Protective Effect Of Hydrogen Saline On Liver Function After Extended Lobectomy And The Mechanism

Liver resection is considered the standard treatment for primary liver safety of hepaticresections improves, more patients with extensive disease requiring extendedlobectomy for complete resection are considered operative candidates. Although themortality rate is low, patients undergoing large resections continue to be at increasedrisk for posthepatectomy liver failure (PHLF) and complications. PHLF remains amajor concern and has been shown to be a predominant cause of hepatectomy-relatedmortality. The resulting small functional remnant liver volumes and ischemia-reperfusion injury are considered to be the main causes of PHLF. Reactive oxygenspecies (ROS) are considered to play a prominent causative role in the developmentof ischemia-reperfusion injury. In recent years, Hydrogen (H2) was reported widelyto selectively reduce the oxidants. Hydrogen has come to the forefront of therapeuticmedical research.Part1. Establishing the model of extended lobectomy in pigsObjective: to establish a feasible, stable and effective model of extendedlobectomy in miniature pigs.Materials and Methods:6Ba-Ma miniature pigs, weighted from25to30kg,underwent70%liver resection in conditions of general anesthesia. The profiles ofintra-operative data were investigated. Animals were followed for7days with dailyserum for biochemistry exam. The survival rate were also calculated respectively.Results: The Intraoperative vital signs of pigs were stable and all animals weresurvived on the post operative day(POD)7without fatal complications. Serum TBILincreased on the POD1, reached the peak on the POD3and then deceased. SerumALT and AST reached the peak on the POD1and then deceased. HGB did not changesignificantly after operation.Conclusions: Porcine model of70%liver resection can be safely established andused for further study of hepatic insufficiency as its good clinical simulation. Part2. Protective effect of hydrogen saline on liver function afterextended lobectomy and the underlying mechanismObjective: to investigate the protective effect of hydrogen saline on liver functionafter extended lobectomy and explore the underlying mechanism.Materials and Methods:6pairs of Ba-Ma miniature pigs were assigned into2groups: NS group, underwent70%liver resection with normal saline as control; H2group, underwent70%liver resection with intravenous injection of hydrogen salineon the first3days after operation. Animals were followed for7days with daily serumfor biochemistry exam. The survival rate were also calculated respectively. Animalswere executed on the POD7, the pathological changes of liver tissue were lightmicroscope and electron microscope. The inflammation of liver tissue were observedthrough immunohistochemical p65staining. The proliferation were observed throughimmunohistochemical ki67staining. The content of malondialdehyde (MDA),superoxide dismutase (SOD) and catalase (CAT) in liver tissue were detected by thecorresponding detection kit. The serum levels of inflammatory factors such as IL-6、TNF-α、 IL-βwere detected by ELISA kits. The mRNA level of inflammationtransduction molecules P65in liver tissue was detected by Real-time PCR. Theexpression of apoptosis related proteins BCL–2and PARP were detected by WesternBlot. The expression of inflammatory transduction molecules p65、IKBα、IKKα、IKKβwere also detected by Western Blot.Results:1. Survival rate: The Intraoperative vital signs of pigs were stable and allanimals were survived on the post operative day(POD)7without fatalcomplications. No significant differences were found between the2groups inoperation time, volume of resected liver, estimated residual volume, warmischemia time and blood loss.（P>0.05）2. Liver function: Compared with the NS group, the ALT and AST in H2group were significant decreased on POD1to7as well as the TBIL.（P<0.05）3. Light Microscopic Examination: Significant liver cell degeneration andinflammatory cells infiltration were observed as well as stripped hepatic sinusendothelial, damage of portal vein endothelial and disorder of hepatic lobule in theNS group, whereas in the H2group, these pathological change were minimal.4. Electron Microscope Examination: Electron microscopic observation showedcondensation of the cytoplasm and nucleuses, margination and fragmentation ofthe chromatin, and formation of apoptotic bodies in the NS group, whereas thesepathological change were minimal in the H2group.5. Oxidative stress in liver: Compared with the NS group, the content of MDAwas significant lower,while the content of SOD and CAT were higher in the H2group.（P<0.05）6. Serum levels of inflammatory cytokines: Compared with the NS group, theserum levels of inflammatory factors such as IL-6、TNF-αand IL-βwere significantlower in the H2group.（P<0.05）7. Immumohistochemical staining: P65staining showed that compared with NSgroup, cytoplasm staining in H2was weakened. Ki67staining showed thatcompared with NS group, ki67positive dyeing cells increased significantly in theH2group.（P<0.05）8. The mRNA level of inflammation transduction molecules P65: Comparedwith the NS group, The mRNA level of P65was significant lower in the H2group.（P<0.05）9. The expression of inflammatory transduction molecules: Compared with theNS group, p65was inhibited but IKBα was more activated in the H2group. Theexpression of IKKα and IKKβ was no significantly different between2groups.10. The expression of apoptosis related proteins: Compared with the NS group,BCL–2was more activated while Cleaved PARP was less activated in the H2group. Conclusions:1. Hydrogen saline can effectively reduce the liver injury after liver resection.2. Hydrogen saline can alleviate oxygen free radical damage to the liver tissuein ischemia-reperfusion.3. Hydrogen saline can reduce tissue inflammation by inhibiting the expressionof inflammation transduction factor.4. Hydrogen saline can reduce cell apoptosis and promote cell proliferation.