ATP—MgC12对缺血后肝组织保护作用的研究

研究ＡＴＰ－ＭｇＣｌ２对缺血后肝组织的作用。方法：家兔全肝缺血３０分钟后，分别给予生理盐水、ＡＴＰ和ＡＴＰ－ＭｇＣｌ２处理。术一第１天、第３天和第５天取血测定谷丙转氨酶（ＡＬＴ０、乳酸脱氢酶（ＬＤＨ０和碱性磷酸酶（ＡＬＰ）并分别检测各 肝组织学的改变。结果：术后第１天ＡＴＰ－ＭｇＣｌ２组和ＡＬＴ明显低于对照组（Ｐ〈０．０５），而ＡＴＰ组与对照组相比无显著差别：ＡＴＰ－ＭｇＣｌ２及ＡＴＰ组的ＬＤＨ和     

三磷酸腺苷氯化镁对兔缺血后肝组织保护作用的研究

家兔全肝缺血３０分钟后，分别给予生理盐水、三磷酸腺苷（ＡＴＰ）和三磷酸腺苷氯化镁（ＡＴＰ－ＭｇＣｌ２），术后第１，３，５天取血测定谷丙转氨酶（ＡＬＴ）、乳酸脱氢酶（ＬＤＨ）和碱性磷酸酶（ＡＬＰ）。术后第１天ＡＴＰ－ＭｇＣｌ２组ＡＬＴ明显低于对照组（Ｐ＜０．０５），而ＡＴＰ组与对照组相比无显著差别；ＡＴＰ－ＭｇＣｌ２及ＡＴＰ组的ＬＤＨ和ＡＬＰ与对照组相比无显著差别。术后２４小时超微结构示：对照组肝明显水肿、细胞器严重损伤和血窦内微小血栓；而ＡＴＰ－ＭｇＣｌ２组细胞轻度水肿，细胞器基本正常，血窦内无微小血栓。实验结果说明ＡＴＰ－ＭｇＣｌ２对缺血肝组织确有保护作用，其机制可能是通过改善微循环和减轻细胞而起作用的     

ATP-MgCl_2对烫伤大鼠肠粘膜保护作用的实验观察

为研究 ATP-MgCl_2对创伤应激动物肠粘膜的保护作用,采用30%TBSAⅢ度烫伤大鼠模型,观察了腹腔注射 ATP-MgCl_2复合液对回肠粘膜丙二醛(MDA)含量及回肠组织形态学改变的影响。结果显示:烫伤后6～24小时,MDA 含量明显升高,并伴有形态学病理改变。经 ATP-MgCl_2注射治疗的大鼠伤后24小时内 MDA 含量维持于正常对照水平,且回肠粘膜病理改变减轻。提示:ATP-MgCl_2能够保护烫伤大鼠肠粘膜,其作用机理可能与降低肠粘膜的脂质过氧化反应有关。     

肝动脉缺血对犬自体移植肝和胆管超微结构的影响

采用自制的简易狗自体肝移植模型，观察肝动脉缺血（ＨＡＩ）对移植肝和胆管超微结构的影响。结果发现：灌注后肝、胆细胞轻度水肿，线粒体基质疏松，嵴较模糊；ＨＡＩ３小时，肝、胆细胞水肿明显加重，胞质疏松，线粒体扩大，嵴断裂或消失，部分空泡变异出现絮状电子致密斑，内质网明显扩张，核糖体解聚。表明ＨＡＩ对供肝及胆管细胞有明显损伤作用。提示肝脏移植后ＨＡＩ是某些并发症，特别是胆道并发症的重要致病因素之一。     

