广泛肝切除联合肝固有动脉切除对正常大鼠及梗阻性黄疸大鼠的影响

目的 探索在正常及高胆红素血症情况下,大鼠70%肝切除联合肝固有动脉切除对肝功能、肝细胞能量代谢以及肝再生和细胞凋亡的影响.方法 雄性成年SD大鼠133只,将其中40只分为2组,每组20只,均行胆总管-十二指肠插管桥接,同时行70%肝切除或70%肝切除联合肝固有动脉切除.另87只行胆总管结扎制备梗阻性黄疸模型.5 d后手术分为70%联合肝切除胆肠再通内引流组,及70%肝切除联合肝固有动脉切除、胆肠再通内引流2组.动态观察术后24 h、72 h、7 d肝功能和肝细胞能量代谢、肝组织HGF和bcl-2 mRNA含量及其蛋白表达、肝细胞增殖指数和凋亡指数的变化,并统计各组死亡率.另取6只作为假手术组,测定术后0 h肝功能和肝细胞能量指标.结果 正常大鼠能够耐受70%肝切除联合肝动脉切除,术后肝细胞能量代谢和肝功能迅速恢复正常,肝再生良好.高胆红素血症时,大鼠术后肝再生受抑制,细胞凋亡增多.较之70%肝切除组,70%肝切除联合肝固有动脉切除组对肝细胞能量代谢的影响更为显著,术后肝功能恶化,肝组织HGF和Bcl-2 mRNA含量显著减少,肝再生明显受抑制,细胞凋亡增多,死亡率显著增高(P<0.05).结论 正常大鼠70%肝切除联合肝动脉切除术后肝再生不受影响,高胆红素血症时,70%肝切除联合肝动脉切除的大鼠死亡率高,因此术前引流减黄应是必要的措施.     

肝切除联合胆肠内引流对梗阻性黄疸大鼠的影响

目的探索在高胆红素血症下,大鼠胆肠内引流联合肝切除对肝功能、肝细胞能量代谢以及肝细胞再生和凋亡的影响。方法雄性成年SD大鼠120只,其中6只大鼠作为假手术(SO)组;20只行70%肝切除,同时行胆总管-十二指肠插管桥接(70%PH组);另94只行胆总管结扎(CBDL)制备梗阻性黄疸模型,建模成功后5d时随机取6只动物作为CBDL组,余下的动物5d后行二次手术,再分为胆肠内引流再通组(BDO-RBF组,20只)、42%PH+BDO-RBF组(20只)及70%PH+BDO-RBF组(25只)。检测CBDL组5d时及二次手术后各组术后24h、72h及7d时肝功能和肝细胞能量代谢、肝组织肝细胞生长因子(HGF)和bcl-2mRNA含量及其蛋白表达、肝细胞增殖指数和凋亡指数的变化。SO组只测定术后0h肝功能和肝细胞能量指标。结果正常大鼠能够耐受70%肝切除,术后肝细胞能量代谢和肝功能迅速恢复正常,肝再生良好、细胞凋亡无明显增多。较之SO组和70%PH组,CBDL导致肝细胞能量代谢恶化、肝功能不良、肝组织HGF和bcl-2mRNA含量减少,肝再生受抑制,细胞凋亡显著增多(P<0.05)。胆肠内引流再通后肝细胞能量代谢可迅速恢复,肝细胞凋亡减少,肝再生活跃。42%+BDO-RBF组及70%PH+BDO-RBF组肝细胞能量代谢同样能够较快恢复,肝再生良好。42%+BDO-RBF组与70%PH+BDO-RBF组间各指标差异均无统计学意义(P>0.05)。结论梗阻性黄疸时,高胆红素血症严重影响肝细胞能量代谢和肝功能,引起肝细胞凋亡增多,肝再生受抑制;及时有效的胆肠内引流能够迅速改善肝细胞能量代谢、肝功能及肝再生,联合肝切除时术前可不需常规胆道引流。     

