The Role Of Mfn2 In Hydrogen-rich Saline Against Hepatic Ischemia/reperfusion Injury

Objective By duplicate partial hepatic ischemia/reperfusion(I/R) injury model in rat, investigate the effects of Hydrogen-rich saline on liver ischemia/reperfusion injury in rats. Analyze the relationship between ischemia/reperfusion injury and mitochondrial damage on molecular level and ultrastructure. Method Fifty-six healthy male Sprague-Dawley rats were randomly divided into seven groups(n=8). There are eight rats in the sham operation group(group N), the rest rats are divided into liver I/R plus normal saline treatment group(NS group) and liver I/R plus Hydrogen-rich saline treatment group(HS group). These groups are as follows: control group 1(NS1) and experimental group 1(HS1), control group 2(NS2) and experimental group 2(HS2), control group 3(NS3) and experimental group(HS3). For NS1、HS1 group, NS2、HS2 group, NS3、HS3 group,the portal triad to the left lobe and the left middle lobe of the liver were completely occluded for 20,40,60 min respectively. The group N was administered by intraperitoneal injection of normal saline and underwent simple laparotomy. The blood and liver samples were collected 24 hours after reperfusion. 1. Liver function was detected using an autoanalyzer. 2. HE staining was used to observe the change of liver morphology and neutrophils accumulation. 3. Observe hepatocyte mitochondrial injury by transmission electron microscope. 4. The expression of Mfn2 mRNA and protein was analyzed by real-time PCR and western blot. Results Compared with group NS, the level of ALT, AST were significantly lower in group HS(P<0.05). Moderate hepatic cell swell, necrosis, and neutrophil cell infiltration were seen in group HS, As compared with group NS, group HS significantly alleviated liver damage. TEM observation of mitochondria, group NS have varying degrees of damage, especially group NS3 of mitochondrial damage is the most important, reduce the number, obvious swelling, cristae blurred, the matrix density is reduced, the majority of mitochondrial membrane integrity, while the group HS less mitochondrial morphological changes. As compared with group NS, the expression of activated Mfn2 RNA and protein were significantly decreased in group HS. Conclusion: 1. Intraperitoneal injection of hydrogen-rich saline can obviously attenuate liver co I/R injury in rats, with the characteristic of lower serum levels of ALT and AST, and less apoptosis cells. 2. Intraperitoneal injection of hydrogen-rich saline can contribute the expression of Mfn2 RNA and protein, which can promote the recovery of mitochondrial structure and function, thereby reducing hepatic ischemia/reperfusion injury.