门静脉、肝动脉同时灌注对供肝动脉缺血损伤的保护作用

目的探讨肝移植门静脉再灌注过程中肝动脉缺血(hepaticarteryischemia,HAI)损伤的严重性和应用肝动脉桥式置管(hepaticarterybridge-conduit,HABC)技术实现门静脉、肝动脉同时灌注对这一损伤的保护作用。方法32只犬按随机数字表法随机均分为4组:正常对照组、HAI30min组、HAI2h组和HABC组。后三组分别建立犬自体原位肝脏移植模型,HABC组应用HABC技术使肝动脉、门静脉同时再灌注。术后取供肝组织与胆管组织电镜观察肝细胞和胆管上皮细胞病理学改变。分别应用硫代巴比妥酸法检测肝组织中丙二醛(MDA)浓度、黄嘌呤氧化酶法检测过氧化物歧化酶(SOD)活性、酶标记法检测肝细胞线粒体琥珀酸脱氢酶(SDH)活性。结果HAI30min即可见供肝肝细胞和胆管上皮细胞水肿、线粒体嵴减少,HAI2h其病理改变进一步加重,胆管上皮尤为明显,而HABC组则见肝细胞和胆管上皮细胞比较完整,胆管上皮绒毛丰富。HAI30min组和HAI2h组肝组织中MDA含量增加,分别为(1.652±0.222)nmol/mgprot和(2.379±0.526)nmol/mgprot,而SOD则降低至(11.15±3.9)U/mgprot和(9.47±3.4)U/mgprot,SDH活性则分别降低至0.362±0.019和0.281±0.029,与正常对照组比较差异有统计学意义(P<0.05,P<0.01);而HABC组MDA含量、SOD和SDH活性与正常对照组比较差异无统计学意义(P>0.05)。结论HABC技术的实施,可为临床肝移植中预防HAI损伤,减少肝移植术后并发症,特别是胆道并发症的发生提供有效的方法。     

缺血后处理对肝脏缺血再灌注中肝窦内皮细胞损伤的保护作用

目的探讨缺血后处理对肝脏缺血再灌注中肝窦内皮细胞损伤的保护作用.方法建立大鼠局部肝脏缺血再灌注模型,将24只健康雄性Wistar大鼠随机分为假手术、缺血再灌注、缺血后处理3组,以缺血再灌前、反复多次的短暂预再灌、停灌作后处理,观察各组血浆肝酶及透明质酸(HA)水平变化和肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、内皮素-1(ET-1)含量,并行肝组织病理形态学检查.结果与缺血再灌注组相比,缺血后处理组肝酶的漏出、血浆HA水平及肝组织中MDA、ET-1的含量明显降低(P＜0.01),而SOD活性则显著升高(P＜0.01),肝组织病理学损伤亦明显减轻.结论缺血后处理可通过抑制再灌注后氧自由基的过量生成而保护肝窦内皮细胞,减轻肝脏缺血再灌注损伤.     

肝动脉缺血对犬自体移植肝和胆管超微结构的影响

采用自制的简易狗自体肝移植模型，观察肝动脉缺血(HAI)对移植肝和胆管超微结构的影响。结果发现：灌注后肝、胆细胞轻度水肿，线粒体基质疏松，嵴较模糊；HAI3小时，肝、胆细胞水肿明显加重，胞质疏松，线粒体扩大，嵴断裂或消失，部分空泡变异出现絮状电子致密斑，内质同明显扩张，核糖体解聚。表明HAI对供肝及胆管细胞有明显损伤作用。提示肝脏移植后HAI是某些并发症，特别是胆道并发症的重要致病因素之一。     

缺血预处理在肝门阻断肝切除中的作用

作者研究2 0 0 1年1月至2 0 0 2年1 2月因各种肝脏病变(包括良性和恶性病变)行肝门阻断、规则性肝切除病例,其中2 1例在持续性肝门阻断肝切除前,先行1 0 m in的肝门阻断后复流1 0 min,称为缺血预处理组;选择同期在年龄、性别、原发病、肝切除体积方面与缺血预处理组匹配,无缺血预处理而直接行肝门阻断肝切除的2 1例病例作为对照。两组患者肝脏均无肝硬化,手术方式基本相同。缺血预处理组的手术时间为31 2±92 m in,持续性肝门阻断时间为5 4±1 9min,其中7例超过6 0 min;对照组手术时间为339±1 1 2min,持续性肝门阻断时间为36±1 4 m in,其…     

