肝脏缺血再灌注损伤的临床与基础研究进展

缺血再灌注损伤(IRI)是指器官在缺血的基础上恢复血流后损伤进一步加重甚至发生不可逆性损伤的现象。在临床上IRI常见于肝、脑和肾等实质性器官。其中, 肝脏缺血再灌注损伤(HIRI)是肝脏部分切除术和肝移植后肝功能受损甚至肝衰竭的主要原因之一, 其与患者的预后密切相关。HIRI的发生发展涉及多种机制, 而氧自由基的积聚和炎症的改变是主要的病理生理过程。此外, HIRI与"双刃剑"补体联系紧密。本文主要论述HIRI的基础和临床两方面研究进展, 为HIRI的防治提供理论和实践的参考。     

肝脏非实质细胞在肝脏缺血再灌注损伤中作用的研究进展

目的总结肝脏非实质细胞与肝脏缺血再灌注损伤(HIRI)之间的关系。方法复习近年来关于肝脏非实质细胞与HIRI关系的研究并进行综述。结果 HIRI过程中的无菌性炎症反应、细胞凋亡等可以对肝细胞造成严重的损伤。肝脏非实质细胞可以释放多种细胞因子与炎症介质促进损伤的加剧,包括Kupffer细胞、肝窦内皮细胞、肝脏星状细胞和树突状细胞,其中部分细胞同时还具有减轻损伤的功能:如Kupffer细胞可以通过表达血红素加氧酶-1而减轻HIRI,肝星状细胞可能参与HIRI损伤的修复过程。肝脏非实质细胞在HIRI中的作用十分复杂,但其同时也具有潜在的治疗应用价值。结论肝脏非实质细胞能够通过多种机制影响HIRI,为临床上减轻HIRI提供了新的目标和策略。     

中药预处理保护肝脏缺血再灌注损伤的研究进展

肝脏缺血再灌注损伤(HIRI)是由多种因素共同作用所引起的复杂的病理生理过程,是肝脏外科中重点关注的一个问题。减轻缺血再灌注损伤(IRI)将有助于肝病手术患者预后。目前临床普遍通过药物预处理来增强肝组织或肝细胞对IRI的耐受,并得到了较理想的效果。本文通过分析总结已证实的HIRI的发生机制,对目前用于干预治疗HIRI的中药制剂作一简单综述,希望能对临床提供一定的参考。     

血红素加氧酶-1与自噬在肝脏缺血-再灌注损伤中的研究进展

血红素加氧酶(HO)-1及其酶解产物可通过减轻活性氧自由基损伤、减少细胞凋亡、抑制炎症反应和维持微循环稳定等作用减轻肝脏缺血-再灌注损伤(HIRI)。自噬是细胞利用溶酶体对自身受损细胞器和大分子物质进行生物学降解的过程,被认为是在应激情况下减轻细胞损伤的适应性反应,可有效减少细胞死亡。近年来越来越多的研究表明自噬与HIRI也有着密切的关系。本文回顾了近年来国内外的研究报道,从HO-1与HIRI、自噬与HIRI、HO-1与自噬在HIRI中的相互作用及其机制等方面进行综述。     

程序性细胞死亡在肝脏缺血-再灌注损伤中的作用研究进展

肝移植是治疗终末期肝病的有效手段,但在肝移植过程中不可避免会发生肝脏缺血-再灌注损伤（HIRI）,可能导致早期移植物功能障碍或加剧排斥反应,其损伤防护机制有待深入研究。程序性细胞死亡是HIRI的重要发生机制,多种新型程序性细胞死亡形式参与了HIRI的病理过程,深入研究程序性细胞死亡有望进一步提高肝移植的治疗效果。本文就细胞凋亡、自噬及自噬依赖性死亡、铁死亡、坏死性凋亡、细胞焦亡、多聚二磷酸腺苷核糖聚合酶（PARP）-1依赖性细胞死亡等常见的程序性细胞死亡方式在HIRI中的研究进展予以综述,以期为提高肝移植手术成功率、改善受者预后提供参考。     

