Hiramatsu M, Hotchkiss R S, Karl I E, Buchman T G
Department of Surgery, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Shock. 1997 Apr;7(4):247-53. doi: 10.1097/00024382-199704000-00002.
Two challenges (intraperitoneal lipopolysaccharide (LPS) administration and cecal ligation and puncture (CLP)) and two strains of mice (LPS-normoresponder (C3H/HeN) and LPS-hyporesponder (C3H/HeJ)) were used to investigate pathways of cell injury. After intraperitoneal administration of LPS, endotoxin was absorbed into the bloodstream (HeN, 10.4 +/- 9.4 x 10(4) EU/mL; HeJ, 14.7 +/- 6.0 x 10(4) EU/mL), but as expected, only C3H/HeN mice produced serum tumor necrosis factor (TNF) (HeN, 2.5 +/- 2.0 x 10(3)pg/mL; HeJ, 87.0 +/- 38.7 pg/mL). Gel electrophoretic analysis of DNA extracted from six organs demonstrated the apoptotic "ladder" only in the thymus and only in the HeN mice. When the mice were challenged with CLP, both HeN and HeJ produced a small amount of serum TNF (HeN, 5.8 +/- 3.5 x 10(2) pg/mL; HeJ, 2.2 +/- 2.5 x 10(2) pg/mL) and both strains had very mild endotoxemia (HeN, 23.4 +/- 3.8 EU/mL; HeJ, 27.9 +/- 10.1 EU/mL). The DNA fragmentation pattern characteristic of apoptosis was observed not only in thymus but also in spleen, lung, and Peyer's patch of gut of both strains. This organ-specific pattern was more pronounced in the thymus of HeN mice; otherwise, the organ-specific patterns were similar for HeN and HeJ mice challenged by CLP but absent in those same organs when those same mice were challenged with LPS. The data suggest the existence not only of an endotoxin-driven activation for thymic apoptosis, but also of an endotoxin-independent, TNF-independent pathway activating widespread apoptosis in the murine CLP model of sepsis.
采用两种挑战方式(腹腔注射脂多糖(LPS)和盲肠结扎穿刺(CLP))以及两种品系的小鼠(LPS无反应型(C3H/HeN)和LPS低反应型(C3H/HeJ))来研究细胞损伤途径。腹腔注射LPS后,内毒素吸收入血(HeN,10.4±9.4×10⁴ EU/mL;HeJ,14.7±6.0×10⁴ EU/mL),但正如预期的那样,只有C3H/HeN小鼠产生血清肿瘤坏死因子(TNF)(HeN,2.5±2.0×10³ pg/mL;HeJ,87.0±38.7 pg/mL)。对从六个器官提取的DNA进行凝胶电泳分析显示,仅在HeN小鼠的胸腺中出现凋亡“梯状条带”。当用CLP对小鼠进行攻击时,HeN和HeJ均产生少量血清TNF(HeN,5.8±3.5×10² pg/mL;HeJ, 2.2±2.5×10² pg/mL),且两个品系均有非常轻微的内毒素血症(HeN,23.4±3.8 EU/mL;HeJ,27.9±10.1 EU/mL)。不仅在胸腺中,而且在两个品系小鼠的脾脏、肺和肠道派尔集合淋巴结中均观察到凋亡特有的DNA片段化模式。这种器官特异性模式在HeN小鼠的胸腺中更为明显;否则,对于接受CLP攻击的HeN和HeJ小鼠,器官特异性模式相似,但在用LPS攻击相同小鼠时,相同器官中不存在这种模式。数据表明,不仅存在内毒素驱动的胸腺细胞凋亡激活,而且存在一种不依赖内毒素、不依赖TNF的途径,在脓毒症小鼠CLP模型中激活广泛的细胞凋亡。