Higami Yoshikazu, Tsuchiya Tomoshi, To Kazuo, Chiba Takuya, Yamaza Haruyoshi, Shiokawa Daisuke, Tanuma Sei-ichi, Shimokawa Isao
Department of Pathology and Gerontology, Nagasaki University Graduate School of Biomedical Science, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan.
Cell Tissue Res. 2004 Jun;316(3):403-7. doi: 10.1007/s00441-004-0890-x. Epub 2004 Apr 29.
Endonuclease-induced DNA fragmentation is a hallmark of apoptosis. DNase gamma (DNase gamma) was recently identified as one of the endonucleases responsible for apoptotic DNA fragmentation. In this study, immunohistochemistry for DNase gamma was performed on paraffin sections of rodent liver in well-defined models of hepatocyte apoptosis induced by Fas antibody (Fas) or cycloheximide (CHX), and necrosis induced by lipopolysaccharide (LPS) or carbon tetrachloride (CCl4). DNase gamma immunoreactivity was compared with TdT-mediated dUTP nick-end labeling (TUNEL) reactivity. Our results showed TUNEL reactivity in both apoptotic and necrotic hepatocytes. DNase gamma immunoreactivity was not detected during LPS-induced or CCl4-induced hepatocyte necrosis. In contrast, it was evident during CHX-induced, but not Fas-induced, apoptotic DNA fragmentation. These findings suggest that DNase gamma plays an important role in Fas-independent apoptotic DNA fragmentation in hepatocytes.
核酸内切酶诱导的DNA片段化是细胞凋亡的一个标志。脱氧核糖核酸酶γ(DNaseγ)最近被确定为负责凋亡性DNA片段化的核酸内切酶之一。在本研究中,对由Fas抗体(Fas)或环己酰亚胺(CHX)诱导的肝细胞凋亡以及由脂多糖(LPS)或四氯化碳(CCl4)诱导的坏死的明确啮齿动物肝脏石蜡切片进行了DNaseγ的免疫组织化学检测。将DNaseγ免疫反应性与末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)反应性进行了比较。我们的结果显示,凋亡和坏死的肝细胞中均有TUNEL反应性。在LPS诱导或CCl4诱导的肝细胞坏死过程中未检测到DNaseγ免疫反应性。相反,在CHX诱导而非Fas诱导的凋亡性DNA片段化过程中,DNaseγ免疫反应性很明显。这些发现表明,DNaseγ在肝细胞中不依赖Fas的凋亡性DNA片段化中起重要作用。