| Backgrouds:Renal ischemia/reperfusion (I/R)-induced tubular epithelial cell injury called ischemic acute renal failure, is associated with high mortality in humans. Protecting the kidney against I/R injury is very important during anesthesia and sedation for susceptible patients.Propofol, a highly lipid-soluble anesthetic, is widely used for the induction and maintenance of general anesthesia, as well as for intubated post-operation sedation for mechanically-ventilated adults in the intensive care unit. Propofol has been proven to ameliorate I/R injury in several organs, including the heart, lung, brain, liver, and testicles.. However, the effects of propofol on renal I/R injury have been rarely reported. We want to find the protect effect and the underling molecular mechanisms of propofol in a celluler model and in a rat in vivo model.Part1:Methods:A reversible ATP depletion (antimycin A) followed by restoration of substrate supply in human kidney proximal tubular epithelial cells(HKC) was used as an in vitro model of renal I-R. Study groups included1) control;2) cells incubated with propofol (20umol/L) alone;3) cells with I-R or4) cells prctreated with propofol20umol/L and I-R injury.Detection of the activity of HK-2cells was done by MTT automatic colorimetry, Cell Apoptosis was evaluated by Hoechst-33258staining, intracellular ROS content by Fluorescence How cytometry,MDA Assays for markers of oxidative stress and lipid peroxidation were determined by TBA assay, Chemiluminescence method for the determination of the total SOD activity in group cells.detection of Caspase-3by Western-blot test, expression of NF-KB by Hlectrophoretic Mobility Shift Assay, Laser Confocal detection HIF-1α and Western-blot quantitative determination of HIF-1α.Result:I-R injury decrease cellviability, induce apoptosis and increased intracellular ROS content and marker of oxidative stress. As the same time of injury the xpression of related apoptosis molecular Caspase-3inflammation relatied NF-κB and HIF-1α increased significantly. Propofol pretreatment before I/R injury significantly increased SOD activity and decrease the ROS,MDA and then apoptosis of HKC. Quantitative measurement of Caspase-3, NF-κB, and HIF-1a in obviously decreased compared to I/R stimulated cells. Propofol alone show no different to control group in HIF-1a expression.Conclusion:Propofol20umol/L can reduce I/R HKC injury damage and apoptosis, This protection was probably due to at least in part by decrease ROS and suppressing caspase-3activity and NF-κB expression mechanism, pretreatment with propofol alone can not induce HIF-1a to protect cell from follow up I/R injury.Part2:Methods:Wistar rats were randomly divided into5groups:(Ⅰ) control (ⅱ) sham-operated control group;(ⅲ)propofol group;(ⅳ) I/R group; and(v) propofol pretreatment and I/R group. Bilateral renal warm ischemia for60mins was performed. After24h reperfusion, blood samples and kidneys were collected for assessment of renal injury, renal tissue ROS activity detective by SOD and MDA, caspas-3activity were analyzed by Western bloting and northen-blotting analysis, NF-K B activity analyzed by EMS A assay,HIF-1a were also measured by Western bloting.Results:Blood serum creatinine and Cystatin C levels in the propofol pretreatment group were significantly lower than that in the I/R group at24h after reperfusion. The mean histological score by Paller’s standard showed that propofol significantly attenuated renal I/R injury. As the same time of injury, the expression of related apoptosis molecular in the renal tissue Caspase-3,inflammation relatied NF-κB and HIF-1a increased significantly, Propofol pretreatment before I/R injury significantly decreased MDA and Caspase-3activity,reduced the expression of NF-K B and HIF-1a. Propofol alone show no different to control group in HIF-1a expression.Conclusion:Propofol can clear the body of oxygen free radicals and oxidative stress in our renal ischemia-reperfusion injury model in vivo, inhibiting apoptosis and inflammatory pathways, and thus pathological damage mitigation, kidney function decline abated. The protection effect of propofol pretreatment is independent with induced expression of HIF-1a... |
PDF Full Text Request