Green C J, Healing G, Lunec J, Fuller B J, Simpkin S
Transplantation. 1986 Feb;41(2):161-5. doi: 10.1097/00007890-198602000-00005.
Rabbit kidneys were stored for 24 or 48 hr at 0 degree C after single-passage vascular flush with 30 ml of cold hypertonic citrate solution or 0.9% isotonic sodium chloride solution. They were then subjected to in vitro biochemical assay for evidence of free-radical damage immediately after storage or after they had been orthotopically autotransplanted and reperfused with blood in vivo for 60 min. Kidney homogenates were incubated at 37 degrees C and assayed for fluorescent conjugated Schiff bases as indicators of lipid peroxidation, as well as for superoxide dismutase activity and reduced and oxidized glutathione. In kidneys flushed with hypertonic citrate, no evidence of peroxidation could be detected immediately after storage for 24 or 48 hr. However, after in vivo reperfusion significantly more peroxidation (P less than 0.01) was evident. Storage in isotonic saline solution produced still higher levels of peroxidation damage whether reperfused or not (P less than 0.001). Schiff base formation was inversely proportional to the reduced and oxidized glutathione levels measured. No changes in superoxide dismutase levels could be detected. It is concluded that lipid peroxidation is important during cold ischemia but most damage occurs during the 60-min of reperfusion in vivo immediately after transplantation.
用30毫升冷的高渗柠檬酸盐溶液或0.9%等渗氯化钠溶液进行单次血管冲洗后，兔肾在0摄氏度下保存24或48小时。然后，在保存后或在原位自体移植并在体内再灌注血液60分钟后，立即对其进行体外生化分析，以检测自由基损伤的证据。将肾匀浆在37摄氏度下孵育，并检测荧光共轭席夫碱作为脂质过氧化的指标，以及超氧化物歧化酶活性和还原型及氧化型谷胱甘肽。在用高渗柠檬酸盐冲洗的肾脏中，保存24或48小时后立即未检测到过氧化的证据。然而，在体内再灌注后，明显有更多的过氧化现象（P<0.01）。无论是否再灌注，保存在等渗盐溶液中都会产生更高水平的过氧化损伤（P<0.001）。席夫碱的形成与所测的还原型和氧化型谷胱甘肽水平成反比。未检测到超氧化物歧化酶水平的变化。得出的结论是，脂质过氧化在冷缺血期间很重要，但大多数损伤发生在移植后立即进行的60分钟体内再灌注期间。
