Ayala A, Evans T A, Chaudry I H
Center for Surgical Research, Brown University School of Medicine, Providence, Rhode Island, USA.
J Surg Res. 1998 May;76(2):165-73. doi: 10.1006/jsre.1998.5314.
Apoptosis (AO) is a process by which cells typically undergo a form of nonnecrotic cellular suicide. AO normally allows the host to selectively delete cells from a given tissue site without producing bystander injury associated with necrosis. However, inappropriate induction of AO has been associated with a variety of acute as well as chronic pathological states and may contribute to the therapeutic nonresponsiveness frequently encountered in the septic animal/patient's organ function. In this respect, while AO has been demonstrated in a variety of immune cell tissues of septic animals it is unclear if it is present in the septic liver. Therefore, it was the aim of our study to determine if AO is evident in hepatocytes of polymicrobial septic animals. To assess this, C3H/HeN male mice were subjected to polymicrobial sepsis (cecal ligation and puncture) or sham- CLP (Sham). Hepatocytes were then harvested at 4 h (early hyperdynamic phase) or 24 h (late hypodynamic state) later, and indices of AO were assessed [cell cycle analysis of Annexin V/propidium iodide (PI) staining for flow cytometric analysis, DNA extracted, and cell death ELISA]. Plasma glutamic pyruvic transaminase (GPT) was also colorimetrically assessed as well as total viable cell yield as an index of hepatocellular necrosis. The results indicate that indices of hepatocellular AO, as determined by cell cycle analysis and cell death ELISA, were markedly increased in polymicrobial septic mice at 24 h. However, while an increase in DNA fragmentation/degradation could be consistently detected, the pattern was typically faint. Similarly, although there was an increase in Annexin V staining it was not dissociated from that of PI (necrotic index). Alternatively, necrosis (as evidenced by increased GPT levels at both 4 and 24 h) preceded the induction of all the indices of AO. Taken together, these data suggest a role for both necrosis and apoptosis in the evolution of hepatocellular injury encountered in the polymicrobial septic animal/patient which may represent a unique pattern of cell death under such conditions.
细胞凋亡（AO）是细胞通常经历的一种非坏死性细胞自杀形式的过程。AO通常允许宿主从给定组织部位选择性地清除细胞，而不会产生与坏死相关的旁观者损伤。然而，AO的不适当诱导与多种急性和慢性病理状态相关，并且可能导致脓毒症动物/患者器官功能中经常遇到的治疗无反应性。在这方面，虽然在脓毒症动物的多种免疫细胞组织中已证实存在AO，但尚不清楚其是否存在于脓毒症肝脏中。因此，我们研究的目的是确定AO在多微生物脓毒症动物的肝细胞中是否明显。为了评估这一点，将C3H/HeN雄性小鼠进行多微生物脓毒症（盲肠结扎和穿刺）或假CLP（假手术）。然后在4小时（早期高动力期）或24小时（晚期低动力状态）后收获肝细胞，并评估AO指标[用于流式细胞术分析的膜联蛋白V/碘化丙啶（PI）染色的细胞周期分析、提取DNA和细胞死亡ELISA]。还通过比色法评估血浆谷丙转氨酶（GPT）以及作为肝细胞坏死指标的总活细胞产量。结果表明，通过细胞周期分析和细胞死亡ELISA确定的肝细胞AO指标在24小时时在多微生物脓毒症小鼠中显著增加。然而，虽然可以持续检测到DNA片段化/降解增加，但其模式通常较微弱。同样，虽然膜联蛋白V染色增加，但其与PI（坏死指标）的染色并无分离。另外，坏死（4小时和24小时时GPT水平升高证明）在所有AO指标诱导之前出现。综上所述，这些数据表明坏死和凋亡在多微生物脓毒症动物/患者中肝细胞损伤的演变中均起作用，这可能代表了在这种情况下独特的细胞死亡模式。
