甲苯对原代培养海马神经细胞的毒性研究  

Study on toxic effects of toluene on primary cultured hippocampus neurons

作　　者：颜士勇[1] 印木泉[1] 韦毅[2] 郭丰涛[3] 王庆蓉[3] 官颖鹏[3] 丛颖[3] 竺青[3] 

机构地区：[1]第二军医大学卫生毒理学教研室,上海200433 [2]海军医学研究所 [3]复旦大学遗传所

出　　处：《卫生研究》2004年第1期18-22,共5页Journal of Hygiene Research

基　　金：海军后勤部指令课题 (No.0 0 - 330 9)

摘　　要：目的　通过在体外对原代培养的海马神经细胞进行染毒 ,观察其对神经细胞的毒作用 ,并探讨其作用机制。方法　取新生的SD大鼠海马神经细胞进行原代培养 ,两周后 ,用甲苯染毒 ,分成 3个染毒组 ,染毒浓度分别为 3、6和 9mmol L ,并设空白对照组和赋形剂组 ,染毒 2 4h ,观察其对细胞形态、细胞存活率、LDH漏出率、细胞内游离钙和细胞凋亡的影响。结果　甲苯染毒后 ,细胞的形态发生改变 ,引起细胞突起萎缩 ,细胞肿胀变圆 ,细胞数量减少 ;神经细胞存活率的显著下降 ,且随着染毒时间的延长及浓度的升高其存活率逐渐降低 ;高剂量组神经细胞LDH漏出率的显著升高 ,与对照组相比有显著性 (P <0 0 5 ) ;细胞细胞内的[Ca2 + ]i浓度显著上升 ,与对照组相比P <0 0 5 ,并呈剂量依赖性的变化 ,加入钙通道拮抗剂硫氮卓酮后 ,则未见 [Ca2 + ]i浓度上升 ;可见明显的细胞凋亡。结论　甲苯体外染毒对神经细胞有较强的毒性。这可能与甲苯有较强的脂溶性、引起细胞钙内流增加 ,促进细胞凋亡有关。Objective Study the cytotoxicity of toluene and its mechanism, the hippocampus neurons were primarily cultured and were exposed to toluene in vitro.Methods The neurons from newborn SD rat's hippocampus were primarily cultured for two weeks, then administered with toluene(3,6,9mmol/L), with blank control group and excipient group being also set up. 24 hours later, Morphology and viability of the cells, the LDH activity, [Ca 2+]i, and cell apoptosis were examined. Results Protuberances of neurons of the toluene-exposed groups were damaged; the bodies of the neurons became round and swollen; the number of the cells decreased; the LDH activity of neurons of high-dose group increased significantly compared with control group (P<0.05). [Ca 2+]i of toluene- exposed groups also increased significantly compared with control group(P<0.05) in a dose-dependent manner; after diltizem as antagonist of calcium tunnel was added, no increase of [Ca 2+]i was found; and evident apoptosis of the exposed cells were also found.Conclusion Toluene was toxic to the neurons after being administered in vitro, which might be ascribed to higher lipid solubility of toluene and it's ability to increase calcium influx, the latter facilitating apoptosis.

关 键 词：甲苯 原代培养 细胞毒性 细胞凋亡 

分 类 号：X550.3[环境科学与工程—环境工程] P46[天文地球—大气科学及气象学]