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Evaluation by Measuring Activation of Calpain µ

Meng Wang, MD; Masato Sakon, MD; Koji Umeshita, MD; Hideyuki Miyoshi, MD; Kazunori Taniguchi, MD; Shin-ichi Kishimoto, MD; Shinobu Imajoh-Ohmi, PhD; Morito Monden, MD

Arch Surg. 1998;133:983-987.

Objective  To determine the safest method of hepatic vascular clamping associated with the least ischemia-reperfusion injury of the liver during liver surgery.

Setting  University laboratories.

Subjects  Sixty-five adult male Wistar rats.

Methods  The hilar area of the left lateral and median lobes of rat liver was clamped for 10 minutes (group 1), 15 minutes (group 2), or 20 minutes (group 3) followed by 5 minutes of reperfusion. The procedure was repeated for a total period of ischemia of 60 minutes in each group. Control rats underwent laparotomy without vascular clamping. In addition to histological examination, we determined calpain µ activity, a marker of liver injury, by Western blotting using specific antibodies against the intermediate (activated) and proactivated forms of calpain µ. Measurements were performed at the end of ischemia and after 2 hours of reperfusion. We also determined the degradation of talin, an intracellular substrate of calpain µ, by Western blotting.

Results  The level of adenosine triphosphate and energy charge at 2 hours after reperfusion did not change after ischemia-reperfusion irrespective of the duration of ischemic cycle. After 60 minutes of intermittent ischemia followed by 2 hours of reperfusion, cell membrane bleb formation, calpain µ activation, and talin degradation were detected in groups 2 and 3 but not in group 1.

Conclusion  The safest method of hepatic vascular clamping that produces a minimum or no ischemia-reperfusion injury is 60 minutes of 6 cycles of 10-minute vascular clamping interrupted by 5 minutes of reperfusion.

From the Department of Surgery II, Osaka University Medical School, Osaka (Drs Wang, Sakon, Umeshita, Miyoshi, Taniguchi, Kishimoto, and Monden), and Institute of Medical Science, University of Tokyo, Tokyo (Dr Imajoh-Ohmi), Japan.

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