肝动脉结扎对梗阻性黄疸大鼠残肝肝细胞凋亡及再生的影响

目的探讨正常肝脏状态下和梗阻性黄疸状态下,肝固有动脉结扎对大鼠肝细胞凋亡和肝脏再生的影响。方法设计动物实验。80只Wistar雄性大鼠分为4组,每组大鼠20只,即:建立梗阻性黄疸模型后3 d施行70%肝切除+肝固有动脉结扎+胆肠内引流组(A组);建立梗阻性黄疸模型后3 d施行70%肝切除+胆肠内引流(B组);假手术后3 d施行70%肝切除+肝固有动脉结扎(C组)和假手术后3 d施行70%肝切除(D组)。各组分别在肝大部切除模型建立后1、2、3和6 d共4个时间点观察动物生理状态并采集血液及残肝组织标本。记录各组术后大鼠的生理状态及死亡情况;检测血清肝功能指标:胆红素(TB)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)及白蛋白(ALB);采用TUNEL法检测残肝组织肝细胞的凋亡;采用Brdu检测残肝再生情况,并进行统计分析。结果 A、B 2组大鼠间及C、D 2组大鼠间术后生存率比较差异无统计学意义(均P0.05)。A组和B组大鼠的术后肝功能差异无统计学意义(P0.05),但术后早期的白蛋白合成A组要弱于B组(术后3 d,P0.05);C组和D组术后肝功能指标差异也无统计学意义(P0.05),但术后早期C组的白蛋白合成要弱于D组(术后1 d和3 d均P0.05)。A、B、C 3个组术后1 d残肝凋亡指数达到最大,而后逐渐下降;而D组残肝凋亡指数在4个时间点上均为最低。术后1 d,C、D 2组肝细胞增殖达到高峰,增殖指数大小为D组C组B组A组,各组之间差异均有统计学意义(均P0.05)。术后6 d,A、B组肝细胞增殖指数无明显增大,C、D组肝细胞增殖指数明显降低,4个组的肝细胞增殖指数均处于较低水平。结论结扎肝动脉将增加残肝肝细胞凋亡,减弱肝细胞再生。但结扎肝动脉并未增加梗阻性黄疸大鼠术后死亡风险。     

乙酰肝素酶siRNA对部分肝切除大鼠术后肝再生的抑制作用

目的探讨乙酰肝素酶(HPSE)siRNA对部分肝切除大鼠术后肝再生的抑制作用。方法设计合成HPSE siRNA,以vivo-jetPEI-Gal为载体转染70%肝切除大鼠,以逆转录-聚合酶链反应(RT-PCR)检测转染后HPSE mRNA表达,以Western blot检测AFP、ALB、CK-18和CK-19蛋白表达,以原位末端标记法(Tunel)检测肝细胞凋亡,以免疫组化检测肝组织增殖细胞核抗原(PCNA)表达,并测定肝再生率。结果与各对照组相比,siRNA干扰组HPSE mRNA表达明显降低,AFP、ALB、CK-18和CK-19蛋白表达明显降低,肝细胞凋亡指数明显增高,增殖指数明显降低,肝再生率明显降低,差别显著(P0.05)。结论 siRNA沉默HPSE表达后,部分肝切除大鼠术后肝再生受到明显抑制,提示HPSE对肝再生具有调控作用。     

阻塞性黄疸大鼠术前胆道内、外引流对肝细胞再生能力的影响

目的　探讨阻塞性黄疸大鼠肝叶切除术前胆道内、外引流对肝细胞再生能力的影响和机制。方法　将大鼠胆总管结扎 5d后 ,分别行胆道内、外引流 5d ,再行 70 %肝叶切除术。结果　胆道外引流组与内引流组和对照组大鼠相比 ,反映肝细胞再生能力的肝细胞核DNA含量、增殖细胞核抗原 (proliferatingcellnuclearantigen ,PCNA)指数、有丝核分裂指数 (mitoticindexMI)明显减低 (P <0 0 5 )。胆道外引流组肝细胞C met/HGF R基因表达也减低 (P <0 0 1)。结论　阻塞性黄疸大鼠肝叶切除术前胆道外引流对肝细胞再生能力有明显抑制作用 ;胆道外引流组肝细胞C met/HGF R基因表达减弱可能是该组肝细胞再生能力下降的重要因素。     

