Protective role of endothelial calpain knockout in lipopolysaccharide-induced acute kidney injury via attenuation of the p38-iNOS pathway and NO/ROS production

To explore the role of calpain and its signaling pathway in lipopolysaccharide (LPS)-induced acute kidney injury (AKI), animal models of endotoxemia were established by administration of LPS to mice with endothelial-specific Capn4 knockout (TEK/Capn4(-/-)), mice with calpastatin (an endogenous calpain inhibitor) overexpression (Tg-CAST) and mice with myeloid-specific Capn4 knockout (LYZ/Capn4(-/-)). Mouse pulmonary microvascular endothelial cells (PMECs) were used as a model of the microvascular endothelium and were stimulated with LPS. Renal function, renal inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) expression, cellular apoptosis, plasma and renal levels of NO and reactive oxygen species (ROS), and phosphorylation of mitogen-activated protein kinase (MAPK) family members (p38, ERK1/2, and JNK1/2) were examined. Moreover, a calpain inhibitor, calpastatin overexpression adenoviruses and MAPK inhibitors were used. Significant renal dysfunction was induced by LPS stimulation, and recovery was observed in TEK/Capn4(-/-) and Tg-CAST mice but not in LYZ/Capn4(-/-) mice. Endothelial Capn4 knockout also abrogated the LPS-induced increases in renal iNOS expression, caspase-3 activity and apoptosis and plasma and renal NO and ROS levels but did not obviously affect renal eNOS expression. Moreover, LPS increased both calpain and caspase-3 activity, and only the expression of iNOS in PMECs was accompanied by increased phosphorylation of p38 and JNK. Inhibiting calpain activity or p38 phosphorylation alleviated the increased iNOS expression, NO/ROS production, and cellular apoptosis induced by LPS. These results suggest that endothelial calpain plays a protective role in LPS-induced AKI by inhibiting p38 phosphorylation, thus attenuating iNOS expression and further decreasing NO and ROS overproduction-induced endothelial apoptosis. Acute kidney injury: enzyme block could combat failure Therapies that inhibit the enzyme calpain could alleviate the effects of acute kidney injury according to researchers in China and Canada. Acute kidney injury is induced by endotoxemia, in which changes in the permeability of the intestine allow lipopolysaccharides (LPS) to pass from gut bacteria into the bloodstream. Calpain is known to be active during this process. Zhifeng Liu at the General Hospital of Guangzhou Military Command and co-workers induced endotoxemia in various mouse models by injecting them with LPS. The LPS induced significant kidney dysfunction and cell death, but these were alleviated in mice that were genetically modified to block calpain activity in the blood vessel lining, and in mice that overexpressed calpastatin, a calpain inhibitor. Blocking calpain reduces the expression of nitric oxide synthases that damage endothelial cells.