蛋白激酶C-ε保护PC12细胞免受甲基苯丙胺诱导的死亡：可能与抑制谷氨酸受体有关。

Protein kinase C-epsilon protects PC12 cells against methamphetamine-induced death: possible involvement of suppression of glutamate receptor.

作者信息

Uemura Koichi, Aki Toshihiko, Yamaguchi Kazuhito, Yoshida Ken ichi

机构信息

Department of Forensic Medicine, Graduate School of Medicine, University of Tokyo, 113-0033, Tokyo, Japan.

出版信息

Life Sci. 2003 Feb 21;72(14):1595-607. doi: 10.1016/s0024-3205(02)02450-5.

DOI:10.1016/s0024-3205(02)02450-5

PMID:12551748

Abstract

The involvement of PKC isoform in the methamphetamine (MA)-induced death of neuron-like PC12 cell was studied. The death and the enhanced terminal dUTP nick end labeling (TUNEL) staining were inhibited by a caspase inhibitor, z-Val-Ala-Asp- (OMe)-CH(2)F (z-VAD-fmk). However, the cell death shows neither morphological nor biochemical features of apoptosis or necrosis. The cell death was suppressed by a protein kinase C (PKC) activator, 12,13-phorbol myristate acetate, but was enhanced by PKC specific inhibitor calphostin C or bisindolylmaleimide, not by PKC inhibitor relatively specific for PKC-alpha (safingol) or PKC-delta (rottlerin). Western blotting demonstrated the expression of PKC-alpha, gamma, delta, epsilon and zeta, of which PKC-epsilon translocated from the soluble to the particulate fraction after MA-treatment. Antisense to PKC-epsilon enhanced MA-induced death. A glutamate receptor antagonist MK801 abrogated the cell death, which is reversed by PKC inhibition. These data suggest that PKC-epsilon promotes PC12 cell survival through glutamate receptor suppression.

摘要

研究了蛋白激酶C（PKC）同工型在甲基苯丙胺（MA）诱导的神经元样PC12细胞死亡中的作用。半胱天冬酶抑制剂z-Val-Ala-Asp-（OMe）-CH(2)F（z-VAD-fmk）可抑制细胞死亡及增强的末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色。然而，细胞死亡既未表现出凋亡也未表现出坏死的形态学或生化特征。蛋白激酶C（PKC）激活剂12,13-佛波醇十四酸酯可抑制细胞死亡，但PKC特异性抑制剂钙泊三醇C或双吲哚马来酰胺可增强细胞死亡，而对PKC-α（沙芬戈）或PKC-δ（罗特勒素）相对特异性的PKC抑制剂则无此作用。蛋白质印迹法显示PKC-α、γ、δ、ε和ζ的表达，其中MA处理后PKC-ε从可溶性组分转位至颗粒性组分。PKC-ε反义寡核苷酸增强了MA诱导的细胞死亡。谷氨酸受体拮抗剂MK801可消除细胞死亡，而PKC抑制可逆转这种作用。这些数据表明PKC-ε通过抑制谷氨酸受体促进PC12细胞存活。