Studies on the intra-cellular response to the stress under the process of neuronal degeneration induced by methamphetamine.

甲基苯丙胺诱导的神经元变性过程中细胞内应激反应的研究。

• 批准号: 11670424
• 负责人: UEMURA Koichi
• 金额: $ 1.6万
• 依托单位: University of Tokyo (2000)Yamaguchi University (1999)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670424/
• 关键词: cell death; PC12; protein kinase C; apoptosis; methamphetamine; PKC; apoptosis; methamphetamine; 覚醒剤; ネクローシス; Protein kinase C; glutamate receptor

Methamphetamine (MAP) induces schizophrenia-like symptoms and neuronal degeneration of dopaminergic neurons. We studied on the mechanism of MAP-induced cell death with a special reference to apoptosis and PKC (protein kinase C).MAP induced dose (1-5mM) and time-dependent (6-48hrs) cell death in PC12 cells. The cell death was associated with the increase in the TUNEL staining, while it was inhibited by a broad spectrum caspase inhibitor z-VAD-fmk. However, the cell death is supposed not to be a typical apoptosis because there were neither morphological changes such as nuclear fragmentation or formation of apoptotic bodies nor DNA laddering on agarose gel. The cell death was suppressed by a PKC activator, PMA, but was enhanced by PKC specific inhibitor, calphostin C or bisindolylmaleimide 1. PKC-a, d are not involved as both safingol and rottlerin inhibitor for PKC-α and δ, respectively, did not aggravate MAP-induced cell death. Western blotting analysis demonstrated the translocation of PKC-ε. Antisense oligodeoxynucleotide to PKC-ε aggravated the MAP-induced cell death. The MAP treatment increased PKC activity in the cell lysate. These observations suggest that PKC-ε suppresses the MAP-induced cell death in PC12 cell.A glutamate receptor may be a PKC target that is involved in the protection against apoptosis after MAP treatment because the antagonist MK801 abrogated all death but the effect was suppressed by PKC inhibition. This effect may be explained by the modulatory effect of PKC on glutamate receptor.

甲基苯丙胺（MAP）可引起精神分裂症样症状和多巴胺能神经元变性。本文从细胞凋亡和蛋白激酶C（PKC）两个方面探讨了MAP诱导细胞死亡的机制。细胞死亡与TUNEL染色的增加相关，而其被广谱半胱天冬酶抑制剂z-VAD-fatal抑制。然而，细胞死亡被认为不是典型的凋亡，因为没有形态学变化，如核碎裂或凋亡小体的形成，也没有DNA梯状在琼脂糖凝胶上。PKC激活剂PMA可抑制细胞死亡，但PKC特异性抑制剂calphostin C或bisindolylmaleimide 1可增强细胞死亡。PKC-α、d不参与，因为沙芬戈和rottlerin分别抑制PKC-α和δ，不加重MAP诱导的细胞死亡。Western blotting分析显示PKC-ε发生了易位。PKC-ε反义寡核苷酸可加重MAP诱导的细胞死亡。MAP处理增加了细胞裂解物中的PKC活性。提示PKC-ε抑制MAP诱导的PC 12细胞凋亡，其作用靶点可能是一个谷氨酸受体。这种作用可能与PKC对谷氨酸受体的调节作用有关。

项目成果

Koichi Uemura, Kazuki Harada, Daikai Sadamitsu, Ryosuke Tsuruta, Mutsuo Takahashi, Toshihiko Aki, Masahiro Yasuhara, Tsuyoshi Maekawa, Ken-ichi Yoshida.: "Apoptotic and necrotic brain lesions in a fatal case of carbon monoxide poisoning."Forensic Sci Int.

Koichi Uemura、Kazuki Harada、Daikai Sadamitsu、Ryosuke Tsuruta、Mutsuo Takahashi、Toshihiko Aki、Masahiro Yasuhara、Tsuyoshi Maekawa、Ken-ichi Yoshida.：“一氧化碳中毒致命病例中的细胞凋亡和坏死性脑损伤。”法医学国际。

Koichi Uemura: "Apoptotic and necrotic brain lesions in a fatal case of carbon monoxide poisoning."Forensic Sci Int. 116. 213-219 (2001)

Koichi Uemura：“一氧化碳中毒致命病例中的细胞凋亡和坏死性脑损伤。”法医科学国际。