Nagano M, Suzuki H, Ui-Tei K, Sato S, Miyake T, Miyata Y
Department of Pharmacology, Nippon Medical School, Tokyo, Japan.
Neurosci Res. 1998 Jun;31(2):113-21. doi: 10.1016/s0168-0102(98)00030-3.
The present study was undertaken to reveal underlying mechanisms of apoptosis in neurons using clonal neuronal cells, ML-DmBG2-c2, derived from Drosophila larval central nervous system 1-(5-Isoquinolinesulfonyl)-2-methylpiperazine (H-7), a protein kinase inhibitor, induced cell death with typical features of apoptosis such as internucleosomal DNA fragmentation, nuclear condensation and apoptotic bodies in the cells. Though H-7 is known to inhibit cAMP-dependent protein kinase (PKA), protein kinase C (PKC), cGMP-dependent protein kinase (PKG), myosin light chain kinase (MLCK), and casein kinase I (CKI), specific inhibitors for these kinases such as H-89, calphostin C, ML-9, or CKI-7 did not induce apoptosis in the cells. Other kinases such as tyrosine kinase. PI3-kinase and Ca2+/CaM kinase II so far examined in the present study were interpreted not to be involved in the apoptotic cascade. Therefore, it is concluded that an H-7-sensitive substance(s) other than these kinases is responsible for the apoptosis in the neuronal cells. Caspase inhibitors prevented apoptosis in the cells treated with H-7. These results suggest that caspase(s) is involved downstream of the H-7-sensitive point in the cascade of the apoptosis.
本研究旨在利用源自果蝇幼虫中枢神经系统的克隆神经元细胞ML-DmBG2-c2揭示神经元凋亡的潜在机制。蛋白激酶抑制剂1-(5-异喹啉磺酰基)-2-甲基哌嗪（H-7）诱导细胞死亡，其具有凋亡的典型特征，如细胞内核小体间DNA片段化、核浓缩和凋亡小体。尽管已知H-7可抑制环磷酸腺苷依赖性蛋白激酶（PKA）、蛋白激酶C（PKC）、环磷酸鸟苷依赖性蛋白激酶（PKG）、肌球蛋白轻链激酶（MLCK）和酪蛋白激酶I（CKI），但这些激酶的特异性抑制剂，如H-89、钙泊三醇C、ML-9或CKI-7并未诱导细胞凋亡。本研究中迄今检测的其他激酶，如酪氨酸激酶、磷脂酰肌醇-3激酶和Ca2+/钙调蛋白激酶II被认为不参与凋亡级联反应。因此，可以得出结论，除这些激酶外，一种对H-7敏感的物质负责神经元细胞的凋亡。半胱天冬酶抑制剂可阻止用H-7处理的细胞发生凋亡。这些结果表明，半胱天冬酶参与了凋亡级联反应中对H-7敏感点的下游过程。
