肝纤维化大鼠部分肝切除后肝星状细胞活化和肝细胞生长因子的表达

目的：研究肝星状细胞在肝纤维化大鼠部分肝切除后的活化情况及其对肝细胞生长因子表达和肝再生的影响。方法：成年雄性SD大鼠，随机分成3组：正常组、肝纤维化组和肝纤维化大鼠部分肝切除组。其中肝纤维化大鼠部分肝切除组根据术后取材时间又分为6小组，分别于术后12h、1、3、5、7和14d取材。计算肝指数评价肝再生情况；用免疫组化、免疫荧光和免疫印迹方法检测各组大鼠肝组织中α平滑肌肌动蛋白和肝细胞生长因子的表达情况。结果：肝纤维化大鼠部分肝切除后肝指数逐渐增加，但递增速度缓慢；肝组织中α平滑肌肌动蛋白的表达呈现出先降低后升高的规律；肝细胞生长因子表达早期下降，而后升到一最高值后开始降低。结论：（1）肝纤维化大鼠部分肝切除后残肝可以再生；（2）活化肝星状细胞术后呈现出先减少后增多的规律性变化；（3）肝星状细胞可能是术后肝细胞生长因子表达和残肝再生的重要影响因素。     

Molecular mechanisms of apoptosis in the liver of rats after portal branch ligation with and without retrorsine

The mechanisms accounting for the atrophy of the portal blood-deprived liver lobes after portal branch ligation (PBL) are still unclear. The first aim of this study was to confirm the role of apoptosis in this process and to determine which apoptotic pathways are involved. The second aim of the study was to evaluate the effect of blocking compensatory hyperplasia of the nonligated lobes with retrorsine on the mechanisms of apoptosis in the ligated lobes. Mitochondrial Bax, Bcl-2 and Bcl-X(L), cytosolic cytochrome c, caspase-3, -8 and -9 activities and TNF-alpha levels were assessed in the liver of rats before and at various time points, ranging from 30 min to 7 days, after PBL. Caspase activities were also measured in rats pretreated with retrorsine. Both the mitochondrial and the death receptor-mediated pathways are activated in the ligated liver lobes after portal branch ligation. Caspase activation is inhibited by retrorsine pretreatment, resulting in fewer apoptotic bodies. Apoptosis accounts for the atrophy of the ligated lobes after PBL. It is inhibited by retrorsine, suggesting an attempt to reduce the loss of liver mass when hyperplasia of the nonligated lobes is impaired     

肝门静脉左支分支分型的数字化研究及编码分析

目的　探究肝门静脉左支新的分型和研究方法,为肝数字解剖研究提供数据和参考。 方法　应用Hisense CAS对110例国人上腹部增强CT数据进行肝血管三维重建,准确显示肝门静脉左支的三维图像。分析肝门静脉左支分支的形态特点及分布规律,进行编码并统计分析。 结果　根据门静脉左支走行特点可分为6型：I型61例（55.45%）,II型29例（26.36%）,Ⅲ型13例（11.82%）,IV型4例（3.64%）,V型2例（1.82%）,VI型1例（0.91%）。 结论　Hisense CAS能够准确地显示肝门静脉三维影像,方便进行肝内管道观察,为肝门静脉分支分型的数字解剖学研究提供了很好的工具;针对较大数据的观测,用编码的方法非常方便统计分析。     

不同浓度肝细胞生长因子和成纤维细胞生长因子 对大鼠肝干细胞的增殖调控

目的：初步探讨不同浓度的肝细胞生长因子(hepatocyte growth factor,HGF)和成纤维细胞生长因子(fibroblast growth factor, FGF)对大鼠肝干细胞增殖的调控作用。方法：先将大鼠肝胎细胞分别种植A、B培养板上进行肝细胞的体外培养。A组：培养液中含有HGF,浓度分为别1,5,10,20,40,80及100 ng/mL。B组：培养液中含有FGF,浓度分别为1,5,10,20,40,80及100 ng/mL。对照组不加细胞因子。由A、B两组剂量效应获得HGF和FGF的最佳浓度。再将细胞接种于C组培养板进行培养,培养液中含有最佳浓度HGF联合上述各浓度FGF, 并用四甲基偶氮唑盐比色法(mononuclear cell direct cytotoxicity assay, MTT)分别测两组的光密度值（optical delnsity, OD）,检测出HGF与FGF的最佳浓度组合。结果：HGF和FGF浓度与肝干细胞增殖效应呈剂量依赖性（dose-dependent）。当HGF浓度为20 ng/mL,FGF浓度为10 ng/mL时,为最佳浓度组合。结论：HGF和FGF均能对肝干细胞的增殖起促进作用,并且两者有一定协同作用。     

Phenotypical and morphological alterations to rat sinusoidal endothelial cells in arterialized livers after portal branch ligation

