生长抑素对鼠大部肝切除术后肝再生的影响

目的　探讨生长抑素对肝切除术后肝再生的影响。　方法　 6 0只SD大鼠随机分为对照组及生长抑素组 ,按Higgins方法行 70 %肝切除术 ,术后给药并分批于术后 6、2 4、48、72、96h处死 ,作如下比较 :①残肝肝重 ;②增殖细胞核抗原 (PCNA)标记指数 ;③图像定量分析测量PCNA阳性产物面积。　结果　与对照组比较 ,生长抑素组残肝肝重在术后 48、72、 96h显著降低 (P <0 .0 5 ) ,PCNA标记指数、PCNA阳性产物面积在术后 2 4、48、72h显著降低 (P <0 .0 5 ) ,且以 2 4、 48h最为明显 (P <0 .0 1)。　结论　生长抑素能抑制肝再生 ,且肝再生程度越强 ,抑制效应越明显     

Quantitative Gene Expression Analysis by cDNA Microarray During Liver Regeneration After Partial Hepatectomy in Rats

Purpose It is speculated that genetic regulation plays an important role during liver regeneration. We conducted this study to analyze quantitative gene expression during liver regeneration after partial hepatectomy.Methods Sixty male Wistar rats were randomly assigned into ten groups of six. One group of rats was killed preoperatively and the other nine groups were killed 2, 4, 6, 12, 24, 48, and 72h, and 5 and 7 days after 70% partial hepatectomy, respectively. The remnant livers were isolated for mRNA extraction. A mass survey of gene expression by cDNA microarray carrying 6144 polymerase chain reaction-amplified cDNA fragments was prepared by an arraying machine. The microarray images were scanned, digitized, and analyzed using a flatbed scanner.Results The variations in gene expression were classified into 72 different patterns including a pattern with a single peak, 2, 4, 6, 12, and 72h, and 5 and 7 days after partial hepatectomy. The other patterns included double peaks, enhancing trend, diminished trend, protruding curve, excavated curve, and various mixed types. Each category of gene expression pattern contained 40–218 different proto-oncogenes.Conclusion The quantitative gene expression profiles have important implications, warranting further investigation of the genetic mechanisms involved in the process of liver regeneration.     

Effects of Organic Apricot on Liver Regeneration After Partial Hepatectomy in Rats

Background

The present study investigated the effects of (sun-dried organic apricot/SDOA) supplementation in chow on liver regeneration after partial hepatectomy/(PH) in rats.

Method

In this study, 28 female rats were randomized into four groups. On the 7th day of the study, group 1 underwent laparoscopic intervention while a PH was performed on the other three cohorts. On day 28, all rats were humanely killed. Blood and liver tissue samples were subjected to biochemical determinations, histological examinations, and measurement of tissue oxidative stress enzyme activity.

Results

Serum levels of alanine transaminase (ALT), alkaline phosphatase (ALP), and liver tissue glutathione (GSH) activities were affected by PH and/or SDOA consumption (P < .05). Moderately staining cell counts in group 4 were significantly different from the other three groups (P < .05). However, no significant differences were detected among all groups in regard to aspartate aminotransferase (AST) serum levels or liver tissue superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT) or glutathione peroxidase (GSHpx) activities (P < .05).

Conclusion

The 5% SDOA supplementation over a 21-day feeding period showed a beneficial effect on liver regeneration in rats, as reflects by Ki-67 finding although there was no change in ALT or ALP or in liver tissue GSH activity.     

Angiogenesis Inhibitor TNP-470 Can Suppress Hepatocellular Carcinoma Growth Without Retarding Liver Regeneration After Partial Hepatectomy

Purpose. We investigated the suppressive effect of the angiogenesis inhibitor TNP-470 on accelerated hepatocellular carcinoma (HCC) growth in the regenerating liver. Methods. After 70% partial hepatectomy (PH), AH-130 cells were injected into the portal vein of Donryu rats. A control group was given the vehicle only, and the treated group was given 10mg/kg TNP-470 subcutaneously every second day, from 24h after tumor implantation, seven times. On day 14, tumor growth was evaluated by the number of foci on the liver surface, liver weight, and the microvessel density of the tumor. Results. The number of foci was significantly less in the treated group (116.5 ± 103.1) than in the control group (319.3 ± 223.1) (P 0.05), as was microvessel density, which was 31.3 ± 14.0/mm² in the treated group and 61.2 ± 18.9/mm² in the control group (P 0.05). The liver tended to weigh less in the treated group (12.15 ± 1.28g) than in the control group (15.22 ± 5.35g). We also assessed whether TNP-470 retards liver regeneration. Seven days after 70% PH, the liver weight in the treated group was similar to that in the control group. Total bilirubin, serum glutamic oxaloacetic transaminase, and serum glutamic pyruvic transaminase were not higher in the treated group than in the control group. Conclusion. TNP-470 can suppress HCC growth without retarding liver regeneration after PH.     

