Hepatocyte transplantation through the hepatic vein: a new route of cell transplantation to the liver

The efficiency of hepatocyte transplantation into the liver varies with the method of administration. This study investigated whether retrograde infusion via the hepatic vein provides a sufficient number of donor cells for the liver. Donor hepatocytes were isolated from dipeptidyl peptidase IV (DPPIV(+)) rats and transplanted into DPPIV(-) rat livers either by antegrade portal vein infusion or retrograde hepatic vein infusion. Hepatocyte engraftment ratios and localization were evaluated by histological DPPIV enzymatic staining at 1 week and 8 weeks after the transplantation. No significant differences in engraftment efficiency were observed at either 1 week or 8 weeks after transplantation by either route. However, the localization of the transplanted hepatocytes differed with the administration route. Portal vein infusion resulted in predominantly periportal engraftment, whereas hepatic vein infusion led to pericentral zone engraftment. Immunohistochemical analysis showed that the transplanted hepatocytes engrafted in the pericentral zone after retrograde infusion displayed intense CYP2E1 staining similar to the surrounding native hepatocytes. CYP2E1 staining was further enhanced by administration of isosafrole, an inducing agent for various cytochrome P450 enzymes, including CYP2E1. This study demonstrates a novel approach of transplanting hepatocytes into the liver through retrograde hepatic vein infusion as the means to target cell implantation to the pericentral zone.     

Hepatocellular transplantation for experimental ischemic acute liver failure in dogs

Recovery from acute liver failure is possible if metabolic support can be provided during the period of exogenous liver regeneration. The ability of transplanted dispersed autologous hepatocytes to alter the course of experimental ischemic acute liver failure in dogs was tested. Liver failure was induced by occlusion of blood flow in the proximal portal vein and hepatic artery(s) 48 hr after creation of a side to side portacaval shunt and immediately after a left lateral hepatic lobectomy. Dogs in Group 1 had ischemic injury with no treatment. Dogs in Group II received intrasplenic autotransplants of hepatocytes (26 ± 4x × 10⁸ intact cells) after the ischemic period. Cells for transplantation were prepared from the excised lobe during the period of liver ischemia. Dogs in Group III received intrasplenic transplants of autologous hepatocytes (26 = 3 × 10⁸ intact cells) after liver ischemia and after ligation of the main splenic artery. Serum bilirubin, serum glutamic oxalocetic transaminase, lactate dehydrogenase, and alkaline phosphatase were measured before and serially after ischemia, and showed that the degree of liver injury in all three groups was similar, although survival in Group III was better. Only 20% of nontransplanted animals (Group I) survived 10 days. Liver histology in animals that died showed hemorrhagic necrosis situation around the terminal hepatic central veins. Transplantation did not improve survival in dogs with arterialized spleens and histological examination of dogs that died showed pulmonary infarcts and additional liver injury from embolization of hepatocytes. In contrast, 70% of the animals undergoing splenic artery ligation before intrasplenic transplantation of hepatocytes were alive at 10 days. Ligation of the splenic artery reduced the tendency for hepatocytes to escape into the splenic vein and the spleen remained viable due to collateral circulation. On histological examination, hepatocytes were readily identified in the splenic parenchyma at 24 hr. 2 and 4 weeks after transplantation. In conclusion, intrasplenic hepatocytes provide sufficient metabolic support for dogs to recover from otherwise lethal ischemically induced, acute liver failure.     

Enhanced proliferation of hepatic progenitor cells in rats after portal branch occlusion.

