Deletion of the kinase domain in death-associated protein kinase attenuates tubular cell apoptosis in renal ischemia-reperfusion injury
Deletion of the kinase domain in death-associated protein kinase attenuates tubular cell apoptosis in renal ischemia-reperfusion injury
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DOI：
10.1097/01.asn.0000131527.59781.f2
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发表时间：
2004-07-01
影响因子：
13.6
通讯作者：
Mune, M
中科院分区：
文献类型：
--
作者：
Kishino, M;Yukawa, K;Mune, M
死亡相关蛋白激酶（DAPK）是一种钙/钙调蛋白依赖性丝氨酸/苏氨酸激酶，定位于肾小管上皮细胞。为了阐明DAPK活性对肾缺血-再灌注（IR）损伤中细胞凋亡的贡献，对野生型（WT）小鼠和表达缺少部分激酶结构域的DAPK缺失突变体的DAPK突变小鼠进行肾蒂夹闭和再灌注。IR后，WT肾脏中的DAPK活性升高，但突变肾脏中未升高（1785.7 +/- 54.1 pmol/ min/mg vs 160.7 +/- 60.6 pmol/min/mg）。此外，IR后WT肾脏中TUNEL阳性细胞核和活化的半胱天冬酶3阳性细胞比突变肾脏中更多（缺血后40 h，24.0 +/- 5.9个细胞核或9.4 +/- 0.6个细胞/高倍视野[HPF] vs 6.3 +/- 2.2个细胞核或4.4 +/- 0.7个细胞/HPF）。此外，IR后WT肾中p53阳性小管细胞的增加大于突变肾中的增加（9.9 +/- 1.4个细胞/HPF对0.8 +/- 0.4个细胞/HPF），这与DAPK活性稳定p53蛋白的理论一致。最后，IR后WT小鼠的血清肌酐水平高于突变小鼠（缺血后40小时为2.54 +/- 0.34 mg/dl vs 0.87 +/- 0.24 mg/dl）。因此，这些结果表明，从DAPK分子中删除激酶结构域可以减轻IR损伤后的肾小管细胞凋亡和肾功能障碍。
Death-associated protein kinase (DAPK) is a calcium/calmodulin-dependent serine/threonine kinase localized to renal tubular epithelial cells. To elucidate the contribution of DAPK activity to apoptosis in renal ischemia-reperfusion (IR) injury, wild-type (WT) mice and DAPK-mutant mice, which express a DAPK deletion mutant that lacks a portion of the kinase domain, were subjected to renal pedicle clamping and reperfusion. After IR, DAPK activity was elevated in WT kidneys but not in mutant kidneys (1785.7 +/- 54.1 pmol/ min/mg versus 160.7 +/- 60.6 pmol/min/mg). Furthermore, there were more TUNEL-positive nuclei and activated caspase 3-positive cells in WT kidneys than in mutant kidneys after IR (24.0 +/- 5.9 nuclei or 9.4 +/- 0.6 cells per high-power field [HPF] versus 6.3 +/- 2.2 nuclei or 4.4 +/- 0.7 cells/HPF at 40 h after ischemia). In addition, the increase in p53-positive tubule cells after IR was greater in WT kidney than in mutant kidneys (9.9 +/- 1.4 cells/HPF versus 0.8 +/- 0.4 cells/HPF), which is consistent with the theory that DAPK activity stabilizes p53 protein. Finally, serum creatinine levels after IR were higher in WT mice than in mutant mice (2.54 +/- 0.34 mg/dl versus 0.87 +/- 0.24 mg/dl at 40 h after ischemia). Thus, these results indicate that deletion of the kinase domain from DAPK molecule can attenuate tubular cell apoptosis and renal dysfunction after IR injury.