摘要
用大鼠胚胎中脑物经细胞作原代微团培养,培养物经不同浓度四氯化碳染毒,研究四氯化碳对细胞分化的影响,并用扫描电镜观察细胞形态和膜结构的变化。结果表明,处于分化前期的胚胎中脑细胞经5天培养形成细胞集落,用神经元特异性烯醇化酶(NSE)免疫化学法鉴定,集落中的细胞是已分化的神经元细胞。四氯化碳可抑制细胞分化,并显示剂量——效应关系,其半数生长抑制浓度(ICV50)和半数分化抑制浓度(ICd50)分别为10.0和1.Oμmol/L,V/D值为10。扫描电镜显示四氯化碳可引起细胞形态的改变和膜结构的损伤。经四氯化碳染毒的细胞脂质过氧化作用增强,并与细胞膜被破坏的程度一致。认为四氯化碳具体外细胞致畸作用。
The micromass culture from rat embryo midbrain were used to study the effect of CCl_4 on neuron differ- entiation. The results showed that the embryo midbrain cells can differentiation after 5 days in micromass culture and form irregular cell foci and neurofibrils.The differentiated neuron can be determined by neuron specific enolase (NSE)immunochemical method. Using inhibition of foci formation and reduction in cell num- her as endpoints for differentiation and cytotoxycity, respectivety, the micromass culture were currently em- ployed as model system for prediction of teratogenic effects. Our studies indicated that CCl, can inhibite differ- entiation of neurons (decrement of counts of foci)and had cytotoxiciy with a clear relation of does-effect re- sponse. The IC50 values for foci formation and cell survival were 1.0 and 10 pmol/L, respectively. The ef- fects of CCl_4 on cell membranes were examined by means of scanning electron microscopy. The degree of lipid peroxidation corresponded with the results of membrane damage. According to the determinatedcriteria, this compound was considered as a potential teratogen in vitro.
基金
国家自然科学基金资助项目(编号:39070727)
关键词
四氯化碳
神经细胞
细胞分化
膜结构
致畸作用
Carbon tetrachloride
Micromass culture
Cell differentiation
Microscopy, electron scanning
Teratogenicity in vitro
