STUDIES ON INSCT NEUROCELL CULTURE AND MODE OF ACTION OF NEUROTOXIC COMPOUNDS

INCT神经细胞培养及神经毒性化合物作用方式的研究

• 批准号: 09460023
• 负责人: HONDA Hiroshi
• 金额: $ 9.86万
• 依托单位: UNIVERSITY OF TSUKUBA
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460023/
• 关键词: Toxic phytochemicals; Anisatine; 9.4-octadecenoic acid; Ion channel; Mode of action; Neurotoxins; Neurocell; Cell culture; ディオスコリン

Neurons cerived from the central nervous system of housefly was cultured in M3 medium, LM3 medium, modified Schneicder's medium and IPL41 medium. The viable term, whether axon elongation was occurred or not and its extent, the term till the beginning of axon elongation were examined. No distinct differences were observed among these media on the various aspects in an axon elongation, but axon elongation and viable term were comparatively better in M3/IPL41 medium and LM3 medium. The ratio of viable culture cell fell off to 20-30% at high density of cell. The viable term was over 90 days in successful case, the longest was 130 days. Positive effect in axon elongation and the viable term of nervous cell was observed when iusulin was incorporated into media but not in serine, glutamic acid and fetal bovine serum. Ethylacetate extract of Sikimi seeds was insecticidally active, with EC50 values of 43.7mugml-1. This fraction suppressed GABA-induced currents irreversibly at 0.42 mugml_<-1>. Sikimi extracts decreased the affinity of GABA for its binding site on the GABA receptor, thereby suppressing GABA-induced currents. From rhizome of a tropical yam, 9,4-octadecadienoic acid was identified as a antifeedant to the Diamondback moth larvae and it also showed potent insecticidal activity to the larvae. Dioscorine and dioscorine N-oxide, which were isolated from the yam, were shown to be an openchannel bocker of acetylcholin receptor channel by a whole cell patch clump method.

在Housefly的中枢神经系统中凝聚的神经元在M3培养基，LM3培养基，改良的Schneicder培养基和IPL41培养基中培养。可行的术语，无论是否发生轴突伸长率，是否发生了轴突伸长率，其范围，直到轴突伸长开始的术语。在这些介质中，在轴突伸长率的各个方面没有观察到明显的差异，但是在M3/IPL41培养基和LM3培养基中，轴突伸长和可行项相对较好。在高密度的细胞密度下，可行培养细胞的比率下降到20-30％。在成功的情况下，可行的期限超过90天，最长的时间是130天。当将iusulin掺入培养基但未放入丝氨酸，谷氨酸和胎儿牛血清中时，观察到轴突伸长和神经细胞的可行项的积极作用。西基米种子的乙酸乙酸酯提取物是杀虫活性的，EC50值为43.7mugml-1。该馏分在0.42 mugml _ <-1>下不可逆地抑制了GABA诱导的电流。 Sikimi提取物降低了GABA对其在GABA受体上的结合位点的亲和力，从而抑制了GABA诱导的电流。从热带山药的根茎中，将9,4-二十二烯酸鉴定为对响尾蛇飞蛾幼虫的抗肌动脉剂，并且还显示出对幼虫的有效杀虫活性。从山药中分离出来的二剑氨酸和二轴氮N-氧化物被整个细胞斑块法被证明是乙酰基胆碱受体通道的开频道Bocker。

项目成果

Banaag,B,Alexie: "Effects on non-alkaloid extracts from tam,Dioscorrea hispida against the diamondback moth,plutella xylostella (Lepidoptera : Yponomeutidae)" Applird Entomology and Zoology. vol.33. 369-373 (1998)

Banaag，B，Alexie：“来自 tam、Dioscorrea hispida 的非生物碱提取物对小菜蛾、小菜蛾（鳞翅目：Yponomeutidae）的影响”Applird 昆虫学和动物学。

Banaag, B,Alexie: "Effects on non-alkaloid extracts from tam, Dioscorrea hispida against the diamondback moth.Plutella xylostella (Lepidoptera : Yponomeutidae)" Applird Entomology and Zoology. vol.33. 369-373 (1998)

Banaag，B，Alexie：“来自 tam、Dioscorrea hispida 的非生物碱提取物对小菜蛾的影响。小菜蛾（鳞翅目：Yponomeutidae）”Applird 昆虫学和动物学。

Keiichi Nagata: "Suppression of voltage-gated sodium channel by dihydropyrazole insecticide,DPX-JW062 inrat dorsal root ganglion neurons." Journal of Pesticide Science. 23. 62-64 (1998)

Keiichi Nagata：“二氢吡唑杀虫剂 DPX-JW062 抑制大鼠背根神经节神经元的电压门控钠通道。”

Tomoko,Ikeda: "Insecticidal activity of sikimi extract and its modulation of the GABA _A receptor channel" Pescticide Science. vol.52. 337-342 (1998)

池田智子：“刺味提取物的杀虫活性及其对 GABA _A 受体通道的调节”《农药科学》。

Tomoko, Ikeda: "Insecticidal activity of sikimi extract and its modulation of the GABA _A receptor channel" Pescticide Science. vol.52. 337-342 (1998)

Tomoko, Ikeda：“刺味提取物的杀虫活性及其对 GABA _A 受体通道的调节”《农药科学》。