重组人肝细胞生长因子在Celisor液中对无心跳供肝的保护作用及其机制

目的 探讨Celsior(CS)液中添加重组人肝细胞生长因子(rhHGF)对无心跳供肝(NHBD)的保护作用及其可能机制.方法 利用大鼠肝移植模型,比较添加rhHGF(10μg/L)的CS液(实验组)与添加等量生理盐水的CS液(对照组)两组门静脉再通后胆汁产生量、肝组织含水量及肝酶水平,观察生存率,TUNEL法检测肝细胞凋亡及常规病理组织学检查.结果 与对照组比较,试验组NHBD供肝移植后在门静脉再通1 h,3 h及6 h胆汁产生量明显增多(P<0.05),ALT与AST水平明显降低(P<0.05);在门静脉再通1 h肝组织含水量及肝细胞凋亡指数明显下降(P<0.05);7 d大鼠生存率明显升高(P<0.05),病理组织学提示肝细胞水肿变性明显减轻,无肝细胞坏死及肝窦淤血.结论 CS灌注保存液中添加外源性rhHGF可明显改善其对NHBD供肝的保存效果,这可能与其减少肝细胞凋亡有关.     

Celsior液中HGF对大鼠肝移植后INF-γ、IL-4与IL-10水平的影响

目的 探讨Celsior(CS)液中添加重组人肝细胞生长因子(recombinant human hepato-cyte growth factor,rhHGF)对大鼠肝移植后INF-γ、IL-4与IL-10水平的影响.方法 利用无心跳供肝(non-heart beating donor,NHBD)大鼠肝移植模型,ELISA法检测实验组(CS添加rhHGF)和对照组(CS添加生理盐水)门静脉再通1h血清INF-γ、IL-4与IL-10水平,逆转录聚合酶链反应(RT-PCR)测定肝组织INF-γ、IL-4与IL-10 mRNA水平,并观察肝组织病理及大鼠牛存率.结果 与对照组相比,试验组NHBD供肝移植后在门静脉再通1 h后血清INF-γ水平及肝组织INF-γ mRNA水平明显低于对照组(P<0.05),血清IL-4与IL-10水平及相应肝组织IL-4与IL-10 mRNA水平明显高于对照组(P<0.05),实验组大鼠生存率明显高于对照组(P＜0.05),病理组织学提示实验组肝细胞水肿变性减轻,无明显肝细胞坏死、肝窦淤血及门静脉充血.结论 CS灌注保存液中添加外源性rhHGF可改善其对NHBD供肝的保存效果,提高术后生存率,这可能与其下调INF-γ,上调IL-4与IL-10的产生有关.     

Celsior液与UW液对无心跳大鼠供肝保存效果的比较

Objective To compare the effects of Celsior (CS) solution vs University of Wisconsin (UW) solution on preservation of non-heart- beating donor (NHBD) livers in rats. Methods The healthy male SD rats were taken as donors and recipients. The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 10 min to induce ischemia as a NHBD model. After flushed with the cold CS or UW solution, NHBD livers were stored at 4℃ for 8 h or 16 h. Then they were transplanted using the two-cuff technique with arterial reconstruction. Accordingly, the recipients were divided into four groups: CS 8 h, UW 8 h, CS 16 h and UW 16 h groups. The bile discharge, liver enzymes (ALT and AST), ET-1, IL-1 and TNF-alpha levels in serum at 1, 3, and 6 h after reperfusion were measured. Histopathological examination was carried out by HE staining, and the 7-day survival rate was observed. Results As compared with a homogenous appearance in the NHBD livers perfused with CS solution, the NHBD livers perfused with UW solution showed a heterogenous appearance, with more or less mottling. In the CS 8 h group there were more bile discharge at 1 h, lower ALT and AST levels at 1, 3 and 6 h, and lower serum ET-1, IL-1 and TNF-alpha ? levels at 1 and 3 h after reperfusion than the UW 8 h group (P<0.05). Histopathological examination showed a well-preserved hepatic architecture, milder portal congestion and inflammatory infiltration in CS 8 h group as compared with UW 8 h group. The 7-day survival rate in CS 8 h and UW 8 h groups was 58.3% and 25.0% respectively (P<0.05). There were no significant differences in bile discharge, ALT, AST, ET-1, IL-1 and TNF-alpha ? levels at different time points between CS 16 h and UW 16 h groups (P>0.05), and histopathological examination showed no much difference in the tissue damage between two groups; all recipients died within 3 days (P>0.05). Conclusion CS was more effective than UW in preserving NHBD livers after 8 h of cold storage, which may be due to the better perfusion of CS and inhibition of Kupffer cells, but it had better preserve NHBD livers with CS no more than 16 h.     

