大鼠肝再生过程中线粒体氧化磷酸化的调控

目的：探讨肝部分切除后肝再生过程中线粒体能量代谢的调控。方法：用雄性Ｗｉｓｔａｒ大鼠施行肝部分切除复制肝再生模型,肝部分切除后分别观察０５、１、２、４和７ｄ５个肝再生组以及５个相应的对照组,差速离心法分离肝线粒体并用氧电极极谱法测定其氧化磷酸化活性。结果：肝部分切除后肝再生过程中线粒体呼吸控制率（ＲＣＲ）明显高于相应对照组,其中肝部分切除后０５ｄ和４ｄ为二个峰值,７ｄ时降至对照组水平,早期ＲＣＲ的升高主要是Ｒ３升高的所致,１ｄ后ＲＣＲ升高是Ｒ４下降所致。磷氧比值（Ｐ／Ｏ）的变化类似于ＲＣＲ。结论：肝部分切除后肝再生过程中线粒体通过氧化磷酸化偶联增强来适应肝再生的能量需求,这种增强机制很可能主要是通过降低线粒体内膜通透性实现的。     

Portal vein arterialization promotes liver regeneration after extended partial hepatectomy in a rat model

In the current study, we sought to establish a novel rat model of portal vein arterialization (PVA) and evaluate its impact on liver regeneration after extended partial hepatectomy (PH). A total of 105 Sprague-Dawley rats were randomly assigned to three groups: 68% hepatectomy (the PH group), portal arterialization after 68% hepatectomy (the PVA group), and right nephrectomy only (the control group). Liver regeneration rate (LRR), 5-bromo-2-deoxyuridine (BrdU) labeling index, and liver functions were assessed on postoperative day 2, 7, 14 and 28. The 28-day survival rates were compared among the three groups. The 28-day survival rates were similar in all groups (P  =  0.331), and the anastomotic patency was 100%. The LRR in the PVA group was significantly higher than that of the PH group within postoperative 14 days (P < 0.05). The PVA and PH group had increased serum alanine aminotransferase levels (232 ± 61 U/L and 212 ± 53 U/L, respectively) compared with the control group (101 ± 13 U/L) on postoperative day 2, whereas from postoperative day 7 to day 28 there were no differences among the three groups. Serum albumin values were higher after the PVA procedure within postoperative day 14, which gradually became comparable on postoperative day 28 among the three groups. The peaks of BrdU labeling index appeared on postoperative day 2 in all rats, and the PVA procedure was associated with increased BrdU labeling index from postoperative day 7 to 28. The 28-day survival of the PVA rats was comparable. Our findings demonstrate that the PVA procedure utilizing portal vein trunk-renal artery microvascular reconstruction promotes remnant liver regeneration and confers beneficial effects on maintaining and even optimizing liver function after extended partial hepatectomy in rats.     

大鼠部分肝切除术后Hedgehog信号分子的表达

目的: 探讨部分肝切除术后Hedgehog(Hh)信号分子的表达及其意义。方法: 切除肝脏左叶和中叶以建立SD大鼠部分肝切除术模型。18只雄性SD大鼠随机均分为3组:假手术组(A)、部分肝切除术组1(B)、部分肝切除术组2(C)。A、B组于术后24 h、C组于术后48 h取肝组织,用免疫组化染色法和Western blotting检测肝组织Ki-67、Sonic Hedgehog(Shh)、Indian Hedgehog(Ihh)和Glioblastoma-2(Gli-2)的表达。结果: HE染色显示B、C组可见肝细胞水肿,点状坏死。C组可见处于不同分裂期的细胞。B、C组Ki-67、Shh、Ihh和Gli-2的表达明显高于A组(P<0.01),C组Ki-67、Shh、Ihh及Gli-2的表达高于B组(P<0.01)。结论: 部分肝切除术后Hh信号通路被激活,可能参与促进肝脏修复再生。     

