Jordán J, Galindo M F, Miller R J
Department of Pharmacological and Physiological Sciences, University of Chicago, Illinois 60637, USA.
J Neurochem. 1997 Apr;68(4):1612-21. doi: 10.1046/j.1471-4159.1997.68041612.x.
We investigated the potential role of different proteases in the death of cultured rat hippocampal pyramidal neurons induced by beta-amyloid (A beta) (25-35). Both A beta(25-35)- and staurosporine-induced death of these neurons appeared to involve apoptosis, as indicated using Hoechst 33342 and terminal dUDP nick end labeling staining, whereas NMDA-induced death appeared more complex. Two irreversible inhibitors of the interleukin-1 beta converting enzyme (ICE) and related proteases, Z-Val-Ala-Asp-CH2F and acetyl-Tyr-Val-Ala-Asp-chloromethyl ketone, blocked neuronal death produced by A beta(25-35), staurosporine, and NMDA to differing extents. Furthermore, MDL 28,170, a selective inhibitor of the calcium-regulated protease calpain, also inhibited death induced by all agents. A beta(25-35) and staurosporine stimulated the breakdown of the protein spectrin, a calpain substrate. Spectrin breakdown was inhibited by MDL 28,170 but not by ICE inhibitors. Leupeptin was only effective in preventing NMDA-induced death. These results support the role of apoptosis in neuronal death due to A beta(25-35) treatment and also suggest a role for calcium-regulated proteases in this process.
我们研究了不同蛋白酶在β-淀粉样蛋白（Aβ）（25-35）诱导的培养大鼠海马锥体神经元死亡中的潜在作用。使用Hoechst 33342和末端dUDP缺口末端标记染色表明，Aβ（25-35）和星形孢菌素诱导的这些神经元死亡似乎都涉及细胞凋亡，而NMDA诱导的死亡似乎更为复杂。白细胞介素-1β转换酶（ICE）及相关蛋白酶的两种不可逆抑制剂Z-Val-Ala-Asp-CH2F和乙酰-Tyr-Val-Ala-Asp-氯甲基酮对Aβ（25-35）、星形孢菌素和NMDA诱导的神经元死亡有不同程度的阻断作用。此外，钙调节蛋白酶钙蛋白酶的选择性抑制剂MDL 28,170也抑制了所有药物诱导的死亡。Aβ（25-35）和星形孢菌素刺激了钙蛋白酶底物血影蛋白的降解。血影蛋白的降解被MDL 28,170抑制，但未被ICE抑制剂抑制。亮抑蛋白酶肽仅在预防NMDA诱导的死亡方面有效。这些结果支持细胞凋亡在Aβ（25-35）处理导致的神经元死亡中的作用，也表明钙调节蛋白酶在此过程中发挥作用。