Celsior液与UW液对无心跳大鼠供肝保存效果的比较

Objective To compare the effects of Celsior (CS) solution vs University of Wisconsin (UW) solution on preservation of non-heart- beating donor (NHBD) livers in rats. Methods The healthy male SD rats were taken as donors and recipients. The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 10 min to induce ischemia as a NHBD model. After flushed with the cold CS or UW solution, NHBD livers were stored at 4℃ for 8 h or 16 h. Then they were transplanted using the two-cuff technique with arterial reconstruction. Accordingly, the recipients were divided into four groups: CS 8 h, UW 8 h, CS 16 h and UW 16 h groups. The bile discharge, liver enzymes (ALT and AST), ET-1, IL-1 and TNF-alpha levels in serum at 1, 3, and 6 h after reperfusion were measured. Histopathological examination was carried out by HE staining, and the 7-day survival rate was observed. Results As compared with a homogenous appearance in the NHBD livers perfused with CS solution, the NHBD livers perfused with UW solution showed a heterogenous appearance, with more or less mottling. In the CS 8 h group there were more bile discharge at 1 h, lower ALT and AST levels at 1, 3 and 6 h, and lower serum ET-1, IL-1 and TNF-alpha ? levels at 1 and 3 h after reperfusion than the UW 8 h group (P<0.05). Histopathological examination showed a well-preserved hepatic architecture, milder portal congestion and inflammatory infiltration in CS 8 h group as compared with UW 8 h group. The 7-day survival rate in CS 8 h and UW 8 h groups was 58.3% and 25.0% respectively (P<0.05). There were no significant differences in bile discharge, ALT, AST, ET-1, IL-1 and TNF-alpha ? levels at different time points between CS 16 h and UW 16 h groups (P>0.05), and histopathological examination showed no much difference in the tissue damage between two groups; all recipients died within 3 days (P>0.05). Conclusion CS was more effective than UW in preserving NHBD livers after 8 h of cold storage, which may be due to the better perfusion of CS and inhibition of Kupffer cells, but it had better preserve NHBD livers with CS no more than 16 h.     

Celsior液与UW液对无心跳大鼠供肝保存效果的比较

Objective To compare the effects of Celsior (CS) solution vs University of Wisconsin (UW) solution on preservation of non-heart- beating donor (NHBD) livers in rats. Methods The healthy male SD rats were taken as donors and recipients. The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 10 min to induce ischemia as a NHBD model. After flushed with the cold CS or UW solution, NHBD livers were stored at 4℃ for 8 h or 16 h. Then they were transplanted using the two-cuff technique with arterial reconstruction. Accordingly, the recipients were divided into four groups: CS 8 h, UW 8 h, CS 16 h and UW 16 h groups. The bile discharge, liver enzymes (ALT and AST), ET-1, IL-1 and TNF-alpha levels in serum at 1, 3, and 6 h after reperfusion were measured. Histopathological examination was carried out by HE staining, and the 7-day survival rate was observed. Results As compared with a homogenous appearance in the NHBD livers perfused with CS solution, the NHBD livers perfused with UW solution showed a heterogenous appearance, with more or less mottling. In the CS 8 h group there were more bile discharge at 1 h, lower ALT and AST levels at 1, 3 and 6 h, and lower serum ET-1, IL-1 and TNF-alpha ? levels at 1 and 3 h after reperfusion than the UW 8 h group (P<0.05). Histopathological examination showed a well-preserved hepatic architecture, milder portal congestion and inflammatory infiltration in CS 8 h group as compared with UW 8 h group. The 7-day survival rate in CS 8 h and UW 8 h groups was 58.3% and 25.0% respectively (P<0.05). There were no significant differences in bile discharge, ALT, AST, ET-1, IL-1 and TNF-alpha ? levels at different time points between CS 16 h and UW 16 h groups (P>0.05), and histopathological examination showed no much difference in the tissue damage between two groups; all recipients died within 3 days (P>0.05). Conclusion CS was more effective than UW in preserving NHBD livers after 8 h of cold storage, which may be due to the better perfusion of CS and inhibition of Kupffer cells, but it had better preserve NHBD livers with CS no more than 16 h.     