大鼠肝部分切除后血清氨基酸的代谢变化

本实验研究了大鼠肝部分切除后72小时血清氨基酸的代谢变化。实验分三纰:正常对照组、假手术组和肝部分切除组。用氨基酸自动分析仪测定了主动脉、下腔脉静,门静脉和肝静脉血清20种氨基酸浓度。结果表明大鼠肝部分切除后72小时,上述血管血清氨基酸浓度均显著升高。肝摄取氨基酸总量显著增加,门静脉区内脏释放氨基酸总量占肝摄取量的87%。二者呈正相关。外周血管血清必需与非必需氨基酸比值和支链与芳香族氨基酸比值均显著下降。外周组织对支链氨基酸摄取严重抑制,而肝脏摄取则显著增加,这不仅为肝再生提供充分支链氨基酸的供应,而且促进肝蛋白质的合成。门静脉区内脏释放和肝脏摄取丙氨酸的量均显著增加,从而为肝糖原异生提供原料的供应。     

Facilitation of liver regeneration after partial hepatectomy by ventromedial hypothalamic lesions in rats

Whether or not the hypothalamus is involved in initiating hepatic DNA synthesis after partial hepatectomy is unclear. To determine the role of the ventromedial hypothalamic nuclei in liver regeneration after partial hepatectomy, we studied hepatic DNA synthesis during liver regeneration in rats with bilateral lesions of these nuclei. Lesions of the ventromedial hypothalamus accelerated the increase in hepatic DNA synthesis and raised the peak level of thymidine incorporation after partial hepatectomy. These effects of hypothalamic lesions were completely inhibited by hepatic vagotomy. Thus, lesions of the ventromedial hypothalamus appear to promote hepatic regeneration by increasing vagal stimulation of the liver.     

索拉菲尼对肝硬化大鼠部分肝切除术后肝脏再生的影响

目的: 在大鼠肝硬化模型的基础上行肝脏部分切除(PH),研究索拉菲尼(sorafenib)对大鼠肝脏再生的影响。方法: 使用二乙基亚硝胺(DEN)诱导Wistar大鼠肝硬化,成功建立30只肝硬化大鼠PH模型后,随机分2组,每组15只。术后第1 d开始,分别给予实验组索拉菲尼(30 mg·kg^-1·d^-1)、对照组生理盐水灌胃10 d后处死。留取PH后及实验结束后的血液及肝脏标本,检测2组肝脏再生率(LRR),增殖细胞核抗原(PCNA),生化指标: 丙氨酸转移酶(ALT)和血清白蛋白(ALB)、血清总胆红素(TBIL)和血清直接胆红素(DBIL)的变化,血管生成相关因子:血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR-2)、血小板源性生长因子受体β(PDGFR-β),以及肝脏微血管密度(MVD)的变化。结果: (1)LRR在实验组及对照组分别为45.43%±3.36%和44.21%±2.77%,无显著差异(P>0.05);(2)免疫组织化学(IHC)没有检测到PCNA;(3)2组的生化指标无显著差异(P>0.05);(4)实验组VEGFR-2和PDGFR-β的表达受到抑制,MVD降低,并且实验组与对照组差异有统计学意义(P<0.01)。结论: 索拉菲尼虽然对肝硬化血管再生相关因子有抑制作用,但是对肝细胞再生和肝功能没有明显影响。     

Expression of epidermal growth factor receptor protein in the liver of db/db mice after partial hepatectomy

Liver regeneration was impaired after partial hepatectomy (PH) in leptin receptor-deficient db/db mice with severe liver steatosis. In the present study, we analyzed the mode of epidermal growth factor receptor (EGFR) protein expression in the liver of 5- and 10-week-old db/db and age-matched control mice. In 5-week-old db/db mice, neither the expression of EGFR protein in the intact liver nor the rate of liver regeneration after PH was significantly different from that in age-matched control mice. However, in 10-week-old db/db mice, the level of EGFR protein expression was very low and liver regeneration was prominently suppressed. Histopathologically, much severer fatty change was observed in the liver of 10-week-old db/db mice than 5-week-old db/db mice. These results suggest that the down-regulation of EGFR protein expression is associated with an impairment of liver regeneration in db/db mice and that the severity of hepatic steatosis plays an indirect role in the impairment of liver regeneration by modifying EGFR expression.     

Reparative regeneration of rat fetal liver after partial hepatectomy

A reproducible model of fetal liver regeneration was created. Resection of 20% liver was carried out in rat fetuses on day 17 of prenatal development. The organ weight was restored after 2 days at the expense of an increase in hepatocyte mitotic activity; cell hypertrophy was minor. After recovery, the cell composition of the operated liver did not differ from the control, i.e. the regeneration was organotypical.     