小鼠肝缺血再灌注后Toll样受体2在缺血肝组织中的激活及其意义

目的　探索小鼠肝缺血再灌注后缺血肝组织中Toll样受体 2 (TLR2 )的激活及其与肝功能损伤之间的关系。方法　缺血再灌注损伤组 (I/R组 ),假手术对照组 (S组 )均采用实时荧光定量多聚酶链反应检测肝组织中TLR2mRNA及TLR2蛋白的表达,同时检测门静脉血浆丙氨酸氨基转移酶(ALT)、肿瘤坏死因子α(TNF α)及门静脉血清内毒素 (endotoxin, EN)水平。结果　肝脏部分缺血1h再灌注 4h后,I/R组与S组小鼠缺血肝组织TLR2 mRNA的表达 (ΔCt值 )分别为 1. 0 6±0. 9 1和5. 0 8±1. 3 2, 两组间差异有显著性 (P < 0. 0 1 ),I/R组缺血肝组织TLR2 蛋白的表达 (OD值 )( 4 3 3. 9 1±2 5. 5 3 )水平较S组 ( 1 0 2. 8 6±1 3. 5 8 )显著升高 (P< 0. 0 1 )。I/R组门静脉血清TNF α[ ( 1 1 2. 5 2±1 4. 4 1 )pg/mL]较S组 [ ( 5. 9 6 ±4. 4 3 )pg/mL]显著升高 (P < 0. 0 1 );I/R组ALT[ ( 8 4 8. 3 3±2 7 1. 3 7 )U/L]较S组 [ ( 4 2. 3 9±1 4. 7 5 )U/L]显著升高 (P < 0. 0 1 );而门静脉血清内毒素水平组间差异无显著性 (P> 0. 0 5 )。结论　TLR2mRNA及蛋白在肝脏缺血再灌注过程中缺血肝组织的表达增强, 此变化伴有TNF α的升高及肝功能的损伤。     

亚低温对肝缺血预处理保护作用的增强效应

目的探讨肝缺血再灌注损伤中亚低温(MH)对缺血预处理(IP)保护作用的增强效应机理.方法观察非缺血对照组、缺血再灌注组、缺血预处理组和亚低温缺血预处理组4组犬肝上下腔静脉血谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)以及丙二醛(MDA)和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化酶(GSH-PX)活性及总抗氧化(TAX)能力变化. 结果肝缺血再灌注后ALT、AST、LDH和MDA含量明显上升(P<0.01), SOD、CAT、GSH-PX活性及TAX能力明显下降(P<0.01); 缺血预处理组与缺血再灌注组比较及亚低温缺血预处理组与缺血预处理组比较,ALT、AST、LDH和MDA含量均明显下降(P<0.01,P<0.05),SOD、CAT、GSH-PX活性及TAX能力明显上升(P<0.01,P<0.05).结论缺血预处理能增强肝组织自身抗氧化能力,减轻肝缺血再灌注后氧自由基对肝脏的损伤; 亚低温对缺血预处理的这种保护作用具有增强效应.     

