The Mechanism Of Hydrogen-rich Saline Protecting Against Ischemia Reperfusion Injury In Liver Transplantation

Objective: To investigate the protective effects of hydrogen-rich saline in rats undergoing orthotopic liver transplantation(OLT) and to explore the mechanism autophagy participated. Methods:1 、 Healthy male Sprague-Dawley rats, weighing 220~250g, were divided randomly into three experimental groups(n=8 each): sham operation group(sham); orthotopic liver transplantation group(OLT); orthotopic liver transplantation + hydrogen-rich saline treatment group(OLT+H). Sham group only underwent simple laparotomy. In group OLT and OLT+H, the orthotopic liver transplantation was performed. After infrahepatic vena cava(IHVA) reperfusion, hydrogen-rich saline and normal saline 6mL/kg were injected respectively in group OLT+H and OLT through IHVA. Blood pressure was monitored in all group during surgery and all rats were sacrificed at 6h after reperfusion. Hepatic function, the level of oxidative stress and pathological changes were detected.2、Rats were divided randomly into four experimental groups(n=8 each): sham operation group(sham); orthotopic liver transplantation group(OLT); orthotopic liver transplantation + hydrogen-rich saline treatment group(OLT+H) and orthotopic liver transplantation + hydrogen-rich saline + 3-methyladenine(OLT+H+3-MA). In group OLT+H+3-MA, 3-MA 2.5mg/kg was injected intraperitioneally 1h before surgery. Blood pressure was monitored in all group during surgery. All rats were sacrificed at 6h after reperfusion. Hepatic function, the level of oxidative stress and pathological changes were detected. Autophagosome was observed by transmission electron microscopy. Apoptosis and autophagy associated proteins and p53 protein were assayed by Western blotting. TUNEL was used to measure the level of apoptosis. Results:1、Blood pressure of rats decreased in anhepatic phase and returned to normal level until 10 min after surgery but no significant difference between group OLT and group OLT+H(P﹥0.05). Compared with group sham, the levels of AST, ALT and MDA, and pathological damage were significantly increased but the level of T-SOD decreased in group OLT and OLT+H(P ﹤ 0.05). Hydrogen-rich saline significantly reduced the level of hepatic function and oxidative stress, and alleviated liver damage compared with group OLT(P﹤0.05).2、Blood pressure of rats in group OLT+H +3-MA had no significant change with group OLT and group OLT+H(P﹥0.05). Compared with group sham, the liver function, the levels of oxidative stress and apoptosis were significantly increased in group OLT、OLT+H and OLT+H+3-MA(P﹤0.05), and the level of autophagy was increased except in group OLT+H+3-MA(P ﹥ 0.05). Hydrogen-rich saline significantly alleviated liver damage and reduced the level of oxidative stress, apoptosis and autophagy but the expression of p-p53 was increased markedly compared to group OLT(P﹤0.05). As compared with group OLT+H, apoptosis and autophagy were alleviated dramatically in group OLT+H+3-MA(P﹤0.05) but the expression of p-p53 did not significantly change(P﹥0.05). Conclusion: The results demonstrate hydrogen-rich saline can alleviate hepatic ischemia reperfusion injury during orthotopic liver transplantation. The possible mechanism of hydrogen-rich saline is inhibiting autophagy through activating the expression of p-p53.