肝硬化部分切除后肝再生及结构变化

背景：肝脏自身具有强大的再生能力,临床资料显示大部分肝癌患者合并有肝硬化,切除肿瘤后肝脏质量可逐渐恢复,提示硬化的肝脏仍具备修复潜能。目的：尝试用肝部分切除方法诱导小鼠硬化肝组织内具再生能力的细胞增殖,观察再生肝脏的结构变化。方法：使用CCl₄皮下注射方法制备小鼠肝硬化模型。对肝硬化小鼠行肝部分切除,于切除后1,3,5,7,10,14及21 d取小鼠肝组织并称质量。观察比较肝部分切除后不同时间小鼠再生肝组织的结构变化;评估肝再生率;免疫组化方法分析肝星形细胞标志分子结蛋白及肝星形细胞活化标志分子α-平滑肌肌动蛋白的表达变化;5-溴脱氧尿核苷标记技术对增殖细胞进行定性、定位。结果与结论：肝硬化小鼠肝部分切除后第1,3,5,7,14,21天肝再生率分别为6.58%,22.03%,21.39%,29.05%,45.22%,50.98%。苏木精-伊红染色和Masson染色显示CCl₄注射8周后呈早期肝硬化病理改变,肝部分切除后肝组织结构逐步改善。免疫组化结果显示肝部分切除后第1天再生肝组织内结蛋白表达减少,但第3天至第14天呈上升趋势,21 d则明显减少;肝再生过程中各时间点α-平滑肌肌动蛋白表达依时递减。5-溴脱氧尿核苷阳性细胞在肝部分切除后1-7 d主要为肝细胞,肝部分切除后14-21 d阳性细胞定位于肝血窦内皮及窦腔内。结果表明早期肝硬化小鼠大部肝切除后3 d出现明显的再生反应,并逐步恢复正常的肝组织结构。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

Impaired liver regeneration after partial hepatectomy in db/db mice

Fatty liver is the most common hepatic disorder in humans and supposed to be a cause of poor prognosis after liver transplantation and hepatic resection which could be resulted from impaired liver regeneration. This study was carried out to analyze the process of liver regeneration in db/db mice which show severe steatosis because of abnormal leptin receptor. We performed 70% partial hepatectomy (PH) on db/db mice and normal +m/+m mice, and then sacrificed the animals 1, 2, 3, 5, 7 and 10 days later. The liver samples were weighed and examined histologically or immunohistochemically. As a result, the liver mass restitution was significantly inhibited in db/db mice compared with +m/+m mice. The BrdU labelling index peaked at 2 days after PH in both strains, although the value was lower in db/db mice. After that, interestingly, it decreased to the control level at 5 days in +m/+m mice while the recovery was delayed in db/db mice. Similar sequence was also observed in the PCNA labelling index. In addition, the peak time of the mitosis index was 2 days and 5 days after PH in +m/+m mice and in db/db mice, respectively. Thus, although not significant, the proliferative response of hepatocytes to PH occurred somewhat more transient and sharply in +m/+m mice while it lasted somewhat longer in db/db mice. This suggests that db/db mice may be valuable as one of the animal models for the investigation of the effects of steatosis on the liver regeneration.     

Effect of physical training on regeneration of the rat liver after partial hepatectomy

Training rats by swimming was shown to cause changes in the functional state of the hepatocytes, accompanied by changes in the distribution of nuclei relative to their DNA ploidy classes. The blood sugar and liver glycogen concentrations of the trained animals 20 and 48 h after partial hepatectomy were significantly higher than in untrained rats. Reparative regeneration after partial hepatectomy in trained rats is characterized by a unique distribution of nuclei among DNA ploidy classes and by the entry of only diploid nuclei into the phase of synthesis 20 h after the operation.Central Research Laboratory, Novosibirsk Medical Institute. Laboratory of Endocrinology and Laboratory of Biophysics, Institute of Clinical and Experimental Medicine, Siberian Division, Academy of Medical Sciences of the USSR, Novosibirsk. (Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 10, pp. 483–486, October, 1978.     

