零下非结冰温度保存大鼠肾脏的实验研究  

Experimental study on subzero nonfreezing preservation of rat kidney

作　　者：杨龙[1] 门贺伟[1] 王雅博[2] 诸凯[2] 张雅敏[3] 

机构地区：[1]天津医科大学一中心临床学院,300070 [2]天津商业大学天津市制冷技术重点实验室 [3]天津市第一中心医院器官移植中心

出　　处：《中华器官移植杂志》2013年第9期559-562,共4页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金资助项目（51076117）;天津市卫生局课题（2010KR05）

摘　　要：目的寻找保存大鼠肾脏的最适零下非结冰温度，探讨零下非结冰保存大鼠肾脏的效果。方法使用热电偶探针和温度巡检仪探测大鼠肾脏不同部位的降温曲线，测定出肾脏的冰点。然后将在体灌注的大鼠肾脏取下，并放人盛有肾脏保存液2．5ml的无菌管中，将其分为6组：-0．8℃组（零下非结冰组）、-0．5℃组（零下非结冰组）、0℃（零度非结冰组）、4℃组（对照组）、-1℃组（零下结冰组）、-4℃组（零下结冰组）。低温保存24h和48h后，取保存液检测LDH和AST含量，取肾脏上极组织观察病理改变及细胞凋亡情况。结果肾脏的冰点温度是-1℃，鼠肾的最适零下非结冰保存温度为-0．8℃。零下非结冰保存较传统器官保存温度明显抑制了组织细胞的基础代谢率，减少了因膜损伤而释放的LDH和AST含量，降低了细胞凋亡率[48h时，-0．8℃组为（40．1±7．0）％，4℃组为（47．1±7．6）％]；保存48h后，-0．8℃组较其他组的病理改变明显减轻。结论与0℃到4℃的常规器官保存温度相比，零下非结冰温度（-0．8℃）可明显抑制组织细胞的基础代谢率，减少细胞的势能消耗，降低低温损伤引起的细胞凋亡。Objective To search for the most appropriate subzero nonfreezing temperature, and explore the effect of subzero nonfreezing preservation of rat kidney by comparing with the kidney preservation with conventional temperature （4 ℃, 0 ℃ ） and freezing temperature （ - 4℃ ）. Method The thermocouple probeand the temperature data logging device were used to detect the temperature decreasing curves in different parts of the rat kidney and determine the freezing point of the kidney. The perfused kidneys in rats were removed and put into the sterile tubes containing 2.5 mL hypertonic citrate adenine. Following 6 group were set up： - 0. 8 ℃ group （subzero nonfreezing）, - 0. 5 ℃ group （subzero nonfreezing）, 0 ℃ group （zero nonfreezing）, - 4 ℃ group （control group）, - 1 ℃ group（subzero freezing）, - 4 ℃ group （subzero freezing）. After the cryopreservation for 24 and 48 h, the preservative fluid was harvested for measurement of the contents of LDH and AST, and the paraffin sections from the upper pole of the kidney were made for observation of the pathological changes and apoptosis. Result The freezing temperature of kidney was - 1℃ and the most appropriate subzero nonfreezing temperature for preserving the rat kidney was -0. 8 ℃. Subzero nonfreezing significantly inhibited the basal metabolic rate of the histiocytes, reduced the contents of LDH and AST released due to the membrane damage, and decreased the apoptosis rate [48 h： - 0. 8 ℃ （40. 1 ± 7. 0） % vs. 4 ℃ （47..1 ± 7. 6） %]. Under the light microscope after preservation for 48 h, the pathological changes in - 0. 8 ℃ group were less than in 4 ℃ group. Conclusion Compared with the organ preservation in conventional temperature （0 ℃ - 4 ℃ ）, the subzero nonfreezing （ - 0. 8 ℃） can further inhibit the basal metabolic rate of histocytes obviously, reduce its energy consumption, and lower the apoptosis caused by low temperature damage.

关 键 词：大鼠 肾脏 器官保存 温度 零下非结冰 

分 类 号：R318.52[医药卫生—生物医学工程]