| Objective The goal of the present study was to confirm the protective effect of hydrogen-rich saline on acute kidney injury after orthotopic live transplantation and to explore the molecular mechanism of autophagy participated in the effect.Methods Healthy male Sprague-Dawley rats, weighting 220- 250 g, were randomly assigned into 4 groups using a random number table taking into consideration of the weight: sham, orthotopic liver transplantation(OLT), hydrogen-rich saline treatment group(HS) and chloroquine pretreatment group(CQ). Laparotomy was perform, and the related blood vessels were isolated in sham. Orthotopic liver transplantation was induced in OLT, HS and CQ groups. Saline or hydrogen-rich saline was injected through inferior vena cava five minutes before anhepatic phase in OLT, HS and CQ groups. Chloroquine was injected intraperitoneally at a dose of 60 mg/kg an hour before surgery. Then the rats were sacrificed 6 h after reperfusion. The renal function and the extent of oxidative stress were Observed. Microscopic examination the pathological changes in renal tissues. Using RT-PCR detected the expression of activated caspase-3 and cyt c. The levels of autophagy and apoptosis relative proteins were assayed by western blot analysis. And transmission electron microscopy observed the formation of autophagosome and autolysome.Results There showed a serious renal injury in OLT group, including tubular dilatation, cellular vacuolization, irregularities in cytoplasm and nuclei of epithelial cells and loss of brush borders, and a significant rise in concentration of BUN, Cr and MDA, while SOD activity decreased compared to sham. The expression of activated caspase-3 and cyt c were up-regulated significantly. Whereas hydrogen-rich saline dramatically ameliorated the renal function, reduced oxidative stress level and histopathology damage after OLT. Simultaneously, hydrogen significantly attenuated the apoptosis by decreasing the apoptotic rate, inhibiting activated caspase-3 and cyt c activity. The expression of p-p53, beclin-1, and LC3-Ⅱ were effectively up-regulated. Nevertheless inhibition of autophagy by chloroquine counteracted the renoprotective effects of hydrogen.Conclusion Hydrogen-rich saline induces autophagy through activating the expression of p53, and autophagy activation exerts a renoprotective role via inhibiting the tubular cell apoptosis and alleviating oxidative stress injury during AKI after liver transplantation. |
