A NEW NEUROTOXICOLOGICAL TEST SYSTEM USING NERVE CELL LINE

使用神经细胞系的新型神经毒理学测试系统

• 批准号: 10670366
• 负责人: SUZUKI Yuji
• 金额: $ 0.38万
• 依托单位: JIKEI UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670366/
• 关键词: Neurotoxicology; micronuclei; apoptosis; nerve cell; action potential; cisplatin; calcium ion; 膜電位; カルシウム; スクリーニング; 小核; キャスパーゼ; タンネル法; DNA; ハイブリド細胞

Many new chemicals have produced in this century from industrial circles. We established a in vitro neurotoxicological test system for the purpose of assessing the neurotoxic potential of these chemicals.(1) Induction of micronuclei and apoptosisHybrid cell line N18DRG-D2 of BALB/c mouse dorsal root ganglion neuron and mouse neuroblastoma cell (N18TG-D2) was used in this study. Induction of micronuclei by propidium iodide peaked at 96 hours of cisplatin treatment and frequency of micronuclei induced by cisplatin increased dose dependent manner. However, number of apoptotic micronuclei (double staining with proidium iodide and TUNEL method) did not increase following cisplatin treatment.(2) Alteration of membrane action potentialMembrane action potential was determined using a fluorophotometer after stained with 3, 3'-dipenty loxacarbocyanine iodide. Fluorescence intensity of N18DRG-D2 was attenuated after exposure to cisplatin.(3) Alteration of intracellular calcium ion concentrationMembrane action potential was determined using a fluorophotometer after stained with fura-2. Fluorescence intensity of N18DRG-D2 was attenuated after exposure to cisplatin.These results suggested that a new neurotoxicological test system using N18DRG-2 cell line might be useful for estimating neurotoxic substances.

本世纪工业界生产了许多新的化学品。我们建立了一个体外神经毒理学试验系统，以评估这些化学品的神经毒性潜力。(1)本研究采用BALB/c小鼠背根神经节神经元和小鼠神经母细胞瘤细胞杂交细胞系N18 TG-D2。碘化丙啶诱发的微核率在顺铂处理96小时达到峰值，顺铂诱发的微核率呈剂量依赖性增加。然而，顺铂处理后，凋亡微核数（碘化丙啶和TUNEL法双重染色）没有增加。(2)细胞膜动作电位的改变用碘化3，3 ′-二戊基氧碳菁染色后用荧光光度计测定细胞膜动作电位。N18 DRG-D2的荧光强度在暴露于顺铂后减弱。(3)细胞内钙离子浓度的变化Fura-2染色后用荧光分光光度计测定细胞膜动作电位。N18 DRG-D2细胞经顺铂处理后荧光强度减弱，提示N18 DRG-2细胞系可用于神经毒性物质的评价。

项目成果

YUICHI MIYAKOSHI: "MICRONUCLEUS TEST USING NEW BORN RAT ASTROCYTES"INDUSTRIAL HEALTH. 37. 95-102 (1999)

Yuichi Miyakoshi：“使用新生大鼠星形胶质细胞进行微核测试”工业健康。

Masateru Ikehata: "Mutagenicity and co-mutagenicity of static magnetic fields detected by bacterial mutation assay."Mutation Research. 427・2. 147-156 (1999)

Masateru Ikehata：“通过细菌突变测定检测静磁场的致突变性和共致突变性。”突变研究427・2（1999）。

Yuichi Miyakoshi: "Micronucleus test using cultured new born rat astrocytes"Industrial Health. 37・2. 95-102 (1999)

Yuichi Miyakoshi：“使用培养的新生大鼠星形胶质细胞的微核测试”工业健康37・2（1999）。

Miyakoshi,Y.: "Micronucleus test using cultured new born rat astrocytes" Industrial Health. 37・1. 95-102 (1999)

Miyakoshi，Y.：“使用培养的新生大鼠星形胶质细胞的微核测试”工业健康37・102（1999）。

MASATERU IKEHATA: "MUTAGENICITY AND CO-MUTAGENICITY OF STATIC MAGENTIC FIELDS DETECTED BY BACTERIAL MUTATION ASSAY"MUTATION RESEARCH. 427. 147-156 (1999)

MASATERU IKEHATA：“细菌突变试验检测到的静磁场的突变性和共突变性​​”突变研究。