The hepatic sinusoids are preferentially supplied with portal venous blood and equipped with fenestrated endothelial cells that are distinct from capillary endothelial cells. We previously observed in rats that sinusoidal capillarization proceeded concurrently with arterial blood supply during hepatocarcinogenesis. This study aimed to clarify the inducing role of arterialization in sinusoidal capillarization by investigating phenotypical, morphological and functional alterations to sinusoidal endothelial cells (SECs) in arterialized rat livers induced by portal branch ligation. At one week, after massive hepatic necrosis following ligation, the livers were restored to their normal architecture without causing post-necrotic fibrosis. At 12-21 weeks, they exhibited a normal histology except for mild pericellular fibrosis which developed along sinusoids or between adjacent hepatocytes. SECs expressed factor VIII-related antigen and showed a decrease in the number of fenestrae and porosity, still lacking any basement membrane but further retaining the functional capacity for carmine dye uptake. Stellate cells, while occasionally associated with large amounts of collagen bundles, contained many lipid droplets and expressed no alpha-smooth muscle actin, indicating a quiescent property. Kupffer cells were commonly found within the sinusoids. The present results indicate that arterialization of the liver induces a partial (but not complete) transition of SECs into capillary-type endothelial cells, suggesting that arterialization might be one of the factors which induce sinusoidal capillarization in the development of hepatocellular carcinoma.     

Thermostable hepatocyte growth factor and energy metabolism in rats after partial hepatectomy

The relationship between energy metabolism in the remaining liver tissue after 60% hepatectomy and the activity of low-molecular-weight thermostable hepatocyte growth factor was studied in rats. The energy status of the liver was markedly reduced 6 h and to a greater degree 12 h after the operation, judging from the levels of ATP, ADP, AMP, and energy potential. The energy status improved (energy potential increased to 95% of the initial level) 24–72 h after the operation. This coincided with a decrease in hexokinase and phosphofructokinase activities and an increase in glucose-6-phosphate dehydrogenase and isocitrate dehydrogenase activities, indicating suppressed glycolysis and activation of the Krebs' cycle. High activity of low-molecular-weight thermostable hepatocyte growth factor was detected 24–72 h after resection of the liver (with maximum activity after 48 h). The activity of the hepatocyte growth factor increased if the range of energy potential surpassed the level sufficient for maintaining protein production, which is an energy-consuming process, but lower than the normal level. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 127, No. 1, pp. 53–56, January, 1999     

肝门静脉、肝右动脉及胆囊动脉变异1例

在肝门部结构中,肝动脉及胆囊动脉的变异较为常见,而肝门静脉的变异较少见,我们在解剖一成年男性标本时见其肝门静脉、肝右动脉及胆囊动脉存在变     

Effects of adenoviral-mediated hepatocyte growth factor on liver regeneration after massive hepatectomy in rats

Resection is the only curative treatment for liver metastasis of colorectal cancers. Despite the supreme regenerative potential of the liver, major hepatectomy sometimes leads to liver failure, and the limitation of resectable liver volumes makes advanced tumors inoperable. This study was attempted to promote liver regeneration using hepatocyte growth factor (HGF) gene transfection by venous-administered adenovirus and to improve the survival of rats after massive hepatectomy. The adenovirus that encodes HGF was administered to rats before 85%-hepatectomy. The administration of HGF gene improved the survival of rats after massive hepatectomy, while the administration of control adenovirus deteriorated their survival. Gene transfection of HGF showed up-regulation of serum HGF, stimulation of hepatocellular proliferation and rapid liver regeneration. Moreover, HGF administration reduced apoptosis of hepatocytes. The administration of HGF gene prevented liver dysfunction after major hepatectomy and may be a new assist for surgery.     

大鼠肝脏缺血再灌注损伤后水通道蛋白4表达的变化

目的: 通过建立大鼠肝脏缺血再灌注损伤(IRI)模型,观察水通道蛋白 4(AQP4)在肝内胆管细胞的表达变化,并同时检测肝脏功能变化,探讨肝脏IRI 与AQP4表达变化的关系。方法: 建立大鼠肝脏30 min IRI模型。随机将配对动物分为对照和缺血再灌注2 h、1 d、3 d、7 d组,留取血清标本行间接胆红素(IB)、直接胆红素(DB)及丙氨酸氨基转移酶(ALT)指标检测;留取肝脏标本用于HE染色光镜下观察肝脏病理形态学变化、免疫组化染色观察AQP4表达变化, RT-PCR 方法测定 AQP4 mRNA表达变化。结果: (1)肝功能变化:缺血再灌注损伤2 h、1 d、3 d组IB、DB及ALT明显升高,与对照组间比较有显著差异(P<0.05),并于术后第1 d达到最高(P<0.05);以后逐渐降低,术后第7 d恢复正常。(2) 肝脏病理学变化:术后HE染色可见缺血再灌注可造成大鼠肝组织变性、坏死。(3) 免疫组织化学检测肝脏AQP4的变化:缺血再灌注损伤2 h、1 d、3 d组AQP4的表达量较对照组降低(P<0.05),于术后第1 d最低(P<0.05),以后逐渐增高,于术后第7 d恢复正常。(4) 肝脏AQP4 mRNA表达变化:缺血再灌注损伤组2 h、1 d、3 d组AQP4 mRNA表达水平较对照组明显降低(P<0.05),于术后第1 d最低,以后又逐渐升高,于术后第7 d恢复正常。结论: 大鼠肝脏缺血再灌注损伤后可明显引起肝内胆管上皮细胞AQP4的表达降低,术后第1 d最明显,至术后7 d逐渐恢复正常,此种损害与包括胆红素在内的肝功能损害相对应。     

