Green C J, Healing G, Simpkin S, Lunec J, Fuller B J
Comp Biochem Physiol B. 1986;83(3):603-6. doi: 10.1016/0305-0491(86)90303-2.
Rabbit kidneys were clamped and rendered warm ischaemic (WI) in situ for 60 and 120 min. They were then either removed immediately after the ischaemic insult or after reperfusion with blood for 60 min or 24 hr. Homogenates were assayed for phospholipid-Schiff base fluorescence (Ex. 360 nm, Em. 435 nm) and for diene conjugate formation by u.v. spectrophotometry (240 nm) as indices of lipid peroxidation. No alteration in tissue levels of Schiff base was evident immediately after WI but when the homogenates were incubated at 37 degrees C for 90 min, the rate of peroxidation was significantly elevated compared to controls (P less than 0.02 after WI of 60 min and P less than 0.001 after 120 min of WI). These values were still further elevated after reperfusion with blood for 60 min and 24 hr (P less than 0.001). Diene conjugates were raised after WI alone and further still after reperfusion. Thus an early index of lipid peroxidation (diene conjugation) suggested peroxidative damage during the warm ischaemic period itself, whilst detection of Schiff bases was only possible after in vitro incubation of the tissue. Both indices of oxygen-derived free radical damage were increased after reperfusion in vivo with blood and may relate to the degree of tissue damage sustained during ischaemia and reflow.
将兔肾原位夹闭，使其处于热缺血（WI）状态60分钟和120分钟。然后在缺血损伤后立即取出，或者在血液再灌注60分钟或24小时后取出。对匀浆进行磷脂 - 席夫碱荧光检测（激发波长360nm，发射波长435nm）以及通过紫外分光光度法（240nm）检测二烯共轭物的形成，以此作为脂质过氧化的指标。热缺血后即刻，组织中席夫碱水平无明显变化，但当匀浆在37℃孵育90分钟时，与对照组相比，过氧化速率显著升高（热缺血60分钟后P＜0.02，热缺血120分钟后P＜0.001）。血液再灌注60分钟和24小时后，这些值进一步升高（P＜0.001）。单独热缺血后二烯共轭物升高，再灌注后进一步升高。因此，脂质过氧化的早期指标（二烯共轭）提示在热缺血期本身就存在过氧化损伤，而席夫碱的检测只有在组织体外孵育后才有可能。体内血液再灌注后，两种氧自由基损伤指标均升高，这可能与缺血和再灌注期间所遭受的组织损伤程度有关。