It is known that hepatic progenitor cells increase in number after liver injury caused by carcinogens, but this injury cannot be reproduced in humans. In order to create a practical source of hepatic progenitor cells, changes in the number of liver epithelial cells (LECs), a type of hepatic progenitor cell, were examined following partial interruption of the portal flow. Efficiency in this isolation procedure was investigated, and isolated LECs were transplanted into livers to demonstrate their differentiation into hepatocytes in vivo.A volume of 70% of Sprague-Dawley rat's livers was exposed to portal vein ligation. LECs, identified as alpha-fetoprotein (AFP)-positive and albumin-negative cells, were counted and LECs isolated from the portal vein ligated-lobe were characterized by immunostaining and Western blotting. Isolated cells were subjected to a 1-week-culture, and the number of colonies formed on dishes was counted. The cells were then transplanted to the livers of genetic analbuminemic rats and identified by immunohistochemistry. The number of LECs in the portal ligated-lobes on day 7 was 14.7 +/- 6.5 cells/1,000 hepatocytes: 18 times higher than numbers in a normal liver. A significant increase was noted from day 3 until day 28. Isolated LECs were AFP-positive, albumin-negative, and cytokeratin-19-positive cells. The number of colonies on the 7th day following portal vein ligation was 42 times higher than in a normal liver. After transplantation of the LECs to the analbuminemic rat, a cluster of albumin-producing cells was present until day 56, suggesting that they differentiate into hepatocytes. We conclude that after portal vein occlusion, the liver can be a good source of hepatic progenitor cell. These results open up the possibility of cellular transplantation for liver functional support in clinical settings.     

硫代乙酰胺诱导的门静脉高压大鼠肝脏超微结构观察

目的 该研究以硫代乙酰胺复制门脉高压大鼠模型,对成模后的大鼠肝脏超微结构进行观察,探讨门脉高压大鼠肝脏超微结构的变化.方法 50只大鼠分为造模组(40只)和正常对照组(10只).造模组采取前5周使用0.03%的硫代乙酰胺,后5周使用0.04%硫代乙酰胺作为其饮用水,共诱导10周成模.对成模大鼠测定肠系膜上静脉压力,取硬化后肝脏标本,对收集后标本行HE染色,同时行电子显微镜观察肝脏超微结构变化.结果 模型组内皮细胞窗孔数量明显少于对照组,同时,门静脉压力明显高于对照组.结论 肝窦毛细血管化在硫代乙酰胺诱导的肝硬化门脉高压发生中起重要作用,肝窦内皮细胞窗孔减少并减小,内皮细胞下基底膜形成使肝细胞与外界物质交换减少,导致细胞缺氧、代谢产物聚集,肝细胞坏死,最终导致门脉高压的发生.     

Hyperbaric oxygenation after portal vein emobilization for regeneration of the predicted remnant liver

BACKGROUND: Liver failure often develops after extensive liver resection. Preoperative portal vein embolization to induce compensatory hypertrophy in the predicted remnant liver decreases clinical complications after hepatectomy. The aim of this study was to examine whether hyperbaric oxygenation (HBO) after portal vein embolization increases compensatory hypertrophy of the predicted liver remnant. We performed portal vein ligation and HBO in rats to investigate whether HBO after portal vein embolization increases compensatory hypertrophy of the predicted remnant liver. METHODS: Rats were divided into four groups that underwent (1) laparotomy only (control group); (2) right portal vein ligation (RPL group); (3) RPL followed by HBO at 2 atm (HBO-2 atm group; 1 h/day, 5 days/week for 2 weeks); or (4) RPL followed by HBO at 3 atm (HBO-3 atm group). Laparotomy was repeated after 2 weeks in each group; serum levels of albumin and hepatocyte growth factor (HGF) were measured, and the ratio of the weights of nonligated to ligated hepatic segments and the percentage of hepatocytes expressing proliferating cell nuclear antigen (PCNA) in ligated hepatic segments were determined. RESULTS: In rats that had received HBO after RPL, serum levels of HGF, weight ratios of nonligated to ligated hepatic segments, and the percentage of PCNA-positive hepatocytes in nonligated liver were significantly higher than those in the control group. Furthermore, rats that had undergone 3-atm HBO after RPL had significantly higher serum levels of HGF and percentages of PCNA-positive hepatocytes in nonligated hepatic segments. CONCLUSIONS: Preoperative HBO after portal vein embolization may be useful for inducing compensatory hypertrophy of the predicted remnant liver.     

