Experimental study on effect of preconditioning to hepatic ischemia/reperfusion injury.

预处理对肝缺血/再灌注损伤影响的实验研究。

• 批准号: 11671228
• 负责人: NAGINO Masato
• 金额: $ 2.18万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671228/
• 关键词: Ischemia reperfusion injury; Preconditioning; Liver failure; Hypercholesterolemia; Leukocyte; Kupffer cell; Adhesion molecule; プレコンディショニング; 虚血再灌流傷害; 肝細胞傷害; 肝微小循環; SWOP

Our goal of this study was to examine the effect of preconditioning to hepatic ischemia/reperfusion (I/R). At first, to establish experimental model for hepatic I/R, experiments on hepatic I/R in hypercholesterolemic mice were conducted. The objective of this series was to determine whether hypercholesterolemia affects leukocyte adhesion and tissue injury after I/R in the mouse liver. The accumulation of rhodamine-6G-labelled leukocytes and number of non-perfused sinusoids (NPS) were monitored (by intravital microscopy) in liver of normal (WT) and low-density lipoprotein receptor-deficient (LDLr^<-/->) mice for 1 hr after a 30 min period of normothermic ischemia. Plasma Alanine Transaminase (ALT) level was measured as an index of liver tissue injury Microvascular leukostasis as well as increases in NPS and plasma ALT level were observed at 60 min after hepatic I/R in both WT and in LDLr^<-/-> mice ; however, these responses were greatly exaggerated in LDLr^<-/-> mice. Pretreatment of L … More DLr^<-/-> mice with gadolinium chloride, which reduces Kupffer cell function, attenuated the hepatic leukostasis, NPS and liver tissue injury elicited by I/R.Similar protection against I/R was observed in LDLr^<-/-> mice pretreated with antibodies directed against either tumor necrosis factor-α, intercellular adhesion molecule-1 (ICAM-1), or P-selectin. These findings indicate that chronic hypercholesterolemia predisposes the hepatic microvasculature to the deleterious effects of I/R.Kupffer cell activation and the leukocyte adhesion receptors ICAM-1 and P-selectin appear to contribute to the exaggerated inflammatory responses observed in the postishemic liver of LDLr^<-/-> mice. Using this established experimental model of mice liver, a small amount of endotoxin was intraperitoneally administered 24 hours prior to 1 hour hepatic ischemia in wild-type mice. Pretreatment of endotoxin attenuated microvascular leukostasis at 60 win after hepatic I/R, and preconditioning effect in mice liver was confirmed. Hereafter, we will examine the preconditioning effect by a short duration of ischemia and elucidate its mechanism. Less

本研究的目的是研究预处理对肝缺血/再灌注（I/R）的影响。首先，建立肝脏I/R实验模型，对高胆固醇血症小鼠进行肝脏I/R实验。本系列的目的是确定高胆固醇血症是否影响小鼠肝脏I/R后的白细胞粘附和组织损伤。在正常（WT）和低密度脂蛋白受体缺陷（LDLr^<-/->）小鼠肝脏缺血30分钟后1小时内（通过活体显微镜）监测罗丹明- 6g标记白细胞的积累和非灌注窦（NPS）的数量。以血浆丙氨酸转氨酶（ALT）水平作为肝组织损伤指标，观察WT和LDLr^<-/->小鼠肝I/R后60min微血管白质停滞、NPS和血浆ALT水平升高；然而，这些反应在LDLr^<-/->小鼠中被大大夸大。氯化钆预处理L…More DLr^<-/->小鼠，可降低Kupffer细胞功能，减轻I/R引起的肝白斑、NPS和肝组织损伤。在LDLr^<-/->小鼠中，用针对肿瘤坏死因子-α、细胞间粘附分子-1 （ICAM-1）或p -选择素的抗体预处理后，观察到类似的抗I/R保护作用。这些发现表明，慢性高胆固醇血症使肝微血管易受I/R的有害影响。Kupffer细胞活化和白细胞粘附受体ICAM-1和p -选择素似乎有助于在LDLr^<-/->小鼠的缺血后肝脏中观察到的夸大的炎症反应。利用所建立的小鼠肝脏实验模型，在野生型小鼠肝缺血1小时前24小时腹腔注射少量内毒素。内毒素预处理可减轻小鼠肝脏I/R后60分钟微血管白质淤积，并证实其预处理作用。接下来，我们将通过短时间缺血来研究预适应效应并阐明其机制。少

项目成果

Hayata A: "Hepatocyte growth factor concentration in rat bile is affected by hepatic resection volume and external biliary drainage"Journal of Surgical Research. 85. 71-76 (1999)

Hayata A：“大鼠胆汁中的肝细胞生长因子浓度受肝切除体积和胆汁外引流的影响”《外科研究杂志》。

Tsugane K: "A possible role nuclear ceramide and sphingosine in hepatocyte apoptosis in rat liver"Journal of Hepatplogy. 31. 8-17 (1999)

Tsugane K：“核神经酰胺和鞘氨醇在大鼠肝脏肝细胞凋亡中的可能作用”肝脏病学杂志。

Naoharu Mori: "Hepatic microvascular responses to ischemia-reperfusion in low-density lipoprotein receptor knockout mice"Am J Gastrointest Liver Physiol. 279. G1257-G1264 (2000)

Naoharu Mori：“低密度脂蛋白受体敲除小鼠的肝脏微血管对缺血再灌注的反应”Am J Gastrointest Liver Physiol。