合并肝动脉切除重建的肝门部胆管癌根治术

目的观察肝动脉切除重建在肝门部胆管癌治疗中的价值。方法1998年1月至2005年12月计收治125例肝门部胆管癌，其中行肝动脉切除13例，对该资料进行分析。结果在行肝动脉切除13例中，同时合并门静脉切除重建3例，其中部分肝固有动脉＋右或左肝动脉切除联合左或右半肝及尾状叶切除10例，局部切除联合肝固有动脉切除1例，部分肝固有动脉＋右或左肝动脉切除联合扩大左或右半肝及尾状叶切除各1例，肝动脉切除后未重建2例。术后胆肠吻合口漏4例，围手术期肝功能衰竭死亡1例，其余12例病人术后随访4个月至6年，平均20个月，其中最长的1例已存活5年5个月。结论肝动脉切除重建可提高肝门部胆管癌的治愈切除率，改善术后病人预后；肝脏大部切除联合肝动脉切除在中、重度黄疸病人须重建动脉血供。     

肝门胆管癌伴重度梗阻性黄疸广泛肝切除安全性的回顾分析

目的探讨重度梗阻性黄疸患者广泛肝切除手术的安全性。方法回顾性分析2000年至今连续21例重度梗阻性黄疸患者术前检查结果、术后并发症及恢复情况。患者平均血清胆红素为341.05μmol/L(191.3~641.6μmol/L)。21例均为肝门胆管癌,1例合并肝内胆管结石。术前均不减黄。切肝方式包括左半肝切除14例,其中2例切除受累肝固有动脉未做动脉重建,1例切除部分门静脉并重建,左半肝加左尾状叶切除4例,右半肝切除1例,扩大右半肝切除1例,右三叶加全尾状叶并门静脉部分切除重建1例。结果手术治愈率95.2%,术后30 d内死亡率4.8%(1/21),术后并发症发生率90.5%(19/21),术后肝功衰发生率4.8%(1/21)但逆转,其余并发症有胆瘘(28.6%)、腹腔感染(19.0%)、腹水(38.1%)、切口并发症(9.5%)、心衰(9.5%)、胸腔积液(4.8%)等。28.6%(6/21)需带腹腔引流管出院。结论重度梗阻性黄疸患者如一般情况较好、无严重合并症,不作术前减黄仍能耐受较大量肝切除。术后死亡率在可接受范围,但并发症发生率高。     

肝叶切除联合胆肠吻合治疗肝内胆管结石

目的 探讨肝内胆管结石的手术治疗方法及其效果.方法 总结1997~2007年我院92例肝内胆管结石患者的外科手术治疗情况.其中A组50例行肝叶(或肝段)切除的各种术式,同时行胆管空肠吻合40例,T管和U形管引流分别为6例和4例;B组42例行非肝叶(或肝段)切除的各种术式.其中胆总管切开取石+T管引流28例,胆总管切开取石+U形管引流5例.胆总管切开取石+胆肠吻合9例.结果 A组无手术死亡;术后残留结石5例,残石率10%;症状复发2倒,复发率4%,无再手术者.B组死亡1例,死亡率2.4%;术后残留结石8例,残石率19.5%;症状复发5例,复发率11.9%,其中有3例再次行肝叶(或肝段)切除治愈.结论 肝叶切除联合胆肠吻合术是治疗肝内胆管结石较为理想而有效的手术方式.     

34例肝门部胆管癌的诊断和治疗

目的:探讨肝门部胆管癌合理选择治疗方式,方法:对34例患者按不同的治疗方式分为根治性切除, 姑息性切除,胆肠内引流,金属支架内引流和外引流5组,分析各种治疗方式与病变分型,生存时间和并发症的关系.结果:生存时间,根治性切除组3.3年,姑息性切除组平均20个月,胆肠内引流组平均13个月,金属支架内引流组平均12个月;外引流组6.3个月;总并发症23.5%,手术死亡率0.结论:肝门部胆管癌的治疗应首选根治性切除,选择性和肝段切除是更适合的,姑息性切除的疗效优于各种内外引流术,胆肠吻合内引流术和金属支架内引流疗效相似.     

