Sun J S, Hang Y S, Huang I H, Lu F J
Department of Orthopedic Surgery, National Taiwan University Hospital, Taipei, ROC.
Free Radic Biol Med. 1996;20(1):107-12. doi: 10.1016/0891-5849(95)02011-x.
This study was designed to evaluate a simple, sensitive, nonstimulated chemiluminescence assay to measure the oxidative stress production in the whole blood of rabbits after an ischemic insult. By using an ultra-sensitive chemiluminescence (CL) analyzer and lucigenin amplification, the assay system can be performed without leukocyte isolation and stimulant administration. The blood CL levels of healthy rabbits were 122 +/- 18 counts/10 s. After 8 h of ischemia, the CL levels of whole blood immediately after reperfusion, 1 h, 2 h, 24 h, 48 h, 7 days, and 14 days of reperfusion were 409 +/- 78 counts/10 s, 283 +/- 55 counts/10 s, 256 +/- 43 counts/10 s, 228 +/- 33 counts/10 s, 185 +/- 32 counts/10 s, 160 +/- 16 counts/10 s, and 119 +/- 15 counts/10 s. The differences were statistically significant between the control and samples obtained up to 24 h after reperfusion. The corresponding creatine phosphate kinase (CPK) levels of the rabbits in this study were 938 +/- 43 U/l for the control blood sample and 5921 +/- 498 U/l, 6948 +/- 427 U/l, 6860 +/- 1115 U/l, 5763 +/- 516 U/l, 1545 +/- 291 U/l, 478 +/- 60 U/l, and 458 +/- 48 U/l for the experimental blood samples. The CPK levels of the blood samples before 24 h and after 7 days of reperfusion were significantly different to that of the control blood sample. The changes in the CL and CPK levels were quite similar before 48 h reperfusion. This assay has proved to be valuable in the quantitative measurement of ischemic insults of skeletal muscles.
本研究旨在评估一种简单、灵敏的非刺激化学发光测定法，以测量家兔缺血性损伤后全血中的氧化应激产生情况。通过使用超灵敏化学发光（CL）分析仪和光泽精放大技术，该测定系统无需分离白细胞和给予刺激剂即可进行。健康家兔的血液CL水平为122±18计数/10秒。缺血8小时后，再灌注即刻、再灌注1小时、2小时、24小时、48小时、7天和14天的全血CL水平分别为409±78计数/10秒、283±55计数/10秒、256±43计数/10秒、228±33计数/10秒、185±32计数/10秒、160±16计数/10秒和119±15计数/10秒。对照组与再灌注后24小时内获取的样本之间差异具有统计学意义。本研究中家兔相应的磷酸肌酸激酶（CPK）水平，对照血样为938±43 U/l，实验血样分别为5921±498 U/l、6948±427 U/l、6860±1115 U/l、5763±516 U/l、1545±291 U/l、478±60 U/l和458±48 U/l。再灌注24小时前和7天后血样的CPK水平与对照血样显著不同。再灌注48小时前CL和CPK水平的变化非常相似。该测定法已被证明在骨骼肌缺血性损伤的定量测量中具有重要价值。