预处理对缺血再灌注肝细胞自噬的影响

大型肝脏手术及肝移植手术时常伴随肝缺血再灌注损伤(hepatic ischemia reperfusion injury, HIRI), 其与围手术期病死率、术后并发症等密切相关。既往研究均已证实预处理可通过调节体内肝细胞自噬水平来减轻HIRI。文章对近年来国内外通过调节肝细胞自噬水平减轻HIRI的预处理方式及其简要的生物学机制进行综述, 为临床上采用合理的预处理方式以减轻HIRI及进一步研究提供参考。     

牛磺熊去氧胆酸对大鼠肝脏缺血再灌注损伤的保护作用

目的：探讨牛磺熊去氧胆酸（TUDCA）抗大鼠肝脏缺血再灌注（HIRI）损伤的作用及机制。 方法：将20只雄性SD大鼠随机均分为假手术组、TUDCA组、HIRI组、TUDCA+HIRI组,分别行假手术、TUDCA+假手术、HIRI造模,TUDCA+HIRI造模;TUDCA（250 mg/kg）于术前1 h灌胃给予,HIRI造模采用Pringle法（缺血60 min,再灌注12 h）。再灌注12 h后处死各组大鼠取材,观察肝组织病理学改变,检测血清谷丙转氨酶（ALT）水平,TUNEL法检测肝细胞凋亡,Western blot技术检测肝组织内质网应激分子糖调节蛋白78（GRP78）、p-真核细胞翻译起始因子2α（p-eIF2α）和C-EBP同源蛋白（CHOP）的表达。 结果：除假手术组与TUDCA组外,HIRI组与TUDCA+HIRI组大鼠肝组织均出现明显肝损伤病理改变,但TUDCA+HIRI组的损伤程度明显轻于HIRI组。与假手术组比较,HIRI组与TUDCA+HIRI组大鼠血清ALT水平明显升高,肝细胞凋亡和内质网应激分子GRP78、p-eIF2a和CHOP蛋白水平均明显升高（均P<0.05）,但TUDCA+HIRI组各项指标升高幅度均明显低于HIRI组（均P<0.05）;TUDCA组各项指标未见改变（均P>0.05）。 结论：TUDCA有抗大鼠肝HIRI的作用,其机制可能与抑制内质网应激反应有关。     

异甘草酸镁对大鼠肝脏缺血再灌注损伤治疗作用研究

肝脏缺血再灌注损伤(hepatic ischemia reperfusion injury,HIRI)常发生于肝脏手术、肝移植及介入治疗时,有效控制HIRI对手术成功率和预后具有重要意义.异甘草酸镁(magnesium isoglycyrrhizinate,MgIG)具抗炎、保护肝细胞膜及改善肝功能的作用,是一种广泛应用的肝细胞保护剂.本实验研究异MgIG对大鼠HIRI的治疗作用,并初步探讨其可能的机制.     

白术多糖预处理对大鼠肝脏缺血再灌注损伤的实验研究

目的:研究白术多糖(AMP)对大鼠肝脏缺血再灌注损伤的影响及作用机制。方法:将72只成年雄性SD大鼠随机分成3组:假手术组(Sham组)、缺血再灌注组(HIRI组)、AMP预处理缺血再灌注组(AMP组)。缺血60 min后分别再灌注1、6、24 h后取材,测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)及一氧化氮(NO)、内皮素(ET-1)水平,光镜及透射电镜下观察各组肝细胞的显微结构及超微结构变化。结果:AMP组及HIRI组ALT、AST水平均高于Sham组;与HIRI组比较,AMP组ALT、AST水平显著降低(P<0.05)。与Sham组比较,HIRI组及AMP组术后各时段的NO水平均明显降低,ET-1水平明显升高,术后6 h时明显(P<0.05);与HIRI组相比较,AMP组术后各时段NO水平升高,而ET-1水平降低,差异有统计学意义(P<0.05)。光镜下HIRI组大鼠肝细胞肿胀、变性坏死,肝血窦变窄、淤血及炎性细胞浸润等,而AMP组明显好转。电镜下HIRI组大鼠肝细胞线粒体细胞核皱缩变形,染色质粗糙,核仁浓缩甚至裂解,部分膜破裂,线粒体嵴疏松溶解等,而AMP组明显好转。结论:AMP可以提高缺血再灌注大鼠体内NO水平,同时降低ET-1水平,改善肝脏微循环障碍,对肝脏起保护作用。     