Efficient hepatocyte engraftment in a nonhuman primate model after partial portal vein embolization

BACKGROUND: Hepatocyte transplantation could be an alternative to whole liver transplantation for the treatment of metabolic liver diseases. However, the results of clinical investigations suggest that the number of engrafted hepatocytes was insufficient to correct metabolic disorders. This may partly result from a lack of proliferation of transplanted hepatocytes. In rodents, portal ligation enhances hepatocyte engraftment after transplantation. We investigated the effects of partial portal ligation and embolization on engraftment and proliferation of transplanted hepatocytes in primates. METHODS: Hepatocyte autotransplantation was performed in Macaca monkeys. The left lateral lobe was resected for hepatocyte isolation. The first group of monkeys underwent surgical ligation of the left and right anterior portal branches; in the second group, the same portal territories were obstructed by embolization with biological glue. To evaluate the proportion of cell engraftment hepatocytes were Hoechst-labeled and transplanted via the portal vein. Cell proliferation was measured by BrdU incorporation. RESULTS: Hepatocyte proliferation was induced by both procedures but it was significantly higher after partial portal embolization (23.5% and 11.2% of dividing hepatocytes on days 3 and 7) than after ligation (3% and 0.8%). Hepatocytes engrafted more efficiently after embolization than after ligation. They proliferated and participated to liver regeneration representing 10% of the liver mass on day seven and their number remained constant on day 15. CONCLUSIONS: These data suggest that partial portal embolization of the recipient liver improves engraftment of transplanted hepatocytes in a primate preclinical model providing a new strategy for hepatocyte transplantation.     

Metabolic studies following intrahepatic autotransplantation of pancreatic islet grafts

A patient with chronic pancreatitis underwent 95% pancreatectomy and islet autotransplantation. The pancreatic tissue was prepared by mincing and collagenase digestion, and then embolized into the liver via the portal vein. The patient has been followed with metabolic studies for 1 year. Fasting normoglycemia returned 3 weeks following operation. Intravenous glucose tolerance tests revealed K values that were similar before operation (1.06), and 3 weeks (0.96) and 4 months (1.09) after operation. Four months after islet transplantation, peripheral, portal, and hepatic vein insulin levels were determined simultaneously following an infusion of glucose (0.25 gm/kg) into the portal vein. During a 10-minute interval, right hepatic vein insulin increased fourfold, left hepatic vein insulin increased twofold, and peripheral vein insulin doubled. During this time portal vein insulin remained constant. In addition, significant levels of pancreatic glucagon were present in both hepatic veins, but were undetectable in the portal vein. At 6 months the patient became hyperglycemic and subsequently has required insulin therapy. Restudy at 10 months following islet transplantation revealed a marked drop in K value (0.34), and no evidence of graft function in the liver. These studies represent convincing evidence of transplanted intrahepatic islet cell function for a 6-month period following operation. The grafts for unknown reasons ceased to function at 6 months.     

Transplantation of Embryonic Small Hepatocytes Induces Regeneration of Injured Liver in Adult Rat

Small hepatocytes as hepatic stem cells or progenitors may be transplanted to treat several end-stage liver diseases. To identify the characteristics of epithelial cells enriched from fetal liver, we used immunocytochemistry and electron micrography. All cells in the colonies were immunocytochemically positive for alpha fetoprotein and cytokeratins (CK) 7, CK8, and CK18, which are markers of hepatic progenitor. Under transmission electron microscopy, we observed the cultured cells to show naive characteristics of stem cells and to be significantly distinct from mature hepatocytes. To identity whether these small hepatocytes were able to proliferate and differentiate into mature hepatocytes, we cultured them in vitro, and, through the portal vein, and transplanted elements whose membrane were stained with red fluorescence using PKH26 linker dye, into the livers of CCl₄-treated rats that had been subjected to two-thirds partial hepatectomy. Significant liver regeneration was observed 30 days later in rats that did or did not receive the cells. The livers of hepatocytes recipients showed sharper edges and smoother surfaces than the control group. Diffused cells labeled with red fluorescence were observed in the portal area, with branch-like red fluorescence in regions near portal areas of some lobules, suggesting that these elements were involved in the repair of liver lobules and differentiation into mature hepatocytes. Our results revealed that small hepatocytes not only have characteristics of hepatic stem cells, but also may be a source of cellular transplantation to treat liver diseases.     

