Experimental Study Of Self-prepared PV Solution On The Liver Cold Preservation In Rats

Objectivity:To discuss the effectiveness and mechanism of self-prepared PV solution for preserving rat's liver at a lower temperature,? then provide theory and experiment evidence for the application of PV solution in donor liver preservation.Materials and methods:1.? The PV solution was produced according to the standard preparation and formulation and then sterilised through aseptic filtering. The PH was controlled at 8.0±0.5 and osmotic pressure was(320±10)mOsm/L。2.?Experiment animal:90 adult, healthy, sanitary and male Wistar rats, of which weight ranged from 200 to 250 gram, were utilised in the experiment. They were randomly categorised into UW solution group﹑PV solution group and NS group based on respective preservation solution with 30 rats in each group. The three groups then divided into 5 subgroups randomly based on variable preservation time, 0 hour﹑6 hour﹑12 hour﹑18 hour and 24 hour respectively, with 6 rats in each subgroup.3.? Animal model: to anaesthetize rats by injecting napental from abdominal cavity, rats'livers were perfused and preserved at a lower temperature by adopting IPRL model.4. Measure content:collected the perfusion solution after 0 hour﹑6 hour﹑12 hour﹑18 hour and 24 hour preservation time respectively and then detected changes of hepatic enzyme(ALT, AST and LDH) and amounts of MDA﹑SOD﹑TNF-αas well as the amount of bile secretion in these solution. The histological variation of liver under microscope and electron microscope was also recorded.Result: 1. Enzymological index: the amount of enzyme increased along with preservation time extended in all the three groups. At the same time, amounts were much lower in PV solution and UW solution group than in NS group(P<0.05). There is no significant difference during the PV solution and UW solution(P>0.05), although there was a trend that the figure would be lower in previous group than in the latter one.2.? Amounts of MDA and SOD; the amounts of MDA in the three groups increased as the preservation time prolonged, but it was the reverse pattern in terms of SOD. The amounts of MDA in both PV solution group and UW solution group were less than NS group in different preservation phase (P<0.01), and the SOD were higher than NS group. After 12 hours, the content of MDA in PV solution group was lower than UW solution group (P<0.01), but there was no significant difference in SOD between those two groups(P>0.05).3.?The contents of TNF-α: the amounts of TNF-αin both PV solution group and UW solution group were less than NS group in different preservation phase (P<0.01). There was also significant difference between PV solution group and UW solution group after 6 hours, which the figure was higher in the previous group than the latter one (P<0.05).4.?Amounts of bile secretion: after the perfusion of liver, there was golden brown bile secreted from liver. As the extension of preservation time, the amounts of bile declined gradually. During different periods, there were larger amounts of bile secretion in either PV solution group or UW solution group in comparison with NS group (P<0.05). After 18 hours, the amounts were lower in PV solution group than UW solution group (P<0.05).5.?Microscope observation: as the extension of preservation time, the impairment of liver became gradually severe. There were cytoplasm loosen, oedema or even necrosis in liver cells, and endothelial cell in sinus hepaticus became swollen and gradually shed to liver sinusoid. Among the three groups, the impairment of liver was the most severe in NS group but the least in UW solution group in terms of swollen. The hepatic lobule and structure of sinus hepaticus was still clear and regular.6.?Electron microscope observation: as the extension of preservation time, liver cell became more swollen and vacuole degeneration and cytoplasm became dissolved. Nevertheless,there are no evident structure variation in mitochondria and endoplasmic reticulum. The liver cell in NS group was more evidently swollen and the cytoplasm was found partially dissolved and karyopyknosis compared to the other two groups. Liver cells in both UW solution group and PV solution group were slightly swollen and no obvious abnormal structure and constitution were detected.Conclusion:?Both PV solution and UW solution are able to maintain the function of the liver of Wistar rat and reduce hepatic ischemia-reperfusion injury. They have comparable effects during short preservation period, but as the extension of preservation time, PV solution is superior than UW solution in terms of oxidation resistance and oxygen radical elimination.