落新妇苷对大鼠肝脏缺血再灌注损伤及自噬的影响

目的探讨落新妇苷对大鼠肝脏缺血再灌注损伤（HIRI）的保护作用及自噬的影响。 方法将54只SD大鼠随机分为假手术组（Sham组）、缺血再灌注组（HIRI组）以及落新妇苷组（40 mg·kg^-1·d^-1,连续7 d）,每组18只。建立大鼠HIRI模型,于再灌注4、8、16 h后取下腔静脉血及肝左外叶组织。全自动生化分析仪检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）水平;光学显微镜下观察肝细胞显微结构变化;Western blotting分析肝组织中自噬相关蛋白LC3、Beclin-1的表达;电镜下观察肝脏组织内自噬小体数量情况。 结果同检测时间点比较,Sham组和落新妇苷组的血清ALT和AST水平均低于HIRI组（P<0.05）,且肝细胞肿胀、炎性细胞浸润、汇管区结构损伤明显较轻。落新妇苷组LC3-Ⅱ/ LC3-Ⅰ灰度值比值及Beclin-1蛋白灰度值在术后4、8、16 h明显低于HIRI组（P<0.05）。落新妇苷组肝组织的自噬小体数量较HIRI组有所减少（3.68±0.42 vs 7.12±0.60,t=36.382,P<0.01）。 结论落新妇苷预处理可减轻大鼠HIRI,其作用机制可能与其抑制自噬有关。     

Ghrelin ameliorates transformation of hepatic ischemia-reperfusion injury to liver fibrosis by blocking Smad and ERK signalling pathways,and promoting anti-inflammation and anti-oxidation effects

Background & aimsGhrelin, a gut hormone with pleiotropic effects, may act as a protective signal in parenchymal cells. Hepatic ischemia-reperfusion injury (HIRI) causes acute-on-chronic liver failure and induces transformation of acute to chronic injury. HIRI model of mice was established by a semi-hepatic blocking method and treated with Ghrelin. This process is involved in inflammation, oxidative stress damage and apoptosis, and is associated with the expansion and activation of fibrotic haematopoietic stem cells (HSCs) which express and secrete high levels of collagen that induces liver fibrosis. Therefore, we investigated the effects of Ghrelin during transformation of HIRI to liver fibrosis, and explored the molecular mechanism of Ghrelin's action based on Smad and ERK pathways.MethodsHepatic injury was detected by plasma ALT levels. The hepatic histology and collagen were elucidated by HE staining and Masson staining, respectively. Liver inflammation levels and inflammatory cell counts were assessed by MPO and HE staining, respectively. The antioxidant capacity of plasma was evaluated based on the levels of SOD, MDA, and XOD. The mRNA or protein expression levels of genes related to apoptosis, fibrosis, Smad, and ERK pathways were assessed by real-time quantitative PCR (RT-qPCR), ELISA, or western blotting.ResultsThe HIRI model was established to investigate the effects of the liver injury transformed to liver fibrosis. Ghrelin exhibited good hepatic protection by ameliorating liver histological changes and decreasing plasma ALT levels. Ghrelin significantly decreased the expression of MPO than that in model group, suggesting that Ghrelin blocked the inflammatory response in the HIRI liver tissue; this supports the anti-inflammatory effects of Ghrelin. Ghrelin significantly decreased apoptosis (enhanced Bcl-2 expression, and down-regulated Bax and Caspase 3). Ghrelin exhibited anti-oxidative effects as it inhibited plasma MDA levels, and promoted plasma SOD and XOD levels. Moreover, Ghrelin inhibited activation of hepatic stellate cells, blocked traditional fibrotic Smad and ERK signalling pathways, and reduced hepatic fibrosis by stimulating degradation of extracellular matrices (ECMs; such as collagen I, collagen III, HA, and LN).ConclusionsThis study demonstrates that Ghrelin delays the transformation of HIRI to liver fibrosis process which is correlated to its anti-apoptotic, anti-inflammatory, and anti-oxidative effects. Moreover, Ghrelin alleviates HIRI-mediated liver fibrosis, inhibits activation of HSCs, and reduces accumulation of ECM via inhibition of Smad and ERK signalling pathways.     

