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CARDIOVASCULAR ANESTHESIA

Hippocampus bcl-2 and bax Expression and Neuronal Apoptosis After Moderate Hypothermic Cardiopulmonary Bypass in Rats

Ting-Jie Zhang, MD, PhD^*, Jian Hang, MD, PhD, Da-Xiang Wen, MD, PhD^*, Yan-Nan Hang, MD^*, and Frederick E. Sieber, MD

*Department of Anesthesiology, Ren Ji Hospital, Shanghai Second Medicine University, China; and Department of Anesthesiology, Johns Hopkins Bayview Medical Center, Johns Hopkins Medical Institutions, Baltimore, Maryland

Address correspondence and reprint requests to Yan-Nan Hang, MD, Department of Anesthesiology, Ren Ji Hospital, 145 Shan Dong (c) Rd., Shanghai, 200001, People’s Republic of China. Address e-mail to prohynnc{at}online.sh.cn.

Using a rat model of moderate hypothermic (26°C–28°C) cardiopulmonary bypass (CPB) with hemodilution, we investigated hippocampal apoptotic gene expression and neuronal apoptosis up to 6 h after CPB. The CPB was performed on male rats (380–400 g) under general anesthesia with isoflurane and fentanyl. The right atrium and tail artery were cannulated, and a peristaltic pump and membrane oxygenator were used for CPB. Two groups were studied: Group 1 consisted of fasted rats (n = 15) subjected to 60 min of moderate hypothermic nonpulsatile CPB; Group 2 consisted of sham-operated rats (n = 15). At 1 h after CPB, in 6 rats per group, hippocampus was processed for the apoptotic gene (bcl-2 and bax) messenger RNAs detection by reverse transcriptase polymerase chain reaction, and messenger RNA expression was determined by the ratio of the polymerase chain reaction product of bcl-2 or bax to the ß-actin gene. At 6 h after CPB, in 6 rats per group, hippocampus expression of Bcl-2 and bax protein was determined by immunohistochemistry, and neuronal apoptosis was detected by TUNEL. At 6 h after CPB, in three rats per group, changes in hippocampal CA1 neuronal ultra structure were determined with electron microscopy. Group 1 had increased ratios of bcl-2/ß-actin, bax/ß-actin, and bax/bcl-2 mRNA at 1 h after CPB (bcl-2/ß-actin, 0.82 ± 0.14 versus 0.63 ± 0.07; P = 0.03; bax/ß-actin, 1.04 ± 0.14 versus 0.56 ± 0.03; P = 0.00; bax/bcl-2, 1.31 ± 0.12 versus 0.84 ± 0.09; P = 0.02; Group 1 versus Group 2, respectively). Group 1 had increased bcl-2 and bax protein expression in hippocampal CA1 region at 6 h after CPB (bcl-2, 0.18 ± 0.05 versus 0.09 ± 0.01; P = 0.02; bax, 0.20 ± 0.06 versus 0.04 ± 0.02; P = 0.01; Group 1 versus Group 2, respectively). Group 1 had increased TUNEL staining in hippocampus CA1 at 6 h after CPB (0.14 ± 0.02 versus 0.03 ± 0.01; P = 0.00; Group 1 versus Group 2, respectively). In Group 1 CA1 hippocampus neurons, ultra-structural changes consistent with apoptosis occurred. In rats, moderate hypothermic CPB with hemodilution is associated with CA1 hippocampus bax and bcl-2 gene expression and neuronal apoptosis during the early post-CPB recovery period.

Anesthesia & Analgesia® is published for the International Anesthesia Research Society® by Lippincott Williams & Wilkins with the assistance of Stanford University Libraries' HighWire Press®. Copyright 2006 by the International Anesthesia Research Society. Online ISSN: 1526-7598   Print ISSN: 0003-2999