Gabaĭ V L, Mosina V A, Makarova Iu M, Maliutina Ia V, Budagova K R, Mosin A F
Biokhimiia. 1995 Aug;60(8):1201-8.
Incubation of rat thymocytes in serum-free media was found to result in their apoptotic death characterized by internucleosomal DNA fragmentation, nuclear pyknosis and subsequent irreversible plasma membrane damage. As in the case of glucocorticoid (hydrocortisone)-induced apoptosis, DNA fragmentation under serum withdrawal was suppressed by endonuclease inhibitors (Zn2+ and spermine). At the same time, protein synthesis inhibitors (cycloheximide and puromycin) failed to block the apoptosis induced by serum withdrawal but inhibited the hydrocortisone-induced apoptosis. Various inhibitors of oxidative phosphorylation (uncoupler, rotenone, oligomycin), causing sharp decrease in cellular ATP did not suppress DNA fragmentation, whereas thymocyte plasma membrane damage accelerated under their effect. The results obtained indicate that intact thymocytes contain all the components of the apoptotic system; however, in the absence of apoptotic stimuli (e.g., hydrocortisone) the system is blocked by some growth factors of serum origin. Serum withdrawal is sufficient by itself to induce apoptosis and does not require the synthesis of special proteins.
研究发现,将大鼠胸腺细胞置于无血清培养基中培养会导致其发生凋亡性死亡,其特征为核小体间DNA片段化、核固缩以及随后不可逆的质膜损伤。与糖皮质激素(氢化可的松)诱导的凋亡情况相同,血清撤除时的DNA片段化受到核酸内切酶抑制剂(Zn2+和精胺)的抑制。与此同时,蛋白质合成抑制剂(环己酰亚胺和嘌呤霉素)未能阻断血清撤除诱导的凋亡,但抑制了氢化可的松诱导的凋亡。各种氧化磷酸化抑制剂(解偶联剂、鱼藤酮、寡霉素)导致细胞ATP急剧减少,并未抑制DNA片段化,而在其作用下胸腺细胞质膜损伤加速。所得结果表明,完整的胸腺细胞含有凋亡系统的所有成分;然而,在没有凋亡刺激(如氢化可的松)的情况下,该系统被一些血清来源的生长因子所阻断。血清撤除本身就足以诱导凋亡,且不需要合成特殊蛋白质。