Activation of GPR81 Aggravates Remote Organ Injury During Hepatic Ischemia-Reperfusion Injury

Hepatic ischemia-reperfusion injury (HIRI) is a serious situation with high morbidity and mortality, which is usually accompanied with hyperlactatemia due to impaired lactate clearance in liver. G-protein-coupled receptor 81 (GPR81) has recently been identified as the bioactive receptor of lactate. GPR81 is profoundly involved in the modulation of metabolism and inflammation, but its significance in HIRI remains unclear. The present study investigated the potential roles of GPR81 in HIRI by using the GPR81 agonist 3-chloro-5-hydroxybenzoic acid (CHBA). The results indicated that treatment with CHBA had no obvious effects on HIRI-induced histologic abnormalities and elevation of serum aspartate aminotransferase, alanine aminotransferase. However, CHBA significantly upregulated the serum level of tumor necrosis factor alpha and interleukin-6 in mice with HIRI. Administration of CHBA also exacerbated HIRI-induced histologic lesions in lung, increased the level of myeloperoxidase in lung tissue and the protein concentration in bronchoalveolar lavage fluid. In addition, the serum levels of brain natriuretic peptide and creatinine also increased in CHBA-treated mice. The results indicate that activation of GPR81 might aggravate HIRI-induced remote organ injury and result in serious outcomes.     

黄芪甲苷对大鼠肝脏缺血再灌注损伤和Caspase途径的影响

目的探讨黄芪甲苷对大鼠肝脏缺血再灌注损伤（HIRI）的保护作用以及相关机制。 方法将75只纯系SD大鼠随机分为Sham组（假手术组）、HIRI组（缺血再灌注组）以及黄芪甲苷低剂量组、中剂量组、高剂量组,建立大鼠HIRI模型,于再灌注4、8、16 h后取下腔静脉血及肝左外叶组织。全自动生化分析仪检测血清谷丙转氨酶（ALT）、谷草转氨酶（AST）水平;光学显微镜下观察肝细胞显微结构变化;Western Blotting及免疫组化染色分析肝组织中Caspase-3、8、9的表达;流式细胞仪检测肝细胞凋亡率。 结果HIRI组的ALT和AST水平升高,而黄芪甲苷组明显降低,且肝细胞损伤明显减轻。高剂量组Caspase-3、8、9蛋白的表达水平以及细胞凋亡率明显低于HIRI组。 结论黄芪甲苷预处理可减轻大鼠HIRI,其作用机制可能与其抑制Caspase-3、8、9的表达,从而抑制细胞凋亡有关。     

Hepatocellular Protein Profiles After Hepatic Ischemia/Reperfusion Injury With or Without Octreotide Preconditioning in a Rabbit Model

Hepatic ischemic/reperfusion injury (HIRI) is a major complication of liver resection and transplantation. Octreotide, a somatostatin analogue, has been used to treat hepatic fibrosis and portal hypertension; however, its function against HIRI remains unclear. To elucidate the effect of octreotide in HIRI, we investigated the hepatocellular protein profiles in response to octreotide preconditioning in a rabbit model by using proteomic analysis. Twenty-four rabbits were divided into 3 groups: the sham operative group (control), the ischemia/reperfusion group (IR), and the ischemia/reperfusion + octreotide group (IR+Oct). They were subjected to 30 minutes of normothermic ischemia followed by 120 minutes of reperfusion by using Pringle's maneuver method. Proteomic studies were then performed to compare the protein profiles of their left liver lobe. A total of 16 differential proteins were successfully identified. These findings suggest that octreotide might exert an effect against HIRI through up-regulating the expression of the anti-injury substances, such as heat-shock proteins 70 and 27 (confirmed by using Western blot analysis); significantly raising the phosphatidylethanolamine-binding protein that alleviates IR-related apoptosis; and down-regulating mitochondrial metabolic enzymes such as NADH2 dehydrogenase and triosephosphate isomerase.     