肝门阻断再灌注对脑兴奋性氨基酸及受体mRNA表达的影响

目的： 探讨无肝期后的再灌注对脑组织兴奋性氨基酸（EAAs）及其受体mRNA表达的影响。方法：分别采用血气分析、高效液相色谱荧光检测和半定量RT-PCR方法，研究大鼠肝门阻断和假手术组再灌注6 h、12 h和24 h后门静脉电解质和pH、脑组织Glu、Asp含量以及大脑皮层中NMDAR mRNA的相对含量表达的差异，观察无肝期后再灌注损伤对肝外远隔脏器脑EAAs及NMDA受体mRNA表达的的影响。结果：肝门阻断期间门静脉pH显著降低，［K⁺］升幅超过1倍，但开放后逐渐恢复；［Ca²⁺］于再灌注后持续下降至12 h；脑皮层中的Glu和Asp再灌注后分别在6 h、24 h出现2个高峰［Glu:(349±145) μg·g^-1wt,(456±203) μg·g^-1wt vs (238±24) μg·g^-1wt,(225±59) μg·g^-1wt;Asp: (134±50) μg·g^-1wt,(166±50) μg·g^-1wt vs (99±24) μg·g^-1wt,(71±20) μg·g^-1wt］；NMDAR亚单位NR1mRNA的表达在再灌注后显著升高后逐渐下降［(1.63±0.06) μg·g^-1wt vs (1.18±0.05) μg·g^-1wt;(1.73±0.06) μg·g^-1wt vs (1.17±0.03) μg·g^-1wt］,而NR2B mRNA在12 h升高(1.75±0.04 vs 1.18±0.05)，2者24 h时均恢复至对照组水平。结论：肝门阻断后肝和肠道再灌注过程可启动神经突触前Glu大量释放和重吸收抑制，引起脑皮层EAAs的堆积，同时刺激NMDARmRNA的表达，通过Glu - NMDAR途径可能引发脑损伤。     

Regulation of spleen growth and portal pressure in hepatic shcistosomiasis.

Growth rate and histology of splenic autotransplants in subcutaneous pockets were compared with those of autotransplants in the extrahepatic portal bed in splenectomized mice infected with cercariae of Schistosoma mansoni and in splenectomized uninfected controls. By the fifteenth week after transplantation (and tenth week after injection of cercariae) subcutaneous transplants gained 6.5 times and omental transplants 8.2 times more weight in infected animals than corresponding transplants in uninfected animals. Portal pressures averaged 11 to 13 cm of water in infected animals with transplants and 17 cm in those with intact spleens (compared to that of 6 to 7 cm in controls). Hyperplasia of white pulp with increase in germinal center activity characterized transplants as well as intact spleens of infected animals. The results suggest that a) During the first 10 weeks of experimentally induced infection, portal congestion is not the predominant mechanism regulating increased spleen growth; and b) An intact enlarged spleen appears to contribute to elevated portal pressure.     

Occurrence of cardiac muscle in the hepatic portal vein wall of the mouse and rat

Cardiac myocytes have been shown to occur in the tunica media and adventitia at the region near the hepatic end of the portal vein of the mouse and rat, and have been studied by electron microscopy in the mouse portal vein. They measured 3-10 microns in breadth at their nuclear level, possessed centrally located nuclei, and were connected with each other by the intercalated disk. In these myocytes in the mouse portal vein, sarcoplasmic reticulum was represented by a rather simple and loose network of the anastomosing tubules. The membrane-bound granules, which closely resemble the atrial specific granules, were found in many of the mouse portal vein myocytes. Transverse tubules, 40-200 nm in diameter, were sometimes detectable at the Z line level. The nexus occupied about 3-5% of the whole junctional area between cardiac myocytes in the tunica media, whereas in the tunica adventitia the corresponding value was about 17%. Blood capillaries with fenestrated endothelium supplied the cardiac myocytes in the adventitia of mouse portal vein. The closest relationship between the adrenergic axon and portal vein cardiac myocytes was observed to be ca. 0.3 micron apart. The significance of these findings is discussed in relation to pulsations of the portal vein.     

The effect of haemorrhage on hepatic artery and portal vein flows in the anesthetized cat

1. In cats anaesthetized with pentobarbitone, the hepatic artery and portal vein flows and pressures were recorded simultaneously.

2. Removal of blood from the animal did not cause a decrease in the hepatic artery flow unless the arterial pressure fell below 80 mm Hg. In contrast, the portal vein flow fell markedly.

3. After restoration of the blood, the hepatic artery flow increased to above the pre-haemorrhage level, while portal vein flow returned only partly towards the control level.

4. It is concluded that haemorrhage causes a vasodilation in the hepatic artery vascular bed and a vasoconstriction in the vascular beds drained by the portal vein. By this means, the oxygen supply to the liver is maintained as far as possible.