Intermittent Ischemic Preconditioning Protects Against Hepatic Ischemia-Reperfusion Injury and Extensive Hepatectomy in Steatotic Rat Liver

ABSTRACT

Background: Hepatic steatosis causes severe liver damage and has deleterious effects when associated with ischemia-reperfusion mechanisms. Ischemic preconditioning (IPC) protects lean liver against prolonged ischemia by improving micro-circulation and reducing lipid peroxidation. We investigated the effect of intermittent IPC on liver ischemia-reperfusion injury (IRI) and extensive hepatectomy in severe hepatic steatosis. Methods: Severe hepatic steatosis was performed by 12–14 weeks of choline-free diet in 108 Wistar rats. We induced 30-minute ischemia-reperfusion manipulations and extensive hepatectomy with or without prior IPC in steatotic livers and after 6 and 24 hours of reperfusion blood transaminases, and IL6, TNFα, NO and Lactate in blood and liver tissue were measured. Results: Steatotic rats subjected to hepatic ischemia-reperfusion alone after extensive hepatectomy, showed severe liver damage with significantly increased values of AST, ALT, TNFα and Lactate and significantly reduced IL6 and NO, while no one rat survived for more than 29 hours. On the contrary, steatotic rats subjected to intermittent IPC, 24 hours before ischemia-reperfusion, presented increased 30-day survival (67%), lower values of AST, ALT, TNFα and Lactate, and increased IL6 and NO levels. Simple and intermittent IPC manipulations, 1 hour before the IRI and extended hepatectomy, did not prolong survival more than 57 and 98 hours, respectively. Simple IPC, 24 hours before IRI and extended hepatectomy had the lowest possible survival (16.7%).Conclusions: Hepatic steatosis and IRI after major liver surgery largely affect morbidity and mortality. Intermittent IPC, 24 hours before IRI and extensive hepatectomy, presents higher 30-day survival and improved liver function parameters.     

Total Blood Exchange Suppresses the Early Stage of Liver Regeneration Following Partial Hepatectomy in Rats

Despite its obscure and short effect, plasma exchange (PE) remains a mainstay in the treatment of liver disease. However, the question still remains as to whether or not PE suppresses the regeneration of the liver because PE deprives patients of hepatotrophic factors. The effect of PE, which could be a total blood exchange (TBE) in a syngeneic setting, on liver regeneration following a 68% partial hepatectomy (PH) was investigated in rats. In Group 1, 20 ml of blood from normal rats was infused while native blood was removed at 6 and 12 h after PH. In Group 2, 20 ml of blood obtained from PH rats at the same time points was infused. The regeneration rate, labeling index of proliferating cell nuclear antigen (PCNA), and plasma hepatocyte growth factor (HGF) level were determined, and standard liver function tests performed at 24, 48, and 72 h. Although all liver function tests improved in Group 1 at 24 and 48 h, the regeneration rate was significantly impaired. Similarly, the PCNA labeling index was significantly lower in Group 1 than that in Group 2. The plasma HGF level was significantly reduced in Group 1 (6 h blood out versus blood in: 1.1 ± 0.5 vs. 0.1 ± 0.1 ng/ml, p < 0.05). TBE with normal blood following PH suppressed the early stage of liver regeneration, in part, because of the reduction of HGF even though the blood was purified.     

S-腺苷蛋氨酸在肝硬变大鼠行部分肝叶切除后对肝功能的保护作用观察

目的　探讨S 腺苷蛋氨酸在肝硬变大鼠行部分肝叶切除后对肝细胞的再生及肝功能的影响。方法　用40 %四氯化碳诱导Wistar大鼠肝硬变模型 ,而后将成模大鼠随机分为肝硬变组 (n =2 0 )、治疗 1组〔S 腺苷蛋氨酸 10mg/ (kg·d) ,n =16〕及治疗 2组〔S 腺苷蛋氨酸 2 0mg/ (kg·d) ,n =16〕 ,另设正常对照组 (n =16)。 4组大鼠均行 3 0 %左右肝叶切除。自手术当天起 ,治疗 1、2组分别肌注S 腺苷蛋氨酸 10mg/ (kg·d)和 2 0mg/ (kg·d) ,对照组肌注生理盐水 5ml/d ,共 15d。 4组大鼠分别于术后 15d和 3 0d处死一半 ,取血检测Alb、ALT、TB、TBA及TNF α ,同时在光镜及电镜下观察肝组织的病理改变及超微结构变化。结果　肝硬变组术后 15d和 3 0d的TB、TBA、ALT及TNF α水平明显高于正常对照组 (P<0 .0 1) ,Alb水平明显低于正常对照组 (P<0 .0 1) ;治疗 1、2组术后 15d的上述 4项指标高于正常对照组 (P<0 .0 5 ) ,但明显低于肝硬变组 (P<0 .0 1) ,Alb水平低于正常对照组 (P<0 .0 5 ) ,却明显高于肝硬变组 (P<0 .0 1) ;治疗 1、2组术后 3 0d上述各指标均接近正常对照组水平。TNF α和TB、TBA、ALT呈显著正相关 (r值分别为 0 .99、0 .97、0 .93 ,P<0 .0 1) ,与Alb呈显著负相关 (r值为 -0 .88,P<0 .0 1)。治疗组光镜     