Celsior液与UW液对无心跳大鼠供肝保存效果的比较

Objective To compare the effects of Celsior (CS) solution vs University of Wisconsin (UW) solution on preservation of non-heart- beating donor (NHBD) livers in rats. Methods The healthy male SD rats were taken as donors and recipients. The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 10 min to induce ischemia as a NHBD model. After flushed with the cold CS or UW solution, NHBD livers were stored at 4℃ for 8 h or 16 h. Then they were transplanted using the two-cuff technique with arterial reconstruction. Accordingly, the recipients were divided into four groups: CS 8 h, UW 8 h, CS 16 h and UW 16 h groups. The bile discharge, liver enzymes (ALT and AST), ET-1, IL-1 and TNF-alpha levels in serum at 1, 3, and 6 h after reperfusion were measured. Histopathological examination was carried out by HE staining, and the 7-day survival rate was observed. Results As compared with a homogenous appearance in the NHBD livers perfused with CS solution, the NHBD livers perfused with UW solution showed a heterogenous appearance, with more or less mottling. In the CS 8 h group there were more bile discharge at 1 h, lower ALT and AST levels at 1, 3 and 6 h, and lower serum ET-1, IL-1 and TNF-alpha ? levels at 1 and 3 h after reperfusion than the UW 8 h group (P<0.05). Histopathological examination showed a well-preserved hepatic architecture, milder portal congestion and inflammatory infiltration in CS 8 h group as compared with UW 8 h group. The 7-day survival rate in CS 8 h and UW 8 h groups was 58.3% and 25.0% respectively (P<0.05). There were no significant differences in bile discharge, ALT, AST, ET-1, IL-1 and TNF-alpha ? levels at different time points between CS 16 h and UW 16 h groups (P>0.05), and histopathological examination showed no much difference in the tissue damage between two groups; all recipients died within 3 days (P>0.05). Conclusion CS was more effective than UW in preserving NHBD livers after 8 h of cold storage, which may be due to the better perfusion of CS and inhibition of Kupffer cells, but it had better preserve NHBD livers with CS no more than 16 h.     

Celsior液与UW液对无心跳大鼠供肝保存效果的比较

Objective To compare the effects of Celsior (CS) solution vs University of Wisconsin (UW) solution on preservation of non-heart- beating donor (NHBD) livers in rats. Methods The healthy male SD rats were taken as donors and recipients. The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 10 min to induce ischemia as a NHBD model. After flushed with the cold CS or UW solution, NHBD livers were stored at 4℃ for 8 h or 16 h. Then they were transplanted using the two-cuff technique with arterial reconstruction. Accordingly, the recipients were divided into four groups: CS 8 h, UW 8 h, CS 16 h and UW 16 h groups. The bile discharge, liver enzymes (ALT and AST), ET-1, IL-1 and TNF-alpha levels in serum at 1, 3, and 6 h after reperfusion were measured. Histopathological examination was carried out by HE staining, and the 7-day survival rate was observed. Results As compared with a homogenous appearance in the NHBD livers perfused with CS solution, the NHBD livers perfused with UW solution showed a heterogenous appearance, with more or less mottling. In the CS 8 h group there were more bile discharge at 1 h, lower ALT and AST levels at 1, 3 and 6 h, and lower serum ET-1, IL-1 and TNF-alpha ? levels at 1 and 3 h after reperfusion than the UW 8 h group (P<0.05). Histopathological examination showed a well-preserved hepatic architecture, milder portal congestion and inflammatory infiltration in CS 8 h group as compared with UW 8 h group. The 7-day survival rate in CS 8 h and UW 8 h groups was 58.3% and 25.0% respectively (P<0.05). There were no significant differences in bile discharge, ALT, AST, ET-1, IL-1 and TNF-alpha ? levels at different time points between CS 16 h and UW 16 h groups (P>0.05), and histopathological examination showed no much difference in the tissue damage between two groups; all recipients died within 3 days (P>0.05). Conclusion CS was more effective than UW in preserving NHBD livers after 8 h of cold storage, which may be due to the better perfusion of CS and inhibition of Kupffer cells, but it had better preserve NHBD livers with CS no more than 16 h.     

