| Objective: In each transplant center at present, due to the donor livertransplantation can’t short time into the receptors, including transportationtime, donor transplant patients because of certain factors not sufficientlycomplete for multiple surgical removal of preoperative preparation, surgicaldifficulty disease of liver, and other factors. Therefore, extend the liver topreservation time, reduce the liver tissue ischemia injury, reduced liver bloodflow after opening of ischemia-reperfusion injury, has become a urgentproblem to be solved. To discuss the protective effect of creatine phosphate oncold storage of rats in vitro liver, and explore the related mechanism, in orderto provide the evidence for clinical application.Methods: Male Wister40rats, regular grade.40rats were randomlydivided into control group (simple UW solution perfused liver)10; Low dosegroup A (with UW solution as the base fluid in CP, concentration of1g/100mlperfusion of liver)10; Dose in group B (with UW solution as the base fluid inCP, concentration of2g/100ml perfusion of liver)10; High dose group C(with UW solution as the base fluid in CP, concentration of3g/100mlperfusion of liver)10. With rat liver pure cold preservation in vitro perfusionmodel of rat preoperative, fasting for12h before operation, water, weighing,the left leg muscle injection heparin (100u/100g) preoperative30min,10%chloral hydrate in0.3ml/100g through abdominal cavity anesthesia, afteranesthesia satisfaction, abdominal "ten" glyph incision into the abdominalcavity, abdominal aorta, abdominal aorta intubation, perfusion4℃HCA(kidney preservation in vitro of purines with citrate salt solution) fluid20ml,free portal, the portal vein intubation, perfusion pure4℃UW solution incontrol group, the experimental group, respectively, according to thecategories perfusion in4℃UW solution for the base of different concentration of CP, line of donor liver in situ low gravity irrigation, irrigationquantity20ml), at the same time open the outflow is cut open rat heart,perfusion wash the residual blood in the liver at the same time, to the liver islight in color khaki, deltoid ligament and the falciform ligament aroundseparation, the left phrenic vein ligation, and break away from the hepaticartery ligation, inferior vena cava, portal vein and bile duct, the perfusion fluidretention. Complete for the liver, the experimental group with perfusion ofliver in the same concentration of CP UW solution of4℃, the control groupfor4℃pure UW solution. Experimental applications including high, mediumand low three different doses of creatine phosphate UW fluid infusion andlow-temperature preserved liver, control group pure UW fluid infusion andkept at low temperature is liver, determination of different time points storedliquid cereal third transaminase (ALT) and lactate dehydrogenase (LDH)activity and liver tissue malondialdehyde (MDA), myeloperoxidase (MPO)activity and liver cell apoptosis index (apop tosis index, AI), the nf-kappa Bpositive expression rate, observe the dirty tissue pathological changes.Result: ALT and LDH were lower than those of the control group afterdifferent dosages in cold storage12h; And after18h malondialdehyde (MDA)content, pulp peroxidase (MPO) experimental group were lower than thecontrol group, No obvious difference between the terminal24h groups; Livercell apoptosis index (AI) and the nf-kappa B positive expression rate in thecold storage12h and18h are lower than those of the control group, no obviousdifference between the terminal24h groups; Pathology showed: liver lobulestructure disorder, liver cell degeneration, swelling and cavity formation,hepatic sinusoids and central vein have different degrees of gore, part ofhepatic sinus endothelial cell necrosis and fall off, with reperfusion timeextension the lesion is aggravating, the changes in the control group, eachdose ease between indifference. High dose group after24h after cold storage,liver tissue and save the MDA content in liquid ALT content were higher thanthose in the control group and the middle and low dose group.Conclusion: This experiment showed that CP of in vitro liver cold storage has better protective effect, the mechanism of protecting livertransplantation may be related to reducing energy metabolic disorders andreduce oxygen free radical injury, so as to reduce the membrane lipidperoxidation injury, the stability of cell membrane, reduce perfusion of liverinjury. Especially for the liver after12h after cold keep have effectiveprotection, but unable to extend that24h after cold storage quality of liver. SoCP in liver transplantation for hepatic preservation may have good applicationprospects, but in clinical liver transplantation application to add CP perfusionliquid so far has not been reported at home and abroad, has need to be furtherresearch. And regarding the quantity of perfusion fluid to add CP drugconcentration needs further research. |
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