异位辅助性分肝移植供肝切取与植入体会

目的　为临床辅助性肝移植的开展,探索供肝切取及植入方法。　方法　猪异位辅助性部分肝移植13例,供肝切取按预分离方法分为A、B两组,A组标准法5例,B组快速灌注法8例。　结果　A组供肝质量优良为100%,B组供肝质量优良占88%。辅肝移植手术时间为122±28min,腔静脉和门静脉吻合时间为40±12min。　结论　如条件允许应首选标准法预分离,血流动力学不稳定时,可行快速灌注法预分离。     

间断低氧预适应对大鼠肝切除缺血再灌注损伤的保护作用

目的探讨间断低氧预适应对大鼠肝大部切除术后残余肝脏合并缺血再灌注引发过氧化损伤的保护作用。方法78只SD大鼠,用SPSS软件将其随机分为4组：假手术组（SO组,n=6）、肝切除组（PH组,n=24）、肝切除合并缺血再灌注损伤组（IR组,n=24）和间断低氧预适应组（IHP组,n=24）。以无创伤血管夹阻断IR组大鼠入肝血流后切除肝脏的左叶和中叶（约占全肝的70％）,20min后开放入肝血流,残余肝脏发生了缺血再灌注损伤。将IHP组大鼠暴露于10％的低氧环境中,每日持续1h,连续进行1周,最后1次低氧暴露后行肝切除术（同IR组）。SO组大鼠在术后2h取材检测,其余各组分别于术后2、6、12及24h进行检测。检测血清转氨酶（ALT、AST）水平和肝匀浆组织中超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量。结果术后2h,PH组、IR组和IHP组大鼠血清ALT和AST水平均高于SO组（尸〈0．05）。在术后6、12和24h,IHP组大鼠血清ALT和AST均高于PH组,但低于IR组（均P〈0．05）。与IR组相比,IHP组大鼠术后各时间点残余肝脏中SOD活性明显升高,而MDA含量则显著降低（均P〈0．05）。结论间断低氧预适应对残余肝脏缺血再灌注损伤具有保护作用,其机理可能与提高肝脏的抗氧化能力有关。     

缺血预处理对大鼠肝脏低温保存损伤的保护作用

目的　探讨缺血预处理 (IPC)对大鼠肝脏低温保存损伤的保护作用。方法　制备大鼠肝脏离体非循环灌注模型 ,对供肝分别作不同时间的IPC (IPC1组缺血 5min、IPC2 组缺血 10min、IPC3 组缺血 15min) ,而后比较各组供肝的损伤程度。结果　流出液中AST和ALT的水平 ,IPC1组分别为 (4 0 .1± 6.3 )U/L和 (17.1± 0 .5 )U /L ,IPC2 组分别为 (5 3 .6± 3 .7)U/L和 (19.7± 0 .5 )U /L ,均显著低于未预处理 (NPC)组的 (64 .5± 8.2 )U/L和 (2 3 .8± 3 .9)U /L (P<0 .0 5 ) ;IPC1组又显著低于IPC2 组和IPC3 组的 (63 .8± 7.2 )U/L和 (2 2 .8± 2 .5 )U /L (P<0 .0 5 )。LDH水平 ,NPC组、IPC1组、IPC2 组和IPC3 组分别为 (10 4.3± 2 0 .6)U/L、(84.1± 19.7)U /L、(90 .5± 2 1.1)U/L和 (10 3 .1± 18.5 )U /L ,4组间差异无统计学意义 (P>0 .0 5 ) ,但均高于正常组〔(71.5± 18.9)U /L〕 (P<0 .0 5 )。胆汁分泌量及肝组织ATP含量 ,IPC1组分别为 (5 3 .5± 10 .2 ) μl和 (6.15± 0 .65 ) μmol/g ,IPC2 组分别为 (4 1.5± 8.1) μl和 (4 .77± 0 .2 1) μmol/g ,均显著高于NPC组的 (2 2 .8± 9.7) μl和 (2 .62± 0 .3 4) μmol/g (P<0 .0 5 ) ;IPC1组又显著高于IPC2 组和IPC3 组的 (2 7.5± 2 .8) μl和 (2 .61     