Auxiliary heterotopic partial liver transplantation in pigs with acute liver failure

Fulminant hepatic failure is usually fatal without liver transplantation; however, orthotopic liver transplantation is often difficult to perform due to the high risk of coagulopathy and the development of multiple organ failure. Auxiliary heterotopic partial liver transplantation (APLT), however, has the potential to provide an effective hepatic support system considering that the host liver is left in situ and the surgical procedure is less invasive. In this report, we describe the beneficial effects of performing 60% APLT on the hepatic function and survival of pigs with acute hepatic failure induced by hepatic artery ligation. The pigs were divided into a control group of nine animals (group 1) that had portal vein and hepatic artery ligation with a side-to-side portacaval shunt, and an APLT group of seven animals (group 2) that had portal vein and hepatic artery ligation with APLT. The two left lateral lobes of the donor liver were resected, reducing the liver weight to about 60%, and the graft was placed in the right subhepatic space. No deaths occurred intraoperatively. In group 1, eight pigs died of massive liver necrosis within 48 h and one died between 48 and 72 h (median surivival 23 h). In group 2, two pigs died within 72 h due to preservation or anesthetic problems, but five survived for more than 3 days (median survival 13.4 days), with a significant difference between the two groups (P<0.05). One animal was killed 30 days after APLT and excellent graft function was demonstrated by the synthesis of clotting factors, ammonia detoxification, and glucohomeostasis. Moreover, evidence of hepatic regeneration was found in the transplanted livers. These results indicate that APLT provides metabolic support and improves survival in animals with induced acute liver failure.     

麦角新碱预处理对减轻大鼠移植肝缺血再灌注损伤的作用

目的 探讨使用外源性药物麦角新碱预处理对减轻大鼠移植肝缺血再灌注损伤的作用.方法 在大鼠的门静脉-左肾静脉搭桥、肝后下腔静脉内置管分流法自体原位肝移植模型中,于肝门阻断前10 min经大鼠尾静脉注射麦角新碱;观察移植肝缺血前和再灌注后5 min、30 min、2 h时血清一氧化氮(NO)和血浆内皮素1(ET1)水平以及NO/ET1的比值变化;测定血清丙氨酸转氨酶(ALT)酶学差异和肝组织内三磷酸腺苷(ATP)和丙二醛(MDA)含量变化;再灌注2 h取肝组织检测肝细胞、肝小叶超微结构.结果 应用麦角新碱预处理的大鼠移植肝缺血前门脉血浆中ET1升高(P＜0.01),但再灌注后5 min、30 min时,血浆中ET1水平降低(P＜0.05);而缺血前NO/ET1比值降低(P＜0.01),再灌注后5 min时,NO/ET1比值升高(P＜0.01);再灌注后ALT的升高有逐渐降低趋势;再灌注后2 h肝细胞内超微结构的损害程度减轻.结论 使用麦角新碱预处理能减轻大鼠移植肝缺血再灌注损伤.移植肝缺血再灌注损伤的靶细胞是肝血窦内皮细胞,NO/ET1比值平衡可能是影响移植肝微循环血流量变化的调节因素.     

Comparative study of two models for experimental cirrhosis in the dog

Two canine models of cirrhosis induced by administration of dimethyl-nitrosamine (DMNA) alone or in combination with hepatic vein ligation, were studied. The criteria used for development of cirrhosis were not only the appearance of portal hypertension, ascites, biochemical disorders or retention of BSP but also the formation of histological lesions such as sclerotic transformation of the liver lobules together with nodular regeneration. According to these criteria, the experimental protocol consisting in hepatic vein ligation and prolonged administration of DMNA was successful in inducing in animals liver cirrhosis similar to human conditions.     

肝动脉和门静脉高位分支双重处理对犬肝脏结构和功能的影响

目的 观察肝动脉和门静脉分支双重处理后犬肝脏结构和功能的改变。方法 杂种犬25只。随机分成假手术对照组和实验组,实验组进行肝动脉左支结扎和门静脉左支缩窄70％。分别于术后1,2,3,7和14d处死动物,观察肝脏大体结构的变化,分别称取左右半肝脏的质量,检测肝脏功能。光学显微镜下观察肝脏显微结构改变,电镜下观察肝脏超微结构改变。结果 肝动脉左支结扎和门静脉左支缩窄70％后,左半肝脏呈进行性萎缩变小,右半肝脏则成比例地代偿性增生,在整个观察过程中,全肝的总质量维持恒定。肝脏功能在术后2-3d时有轻度异常,很快即恢复正常。光镜下左半肝脏在早期出现少量的点、片状坏死。坏死周围出现大量的肝细胞凋亡,右半肝脏的肝小叶结构保持完整,术后肝细胞的增殖明显,有较多的分裂相存在。电镜下左半肝脏可见到典型的肝细胞凋亡,晚期呈明显纤维化改变。结论 肝动脉左支结扎和门静脉左支缩窄70％后。左半肝脏由于肝细胞大量凋亡呈进行性萎缩变小,右半肝脏则成比例地代偿性增生肥大,全肝的总质量和肝脏功能保持正常。     