远隔缺血预处理对肝切除术患者干预效果的系统评价与Meta分析

背景与目的：尽管大量的动物实验已证实远隔缺血预处理（RIPC）可显著改善肝切除术后残余肝早期的组织灌注和氧合作用,减少肝缺血-再灌注损伤（HIRI）,改善手术预后,提高生存率。然而,在临床研究中关于RIPC减少HIRI的作用仍存在争议。因此,本研究通过Meta分析评价RIPC在肝切除术中的有效性和安全性,为临床提供循证参考。方法：检索多个国内外文献数据库,收集评估RIPC策略有效性和安全性的随机对照试验（RCT）,检索时间段为建库至2022年5月。对纳入的RCT进行质量评价,采用Revman 5.3统计软件进行分析。主要评价指标为术后肝功能指标、手术时间、术中出血量、住院时间、主要手术并发症。结果：共纳入11项RCT,包含851例患者,其中RIPC组422例,对照组429例。Meta分析结果显示,两组术前各项肝功能指标差异均无统计学意义（均P>0.05）;术后第1天,RIPC组的丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）水平明显低于对照组（WMD=-74.92,95%CI=-126.18～-23.67,P=0.004;WMD=-66.37,95%CI=-106.4...     

丙泊酚与肝脏缺血/再灌注损伤

缺血/再灌注对肝脏造成损伤.众多资料显示丙泊酚对肝脏缺血/再灌注损伤有保护作用,这一保护作用与其抗氧化,阻断钙超载,减轻炎性细胞导致的损伤有关.肝脏缺血/再灌注也影响了丙泊酚的代谢.     

异甘草酸镁联合还原型谷胱甘肽对肝细胞癌肝切除患者的应用价值

背景与目的:目前,手术切除仍然是肝细胞癌(肝癌)治疗的首选方法,然而,肝脏缺血再灌注损伤(HIRI)是肝脏切除难以避免的并发症,并且严重影响患者疗效和预后。由于氧化应激和炎症是诱导HIRI的重要机制,故本研究探讨抗炎类护肝药异甘草酸镁联合抗氧化剂还原型谷胱甘肽在肝癌肝切除术患者中的应用效果。方法:选择2016年3月-2019年3月间90例行肝切除手术治疗的肝癌患者随机分为对照组与观察组,每组45例。对照组术后采用静脉滴注还原型谷胱甘肽,观察组术后采用静脉滴注异甘草酸镁联合还原型谷胱甘肽,均为每天1次,连续7 d。比较两组患者的相关临床指标、术前与术后3、7 d的氧化应激与炎性因子水平以及术后1个月内的并发症发生情况。结果:两组患者术前一般资料、手术方式、肝切除范围以及肝功能、氧化应激与炎症因子指标均无统计学差异(均P>0.05)。所有患者手术均顺利完成,两组肝门阻断时长、手术时间和术中出血量差异均无统计学意义(均P>0.05)。与对照组比较,观察组术后3、7 d的肝功能指标明显优于对照组(均P<0.05);超氧化物岐化酶、还原型谷胱甘肽过氧化物酶水平明显高于对照组,而丙二醛水平明显低于对照组(均P<0.05);C反应蛋白、白细胞介素6、肿瘤坏死因子α低于对照组(均P<0.05)。两组均未见皮疹、瘙痒或过敏症状发生。观察组术后1个月内肝功能衰竭发生率明显低于对照组(4.4% vs.11.1%,P<0.05)。结论:异甘草酸镁联合还原型谷胱甘肽对肝癌肝切除术患者的肝功能具有明显保护作用,且优于单纯抗氧化治疗,该作用可能与抗炎抗氧化双重作用后有效抑制HIRI有关。