Quoilin 2014 Dissertation
|Quoilin C (2014) Les effets délétères métaboliques et oxydants induits lors d'un sepsis sur la fonction rénale. Dissertation 1-195.|
Abstract: Acute kidney injury (AKI) is a frequent complication of sepsis that can increase mortality as high as 70%. The pathophysiology of this kidney failure was previously believed to be secondary to decreased global renal perfusion causing hypoxia-induced injury. However, new research suggests this paradigm is overly simplistic, and injury is now considered multifactorial in origin. Mechanisms that contribute to kidney injury mainly include inflammation, alterations in microvascular renal blood flow and changes in bioenergetics.
To study the mechanism of oxygen regulation in acute kidney injury during sepsis, we developed a sepsis-induced in vitro model using proximal tubular epithelial cells (HK-2) exposed to a bacterial endotoxin (lipopolysaccharide, LPS). Our first investigation, by using both high-resolution respirometry and electron spin resonance spectroscopy, showed that HK-2 cells exhibit a decreased oxygen consumption rate when treated with LPS. Surprisingly,this cellular respiration alteration persists even after the stress factor is removed. We suggested that this irreversible decrease in renal oxygen consumption after LPS challenge is related to a pathologic metabolic down-regulation such as a lack of oxygen utilization by cells for ATP production. In the long term, this metabolic disturbance leads cells to a predominantly apoptotic death.
• O2k-Network Lab: BE Liege Votion DM
Labels: MiParea: Respiration, Patients Pathology: Sepsis Stress:Cell death, Ischemia-reperfusion, Oxidative stress;RONS
Tissue;cell: Kidney, Endothelial;epithelial;mesothelial cell
Enzyme: Complex IV;cytochrome c oxidase
To confirm this hypothesis of cytopathic hypoxia, we demonstrated that this alteration in the renal respiratory function is mainly due to an impairment in the metabolic activity of HK-2 cell mitochondria. Following LPS treatment,the oxidative phosphorylation is interrupted because of the inhibition of cytochrome c oxidase activity. As a consequence, disruptions in the electron transport and the proton pumping across the system occurr, leading to a decrease of the mitochondrial membrane potential, the release of apoptotic-inducing factors and a decrease in ATP production.To clarify the mechanism by which the LPS induces mitochondrial alterations, we studied the oxidative stress generation in HK-2 cells. Interestingly, we revealed that the induction of a cytosolic oxidative stress is an event that appears before mitochondrial dysfunction in the LPS-treated HK-2 cells. This primary redox state is notably due to the activation of the two enzymes NADPH oxidase 4 and inducible NO synthase. The simulta neous production of anion superoxide and nitric oxide strongly suggests the formation of peroxynitrite, a relative stable powerful oxidant, that can diffuse through mitochondrial compartments and undergo cytotoxic reactions. To our knowledge, our model reveals for the first time the role of NADPH oxidase-derived cytosolic ROS in triggering tubular cell damage. Moreover, after being first target of the oxidative stress, mitochondria become in turn producer of reactive oxygen species that carry on mitochondrial dysfunction. It seems thus that a mechanism of oxidative stress-induced redox cycling is a main cause of the mitochondrial dysfunction of LPS-treated HK-2 cells. The role of oxidants in mitochondrial dysfunction was further confirmed by the use of iNOS inhibitors or antioxidants that preserve cytochrome c oxidase activity and block mitochondrial membrane potential dissipation.
Overall, these results suggest that sepsis-induced AKI should not only be regarded as failure of energy status but also as an integrated response, including transcriptional events, ROS signaling, mitochondrial activity and metabolic orientation such as apoptosis.