Chatterjee Prabal K, Todorovic Zoran, Sivarajah Ahila, Mota-Filipe Helder, Brown Paul A J, Stewart Keith N, Mazzon Emanuela, Cuzzocrea Salvatore, Thiemermann Christoph
Centre for Experimental Medicine, Nephrology and Critical Care, William Harvey Research Institute, Queen Mary-University of London, London, UK.
Biochem Pharmacol. 2005 Apr 1;69(7):1121-31. doi: 10.1016/j.bcp.2005.01.003.
Calpain activation has been implicated in the development of ischemia-reperfusion (I-R) injury. Here we investigate the effects of two inhibitors of calpain activity, PD150606 and E-64, on the renal dysfunction and injury caused by I-R of rat kidneys in vivo. Male Wistar rats were administered PD150606 or E-64 (3mg/kg i.p.) or vehicle (10%, v/v, DMSO) 30min prior to I-R. Rats were subjected to bilateral renal ischemia (45min) followed by reperfusion (6h). Serum and urinary biochemical indicators of renal dysfunction and injury were measured; serum creatinine (for glomerular dysfunction), fractional excretion of Na(+) (FE(Na), for tubular dysfunction) and urinary N-acetyl-beta-d-glucosaminidase (NAG, for tubular injury). Additionally, kidney tissues were used for histological analysis of renal injury, immunohistochemical analysis of intercellular adhesion molecule-1 (ICAM-1) expression and nitrotyrosine formation. Renal myeloperoxidase (MPO) activity (for polymorphonuclear leukocyte infiltration) and malondialdehyde (MDA) levels (for tissue lipid peroxidation) were determined. Both PD150606 and E-64 significantly reduced the increases in serum creatinine, FE(Na) and NAG caused by renal I-R, indicating attenuation of renal dysfunction and injury and reduced histological evidence of renal damage caused by I-R. Both PD150606 and E-64 markedly reduced the evidence of oxidative stress (ICAM-1 expression, MPO activity, MDA levels) and nitrosative stress (nitrotyrosine formation) in rat kidneys subjected to I-R. These findings provide the first evidence that calpain inhibitors can reduce the renal dysfunction and injury caused by I-R of the kidney and may be useful in enhancing the tolerance of the kidney against renal injury associated with aortovascular surgery or renal transplantation.
钙蛋白酶激活与缺血再灌注（I-R）损伤的发展有关。在此，我们研究钙蛋白酶活性的两种抑制剂PD150606和E-64对大鼠肾脏体内I-R所致肾功能障碍和损伤的影响。雄性Wistar大鼠在I-R前30分钟腹腔注射PD150606或E-64（3mg/kg）或溶剂（10%，v/v，二甲亚砜）。大鼠经历双侧肾脏缺血（45分钟），随后再灌注（6小时）。测量肾功能障碍和损伤的血清及尿液生化指标；血清肌酐（用于评估肾小球功能障碍）、钠分数排泄率（FE(Na)，用于评估肾小管功能障碍）和尿N-乙酰-β-D-氨基葡萄糖苷酶（NAG，用于评估肾小管损伤）。此外，肾脏组织用于肾损伤的组织学分析、细胞间黏附分子-1（ICAM-1）表达的免疫组织化学分析以及硝基酪氨酸形成的分析。测定肾髓过氧化物酶（MPO）活性（用于评估多形核白细胞浸润）和丙二醛（MDA）水平（用于评估组织脂质过氧化）。PD150606和E-64均显著降低了肾脏I-R所致血清肌酐、FE(Na)和NAG的升高，表明肾功能障碍和损伤得到减轻，且I-R所致肾损伤的组织学证据减少。PD150606和E-64均显著减轻了I-R大鼠肾脏的氧化应激证据（ICAM-1表达、MPO活性、MDA水平）和亚硝化应激（硝基酪氨酸形成）。这些发现首次证明钙蛋白酶抑制剂可减轻肾脏I-R所致的肾功能障碍和损伤，可能有助于提高肾脏对与主动脉血管手术或肾移植相关的肾损伤的耐受性。