Reversal of experimental acute hepatic failure in the rat

A suitable animal model of acute fulminant hepatic failure was developed in the rat by the administration of the selective hepatotoxin, galactosamine hydrochloride. A dosage of 2.6 g/kg given intraperitoneally to Lewis rats resulted in reproducible, potentially reversible hepatic necrosis and high mortality. The ability of dispersed single-cell suspensions of syngeneic hepatocytes, allogeneic hepatocytes, and bone marrow to provide “support” so that liver recovery could occur was explored in this model. Syngeneic hepatocytes (4 × 10⁷ cells) given intraperitoneally 48 and 60 hr after toxin administration increased survival to 70 and 66%, respectively, compared to a control of 0%. Hepatocytes prepared from Fischer strain animals which differ from Lewis strain at minor histocompatibility loci similarly improved survival (62.5%). Syngeneic bone marrow cells (4 × 10⁷ cells) given intraperitoneally 24 hr after toxin administration improved survival to 62.5% compared to a control group of 0%. This effect is not mediated by a mature macrophage or lymphoid cell population, but is specific to the cells derived from bone marrow, since single-cell suspensions of splenocytes, thymocytes, and peritoneal exudate cells had no effect on increasing survival. The mechanism of action of hepatocytes and bone marrow cells and whether they act by the same or different mechanisms remain to be defined. Cellular transplantation may offer a new simple measure in the treatment of acute hepatic failure.     

门静脉转流下大鼠门静脉缺血对肝脏能量代谢的影响

目的探讨在门静脉转流下大鼠门静脉缺血后肝脏能量代谢变化。方法在门静脉转流下阻断门静脉不同时间后,观察大鼠存活率、肝细胞线粒体呼吸活性、肝组织ATP含量及动脉血酮比值。结果在门静脉缺血60、90及120min后7d大鼠存活率分别为100%、100%及40%;缺血后肝脏能量代谢功能明显受损,再灌注后24h,门静脉缺血60、90min两组大鼠肝脏能量代谢功能已有明显恢复,而门静脉缺血120min组仍维持在低水平。结论在门静脉转流下大鼠门静脉缺血90min以内肝脏能量代谢损害可逆,而门静脉缺血120min则不可逆。     

Long-term fate of heterotopic liver transplants in rats with portal vein inflow only

Previous studies have suggested that hepatic arterial flow in heterotopic partial liver transplants is necessary to ensure graft survival and regenerative capacity. This report presents findings in a syngeneic rat strain (Lewis) that partial liver transplants can be successfully heterotopically transplanted in the long term with the only inflow coming from the portal vein. When the host liver undergoes a nearly complete resection at 3-4 weeks, the transplanted liver regenerates to maintain the health of the host. Moderate to massive hepatocellular necrosis occurs in the first 3 months postoperatively, with recovery by 4-5 months. Liver transplants 8-10 months postoperatively appear architecturally normal. No host liver tissues were found to be regenerating after subtotal host liver resection. We conclude that portal vein reconstruction without hepatic arterial inflow can sustain a partial liver transplant in the long term, replacing the function of the host liver.     