冷缺血损伤后大鼠肝脏移植后肝脏再生的分子机制

目的： 研究不同冷缺血损伤条件下，大鼠肝脏移植后肝脏再生的分子机制。方法: 建立大鼠原位肝移植模型。实验分为: 冷缺血1 h、8 h和16 h组。观察各组的生存率，并在术后90 min、1、2、4和7 d收集标本，观察白介素-6(IL-6)、肿瘤坏死因子（TNF-α）、信号转导激活蛋白3 (STAT3)等表达情况。免疫组化检测细胞周期素D1的表达和肝细胞摄取溴脱氧尿核苷情况。比较实验各组肝脏移植后TNF-α和IL-6的表达水平。分析实验各组移植术后48 h 溴脱氧尿核苷染色阳性的肝细胞数。结果: 冷缺血1 h、8 h和16 h后肝脏移植物的存活率均为100％（>14 d）。移植术后90 min，与冷缺血1 h组相比，冷缺血8 h和16 h组大鼠移植肝内TNF-α等因子表达明显增加（P<0.05）。移植术后90 min，与冷缺血1 h和8 h组相比，16 h组IL-6的表达亦明显增强（P<0.05）。冷缺血8 h和16 h组STAT3活性明显增强。冷缺血8 h组细胞周期素D1在胞浆和细胞核内均有表达。冷缺血16 h组细胞周期素D1仅在核内表达。肝细胞复制活跃。与冷缺血1 h和8 h组相比，冷缺血16 h组术后48 h溴脱氧尿核苷染色阳性的肝细胞计数也明显增多（P<0.05）。结论: 大鼠肝脏移植物经受16 h冷缺血损伤后仍然能够启动，完成肝脏再生，修复组织损伤。大鼠肝脏移植后的肝脏再生可能通过TNF-α/IL-6／STAT3/cyclin D1/DNA合成的途径调节。     

胆汁酸对肝硬化大鼠肝切除术后法尼酯X受体、白细胞介素－6的表达及肝功能恢复的影响

目的探讨胆汁酸对肝硬化大鼠肝切除术后法尼酯X受体(FXR)、白细胞介素-6(IL-6)的表达和肝功能恢复的影响作用。 方法将24只SD大鼠采用0.03%硫代乙酰胺溶液喂养方法制备肝硬化模型,行肝切除术后采用简单随机法分为对照组和实验组,每组各12只。其中对照组用普通标准饲料喂养,而实验组用含0.2%胆汁酸的饲料喂养。1周后抽取下腔静脉血液行肝功能检查,随后处死实验动物,迅速切取残肝组织备用。分别采用蛋白质印迹法和全自动化学发光分析等方法检测FXR、IL-6的表达及肝功能恢复情况。数据比较采用t检验。 结果实验组中天冬氨酸基转移酶(AST)、丙氨酸转氨酶(ALT)和体内胆汁酸水平分别为(156.4±32.5) U/L、(203.3±38.6) U/L、(6.68±3.90) μmol/L,均低于对照组(215.6±42.3) U/L、(273.8±42.1) U/L、(6.51±4.30) μmol/L,差异均有统计学意义(t＝4.432、2.566、2.114,P＝0.004、0.025、0.035);实验组中FXR的表达量为0.45±0.16,而对照组的表达量为0.28±0.08,差异有统计学意义(t＝4.47,P＝0.003);实验组中肝脏再生指标肝细胞有丝分裂指数(MI)、肝细胞增殖细胞核抗原(PCNA)标记指数和细胞核DNA含量分别为(2.89±0.67)%,(4.76±0.89)%和(8.82±2.12)%,均高于对照组(1.12±0.34)%,(2.46±0.68)%和(3.98±1.16)%,差异均有统计学意义(t＝4.688、5.069、6.858,P＝0.003、0.006、0.005);而实验组中IL-6的水平为(1.34±0.56) ng/mL,对照组为(1.68±0.55) ng/mL,实验组明显低于对照组,差异有统计学意义(t＝2.29,P＝0.02)。 结论胆汁酸可以通过启动FXR、减少IL-6炎性介质、促进大鼠肝功能恢复和肝脏再生。     

Proliferation kinetics of rat Kupffer cells after partial hepatectomy. Immunohistochemical and ultrastructural analysis