大鼠肝、胰十二指肠联合切除残肝肝细胞生长因子表达的研究

目的 用实验动物模型同时切除肝脏、胰腺及十二指肠，以探讨肝细胞生长因子(HGF)对残肝再生的影响。方法 60只SD大鼠被随机分为4组：①68％肝切除组；②68％肝叶切除加50％胰腺切除组；③68％肝叶切除加十二指肠切除组；④68％肝叶切除、加50％胰腺切除加近端十二指肠切除组。每组中3只大鼠分别于术后12、24、48、72和168h处死并采集肝脏样本，计算肝再生率。切除的肝脏组织用RT-PCR技术检测HGF在术后不同时间的表达，并与免疫组化法检测的肝细胞增殖细胞核抗原(PCNA)进行对比研究。结果 68％肝切除组残肝HGF的表达逐渐增强，24h达到高峰，随后逐渐下降，168h时达最低水平。而肝、胰切除，肝十二指肠切除和肝、胰十二指肠切除可显著减少残肝细胞HGF的表达，并使HGF的表达延迟至术后48h。结论 HGF可促进肝脏切除术后残肝的再生反应，而起源于胰腺和十二指肠外分泌腺的其他因素亦对HGF的表达起显著的调节作用，从而影响肝细胞的增殖。     

Delayed liver regeneration with negative regulation of hepatocyte growth factor and positive regulation of transforming growth factor-beta1 mRNA after portal branch ligation in biliary obstructed rats

BACKGROUND: The influence of obstructive jaundice on liver regeneration is still controversial. The aim of this study was to investigate liver regeneration after portal branch ligation (PBL) in the jaundiced rat, focusing on hepatocyte growth factor (HGF) and transforming growth factor-beta1 (TGF-beta 1). METHODS: Male Wistar rats underwent PBL or a sham operation 7 days after a common bile duct ligation. Liver wet weight, proliferating cell nuclear antigen labeling, HGF and TGF-beta 1 mRNA expression, and immunohistochemical staining with alpha-smooth muscle actin antibody were studied. RESULTS: The rate of liver regeneration in jaundiced liver was decreased as compared to a non-jaundiced liver. DNA synthesis in the jaundiced non-ligated lobe was significantly lower than in the non-jaundiced liver as was the peak level of HGF mRNA expression after PBL. In contrast, the level of TGF-beta 1 mRNA expression was higher in the jaundiced liver, and alpha-smooth muscle actin staining showed that hepatic stellate cells were gradually activated into myofibroblast-like cells. CONCLUSIONS: Obstructive jaundice decreased the expression of HGF mRNA and increased the expression of TGF-beta 1 mRNA, resulting in delayed liver regeneration after PBL. We suggest that hepatic stellate cells activated in obstructive jaundice may affect the expression of these growth factors.     

Inhibition of hepatic DNA synthesis after partial hepatectomy in the rats with obstructive jaundice with special reference to the enhancement of hepatic protein synthesis

We studied the liver regeneration after partial (68%) hepatectomy in rats with obstructive jaundice followed by the relief of obstruction. Rats received bile duct ligation, then 5 or 14 days later choledocho-duodenostomy was performed. Partial hepatectomy was done at various intervals after the relief of obstruction. DNA synthesis of the regenerating liver, hepatic protein synthesis and mitochondrial swelling induced by exogenous phospholipase A2 (PLA2) were determined. Hepatic DNA synthesis was significantly inhibited in obstructive jaundiced rats compared to controls. While the inhibition disappeared 5 days after the relief of obstruction in 5-day-obstructed group, it was still detectable as late as 21 days after the drainage in 14-day-obstructed group. Hepatic protein synthesis was markedly increased by obstructive jaundice, and this increase continued until 10 days after drainage in 14-day-obstructed group. Partial hepatectomy also increased the hepatic protein synthesis significantly in normal rats, but failed to show any significant changes in obstructive jaundiced rats. Any difference could not be found in PLA2-induced hepatic mitochondrial swelling between obstructive jaundiced rats and normal rats. We concluded the preceding energy-requiring responses in obstructive jaundiced liver resulted in the reduction of hepatic DNA synthesis and in the lack of additional increase of hepatic protein synthesis as the responses to a further insult of partial hepatectomy.     