Celsior液与UW液对无心跳大鼠供肝保存效果的比较

Objective To compare the effects of Celsior (CS) solution vs University of Wisconsin (UW) solution on preservation of non-heart- beating donor (NHBD) livers in rats. Methods The healthy male SD rats were taken as donors and recipients. The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 10 min to induce ischemia as a NHBD model. After flushed with the cold CS or UW solution, NHBD livers were stored at 4℃ for 8 h or 16 h. Then they were transplanted using the two-cuff technique with arterial reconstruction. Accordingly, the recipients were divided into four groups: CS 8 h, UW 8 h, CS 16 h and UW 16 h groups. The bile discharge, liver enzymes (ALT and AST), ET-1, IL-1 and TNF-alpha levels in serum at 1, 3, and 6 h after reperfusion were measured. Histopathological examination was carried out by HE staining, and the 7-day survival rate was observed. Results As compared with a homogenous appearance in the NHBD livers perfused with CS solution, the NHBD livers perfused with UW solution showed a heterogenous appearance, with more or less mottling. In the CS 8 h group there were more bile discharge at 1 h, lower ALT and AST levels at 1, 3 and 6 h, and lower serum ET-1, IL-1 and TNF-alpha ? levels at 1 and 3 h after reperfusion than the UW 8 h group (P<0.05). Histopathological examination showed a well-preserved hepatic architecture, milder portal congestion and inflammatory infiltration in CS 8 h group as compared with UW 8 h group. The 7-day survival rate in CS 8 h and UW 8 h groups was 58.3% and 25.0% respectively (P<0.05). There were no significant differences in bile discharge, ALT, AST, ET-1, IL-1 and TNF-alpha ? levels at different time points between CS 16 h and UW 16 h groups (P>0.05), and histopathological examination showed no much difference in the tissue damage between two groups; all recipients died within 3 days (P>0.05). Conclusion CS was more effective than UW in preserving NHBD livers after 8 h of cold storage, which may be due to the better perfusion of CS and inhibition of Kupffer cells, but it had better preserve NHBD livers with CS no more than 16 h.     

Celsior液与UW液对无心跳大鼠供肝保存效果的比较

Objective To compare the effects of Celsior (CS) solution vs University of Wisconsin (UW) solution on preservation of non-heart- beating donor (NHBD) livers in rats. Methods The healthy male SD rats were taken as donors and recipients. The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 10 min to induce ischemia as a NHBD model. After flushed with the cold CS or UW solution, NHBD livers were stored at 4℃ for 8 h or 16 h. Then they were transplanted using the two-cuff technique with arterial reconstruction. Accordingly, the recipients were divided into four groups: CS 8 h, UW 8 h, CS 16 h and UW 16 h groups. The bile discharge, liver enzymes (ALT and AST), ET-1, IL-1 and TNF-alpha levels in serum at 1, 3, and 6 h after reperfusion were measured. Histopathological examination was carried out by HE staining, and the 7-day survival rate was observed. Results As compared with a homogenous appearance in the NHBD livers perfused with CS solution, the NHBD livers perfused with UW solution showed a heterogenous appearance, with more or less mottling. In the CS 8 h group there were more bile discharge at 1 h, lower ALT and AST levels at 1, 3 and 6 h, and lower serum ET-1, IL-1 and TNF-alpha ? levels at 1 and 3 h after reperfusion than the UW 8 h group (P<0.05). Histopathological examination showed a well-preserved hepatic architecture, milder portal congestion and inflammatory infiltration in CS 8 h group as compared with UW 8 h group. The 7-day survival rate in CS 8 h and UW 8 h groups was 58.3% and 25.0% respectively (P<0.05). There were no significant differences in bile discharge, ALT, AST, ET-1, IL-1 and TNF-alpha ? levels at different time points between CS 16 h and UW 16 h groups (P>0.05), and histopathological examination showed no much difference in the tissue damage between two groups; all recipients died within 3 days (P>0.05). Conclusion CS was more effective than UW in preserving NHBD livers after 8 h of cold storage, which may be due to the better perfusion of CS and inhibition of Kupffer cells, but it had better preserve NHBD livers with CS no more than 16 h.     