门静脉分支结扎后门静脉压力变化与肝再生的关系

目的 探讨90%门静脉分支结扎后大鼠门静脉压力变化与肝再生的关系.方法 45只雄性SD大鼠行90%门静脉分支结扎术,其中5只进行假手术作为对照.观察不同时相点门静脉压力和非结扎侧肝脏质量变化,光学显微镜下观察非结扎侧肝细胞的形态学变化,免疫组织化学方法检测未结扎侧肝细胞的增殖细胞核抗原(PCNA),TUNEL法检测未结扎侧肝细胞的凋亡情况,并进行定最分析.采用Pearson相关分析和t检验分析数据.结果 95%(38/40)的大鼠存活.结扎侧肝叶进行性萎缩,非结扎侧肝叶占全肝质量的比例随时问推移而增加,12 h内增加较缓慢,仅为10.75%;而1～5 d则增加速度明显加快,达到27.57%;7～28 d达到平台期,缓慢增加到32.37%.术前门静脉压力为(9.1±1.8)cm H_2O(1 cm H_2O=0.098 kPa);结扎后立即升高,12 h达到高峰(15.8±2.7)cm H_2O,与术前比较差异有统计学意义(t=6.847,P<0.05);1～28 d由(13.6±2.3)cm H_2O逐渐下降为(9.3±2.0)cm H_2O.术前大鼠PCNA阳性细胞计数为7%±3%,术后12 h至3 d由14%±5%上升至21%±6%,第5天达到高峰为26%±7%,与术前比较差异有统计学意义(t=9.129,P<0.05),随后逐渐恢复正常.TUNEL法检测结果显示,术前大鼠肝脏和术后各时相点大鼠未结扎侧肝脏仅见极少量凋亡细胞.大鼠门静脉压力与非结扎侧肝叶肝细胞PCNA的表达在术后1、3、5 d呈正相关(r=0.913,0.896,0.908,P<0.05),在术后14 d时相点呈负相关(r=-0.926,P<0.05).结论 大鼠90%门静脉分支结扎术后,引起未结扎侧肝细胞的活跃再生,再生后的肝脏可恢复原来的质量;肝再生以肝细胞增殖加速为主,而非肝细胞凋亡减少;门静脉压力变化在肝再生过程中可能发挥重要作用.     

Induction of endogenous antitumor activities in the portal vein by using a subcutaneously-imbedded pediculated spleen in the rat; experimental study of adjuvant immunotherapy in liver metastases

For the purpose of inducing the antitumor effect in the portal vein, the intrasplenic serial biological response modifier (BRM) administration was performed by using our original subcutaneously-imbedded pediculated spleen in the rats. In a view point of endogenous tumor necrosis factor (TNF) production, lipopolysaccharide, BCG and OK-432 were chosen and injected into the spleen frequently. The study of the portal blood serum revealed that intrasplenic (i.s.) BRM administration group gained higher TNF and interferon (IFN) activity than control group. On the other hand, the study of the mononuclear cells in the portal vein and splenocyte after i.s. BRM administration showed higher cytotoxic activity against YAC-1 cells than control group significantly. Compared with intravenous and intraperitoneal administration groups, i.s. group showed more effective antitumor effects in the portal vein significantly. The experimental liver metastases by intraportal transplantation of AH60C could be cured with i.s. BRM administration, which could increase % survival significantly. According to the result of this study, it is prospective that i.s. serial BRM administration could be new process for suppression of transportal hepatic metastases.     

A single-center experience with retrograde reperfusion in liver transplantation

Poor graft function secondary to injury by ischemia and reperfusion remains a major problem with regard to morbidity and mortality in clinical liver transplantation (LTX). Up to one fifth of patients suffer from poor initial liver function due to severe damage to hepatocytes. This situation leads either to primary nonfunction described in approximately 6% of LTX or to slow recovery. We present a new method of reperfusion during LTX. From July 1998 to July 2002, 42 LTX in 39 recipients, (10 female, 52 years old (26–70) were performed. LTX was carried out in piggy-back technique. After completing the piggy-back anastomosis, the caval vein was declamped immediately, and retrograde low pressure reperfusion of the graft with low oxygenated venous blood was established. Portal anastomosis was performed using a running suture. In order to provide optimal retrograde liver perfusion, no clamping of the donor portal vein was done. After completing portal anastomosis, the recipient portal vein was declamped immediately. During arterial anastomosis, the transplanted liver was antegradely perfused via the portal vein. After completing hepatic artery anastomosis, declamping of the hepatic artery was done and arterial perfusion started. No backtable or in-situ-flushing except the described reperfusion technique was performed. Forty-two LTX in 39 recipients using piggy-back technique and retrograde reperfusion via the caval vein followed by antegrade reperfusion via the portal vein were performed; 38 out of 39 patients (97.44%) were alive and well at day 8 after LTX. One patient (2.56%) died of a pre-existing portal vein thrombosis on day 2 after LTX. Three patients had to undergo retransplantation for hepatic artery thrombosis (7.14%). Liver enzymes, bilirubine, prothrombine time and AT III on day 1, 3, 5 and 8 after LTX showed favourable values. Median aspartate aminotransferase (ASAT) was 219 U/l on day 1 after LTX. One-month survival rate was 95.23%, and 1-year survival rate 87.88%. Two patients died of liver-associated causes (5.12%). One patient died of a late hepatic artery thrombosis, and one more of rejection. No other severe case of rejection appeared. We can conclude that retrograde reperfusion might be highly sufficient method of removing perfusion fluid from the transplanted liver. Low pressure perfusion with low oxygenated blood might reduce the production of free oxygen radicals. Retrograde reperfusion via the caval vein and antegrade reperfusion via the portal vein seemed to lower postoperative liver enzyme values and to improve initial liver function after LTX.     

