCE) and to explore the possible signal pathway of TCE induced KC apoptosis. Methods The KCs isolated and cultured in vitro were exposed to different concentrations (0.125,0.250,0.500,1.000,2.000 mmol / L) of TCE for 4, 8, 12 and 24 hours, respectively. Caspase-3 inhibitor (Z-DEVD-FMK ) Pretreatment group, cells were pretreated with 100μmol / L Z-DEVD-FMK for 1 h and then treated with 2.000 mmol / L TCE for 12 h. Caspase-3 activity was detected by spectrophotometry. Apoptosis was detected by Annexin-V / PI double staining and flow cytometry. Results Compared with blank control group, Caspase-3 activity in TCE group did not change significantly (P> 0.05) at 4 h after TCE exposure. Caspase-3 activity in 2.000 mmol / L TCE group Caspase-3 activity in TCE group was significantly lower than that in control group (P <0.05). Caspase-3 activity in TCE group was significantly higher at 12 and 24 h Significant (P <0.05), and showed a dose-response relationship. The TCE dose group, TCE concentration reached 0.250 mmol / L, the apoptosis rate compared with the control group, the difference was significant (P <0.05), and showed a dose-response relationship. Z-DEVD-FMK pretreatment group, compared with 2.000 mmol / L TCE group, Caspase-3 activity and apoptosis rate were significantly inhibited (P <0.01), but no significant difference compared with the blank control group ). Conclusions Caspase-3 activation may play an important role in TCE-induced KC apoptosis in vitro.