Celsior液与UW液对无心跳大鼠供肝保存效果的比较

Objective To compare the effects of Celsior (CS) solution vs University of Wisconsin (UW) solution on preservation of non-heart- beating donor (NHBD) livers in rats. Methods The healthy male SD rats were taken as donors and recipients. The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 10 min to induce ischemia as a NHBD model. After flushed with the cold CS or UW solution, NHBD livers were stored at 4℃ for 8 h or 16 h. Then they were transplanted using the two-cuff technique with arterial reconstruction. Accordingly, the recipients were divided into four groups: CS 8 h, UW 8 h, CS 16 h and UW 16 h groups. The bile discharge, liver enzymes (ALT and AST), ET-1, IL-1 and TNF-alpha levels in serum at 1, 3, and 6 h after reperfusion were measured. Histopathological examination was carried out by HE staining, and the 7-day survival rate was observed. Results As compared with a homogenous appearance in the NHBD livers perfused with CS solution, the NHBD livers perfused with UW solution showed a heterogenous appearance, with more or less mottling. In the CS 8 h group there were more bile discharge at 1 h, lower ALT and AST levels at 1, 3 and 6 h, and lower serum ET-1, IL-1 and TNF-alpha ? levels at 1 and 3 h after reperfusion than the UW 8 h group (P<0.05). Histopathological examination showed a well-preserved hepatic architecture, milder portal congestion and inflammatory infiltration in CS 8 h group as compared with UW 8 h group. The 7-day survival rate in CS 8 h and UW 8 h groups was 58.3% and 25.0% respectively (P<0.05). There were no significant differences in bile discharge, ALT, AST, ET-1, IL-1 and TNF-alpha ? levels at different time points between CS 16 h and UW 16 h groups (P>0.05), and histopathological examination showed no much difference in the tissue damage between two groups; all recipients died within 3 days (P>0.05). Conclusion CS was more effective than UW in preserving NHBD livers after 8 h of cold storage, which may be due to the better perfusion of CS and inhibition of Kupffer cells, but it had better preserve NHBD livers with CS no more than 16 h.     

Improved machine perfusion preservation of the non-heart-beating donor rat liver using Polysol: a new machine perfusion preservation solution.

Waiting lists for transplantation have stimulated interest in the use of non-heart-beating donor (NHBD) organs. Recent studies on organ preservation have shown advantages of machine perfusion (MP) over cold storage (CS). To supply the liver with specific nutrients during MP, the preservation solution Polysol was developed. The aim of our study was to compare CS in University of Wisconsin solution (UW) with MP using UW-gluconate (UW-G) or Polysol in an NHBD model. After 30 minutes of warm ischemia, livers were harvested from rats for preservation by either CS, MP-UW-G, or MP-Polysol. After 24 hours of preservation, livers were reperfused with Krebs-Henseleit buffer (KHB). Perfusate samples were analyzed for liver damage and function. Biopsies were examined by hematoxylin and eosin staining and transmission electron microscopy. Liver damage was highest after CS compared with the MP groups. MP using Polysol compared with UW-G resulted in less aspartate aminotransferase (AST) and alanine aminotransferase (ALT) release. Perfusate flow, bile production, and ammonia clearance were highest after MP-Polysol compared with CS and MP-UW-G. Tissue edema was least after MP-Polysol compared with CS and MP-UW-G. In conclusion, preservation of the NHBD rat liver by hypothermic MP is superior to CS. Furthermore, MP using Polysol results in better-quality liver preservation compared with using UW-G.     

SX-1液对肝脏保存效果的实验研究

目的 应用改良后的Kamada二袖套法大鼠原位肝移植模型,检验SX-1液、HC-A液和UW液对肝脏保存的效果.方法 在无菌条件下配制肝脏保存液.建立大鼠原位肝移植模型.使用SX-1液、UW液和HC-A液保存大鼠肝脏2、8、24 h后行大鼠原位肝移植,于移植后6 h比较各项肝脏功能.结果 对于ALT、AST,SX-1液组(2、8、24 h)与UW液组同步升高,分析无统计学意义(P＞0.05),与HC-A液组相比,差异有统计学意义(P＜0.05).对于LDH,SX-1液组(2 h、8 h、24 h)与HC-A液组同步升高,差异无统计学意义(P＞0.05),与UW液组相比,差异有统计学意义(P＜0.05).对于分泌胆汁的肝脏个数,各组别与分泌胆汁的肝脏个数无差别(P＞0.05).本组内各时间点分泌胆汁个数有差别(P＜0.05).随肝脏保存时间增长,分泌胆汁的肝脏个数减少.结论 经大鼠原位肝移植模型证实,SX-1液在肝脏酶学方面与UW液作用相当,超过HC-A液水平.肝移植后6 h肝脏分泌胆汁的个数方面,SX-1液与HC-A液、UW液间无明显差别.     