In an effort to settle the conflicting views on the proliferation kinetics of Kupffer cells (Kc), we performed 2/3 partial hepatectomy on rats injected with Pelikan ink. Using an anti-rat macrophage monoclonal antibody, ED 2, we evaluated the numerical changes in total, carbon-positive ED 2+ cells and carbon-negative ED 2+ cells in the portal and central area. We also analyzed the ultrastructure and peroxidase cytochemistry of various types of cells observed during regeneration. The total numbers of ED 2+ cells in the remaining liver increased rapidly from day 2 to 5, and the number of dividing ED 2+ cells reached a maximum on day 2. Thus, the numerical increase in ED 2+ cells corresponded to the division phase. In contrast, the carbon-labeling experiment showed a continuous increase of carbon-negative ED 2+ cells from day 2 to 7. In the central area where division was less frequent, the proportion of carbon-positive cells decreased markedly to 50% on day 7, as against 97% in control rats. These findings suggest the possibility of an influx of carbon-negative Kc in addition to cell division. Ultrastructurally, the presence of carbon-negative "small Kc" and "immature Kc" with morphological features different from those of carbon-positive Kc was demonstrated. Such carbon-negative Kc with a high nucleus-to-cytoplasm ratio and rather few phagosomes, were not observed in control rats. Furthermore, we demonstrated two types of possible precursor cell, i.e. "transitional" forms between monocytes and Kc, and "immature macrophages". The former showed peroxidase activity in some lysosomes as well as in the rough endoplasmic reticulum and nuclear envelope. Our result indicated that the proliferation kinetics of Kc depend upon both local proliferation and influx.     

Regression analysis of relationship between liver weight and body weight after partial hepatectomy in rats

A method for evaluation of the initial weight of the liver is developed for calculating the regeneration coefficient after partial hepatectomy on the basis of linear regression analysis. Experiments on rats showed that the model reflecting the relationship between liver weight and body weight for calculation of regeneration coefficient after partial resection of the liver expressed by the equation P ₀=0.033×M can be used for screening of potential hepatoprotectors for quantitative evaluation of the regeneration processes. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 141, No. 2, pp. 234–236, February, 2006     

法尼酯X受体在肝硬化大鼠肝切除术后肝再生中的作用

目的 探讨法尼酯X受体(FXR)在肝硬化大鼠肝切除术后肝再生中的作用.方法 30只肝硬化大鼠肝切除术后按完全随机法分为对照组、胆酸组和消胆胺组饲养1周,每组10只.对照组给以标准饲料饲养,胆酸组给以含0.2%胆酸的饲料饲养,消胆胺组以含2%消胆胺的饲料饲养.1周后检测各组大鼠胆汁分泌速度及总胆汁酸(TBA)含量,检测肝功能、有丝分裂指数(MI)、增殖细胞核抗原(PCNA)标记指数和细胞核DNA含量,并应用RT-PCR和Western免疫印迹分别检测肝组织FXR、胆固醇7α羟化酶(CYP7A1)两者mRNA和蛋白的表达.结果 胆酸组胆汁分泌速度、TBA含量、血清白蛋白(ALB)含量、MI、PCNA标记指数、细胞核DNA含量均高于对照组(P<0.05),血清谷草转氨酶(AST)、谷丙转氨酶(ALT)均低于对照组(P<0.05).而消胆胺组胆汁分泌速度、胆汁TBA含量、血清ALB含量、MI、PCNA标记指数、细胞核DNA含量均低于对照组和胆酸组(P<0.05),血清谷草转氨酶(AST)、谷丙转氨酶(ALT)均高于对照组和胆酸组(P<0.05).胆酸组肝组织FXR mRNA和蛋白表达均高于对照组(mRNA:0.671±0.027比0.528±0.017,蛋白:0.702±0.039比0.566±0.020,P<0.05).肝组织CYP7A1mRNA和蛋白表达含量均低于对照组(mRNA:0.237±0.020比0.325±0.076,蛋白:0.264±0.015比0.325±0.084,P<0.05).而消胆胺组肝组织FXR mRNA(0.468±0.023)和蛋白表达(0.502±0.021)均低于对照组和胆酸组(P<0.05),肝组织CYP7A1 mRNA(0.411±0.021)和蛋白表达(0.476±0.018)则高于对照组和胆酸组(P<0.05).结论 胆汁酸与其受体FXR结合激活细胞内信号转导对肝硬化大鼠肝切除术后肝再生起重要作用,升高胆汁酸水平促进肝再生,降低胆汁酸水平则抑制肝再生.