Effect of Ischemic Preconditioning and Postconditioning on Liver Regeneration in Prepubertal Rats

Background

Liver regeneration has great importance for transplantation, especially in children; however, it has not been studied sufficiently in development animals. Ischemia-reperfusion injury is a problem, and strategies such as ischemic preconditioning and postconditioning are not well defined regarding regeneration.

Objective

This study sought to evaluate liver regeneration with modulation by ischemic preconditioning and postconditioning in prepubertal rats subjected to total ischemia and reperfusion.

Methods

Thirty-five 5-week-old female Wistar rats were divided into groups of 7 animals each: control group (SHAM), 70% hepatectomy (HEP), total ischemia 30 minutes before hepatectomy (IR), ischemic preconditioning 10/10 minutes before ischemia (PRE), and two 30/30-second ischemic postconditioning cycles after ischemia and hepatectomy (POS). All animals were subjected to 24-hour reperfusion. Aspartate aminotransferase and alanine aminotransferase activity were measured to evaluate liver damage, and histological analysis, proliferating cell nuclear antigen (PCNA) and regenerated mass liver were used to evaluate liver regeneration. Statistical analyses were performed using ANOVA and Kruskal-Wallis test.

Results

Alanine aminotransferase and aspartate aminotransferase levels were significantly lower in conditioned groups than in the IR group. Regarding mitotic index, IR > control group and HEP (P < .05), PRE and POS were not significantly different from IR, and POS > HEP (P < .05). PCNA analysis showed that IR > HEP (P < .01), PRE < IR (P < .01), and no significant differences were observed between POS and IR groups. No significant differences in regenerated mass liver were observed between conditioned groups and HEP.

Conclusions

Ischemic postconditioning prevented ischemic injury, promoted greater liver regeneration, and should be further investigated as an alternative better than ischemic preconditioning.     

重组人生长激素在治疗肝硬变大鼠肝部分切除术后低白蛋白血症中的作用

目的探讨重组人生长激素(rhGH)在治疗肝硬变大鼠肝部分切除术后低白蛋白血症中的作用。方法30只大鼠按编号法随机分为正常对照组、肝硬变组(LC组)、肝硬变切肝组(LCH组)、PN组(于肝叶切除术后行PN治疗)及rhGH PN组(于肝叶切除术后行rhGH PN治疗)．每组6只。检测大鼠肝功能、血糖．用RT-PCR法检测肝组织ALB mRNA的表达．肝组织行Ki67免疫组化染色。结果与PN组比较．rhGH PN组血清ALP降低，ALB、血糖升高．肝组织ALBmRNA表达水平及肝组织Ki67指数均升高。结论重组人生长激素可以改善肝硬变大鼠肝部分切除后低白蛋白血症。     

大鼠门静脉分支结扎后肝再生的实验研究

目的 采用手术方法结扎门静脉分支，观察对侧肝脏的再生状态。方法 健康Wistar雄性大鼠60只，随机均分成结扎组和假结扎组，在乙醚麻醉下行门静脉左支结扎和假结扎手术，分别在术后第1、2、3、7及14d用乙醚麻醉动物，心脏采血检测血清ALT值，取出肝脏后称重量；在光镜下观察肝组织的病理变化，并计数肝细胞核分裂指数；采用免疫组化的方法计数增殖细胞核抗原（PCNA）指数；在电镜下观察肝细胞超微结构变化。结果 ①血清ALT值结扎组与假结扎组比较在术后第1d升高（P〈0．01），但在第2d开始恢复正常；②肝脏重量两组间差异无统计学意义（P〉0．05）；③肝细胞核分裂指数结扎组与假结扎组比较，术后第1～3d明显升高，第2d达高峰，第3d有所下降，但仍高于假结扎组（P〈0．01），以后逐渐恢复正常；④PCNA阳性细胞计数结扎组与假结扎组比较，术后第1～3d明显增多，第2d达高峰，第3d有所减少，但仍高于假结扎组（P〈0．01），以后逐渐恢复正常。结论 ①大鼠门静脉左支结扎后，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重量；②大鼠75％肝叶的门静脉分支结扎不影响肝功能，是安全可行的；③大鼠门静脉分支结扎可以作为研究肝脏再生动物模型的方法。