自制HYD液对大鼠肝脏低温保存后生化功能影响的研究

目的 研制自制ＨＹＤ液对大鼠肝脏低温保存后生化功能的影响。方法 采用大鼠肝脏非循环离体灌注模型，比较ＨＹＤ液，ＵＷ液和乳酸林格液（ＬＲ液）对大鼠肝脏６，１２，２４，３０，３６ｈ保存后生化功能的影响。结果 保存１２ｈ的肝脏，其各项生化功能ＨＹＤ组明显优于ＬＲ组。三磷酸腺苷，磷酸腺苷含量及Ａｔｋｉｎｓｏｎ能荷（ＡＥＣ），ＨＹＤ组略高于ＵＷ组，且保存３６ｈ差异有显著性。     

Advantages of Celsior solution in graft preservation from non-heart-beating donors in a canine liver transplantation model.

BACKGROUND: The optimal method for preserving livers from non-heart-beating donors (NHBD) is still unknown. We compared the Celsior solution, a new extracellular-type, low-potassium, low-viscosity preservation solution, with the University of Wisconsin (UW) solution in a canine orthotopic liver transplantation from NHBD. MATERIALS AND METHODS: Fourteen adult mongrel dogs, weighing 9 to 17 kg, were divided into two groups: the Celsior or the UW group (n = 7 each). The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 20 min to induce warm ischemia as a NHBD model. The liver was flushed with the respective cold preservation solution and then stored at 4 degrees C for 4 h. The grafts were transplanted using the piggy-back technique under portal decompression by leaving the native right lobe as a temporary shunt. RESULTS: The duration of liver flushing out (min) was shorter (P < 0.05), and the serum glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, lactate dehydrogenase, lactate, and alpha-glutathione S-transferase concentrations 2 and 6 h after reperfusion of the graft (RPF) were lower (P < 0.05) in the Celsior group than in the UW group. Hepatic tissue blood flow (HTBF) did not deteriorate as much (P < 0.05) in the Celsior group. The serum endothelin-1 level was lower (P < 0.05) in the Celsior group 2 h after RPF. Histopathology of liver specimens revealed portal congestion and hepatocyte necrosis with neutrophil infiltration in the UW group, while these findings were mild in the Celsior group. CONCLUSIONS: The Celsior solution improves vascular endothelial injury in livers from NHBDs and may have advantages in graft flush and preservation of grafts from NHBDs.     

A preservation solution with polyethylene glycol and calcium: a possible multiorgan liquid

The addition of polyethylene glycol (PEG) to hepatocyte storage medium is known to decrease lipid peroxidation and swelling and to protect the cell cytoskeleton from cold. We therefore decided to investigate the effect of substituting PEG for hydroxyethyl starch (HES) in an extracellular-like UW solution, with and without Ca++, on rat liver preservation. Isolated perfused rat livers were used to assess graft injury after 24h of cold storage. Four groups of preserved livers ( n=6 for each group) were compared to controls (non preserved livers, n=11). For this purpose, Belzer solution (K+-UW, group 1) was stepwise modified. Group 2 (Na+-UW) was treated with the same liquid, however with inverted concentrations of Na+ and K+. Group 3 was preserved in the first experimental solution (EPS-1) with Ca++ (0.5mM) added to the Na+-UW solution. In the EPS-2 (group 4), PEG-35 (0.03mM) was substituted for HES. The last group, EPS-3 (group 5) was treated with the same compounds as EPS-2, but without Ca++. After 24h of cold storage and 120min normothermic reperfusion, there was no statistical difference in transaminases (ALT and AST) release between the control and the Na+-UW groups. Furthermore, rat livers preserved in Na+-UW solution released less ( P<0.05) ALT and AST and excreted more ( P<0.05) indocyanine green (ICG) than livers preserved in K+-UW solution. The addition of 0.5mM Ca++ to Na+-UW solution (EPS-1) dramatically increased ( P<0.05) parenchymal (ALT, AST) and non parenchymal (creatine kinase-BB) cellular injury. The substitution of PEG (0.03mM) for HES (EPS-2) reduced ( P<0.05) membrane injuries due to Ca++ while bile flow was statistically increased ( P<0.05). Finally, the omission of Ca++ from EPS-2, that is EPS-3, has no statistically significant effect on the studied parameters. PEG effectively protected the rat liver grafts from the onset of hypothermic ischemia-reperfusion and Ca++ damages and thus may be a valuable additive to preservation solutions.     