Impact of hepatic vein deprivation on liver regeneration and function after major hepatectomy

Background and aims In extended liver resections, the preservation of vascular and biliary structures of the entire remnant liver is of paramount importance. The impact of venous outflow impairment and its consequences for liver regeneration and function are still a matter of debate. Materials and methods Rats (n = 75) were subjected to a 90% partial hepatectomy (PH), to a 70% liver resection with narrowing of the hepatic outflow of an additional 20% parenchyma (70%+ PH) or to an anatomic 70% PH. Postoperatively hepatocyte proliferation (Ki-67), liver function and survival were assessed. Gene expression analysis for markers of regeneration was determined by in-house complementary (DNA) arrays and quantitative real-time polymerase chain reaction (RT-PCR). Results Ninety percent PH led to a greater regenerative response as shown Ki-67 compared to animals with a 70%+PH (p < 0.05). However, liver function was equally impaired in both groups. Rats with 70% PH showed a greater proliferation index with less hepatic injury and better liver function. While mortality was 0% in the group of 70% PH, rats with 90% PH and 70+PH had a reduced survival of 75% (p < 0.05) Conclusion Venous outflow obstruction leads to an impairment of liver regeneration and liver function. In cases with critically small liver remnants, restoration of an adequate venous outflow may be mandatory.     

Decrease in regeneration capacity of rat liver after external biliary drainage.

In order to discover the effect of external biliary drainage on liver regeneration, we have produced a model system carrying cannula in the common bile duct of rat liver and examined the regeneration capacity of liver after partial hepatectomy under various conditions. Previously we have shown that hepatic cells proliferate by obstructive jaundice alone without partial hepatectomy [Terasaki et al; Jpn J Cancer Res 1991;82:170-175]. In the present study, we showed that DNA polymerase-alpha was induced by partial hepatectomy of rats suffering from obstructive jaundice and the induced level was similar to that of the normal regenerating liver. The level of DNA polymerase-alpha activity corresponded well to the liver regeneration capacity estimated by mitotic index. Contrary to our expectation, external biliary drainage for obstructive jaundice markedly suppressed the regeneration capacity of the remaining liver which was estimated by DNA polymerase-alpha activity, mitotic index and [3H]thymidine incorporation. The suppression may be due to the external biliary drainage itself because the liver regeneration of normal rats without jaundice was also suppressed by the biliary drainage. These results suggest that the external biliary drainage seriously suppresses the regeneration capacity of liver at least at the early stage of obstructive jaundice.     

Effects of ischemic preconditioning on regenerative capacity of hepatocyte in the ischemically damaged rat livers

BACKGROUND: Liver regeneration after partial hepatectomy is regulated by several factors that activate or inhibit hepatocyte proliferation. A short period of ischemia-reperfusion (IR), called ischemic preconditioning (IPC), protects the liver against subsequent sustained ischemic insults. The present study investigated the effects of IPC on liver regeneration after partial hepatectomy under IR in rats. MATERIALS AND METHODS: Male Wistar rats were subjected to 45 min of total hepatic ischemia, and 70% hepatectomy was performed just before reperfusion. Animals were pre-treated with either IPC (10/15 min) (IPC + PHx group) or not (ischemia + PHx). The survival rate, serum transaminases, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-6 levels, hepatocyte proliferation and histological change of the remnant liver were measured in both groups and compared with non-ischemic controls subjected to 70% hepatectomy alone (PHx group). RESULTS: The survival rate was significantly better in the IPC + PHx group than in the ischemia + PHx group. Furthermore, IPC reduced liver injury determined by liver histology and serum transaminases. There was an early rise in serum TNF-alpha and IL-6 levels in the ischemia + PHx group. Compared with non-ischemic controls, IPC significantly decreased TNF-alpha, but not IL-6 during the late (24 and 48 h) phases of reperfusion. Rats subjected to 70% hepatectomy and 45 min of hepatic ischemia showed significantly reduced hepatocyte proliferation (mitotic index, proliferating cell nuclear antigen, and relative liver weight) when compared with animals subjected to hepatectomy alone. However, hepatocyte proliferation was markedly increased in rats pretreatment with IPC when compared with ischemic controls. CONCLUSION: These results suggest that ischemic pre-conditioning ameliorates the hepatic injury associated with ischemia-reperfusion and has a stimulatory effect on liver cell regeneration that may make it valuable as a hepatoprotective modality. Il-6 appears to be key mediator in promoting regeneration after combined ischemia and hepatic resection.     

