三氯乙烯诱导人角质形成细胞凋亡中Caspase-3活力的研究  

Study on Caspase-3 activity on trichloroethylene-induced human keratinocyte apoptosis

作　　者：汪立杰[1] 叶良平[1] 沈彤[2] 朱启星[1,2] 

机构地区：[1]安徽医科大学公共卫生学院劳动卫生与环境卫生学系,安徽合肥230032 [2]安徽医科大学皮肤病研究所,230032

出　　处：《中国职业医学》2009年第1期7-10,共4页China Occupational Medicine

基　　金：国家自然科学基金资助项目(30671787)

摘　　要：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 on Caspase-3 activity and apoptosis of human keratinoeytes （KC） induced by trichloroethylene （TCE）, and to explore the KC apoptosis signaling path way induced by ＇ICE. Methods Cultured human KC were exposed to 0, 0. 125, 0. 250, 0. 500, 1. 000, 2. 000 mmol/L TCE for 4, 8, 12 or 24 hours. In inhibition group, cell were pretreated with 100 μmol/L Z-DEVD-FMK for 1 hour and then exposured to 2. 000 mmol/L TCE for 12 hours, Caspase-3 activities were calculated according to the changes of optical density after reaction with Caspase-3 substrate. Cell apoptosis was detected by flow cytometry （FCM） after double-stained with Annexin-V/PI. Results In the 4-hour group, there were no differences of Caspase-3 activity between TCE-treated group and control group, Caspase-3 activities elevated after 1. 000 and 2. 000 mmol/L TCE treatment for 8 hours （compared with control, P 〈 0. 05）. In 12- and 24-hour group, C, aspase-3 activities elevated with the doses of TCE. FCM results also demonstrated that the rate of apoptosis increased with the doses of rICE, and had significant correlation with Caspase-3 activities. When pretreated with Z-DEVD-FMK,compared with 2. 000 mmol/L TCE group, both Caspase-3 activity and the rate of apoptosis decreased （P 〈 0. 01 ） ,and there was no statistical significance in comparison to the control group（P 〉 0. 01 ）. Conclusion Caspase-3 activation may play an important role in TCE-induced apoptosis of KC.

关 键 词：三氯乙烯 角质形成细胞 CASPASE-3 细胞脱噬作用 

分 类 号：R135.1[医药卫生—劳动卫生]