肝移植对肝硬变大鼠脾脏功能的影响

目的探讨肝移植对肝硬变大鼠脾脏功能的影响。方法制备四氯化碳中毒性肝硬变大鼠模型,采用“二袖套”法进行肝移植。观察肝移植前、后大鼠脾指数及脾脏组织形态学的变化;应用免疫荧光染色技术及流式细胞仪检测肝移植前、后大鼠脾脏T淋巴细胞亚群的变化。结果肝移植前,肝硬变大鼠脾指数增高,从(2.42±0.11)mg/g增至(3.62±0.14)mg/g,P<0.01;脾脏白髓面积缩小,从(23.47±2.30)%缩至(7.70±2.01)%,P<0.01;脾小梁面积扩大,从(1.75±0.61)%增加至(4.46±0.71)%,P<0.01;脾脏T淋巴细胞亚群CD4/CD8比值明显下降,从2.67±0.15降至1.18±0.15,P<0.01。肝移植后,随着时间的延长,大鼠脾脏白髓面积有所扩大,从(7.70±2.01)%增至(15.07±1.97)%,P<0.01;脾小梁面积有所缩小,从(4.46±0.71)%降至(3.11±0.51)%,P<0.05;脾指数逐渐降低,从(3.62±0.14)mg/g降至(2.62±0.11)mg/g,P<0.01;脾脏T淋巴细胞亚群中CD4/CD8比值也有所升高,从1.18±0.15升至2.32±0.11,P<0.01。结论肝硬变大鼠行肝移植后,因肝功能严重损害而导致的脾脏功能异常可得到改善。     

肝细胞凋亡与肝移植中热缺血损伤关系的实验研究

目的：探讨肝细胞凋亡是否参与肝移植中热缺血再灌注这一病理损伤过程。方法：实验以大鼠供肝获取前供体所经历的心脏停搏时间0、30和60min将实验动物分为3组：HB组、NHBD－30组和NHBD－60组，而后行大鼠原位肝移植，每组各20对。于术后1、3、6和24h处死取材。移植肝标本采用HE染色、TUNEL技术和透射电镜进行观察，并采集血样以测定TNF－α。结果：HB组几乎未出现肝细胞凋亡。移植术后3－6h，NHBD－30组出现了少量肝细胞凋亡，而NHBD－60组则出现了大量肝细胞凋亡。移植术后24h，NHBD－30组和NHBD－60组则分别出现了局灶性和大片肝细胞坏死。随着供肝遭遇的热缺血时间不断延长，术后TNF－α呈上升趋势，在术后3h达到峰值。结论：肝细胞凋亡参与肝移植中热缺血再灌注损伤，肝细胞凋亡可能是坏死的原因，并且肝细胞凋亡与TNF－α表达关系密切。     

Quantitative Assessment of Hepatic Function and its Relevance to the Liver Surgeon

Background  Standard evaluation of patients undergoing hepatic surgery has been through radiological and quantitative determination of liver function. As more complex and extensive surgery is now being performed, often in the presence of cirrhosis/fibrosis or following administration of chemotherapy, it is questioned whether additional assessment may be required prior to embarking on such surgery. The aim of this review was to determine the current knowledge base in relation to the performance of quantitative assessment of hepatic function both pre- and post-operatively in patients undergoing hepatic resectional surgery and liver transplantation. Methods  An electronic search was performed of the medical literature using the MEDLINE database to identify relevant articles with cross-referencing of all identified papers to ensure full literature capture. Results and Conclusions  The review has identified a number of different methods of dynamically assessing hepatic function, the most frequently performed being through the use of indocyanine green clearance. With the recent and further anticipated developments in hepatic resectional surgery, it is likely that quantitative assessment will become more widely practiced in order to reduce post-operative hepatic failure and improve outcome.     

Protective Effect of MESNA (2-Mercaptoethane Sulfonate) Against Hepatic Ischemia/Reperfusion Injury in Rats

