Evaluation and maintenance of donor liver quality in rats from donated after cardiac death by hypothermic oxygenated machine perfusion and cold storage
Fu Zhen1, Fan Lin1, Ye Shao Jun1, Ye Qi Fa1,2.
1Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Wuhan,Hubei, People's Republic of China; 2The 3rd Xiangya Hospital of Central South University, Research Center of National Health Ministry on Transplantation Medicine Engineering and Technology, Changsha,Hunan, People's Republic of China
Objective: To investigate the effects of hypothermic oxygenated machine perfusion (HOPE) and cold storage (CS) on liver quality in rats from donated after cardiac death after liver transplantation, it would to provide effective ideas for clinical evaluation and repair of DCD liver.
Methods: Twenty male SD rats were randomly divided into three groups (n=5) : normal group is untreated group, cold storage group (CS group) and hypothermic oxygenated machine perfusion group(HOPE group). The DCD model was established by cardiac arrest caused by asphyxia. The rat liver was induced 30min of in-situ warm ischemia and then to be placed in 4℃ UW solution for 4h(CS group), and the liver was placed in 4℃ UW solution for 3h and hypothermic oxygenated machine perfusion for 1h, it was the HOPE group. The liver of DCD of each group received orthotopic liver transplantation. Blood and tissue samples were collected 30min, 1h, 6h, 12h and 7 days after the operation, respectively. The levels of ALT, AST and LDH in serum were detected by automatic biochemical analyzer, the levels of IL-1β and TNF-A in serum were detected by ELISA, and the morphological changes of liver were observed by HE staining, and the postoperative survival rate was evaluated.
Results: The levels of ALT, AST, LDH, IL-1β and TNF-a in CS group were higher than HOPE group, and the differences were statistically significant (p<0.05). The peak values of ALT appeared at 6h after reperfusion, with an average of 4000±385U/L. The peak values of AST and LDH both appeared at 1h after reperfusion. The mean values were 1700±743U/L and 75,000 ±2100U/L, respectively. One hour after transplantation, HE section of liver showed obvious hepatic cell edema, closed hepatic portal area, red blood cell aggregation in hepatic sinusoids, punctured necrosis of some tissues, structural destruction, and a large number of inflammatory cells infiltration. In the HOPE group, the hepatic sinus was enlarged, the structure of the portal area was relatively intact, and the infiltration of red blood cells and inflammatory cells was significantly less than that in the cold preservation group. The 7-day survival rate of normal group, CS group and HOPE group was 100% (5/5), 20% (1/5) and 40% (2/5), respectively. The main cause of postoperative death was pulmonary infarction (<24h).
Conclusion: HOPE has obvious advantages over CS in repairing DCD donor liver. HOPE can improve the microcirculation of DCD donor liver and reduce the incidence of pulmonary infarction, thus promoting the repair of DCD liver function.Key words: hypothermic oxygenated machine perfusion, cold storage, donated after cardiac death, liver.
This work was supported by the the Science and Technology Innovation Cultivation Fund of Zhongnan Hospital of Wuhan University (No. ZNPY2019041).
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