Topp Stefan A, Upadhya Gundumi A, Strasberg Steven M
Department of Surgery, Washington University in Saint Louis, St. Louis, MO 63110, USA.
Liver Transpl. 2004 Aug;10(8):1041-8. doi: 10.1002/lt.20216.
Cold preservation of rat sinusoidal endothelial cells causes actin disassembly, cell rounding, matrix metalloproteinase (MMP) secretion, and platelet adhesiveness. Studies in rats suggest that gelatinases MMP2 and MMP9 are the key mediators of the injury. We created a model of cold preservation injury in mouse sinusoidal endothelial cell (MSEC) to examine the effect of cold on MSEC, specifically on MSEC from genetically deleted mice (MMP9/KO) mice. MSEC were isolated from wild-type and MMP9/KO mice and cold preserved for up to 24 hours. MMP activity was measured in culture supernatants and in effluents from preserved whole mouse livers. Cellular and actin morphology were studied by light and fluorescence microscopy. A platelet-MSEC adhesion assay was performed. Yield, growth, and appearance of MSEC were similar in wild-type and MMP9/KO mice. Cold-preserved wild-type MSEC exhibited actin disassembly and cell rounding as in the rat but at a much slower rate. These morphologic cell changes were attenuated in MSEC from MMP9/KO mice. Both MMP2 and MMP9 were present in liver effluents of wild-type mice, but MMP9 was absent in effluents from MMP9/KO mice. Total MMP activity in culture supernatants was greater after preservation in wild-type than in MMP9/KO mice. There was significantly more platelet adhesion to wild-type MSEC than to MSEC from MMP9/KO mice. In conclusion, MSEC is an excellent model system for the study of cold preservation injury. Injury is similar to rat sinusoidal endothelial cells but delayed. MMP9 is a key mediator of the cold preservation injury.
大鼠肝血窦内皮细胞的低温保存会导致肌动蛋白解聚、细胞变圆、基质金属蛋白酶（MMP）分泌以及血小板黏附性增加。对大鼠的研究表明，明胶酶MMP2和MMP9是损伤的关键介质。我们建立了小鼠肝血窦内皮细胞（MSEC）低温保存损伤模型，以研究低温对MSEC的影响，特别是对基因敲除小鼠（MMP9/KO）的MSEC的影响。从野生型和MMP9/KO小鼠中分离出MSEC，并进行长达24小时的低温保存。测定培养上清液和保存的整个小鼠肝脏流出液中的MMP活性。通过光学和荧光显微镜研究细胞和肌动蛋白形态。进行血小板-MSEC黏附试验。野生型和MMP9/KO小鼠中MSEC的产量、生长和外观相似。低温保存的野生型MSEC如在大鼠中一样出现肌动蛋白解聚和细胞变圆，但速度要慢得多。这些细胞形态变化在MMP9/KO小鼠的MSEC中减弱。野生型小鼠肝脏流出液中同时存在MMP2和MMP9，但MMP9/KO小鼠流出液中不存在MMP9。野生型小鼠保存后培养上清液中的总MMP活性高于MMP9/KO小鼠。与MMP9/KO小鼠的MSEC相比，野生型MSEC上的血小板黏附明显更多。总之，MSEC是研究低温保存损伤的优秀模型系统。其损伤与大鼠肝血窦内皮细胞相似，但延迟出现。MMP9是低温保存损伤的关键介质。