自制CMU-1液保存大鼠肝脏的实验研究

目的　探讨CMU 1液保存大鼠肝脏的效果。方法　根据灌注液和保存液的种类将Wistar大鼠分为两组 :UW组和CMU 1组 ,每组分 6h、12h、2 4h 3个保存时限 ,每亚组 6只大鼠。采用离体循环灌注模型 ,研究CMU 1保存液对保存肝脏能量代谢、生化功能、胆汁分泌及形态学方面的影响。结果　随着保存时间延长 ,肝组织TAN含量及AEC逐渐降低 ,CMU 1组较UW组下降略缓慢 ,保存 2 4h后高于UW组 (P <0 0 5 )。再灌注 12 0min后CMU 1组的肝脏分泌胆汁量较UW组多 (P <0 0 5 )。相同时限相比 ,灌出液中ALT、LDH值两组之间无显著差异 (P >0 0 5 )。肝脏组织学变化两组间无明显差异。保存 6h后 ,保存液pH值无明显变化 ;保存 12h后pH值下降 ,两组无明显差异 ;保存2 4h后 ,UW组pH值下降较CMU 1组明显。结论　CMU 1保存液保存大鼠肝脏效果与UW液相似 ,在改善保存肝脏能量代谢、预防细胞内酸中毒、胆汁分泌方面略优于UW液。     

Machine perfusion preservation of the non-heart-beating donor rat livers using polysol, a new preservation solution

AIMS: The increasing shortage of donor organs has led to a focus on extended criteria donors, including the non-heart-beating donor (NHBD). An optimal preservation method is required to facilitate successful transplantation of these ischemically damaged organs. The recent literature has shown clear advantages of hypothermic machine perfusion (MP) over cold storage (CS). For MP, modified University of Wisconsin perfusion solution (UW-G) is often used, which, however, is known to cause microcirculatory obstruction, is difficult to obtain, and is expensive. Therefore, Polysol was developed as a MP preservation solution that contains specific nutrients for the liver, such as amino acids, energy substrates, and vitamins. The aim of this study was to compare Polysol with UW-G in a NHBD rat liver model. METHODS: After 24 hours hypothermic MP of NHBD rat livers using UW-G or Polysol, liver damage and function parameters were assessed during 60 minutes of reperfusion with Krebs-Henseleit buffer. Control livers were reperfused after 24 hours CS in UW. RESULTS: Liver enzyme release was significantly higher among the CS-UW group compared to MP using UW-G or Polysol. Flow during reperfusion was significantly higher when using Polysol compared to UW-G. Bile production and ammonia clearance were highest when using Polysol compared to UW-G. There was less cellular edema after preservation with Polysol compared to UW-G. CONCLUSIONS: MP of NHBD rat livers for 24 hours using UW-G or Polysol resulted in less hepatocellular damage than CS in UW. MP of NHBD livers for 24 hours using Polysol is superior to MP using UW-G.     

Extended preservation of non-heart-beating donor livers with normothermic machine perfusion

BACKGROUND: Non-heart-beating donor (NHBD) livers represent an important organ pool, but are seldom utilized clinically and require rapid retrieval and implantation. Experimental work with oxygenated perfusion during preservation has shown promising results by recovering function in these livers. This study compared sanguinous perfusion with cold storage for extended preservation of the NHBD liver in a porcine model. METHODS: Porcine livers were subjected to 60 min of in vivo total warm ischaemia before flushing, after which they were preserved by one of two methods: group 1 (n = 4), University of Wisconsin (UW) solution by standard cold storage for 24 h; group 2 (n = 4), oxygenated autologous blood perfusion on an extracorporeal circuit for 24 h. All livers were subsequently tested on the circuit during a 24-h reperfusion phase. RESULTS: Livers in group 1 showed no evidence of viability during the reperfusion phase with no bile production or glucose utilization; they also displayed massive necrosis. Livers in group 2 demonstrated recovery of function by synthetic function, substrate utilization and perfusion haemodynamics; these livers displayed less cellular injury by hepatocellular enzymes. All differences in parameters between the two groups were statistically significant (P < 0.05). These findings were supported by histological examination. CONCLUSION: Warm ischaemia for 1 h and simple cold storage (UW solution) for 24 h renders the liver non-viable. Oxygenated, sanguinous perfusion as a method of preservation recovers liver function to a viable level after 24 h of preservation.