Purpose Reoxygenation of ischemic tissue generates various reactive oxygen metabolites (ROMs), which have a deleterious effect on various cellular functions. We evaluated the possible protective effect of 2-mercaptoethane sulfonate (MESNA) on hepatic ischemia/reperfusion (I/R) injury.Methods Wistar albino rats were subjected to 45-min hepatic ischemia, followed by 60-min reperfusion. 2-Mercaptoethane sulfonate, 150 mg/kg, or saline was given intraperitoneally (i.p.) twice, 15 min before ischemia and immediately before reperfusioin. We measured serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels to assess liver function. Liver tissue samples were taken to measure the levels of malondialdehyde (MDA), an end-product of lipid peroxidation; glutathione (GSH), a key antioxidant; and myeloperoxidase (MPO) activity, as an indirect index of neutrophil infiltration. We also measured hepatic collagen content, as a fibrosis marker.Results Plasma ALT and AST levels were higher in the I/R group than in the control group, but this increase was significantly decreased by MESNA treatment. Hepatic GSH levels, which were significantly depressed by I/R, increased back to the control levels in the MESNA-treated I/R group. Increases in tissue MDA levels and MPO activity caused by I/R injury decreased back to the control levels after MESNA treatment. Similarly, the increased hepatic collagen content in the I/R group decreased to the level of the control group after MESNA treatment.Conclusion The fact that MESNA alleviated I/R-induced injury of the liver and improved hepatic structure and function suggests that its antioxidant and oxidant scavenging properties may be of therapeutic value in protecting the liver against oxidative injury caused by I/R.     

Remote Renal Injury Following Partial Hepatic Ischemia/Reperfusion Injury in Rats

Liver ischemia/reperfusion has been shown to result in injury of remote organs such as the heart and lungs. Whether or not acute liver injury also results in kidney injury has so far not been adequately addressed. In anesthetized Wistar rats, partial (70%) normothermic hepatic ischemia was applied for 75 min. After 24 h of reperfusion, renal injury was assessed by histology, creatinine and blood urea nitrogen (BUN) serum concentrations, renal expression of proinflammatory genes [quantitative real-time polymerase chain reaction (qRT-PCR)], caspase-3 activation (Western blot), and neutrophil accumulation (myeloperoxidase assay). Twenty-four hours after hepatic ischemia, creatinine (0.57 ± 0.06 vs. 0.32 ± 0.04 mg/dL) and BUN (40.7 ± 15.3 vs. 14.3 ± 2.0 mg/dL) were increased when compared to sham. qRT-PCR revealed higher renal intercellular adhesion molecule-1 gene expression following hepatic ischemia (166 ± 45% when compared to sham) but no differences in renal monocyte chemoattractant protein-1, macrophage inflammatory protein-2, and inducible NO synthase expression. In both groups, kidneys showed no morphological damage and no increase in caspase-3 and myeloperoxidase activity. Severe hepatic ischemia results in a moderate impairment of renal function in rats but does not trigger an inflammatory response in the kidney and does not result in morphological damage of the kidney. A part of this study has been presented as a poster presentation at the 2006 Congress of the American Hepato-Pancreato-Biliary Association in Miami Beach, FL, March 9–12.     

缺血后处理对兔肝脏热缺血再灌注损伤的保护

目的探讨缺血后处理对肝脏热缺血再灌注损伤的保护作用。方法阻断肝十二指肠韧带建立兔肝脏热缺血再灌注模型,将30只健康新西兰大耳白兔随机分为假手术(S组)、缺血再灌注(IR组)、缺血后处理(IP组)3组,每组10只。S组只分离肝十二指肠韧带,不阻断;IR组分离并阻断肝十二指肠25min后再恢复灌注;IP组分离并阻断肝十二指肠韧带,于肝脏缺血25min后恢复血灌前阻断、复灌各1min共3个循环进行缺血后处理。观察麻醉诱导后20min(T0)、阻断肝十二指肠韧带25min(T1)、开放30min(T2)、开放1h(T3)、开放2h(T4)、开放4h(T5)血流动力学、谷丙转氨酶(ALT)变化,并检测肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)变化。结果与IR组比较,IP组T2时HR,T2、T5时SBP及T5时DBP较IR组高;复灌后(T2～T5)血浆ALT明显降低,肝组织中MDA明显下降,而SOD明显升高(P<0.05)。结论缺血后处理可减轻肝脏缺血再灌注损伤。     

Effect of Leptin and Apelin Preconditioning on Hepatic Ischemia Reperfusion Injury in Rats