Hepatic Arterial Perfusion Is Essential for the Spontaneous Recovery From Focal Hepatic Venous Outflow Obstruction in Rats

We previously observed that focal hepatic venous outflow obstruction recovered spontaneously by the formation of sinusoidal canals in a rat model of portal hyperperfusion. We aimed to investigate whether the lack of hepatic arterial perfusion aggravates parenchymal damage, decelerates recovery and influences the formation of sinusoidal canals after focal hepatic venous outflow obstruction. Rats were subjected to arterialized versus nonarterialized syngeneic liver transplantation after ligating the right median hepatic vein in the donor. Hepatic damage, microcirculation, regeneration and vascular remodeling were evaluated. In arterialized‐recipients, confluent necrosis interspersed with viable periportal islands of hepatocytes, and vascularized sinusoidal canals with visible blood flow, surrounded by normal sinusoidal structure, were visible on postoperative day (POD) 2. Complete parenchymal recovery was consequently established by resorption of necrosis and hepatocyte proliferation, detected in viable portal islands and border zone. Lack of hepatic arterial perfusion caused complete necrosis in the obstruction zone without viable hepatocytes in the periportal area on POD2. Hepatocyte proliferation was only visible in the border zone. On POD28, perfused vascular structures, without neighboring normal sinusoidal structures, were observed in the scar‐like area. Hepatic arterial perfusion determined the extent of hepatic necrosis, the formation of vascularized sinusoidal canals and the parenchymal recovery, after focal hepatic venous outflow obstruction.     

梗阻性黄疸鼠肝脏血红素氧化酶-1及一氧化碳含量的研究

目的 探讨梗阻性黄疸时肝脏血红素氧化酶-1(HO-1)及血浆中一氧化碳(CO)含量的变化。方法 48只 Wistar 大鼠随机分为 4组:假手术对照组(CG)、梗阻性黄疸 7 d组(7 d)、梗阻性黄疸 14 d组(14 d)和梗阻性黄疸 21 d组(21 d),采用免疫组织化学法对肝细胞中 HO-1表达进行分析,应用双波长分光光度计法测定大鼠肝静脉、门静脉及下腔静脉血浆中CO含量。结果 梗阻性黄疸时 HO-1不仅在 Kupffer细胞中表达,而且在肝实质细胞中呈弥漫性表达上调,14 d组和 21 d组肝实质细胞中HO-1表达均较7 d组增加(P<0.01)。梗阻性黄疸各组肝静脉血浆中CO含量较CG组增高(P<0.05,P<0.01);14 d组门静脉血浆中CO含量升高明显,与CG组比较差异有显著性(P<0.05);梗阻性黄疸各组肝静脉血浆中 CO含量与同时间组门静脉血浆中 CO含量相比均显著升高(P<0.01)。梗阻性黄疸各时间组肝静脉CO含量的增高与肝实质细胞中HO-1表达的变化呈显著正相关(P<0.01)。结论 梗阻性黄疸时肝细胞HO-1表达上调致CO产生增多,从而有助于增加肝血流量并减少肝脏功能损害。