Abstract:

Objective To explore the protective effect of hydrogen-rich saline on ischemia-reperfusion induced liver injury in rats and the mechanism of this effect. Methods A total number of 24 healthy maleSprague-Dawley（SD）rats age ranging from 8-10 weeks weighting 220-250 g were randomly divided into three groups（n ＝ 8 in each group）using a random number table ：sham operated group（Sham group），cold ischemia- reperfusion induced liver injury model group（I/R group）and hydrogen-rich saline treated group（HS group）. Laparotomy was performed and the related blood vessels were isolated in Sham group. The liver cold ischemia-reperfusion were induced in I/R and HS groups. Saline or hydrogen-rich saline were injected through inferior vena cava 5 min before portal vein was clamped in I/R or HS group. After the total hepatic ischemic in I/R andHS group，the liver was reperfused with ringer lactate solution（pre-cooled，4℃）at a speed of 6-8 ml/min and a1 mm incision was cut on the infrahepatic vena cava（IHVC）as an outflow tract. The whole anhepatic phase lastedfor 30 min. At 6 h of reperfusion，blood samples were obtained from the inferior vena cava to detect the levels of alanine amino-transferase（ALT），aspartate amino-transferase（AST），tumor necrosis factor-alpha（TNF-α）and interleukin-10（IL-10）in serum. Hepatic specimens were obtained for the determination of malondialdehyde （MDA）and superoxide dismutase（SOD）contents，expression of caspase-3，Bcl-2 and Bax mRNA，and microscopic examination of the pathological changes in hepatic tissues by light microscopy. Results Compared with Shamgroup，the levels of ALT，AST，TNF-α and content of MDA，Bax/Bcl-2，and expression of caspase-3 mRNA in serum were significantly increased，and the level of serum IL-10 and the activity of SOD were decreased in I/R and HS groups〔ALT（U/L）：467.3±203.6、275.7±74.0 vs. 73.9±30.1，AST（U/L）：929.6±442.7、581.4±143.3 vs. 194.6±72.2，TNF-α（μg/L）：0.081 3±0.029 8、0.052 0±0.009 9 vs. 0.027 3±0.004 4，MDA（nmol/mg）：1.13±0.10、1.00±0.11 vs. 0.84±0.06，caspase-3（2-ΔΔCt）：2.77±0.98、1.46±0.33 vs. 1.00±0.00，Bax/Bcl-2（2-ΔΔCt）：1.59±0.34、1.12±0.16 vs. 1.02±0.10，IL-10（μg/L）：0.141 5±0.023 3、0.320 6±0.064 8 vs. 0.205 3±0.049 7，SOD（U/mg）：194.9±20.4、218.1±16.0 vs. 240.8±16.9，all P ＜ 0.05〕. While compared with I/R group，the levels of ALT，AST，TNF-α in serum and content of MDA，Bax/Bcl-2，and expression of caspase-3 mRNA weredecreased，the level of IL-10 and the activity of SOD were increased in HS group〔ALT（U/L）：275.7±74.0 vs. 467.3±203.6，AST（U/L）：581.4±143.3 vs. 929.6±442.7，TNF-α（μg/L）：0.052 0±0.009 9 vs. 0.081 3±0.029 8，MDA（nmol/mg）：1.00±0.11 vs. 1.13±0.10，caspase-3（2-ΔΔCt）：1.46±0.33 vs. 2.77±0.98，Bax/Bcl-2（2-ΔΔCt）：1.12±0.16 vs. 1.59±0.34，IL-10（μg/L）：0.320 6±0.064 8 vs. 0.141 5±0.023 3，SOD（U/mg）：218.1±16.0 vs.194.9±20.4，all P ＜ 0.05〕. Conclusion Hydrogen-rich saline can alleviate cold ischemia reperfusion induced liver injury through its selective antioxidant，inhibition of inflammatory response and anti-apoptotic properties.