Leptin and apelin are important adipocytokines involved in a variety of endocrine and paracrine functions. The aim of this study was to evaluate the effect of exogenous leptin and apelin preconditioning on hepatic ischemia reperfusion (I/R) injury in rats. Forty mice were assigned to four groups (n = 10): sham-operated control (sham), I/R injury, I/R + leptin (I/R + L), and I/R + apelin (I/R + A). Leptin 100 μg/kg/day and apelin 2 μg/kg/day were delivered intraperitoneally starting 3 days prior to surgical procedure in I/R + L and I/R + A groups, respectively. All I/R groups underwent 45 min of warm ischemia, followed by 30 min of reperfusion. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), liver malondialdehyde (MDA) and glutathione (GSH), and liver histopathology were compared between groups. MDA was elevated in I/R, but stayed similar in I/R + L and I/R + A compared to sham. I/R + A had significantly lower MDA compared to I/R. GSH levels did not differ significantly between the groups. ALT and AST were elevated in all I/R groups, but significant reduction was observed in I/R + L and I/R + A compared to I/R. Liver histopathology was mostly mild in I/R + L and I/R + A, in contrast to severe injury observed in the I/R group. Leptin and apelin preconditioning significantly reduced hepatic I/R injury in rats.     

Effect of Hepatic Artery Flow on Bile Secretory Function After Cold Ischemia

These studies evaluated the influence of hepatic arterial flow on biliary secretion after cold ischemia. Preparation of livers for transplantation or hepatic support impairs biliary secretion. The earliest indication of cold preservation injury during reperfusion is circulatory function. Arterial flow at this time may be critical for bile secretion. Porcine livers were isolated, maintained at 4 degrees for 2 h and connected in an extracorporeal circuit to an anesthetized normal pig. The extracorporeal livers were perfused either by both the hepatic artery and portal vein (dual) or by the portal vein alone (single). Incremental doses of sodium taurocholate were infused into the portal vein of both the dual and single perfused livers, and the bile secretion was compared. Most endogenous bile acids are lost during hepatic isolation. After supplementation, the biliary secretion of phosphatidyl choline and cholesterol was significantly better in the dual than single vessel-perfused livers; however, no difference was seen in bilirubin output. Single perfused livers were completely unable to increase biliary cholesterol in response to bile acid. The dependence of bile cholesterol secretion on arterial flow indicates the importance of this flow to the detoxification of compounds dependent on phosphatidyl choline transport during early transplantation.     

携带IL-13基因肝干细胞对冷缺血大鼠移植肝脏保护作用的研究

目的以基因工程干细胞治疗的方式来改善缺血-再灌注状况,使移植肝脏更好地耐受缺血-再灌注,以减少移植术后的并发症,延长移植器官存活期。方法利用已建立的Wistar大鼠冷缺血原位肝移植(OLT)模型,术中经受体大鼠门静脉输入IL-13基因修饰的或未修饰的肝卵圆细胞(HOC)即转染组和未转染组,分别于术后1、3、7及14d检测受体大鼠肝脏功能变化、组织学变化、胆管上皮细胞(BEC)的增殖情况、α-平滑肌肌动蛋白(α-SMA)的表达、肝组织中HO-1 mRNA的表达,并观察移植大鼠术后的存活情况。结果转入IL-13基因的HOC对冷保存损伤的肝脏有一定的保护作用;术后7d,肝移植组和未转染组的α-SMA表达较转染组明显(P0.05);术后3d,未转染组和转染组的BEC增殖指数明显低于肝移植组(P0.05)。转染组术后各时相点的HO-1 mRNA表达水平均明显高于相应时相点的其他各组的水平(P0.05);移植术中经门静脉输入HOC对缺血性胆管损伤所诱导的BEC增殖有一定的抑制作用,对BEC具有一定的保护作用;肝移植组生存率明显低于假手术组和转染组(P0.05)。结论IL-13的持续表达能促进术后移植肝内HO-1 mRNA的表达,这有利于对供肝的保护,有利于受体大鼠术后肝功能的恢复。促进HO-1 mRNA的表达是IL-13对BEC的具有保护作用的机理之一。