摘要
[目的]研究体外培养条件下有机溶剂四氯乙烯（perchloroethylene，PERC）对正常人表皮角质形成细胞（normal human epidermis keratinocyte，NHEK）的细胞毒性作用。[方法]用0．25％的胰蛋白酶经冷温两步消化皮肤，分离得到NHEK，进行体外无血清培养；用中性红吸附试验（NRU）测定PERC对NHEK的中性红吸附减少50％的浓度（NR50值），并据此确定PERC细胞毒性试验的染毒剂量；测定乳酸脱氢酶（LDH）活力反映其对细胞膜的损害，测定丙二醛（MDA）含量和超氧化物歧化酶（SOD）活力反映细胞脂质过氧化作用。[结果]PERC可引起NHEK细胞活力剂量依赖性降低，PERC对NHEK的NR50值为2．16mmol／L（95％可信区间为1．27，3．07）；0．05、0．10、0．20、0．40、0．80mmol／LPERC处理NHEK1、2、3、4h后，LDH的释放呈明显剂量-反应和时间-反应关系；以上浓度的PERC处理NHEK4h，可引起MDA含量增加和SOD活力抑制，且均显示剂量-反应关系，与空白对照组、溶剂对照组相比，引起差异有显著性的最低浓度分别为0。20mmol／L（MDA升高）和0．10mmol／L（SOD降低），均为P，〈0．05。[结论]在体外培养条件下，PERC可能通过氧化应激和脂质过氧化作用对人角质形成细胞产生细胞毒性作用。
[ Objective ] To study the eytotoxicity of perchloroethylene （PERC）on the normal human epidermal keratinocytes （ NHEK ） in vitro. [ Methods ] Administered dose of PERC to NHEK were determined according to neutral red uptake （ NRU ）assay with free serum culture in vitro. Membrane damage and lipid peroxidation （ LPO ）were assessed by lactate dehydrogenase （LDH） release test, measurement of malondialdehyde （ MDA ） content and superoxide dismutase （ SOD ） activity were also assessed. [ Results ] PERC could cause a dose-dependent decrease in cell viability. NR50 values of PERC On NHEK^＋ was observed to be 2.16 mmol/L（ 95% CI： 1.27-3.07 ）. A time-and concentration-dependent release of LDH were observed at 1, 2, 3, 4 h after cells exposed to 0.05, 0.10, 0.20, 0.40, 0.80 mmol/L PERC. When NHEK treated with the above concentrations of PERC for 4 hours, it caused an increase of MDA content and an inhibition of SOD activity in a concentration-dependent manner. The lowest concentrations to induce significant difference in increasing MDA and decreasing SOD were 0.20 and 0.10 mmol/L treatment with PERC respectively. [Conclusion ] These results suggest that PERC could induce cytotoxicity to cultured NHEK in vitro, which was associated with lipid peroxidation and oxidative stress.
出处
《环境与职业医学》
CAS
北大核心
2006年第6期479-481,487,共4页
Journal of Environmental and Occupational Medicine
基金
安徽省自然科学基金项目(编号:03043801)
关键词
四氯乙烯
角质形成细胞
细胞毒性作用
perchloroethylene
human epidermis keratinoeytes
eytotoxicity