Cholestasis induces murine hepatocyte apoptosis and DNA synthesis with preservation of the immediate-early gene response

BACKGROUND: Major hepatic resection in patients with unrelieved obstructive jaundice carries an increased risk of postoperative liver failure. We hypothesized that cholestasis induces hepatocyte apoptosis and impairs hepatic regeneration by inhibiting up-regulation of the known immediate-early response genes, nuclear factor kappa B (NF-kappaB) and activating protein-1 (AP-1). The aim of this study was to determine whether the immediate-early gene response in hepatic regeneration remains intact in extrahepatic cholestasis. METHODS: Eight-week-old BALB/c mice underwent either sham operation (SO) or common bile duct ligation (BDL). Two-thirds partial hepatectomy (PH) was performed at 4 and 7 days, with remnant liver harvested 0, 15, 30, or 60 minutes after PH. Serum analysis for markers of cholestasis and histopathology was obtained. Proliferating cell nuclear antigen and terminal deoxyuridine triphosphate nick end labeling (TUNEL) immunohistochemistry for detection of DNA synthesis and apoptosis, respectively, was performed 4, 7, or 10 days after SO or BDL. Liver samples from 0, 15, 30, or 60 minutes after PH were analyzed for NF-kappaB and AP-1 DNA binding activity by using electrophoretic mobility shift assays. RESULTS: Increased serum bilirubin level and hematoxylin-eosin-stained liver sections confirmed cholestasis in BDL mice. BDL induced marked DNA synthesis and hepatocyte apoptosis in prehepatectomy liver at both 4 and 7 days. Substantially higher basal levels of both NF-kappaB and AP-1 binding activity were present in BDL compared with SO mice. Fold induction of NF-kappaB and AP-1, however, was similar between BDL and SO mice. Cholestasis induced hepatocyte DNA synthesis and apoptosis. Basal NF-kappaB and AP-1 DNA binding activity was increased in BDL mice, but fold induction of these immediate-early genes did not differ from controls. CONCLUSIONS: Although basal NF-kappaB and AP-1 DNA binding is increased in cholestasis, the immediate-early gene response to PH remains intact in BDL mice.     

肠道缺乏胆汁酸对大鼠肝再生的影响

目的 探索肠道缺乏胆汁酸对肝再生的影响.方法 通过喂养大鼠0.2%胆酸(胆酸负荷组)、2%考来烯胺(胆酸缺乏组)建立干扰肠道胆汁酸代谢的动物模型,以喂养标准饲料作为对照组,所有大鼠均行70%肝部分切除术(PH).比较PH后0、1、2、3、7 d三组大鼠的肝脏再生情况,并检测法尼酯衍生物X受体FXR及其靶基因胆汁酸合成限速酶CYP7a1的mRNA表达.结果 胆酸缺乏组的肝再生率在PH后3、7 d显著低于胆酸负荷组和对照组(P＜0.05),1 d时PCNA和Ki-67标记指数(22.21%±2.31%、17.25%±6.50%)显著低于胆酸负荷组(44.4%±4.92%、30.83%±3.91%)和对照组(38.74%±6.42%、27.04%±7.22%),且标记指数高峰延迟.在PH后,胆酸缺乏组FXR mRNA表达显著低于其他两组,而FXR的靶基因CYP7al的mRNA则随时间升高,明显高于其他两组.结论 肠道缺乏胆汁酸可延迟肝脏再生,并伴随FXR mRNA表达下降.     

胆道梗阻大鼠肝切除术后残肝细胞能量代谢状况的研究

目的 探讨胆道梗阻大鼠肝切除术后残肝细胞能量代谢状况。方法 实验大鼠分实验组（３０只）、对照组（２０只）２组。对正常大鼠和胆道梗阻５ｄ大鼠行７０％肝切除和胆肠引流术、观察术后２４ｈ残肝细胞线粒体呼吸功能（线粒体ＲＳ３、ＰＣＲ和ＡＤＰ／Ｏ比值）和ＡＴＰ量改变。结果 正常大鼠肝切除后肝细胞线粒体功能代偿性增加。胆道梗阻大鼠肝切除术后肝线粒体代偿能力减弱（Ｐ〈０．０５）,肝细胞能量代谢障碍（Ｐ〈０．０１